Solubilization and purification of melatonin receptors from lizard brain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rivkees, S.A.; Conron, R.W. Jr.; Reppert, S.M.

Melatonin receptors in lizard brain were identified and characterized using {sup 125}I-labeled melatonin (({sup 125}I)MEL) after solubilization with the detergent digitonin. Saturation studies of solubilized material revealed a high affinity binding site, with an apparent equilibrium dissociation constant of 181 +/- 45 pM. Binding was reversible and inhibited by melatonin and closely related analogs, but not by serotonin or norepinephrine. Treatment of solubilized material with the non-hydrolyzable GTP analog, guanosine 5'-(3-O-thiotriphosphate) (GTP-gamma-S), significantly reduced receptor affinity. Gel filtration chromatography of solubilized melatonin receptors revealed a high affinity, large (Mr 400,000) peak of specific binding. Pretreatment with GTP-gamma-S before solubilization resultedmore » in elution of a lower affinity, smaller (Mr 150,000) peak of specific binding. To purify solubilized receptors, a novel affinity chromatography resin was developed by coupling 6-hydroxymelatonin with Epoxy-activated Sepharose 6B. Using this resin, melatonin receptors were purified approximately 10,000-fold. Purified material retained the pharmacologic specificity of melatonin receptors. These results show that melatonin receptors that bind ligand after detergent treatment can be solubilized and substantially purified by affinity chromatography.« less

Scale converters. [Dissolution of scale deposits

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rybacki, R.L.

1979-04-03

A water-soluble maleate salt, preferably disodium maleate, is described as a scale solubilizer. Such salts convert scale into a water-soluble compound without the conventional step of employing either an acid wash or a sequestering agent. 10 claims.

Techno-economic evaluation of an inclusion body solubilization and recombinant protein refolding process.

PubMed

Freydell, Esteban J; van der Wielen, Luuk A M; Eppink, Michel H M; Ottens, Marcel

2011-01-01

Expression of recombinant proteins in Escherichia coli is normally accompanied by the formation of inclusion bodies (IBs). To obtain the protein product in an active (native) soluble form, the IBs must be first solubilized, and thereafter, the soluble, often denatured and reduced protein must be refolded. Several technically feasible alternatives to conduct IBs solubilization and on-column refolding have been proposed in recent years. However, rarely these on-column refolding alternatives have been evaluated from an economical point of view, questioning the feasibility of their implementation at a preparative scale. The presented study assesses the economic performance of four distinct process alternatives that include pH induced IBs solubilization and protein refolding (pH_IndSR); IBs solubilization using urea, dithiothreitol (DTT), and alkaline pH followed by batch size-exclusion protein refolding; inclusion bodies (IBs) solubilization using urea, DTT, and alkaline pH followed by simulated moving bed (SMB) size-exclusion protein refolding, and IBs solubilization using urea, DTT and alkaline pH followed by batch dilution protein refolding. The economic performance was judged on the basis of the direct fixed capital, and the production cost per unit of product (P(C)). This work shows that (1) pH_IndSR system is a relatively economical process, because of the low IBs solubilization cost; (2) substituting β-mercaptoethanol for dithiothreithol is an attractive alternative, as it significantly decreases the product cost contribution from the IBs solubilization; and (3) protein refolding by size-exclusion chromatography becomes economically attractive by changing the mode of operation of the chromatographic reactor from batch to continuous using SMB technology. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Pilot-Scale Batch Alkaline Pretreatment of Corn Stover

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kuhn, Erik M.; O’Brien, Marykate H.; Ciesielski, Peter N.

The goal of biomass pretreatment is to increase the enzymatic digestibility of the plant cell wall polysaccharides to produce sugars for upgrading to biofuels. Alkaline pretreatment has the ability to solubilize much of the lignin in biomass while the carbohydrates remain insoluble. With an increased research focus to produce high-value products from lignin, a low molecular weight, lignin-rich stream in a biorefinery is desirable. Here, this work reports on batch alkaline pretreatment of corn stover conducted using a three-factor, two-level central composite experimental design in a pilot-scale reactor to determine the relationship between sodium hydroxide (NaOH) loading, temperature, and anthraquinonemore » (AQ) charge on solids solubilization, component yields, and enzymatic digestibility of the residual solids. Operating conditions were 100 to 140 °C, 40 to 70 mg NaOH/g dry corn stover, and 0.05% to 0.2% (w/w) AQ loading. An enzymatic hydrolysis screening study was performed at 2% cellulose loading. Empirical modeling results showed that NaOH loading and temperature are both significant factors, solubilizing 15% to 35% of the solids and up to 54% of the lignin. Enzymatic hydrolysis of the residual solids produced good monomeric glucose (>90%) and xylose (>70%) yields at the more severe pretreatment conditions. We also found that the AQ charge was not a significant factor at the conditions studied, so efforts to reduce xylan and increase lignin solubilization using this compound were not successful. Lastly, while good lignin solubilization was achieved, effectively recovering this stream remains a challenge, and demonstrating performance in continuous reactors is still needed.« less

Pilot-Scale Batch Alkaline Pretreatment of Corn Stover

DOE PAGES

Kuhn, Erik M.; O’Brien, Marykate H.; Ciesielski, Peter N.; ...

2015-12-18

The goal of biomass pretreatment is to increase the enzymatic digestibility of the plant cell wall polysaccharides to produce sugars for upgrading to biofuels. Alkaline pretreatment has the ability to solubilize much of the lignin in biomass while the carbohydrates remain insoluble. With an increased research focus to produce high-value products from lignin, a low molecular weight, lignin-rich stream in a biorefinery is desirable. Here, this work reports on batch alkaline pretreatment of corn stover conducted using a three-factor, two-level central composite experimental design in a pilot-scale reactor to determine the relationship between sodium hydroxide (NaOH) loading, temperature, and anthraquinonemore » (AQ) charge on solids solubilization, component yields, and enzymatic digestibility of the residual solids. Operating conditions were 100 to 140 °C, 40 to 70 mg NaOH/g dry corn stover, and 0.05% to 0.2% (w/w) AQ loading. An enzymatic hydrolysis screening study was performed at 2% cellulose loading. Empirical modeling results showed that NaOH loading and temperature are both significant factors, solubilizing 15% to 35% of the solids and up to 54% of the lignin. Enzymatic hydrolysis of the residual solids produced good monomeric glucose (>90%) and xylose (>70%) yields at the more severe pretreatment conditions. We also found that the AQ charge was not a significant factor at the conditions studied, so efforts to reduce xylan and increase lignin solubilization using this compound were not successful. Lastly, while good lignin solubilization was achieved, effectively recovering this stream remains a challenge, and demonstrating performance in continuous reactors is still needed.« less

VISUALIZATION AND SIMULATION OF NON-AQUEOUS PHASE LIQUIDS SOLUBILIZATION IN PORE NETWORKS

EPA Science Inventory

The design of in-situ remediation of contaminated soils is mostly based on a description at the macroscopic scale using a averaged quantities. These cannot address issues at the pore and pore network scales. In this paper, visualization experiments and numerical simulations in ...

ABC-transporters are localized in caveolin-1-positive and reggie-1-negative and reggie-2-negative microdomains of the canalicular membrane in rat hepatocytes.

PubMed

Ismair, Manfred G; Häusler, Stephanie; Stuermer, Claudia A; Guyot, Christelle; Meier, Peter J; Roth, Jürgen; Stieger, Bruno

2009-05-01

The canalicular plasma membrane is constantly exposed to bile acids acting as detergents. Bile acids are essential to mediate release of biliary lipids from the canalicular membrane. Membrane microdomains (previously called lipid rafts) are biochemically defined by their resistance to detergent solubilization at cold temperature. We aimed to investigate the canalicular plasma membrane for the presence of microdomains, which could protect this membrane against the detergent action of bile acids. Highly purified rat liver canalicular plasma membrane vesicles were extracted with 1% Triton X-100 or 1% Lubrol WX at 4 degrees C and subjected to flotation through sucrose step gradients. Both detergents yielded detergent-resistant membranes containing the microdomain markers alkaline phosphatase and sphingomyelin. However, cholesterol was resistant to Lubrol WX solubilization, whereas it was only marginally resistant to solubilization by Triton X-100. The microdomain marker caveolin-1 was localized to the canalicular plasma membrane domain and was resistant to Lubrol WX, but to a large extent solubilized by Triton X-100. The two additional microdomain markers, reggie-1 and reggie-2, were localized to the basolateral and canalicular plasma membrane and were partially resistant to Lubrol WX but resistant to Triton X-100. The canalicular transporters bile salt export pump, multidrug resistance protein 2, multidrug resistance-associated protein 2, and Abcg5 were largely resistant to Lubrol WX but were solubilized by Triton X-100. These results indicate the presence of two different types of microdomains in the canalicular plasma membrane: "Lubrol-microdomains" and "Triton-microdomains". "Lubrol-microdomains" contain the machinery for canalicular bile formation and may be the starting place for canalicular lipid secretion.

Comparative study to develop a single method for retrieving wide class of recombinant proteins from classical inclusion bodies.

PubMed

Padhiar, Arshad Ahmed; Chanda, Warren; Joseph, Thomson Patrick; Guo, Xuefang; Liu, Min; Sha, Li; Batool, Samana; Gao, Yifan; Zhang, Wei; Huang, Min; Zhong, Mintao

2018-03-01

The formation of inclusion bodies (IBs) is considered as an Achilles heel of heterologous protein expression in bacterial hosts. Wide array of techniques has been developed to recover biochemically challenging proteins from IBs. However, acquiring the active state even from the same protein family was found to be an independent of single established method. Here, we present a new strategy for the recovery of wide sub-classes of recombinant protein from harsh IBs. We found that numerous methods and their combinations for reducing IB formation and producing soluble proteins were not effective, if the inclusion bodies were harsh in nature. On the other hand, different practices with mild solubilization buffers were able to solubilize IBs completely, yet the recovery of active protein requires large screening of refolding buffers. With the integration of previously reported mild solubilization techniques, we proposed an improved method, which comprised low sarkosyl concentration, ranging from 0.05 to 0.1% coupled with slow freezing (- 1 °C/min) and fast thaw (room temperature), resulting in greater solubility and the integrity of solubilized protein. Dilution method was employed with single buffer to restore activity for every sub-class of recombinant protein. Results showed that the recovered protein's activity was significantly higher compared with traditional solubilization/refolding approach. Solubilization of IBs by the described method was proved milder in nature, which restored native-like conformation of proteins within IBs.

Solubilization Behavior of Polyene Antibiotics in Nanomicellar System: Insights from Molecular Dynamics Simulation of the Amphotericin B and Nystatin Interactions with Polysorbate 80.

PubMed

Mobasheri, Meysam; Attar, Hossein; Rezayat Sorkhabadi, Seyed Mehdi; Khamesipour, Ali; Jaafari, Mahmoud Reza

2015-12-24

Amphotericin B (AmB) and Nystatin (Nys) are the drugs of choice for treatment of systemic and superficial mycotic infections, respectively, with their full clinical potential unrealized due to the lack of high therapeutic index formulations for their solubilized delivery. In the present study, using a coarse-grained (CG) molecular dynamics (MD) simulation approach, we investigated the interaction of AmB and Nys with Polysorbate 80 (P80) to gain insight into the behavior of these polyene antibiotics (PAs) in nanomicellar solution and derive potential implications for their formulation development. While the encapsulation process was predominantly governed by hydrophobic forces, the dynamics, hydration, localization, orientation, and solvation of PAs in the micelle were largely controlled by hydrophilic interactions. Simulation results rationalized the experimentally observed capability of P80 in solubilizing PAs by indicating (i) the dominant kinetics of drugs encapsulation over self-association; (ii) significantly lower hydration of the drugs at encapsulated state compared with aggregated state; (iii) monomeric solubilization of the drugs; (iv) contribution of drug-micelle interactions to the solubilization; (v) suppressed diffusivity of the encapsulated drugs; (vi) high loading capacity of the micelle; and (vii) the structural robustness of the micelle against drug loading. Supported from the experimental data, our simulations determined the preferred location of PAs to be the core-shell interface at the relatively shallow depth of 75% of micelle radius. Deeper penetration of PAs was impeded by the synergistic effects of (i) limited diffusion of water; and (ii) perpendicular orientation of these drug molecules with respect to the micelle radius. PAs were solvated almost exclusively in the aqueous poly-oxyethylene (POE) medium due to the distance-related lack of interaction with the core, explaining the documented insensitivity of Nys solubilization to drug-core compatibility in detergent micelles. Based on the obtained results, the dearth of water at interior sites of micelle and the large lateral occupation space of PAs lead to shallow insertion, broad radial distribution, and lack of core interactions of the amphiphilic drugs. Hence, controlled promotion of micelle permeability and optimization of chain crowding in palisade layer may help to achieve more efficient solubilization of the PAs.

The effect of pH on solubilization of organic matter and microbial community structures in sludge fermentation.

PubMed

Maspolim, Yogananda; Zhou, Yan; Guo, Chenghong; Xiao, Keke; Ng, Wun Jern

2015-08-01

Sludge fermentation between pH 4 and 11 was investigated to generate volatile fatty acids (VFA). Despite the highest sludge solubilization of 25.9% at pH 11, VFA accumulation was optimized at pH 8 (12.5% out of 13.1% sludge solubilization). 454 pyrosequencing identified wide diversity of acidogens in bioreactors operated at the various pHs, with Tissierella, Petrimonas, Proteiniphilum, Levilinea, Proteiniborus and Sedimentibacter enriched and contributing to the enhanced fermentation at pH 8. Hydrolytic enzymatic assays determined abiotic effect to be the leading cause for improved solubilization under high alkaline condition but the environmental stress at pH 9 and above might lead to disrupt biological activities and eventually VFA production. Furthermore, molecular weight (MW) characterization of the soluble fractions found large MW aromatic substances at pH 9 and above, that is normally associated with poor biodegradability, making them disadvantageous for subsequent bioprocesses. The findings provided information to better understand and control sludge fermentation. Copyright © 2015 Elsevier Ltd. All rights reserved.

Microbial solubilization of coal

DOEpatents

Strandberg, G.W.; Lewis, S.N.

1988-01-21

The present invention relates to a cell-free preparation and process for the microbial solubilization of coal into solubilized coal products. More specifically, the present invention relates to bacterial solubilization of coal into solubilized coal products and a cell-free bacterial byproduct useful for solubilizing coal. 5 tabs.

In-situ capture and conversion of atmospheric CO2 into nano-CaCO3 using a novel pathway based on deep eutectic choline chloride-calcium chloride.

PubMed

Karimi, Mohammad; Jodaei, Akbar; Khajvandi, Asiyeh; Sadeghinik, Amirhosein; Jahandideh, Rahim

2018-01-15

This study presents a newfangled method and provides a new approach toward capturing-entrapping the toxic and harmful greenhouse gas CO 2 and subsequently employing it as a useful precursor for the production of value-added calcite (CaCO 3 ) nanoparticles. All the processes are done in a single system (one-pot and in-situ processing) based on deep eutectic choline chloride-calcium chloride (Calcoline) at mild temperature of 50 °C in direct contact with air. The Calcoline eutectic mixture shows the calcite production yield of 6.2 mg/ml that is reduced up to 22.6% upon recycling. The eutectic mixture provides an interesting all-in-one system acting as carbon dioxide trapper/adsorbent, solvent to solubilize CO 2 , and reagent to convert solubilized CO 2 into pure calcite nanoparticles with average particle size of 30 nm. The featured properties of the developed method including simplicity, recyclability, mobility, affordability, and sustainability make it feasible for large scale applications by which the simultaneous elimination of CO 2 from the environment and conversion of it into value added nano-products will be practicable. Copyright © 2017 Elsevier Ltd. All rights reserved.

CHOBIMALT: A Cholesterol-Based Detergent†

PubMed Central

Howell, Stanley C.; Mittal, Ritesh; Huang, Lijun; Travis, Benjamin; Breyer, Richard M.; Sanders, Charles R.

2010-01-01

Cholesterol and its hemisuccinate and sulfate derivatives are widely used in studies of purified membrane proteins, but are difficult to solubilize in aqueous solution, even in the presence of detergent micelles. Other cholesterol derivatives do not form conventional micelles and lead to viscous solutions. To address these problems a cholesterol-based detergent, CHOBIMALT, has been synthesized and characterized. At concentrations above 3–4μM, CHOBIMALT forms micelles without the need for elevated temperatures or sonic disruption. Diffusion and fluorescence measurements indicated that CHOBIMALT micelles are large (210 ± 30 kDa). The ability to solubilize a functional membrane protein was explored using a G-protein coupled receptor, the human kappa opioid receptor type 1 (hKOR1). While CHOBIMALT alone was not found to be effective as a surfactant for membrane extraction, when added to classical detergent micelles CHOBIMALT was observed to dramatically enhance the thermal stability of solubilized hKOR1. PMID:20919740

Use of an isoelectric solubilization/precipitation process to modify the functional properties of PSE (pale, soft, exudative)-like chicken meat protein: A mechanistic approach.

PubMed

Zhao, Xue; Bai, Yun; Xing, Tong; Xu, Xing-Lian; Zhou, Guanghong

2018-05-15

The functionality of pale, soft, exudative (PSE)-like chicken protein was improved by isoelectric solubilization/precipitation (ISP) treatment. PSE-like chicken proteins were solubilized at an acidic pH 3.5 or an alkaline pH 11.0, followed by precipitating at pH 5.5 and 6.2. PSE-like meat paste was treated as control (CON). Precipitated at pH 6.2 led to a more elastic gel than at pH 5.5. Water distribution of ISP-isolated protein was affected by precipitation pH. More tryptophan residues exposed and -SH was partially oxidized to disulfide bond after ISP treatment, which led to large aggregates formation and higher viscosity of ISP isolated proteins than of CON. Absolute zeta potential of alkali-treated protein was higher than other counterparts, indicating stronger electric repulsion. ISP treatments could convert α-helix structure to relatively irregular structures. Overall, solubilizing at pH 11.0, combined with a precipitation pH 6.2 ISP treatment offers a potential for enhanced functionality of PSE-like chicken protein. Copyright © 2017 Elsevier Ltd. All rights reserved.

Accelerated oral nanomedicine discovery from miniaturized screening to clinical production exemplified by paediatric HIV nanotherapies

NASA Astrophysics Data System (ADS)

Giardiello, Marco; Liptrott, Neill J.; McDonald, Tom O.; Moss, Darren; Siccardi, Marco; Martin, Phil; Smith, Darren; Gurjar, Rohan; Rannard, Steve P.; Owen, Andrew

2016-10-01

Considerable scope exists to vary the physical and chemical properties of nanoparticles, with subsequent impact on biological interactions; however, no accelerated process to access large nanoparticle material space is currently available, hampering the development of new nanomedicines. In particular, no clinically available nanotherapies exist for HIV populations and conventional paediatric HIV medicines are poorly available; one current paediatric formulation utilizes high ethanol concentrations to solubilize lopinavir, a poorly soluble antiretroviral. Here we apply accelerated nanomedicine discovery to generate a potential aqueous paediatric HIV nanotherapy, with clinical translation and regulatory approval for human evaluation. Our rapid small-scale screening approach yields large libraries of solid drug nanoparticles (160 individual components) targeting oral dose. Screening uses 1 mg of drug compound per library member and iterative pharmacological and chemical evaluation establishes potential candidates for progression through to clinical manufacture. The wide applicability of our strategy has implications for multiple therapy development programmes.

Expression, purification, and refolding of active recombinant human E-selectin lectin and EGF domains in Escherichia coli.

PubMed

Kawano, Susumu; Iyaguchi, Daisuke; Okada, Chiaki; Sasaki, Yusuke; Toyota, Eiko

2013-06-01

Attempts to obtain active E-selectin from Escherichia coli (E. coli) have not yet been successful. In this study, we succeeded in expressing the recombinant lectin and epidermal growth factor domain fragments of human E-selectin (rh-ESLE) in E. coli on a large-scale. The rh-ESLE protein was expressed as an inactive form in the inclusion bodies. The inactive form of rh-ESLE was denatured and solubilized by 6 M guanidine hydrochloride and then purified by Ni(2+) affinity chromatography under denaturing conditions. Denatured rh-ESLE was then refolded by a rapid-dilution method using a large amount of refolding buffer, which contained arginine and cysteine/cystine. The refolded rh-ESLE showed binding affinity for sLe(X) (K(d) = 321 nM, B(max) = 1.9 pmol/μg protein). This result suggests that the refolded rh-ESLE recovered its native and functional structure.

Effects of ethanol addition on micellar solubilization and plume migration during surfactant enhanced recovery of tetrachloroethene.

PubMed

Taylor, Tammy P; Rathfelder, Klaus M; Pennell, Kurt D; Abriola, Linda M

2004-03-01

Alcohol addition has been suggested for use in combination with surfactant flushing to enhance solubilization kinetics and permit density control of dense non-aqueous phase liquid (DNAPL)-laden surfactant plumes. This study examined the effects of adding ethanol (EtOH) to a 4% Tween 80 (polyoxyethylene (20) sorbitan monooleate) solution used to flush tetrachloroethene (PCE)-contaminated porous media. The influence of EtOH concentration, subsurface layering and scale on flushing solution delivery and PCE recovery was investigated through a combination of experimental and mathematical modeling studies. Results of batch experiments demonstrated that the addition of 2.5%, 5% and 10% (wt.) EtOH incrementally increased the PCE solubilization capacity and viscosity of the surfactant solution, while reducing solution density from 1.002 to 0.986 g/cm3. Effluent concentration data obtained from one-dimensional (1-D) column experiments were used to characterize rate-limited micellar solubilization of residual PCE, which was strongly dependent upon flow velocity and weakly dependent upon EtOH concentration. Two-dimensional (2-D) box studies illustrated that minor differences (0.008 g/cm3) between flushing and resident solution density can strongly influence surfactant front propagation. A two-dimensional multiphase simulator, MISER, was used to model the influence of EtOH composition on the aqueous flow field and PCE mass recovery. The ability of the numerical simulator to predict effluent concentrations and front propagation was demonstrated for both 1-D columns and 2-D boxes flushed with EtOH-amended Tween 80 solutions. Results of this study quantify the potential influence of alcohol addition on surfactant solution properties and solubilization capacity, and demonstrate the importance of considering small density variations in remedial design.

Cyclodextrin-enhanced solubilization of organic contaminants with implications for aquifer remediation

DOE Office of Scientific and Technical Information (OSTI.GOV)

McCray, J.E.; Boving, T.B.; Brusseau, M.L.

2000-12-31

Reagents that enhance the aqueous solubility of nonaqueous phase organic liquid (NAPL) contaminants are under investigation for use in enhanced subsurface remediation technologies. Cyclodextrin, a glucose-based molecule, is such a reagent. In this paper, laboratory experiments and numerical model simulations are used to evaluate and understand the potential remediation performance of cyclodextrin. Physical properties of cyclodextrin solutions such as density, viscosity, and NAPL-aqueous interfacial tension are measured. Their analysis indicates that no serious obstacles exist related to fluid properties that would prevent the use of cyclodextrin solutions for subsurface NAPL remediation. Cyclodextrin-enhanced solubilization for a large suite of typical groundmore » water contaminants is measured in the laboratory, and the results are related to the physiochemical properties of the organic compounds. The most-hydrophobic contaminants experience a larger relative solubility enhancement than the less-hydrophobic contaminants but have lower aqueous-phase apparent solubilities. Numerical model simulations of enhanced-solubilization flushing of NAPL-contaminated soil demonstrate that the more-hydrophilic compounds exhibit the greatest mass-removal relates due to their greater apparent solubilities, and thus are initially more effectively removed from soil by enhanced-solubilization-flushing reagents. However, the relatively more hydrophobic contaminants exhibit a greater improvement in contaminant mass-removal (compared with water flushing) than that exhibited for the relatively hydrophilic contaminants.« less

Large-scale production and study of a synthetic G protein-coupled receptor: Human olfactory receptor 17-4

PubMed Central

Cook, Brian L.; Steuerwald, Dirk; Kaiser, Liselotte; Graveland-Bikker, Johanna; Vanberghem, Melanie; Berke, Allison P.; Herlihy, Kara; Pick, Horst; Vogel, Horst; Zhang, Shuguang

2009-01-01

Although understanding of the olfactory system has progressed at the level of downstream receptor signaling and the wiring of olfactory neurons, the system remains poorly understood at the molecular level of the receptors and their interaction with and recognition of odorant ligands. The structure and functional mechanisms of these receptors still remain a tantalizing enigma, because numerous previous attempts at the large-scale production of functional olfactory receptors (ORs) have not been successful to date. To investigate the elusive biochemistry and molecular mechanisms of olfaction, we have developed a mammalian expression system for the large-scale production and purification of a functional OR protein in milligram quantities. Here, we report the study of human OR17-4 (hOR17-4) purified from a HEK293S tetracycline-inducible system. Scale-up of production yield was achieved through suspension culture in a bioreactor, which enabled the preparation of >10 mg of monomeric hOR17-4 receptor after immunoaffinity and size exclusion chromatography, with expression yields reaching 3 mg/L of culture medium. Several key post-translational modifications were identified using MS, and CD spectroscopy showed the receptor to be ≈50% α-helix, similar to other recently determined G protein-coupled receptor structures. Detergent-solubilized hOR17-4 specifically bound its known activating odorants lilial and floralozone in vitro, as measured by surface plasmon resonance. The hOR17-4 also recognized specific odorants in heterologous cells as determined by calcium ion mobilization. Our system is feasible for the production of large quantities of OR necessary for structural and functional analyses and research into OR biosensor devices. PMID:19581598

Large-scale production and study of a synthetic G protein-coupled receptor: human olfactory receptor 17-4.

PubMed

Cook, Brian L; Steuerwald, Dirk; Kaiser, Liselotte; Graveland-Bikker, Johanna; Vanberghem, Melanie; Berke, Allison P; Herlihy, Kara; Pick, Horst; Vogel, Horst; Zhang, Shuguang

2009-07-21

Although understanding of the olfactory system has progressed at the level of downstream receptor signaling and the wiring of olfactory neurons, the system remains poorly understood at the molecular level of the receptors and their interaction with and recognition of odorant ligands. The structure and functional mechanisms of these receptors still remain a tantalizing enigma, because numerous previous attempts at the large-scale production of functional olfactory receptors (ORs) have not been successful to date. To investigate the elusive biochemistry and molecular mechanisms of olfaction, we have developed a mammalian expression system for the large-scale production and purification of a functional OR protein in milligram quantities. Here, we report the study of human OR17-4 (hOR17-4) purified from a HEK293S tetracycline-inducible system. Scale-up of production yield was achieved through suspension culture in a bioreactor, which enabled the preparation of >10 mg of monomeric hOR17-4 receptor after immunoaffinity and size exclusion chromatography, with expression yields reaching 3 mg/L of culture medium. Several key post-translational modifications were identified using MS, and CD spectroscopy showed the receptor to be approximately 50% alpha-helix, similar to other recently determined G protein-coupled receptor structures. Detergent-solubilized hOR17-4 specifically bound its known activating odorants lilial and floralozone in vitro, as measured by surface plasmon resonance. The hOR17-4 also recognized specific odorants in heterologous cells as determined by calcium ion mobilization. Our system is feasible for the production of large quantities of OR necessary for structural and functional analyses and research into OR biosensor devices.

EPA RREL'S MOBILE VOLUME REDUCTION UNIT -- APPLICATIONS ANALYSIS REPORT

EPA Science Inventory

The volume reduction unit (VRU) is a pilot-scale, mobile soil washing system designed to remove organic contaminants from the soil through particle size separation and solubilization. The VRU removes contaminants by suspending them in a wash solution and by reducing the volume of...

Cocrystal solubilization in biorelevant media and its prediction from drug solubilization

PubMed Central

Lipert, Maya P.; Roy, Lilly; Childs, Scott L.

2015-01-01

This work examines cocrystal solubility in biorelevant media, (FeSSIF, fed state simulated intestinal fluid), and develops a theoretical framework that allows for the simple and quantitative prediction of cocrystal solubilization from drug solubilization. The solubilities of four hydrophobic drugs and seven cocrystals containing these drugs were measured in FeSSIF and in acetate buffer at pH 5.00. In all cases, the cocrystal solubility (Scocrystal) was higher than the drug solubility (Sdrug) in both buffer and FeSSIF; however, the solubilization ratio of drug, SRdrug = (SFeSSIF/Sbuffer)drug, was not the same as the solubilization ratio of cocrystal, SRcocrystal = (SFeSSIF/Sbuffer)cocrystal, meaning drug and cocrystal were not solubilized to the same extent in FeSSIF. This highlights the potential risk of anticipating cocrystal behavior in biorelevant media based on solubility studies in water. Predictions of SRcocrystal from simple equations based only on SRdrug were in excellent agreement with measured values. For 1:1 cocrystals, the cocrystal solubilization ratio can be obtained from the square root of the drug solubilization ratio. For 2:1 cocrystals, SRcocrystal is found from (SRdrug)2/3. The findings in FeSSIF can be generalized to describe cocrystal behavior in other systems involving preferential solubilization of a drug such as surfactants, lipids, and other drug solubilizing media. PMID:26390213

Effect of Detergents on the Thermal Behavior of Elastin-like Polypeptides

PubMed Central

Thapa, Arjun; Han, Wei; Simons, Robin H.; Chilkoti, Ashutosh; Chi, Eva Y.; López, Gabriel P.

2012-01-01

Elastin-like polypeptide (ELP) fusions have been designed to allow large scale, non-chromatographic purification of many soluble proteins using the inverse transition cycling (ITC) method; however, the sensitivity of the aqueous lower critical solubility phase transition temperature (Tt) of ELPs to the addition of cosolutes, including detergents, may be a potential hindrance in purification of proteins with surface hydrophobicity in such a manner. To identify detergents that are known to solubilize such proteins (e.g., membrane proteins) and that have little effect on the Tt of the ELP, we screened a number of detergents with respect to their effects on the Tt and secondary structures of a model ELP (denoted here as ELP180). We found that mild detergents (e.g., DDM, Triton-X100, and CHAPS) do not alter the phase transition behavior or structure (as probed by circular dichroism) of ELP180. This result is in contrast to previous studies that showed a strong effect of other detergents (e.g., SDS) on the Ttof ELPs. Our results clearly indicate that mild detergents do not preclude ITC-based separation of ELPs, and thus that ELP fusions may prove to be useful in the purification of detergent-solubilized recombinant hydrophobic proteins, including membrane proteins, which are otherwise notoriously difficult to extract and purify by conventional separation methods (e.g., chromatography). PMID:23097230

Contribution of Zinc Solubilizing Bacteria in Growth Promotion and Zinc Content of Wheat.

PubMed

Kamran, Sana; Shahid, Izzah; Baig, Deeba N; Rizwan, Muhammad; Malik, Kauser A; Mehnaz, Samina

2017-01-01

Zinc is an imperative micronutrient required for optimum plant growth. Zinc solubilizing bacteria are potential alternatives for zinc supplementation and convert applied inorganic zinc to available forms. This study was conducted to screen zinc solubilizing rhizobacteria isolated from wheat and sugarcane, and to analyze their effect on wheat growth and development. Fourteen exo-polysaccharides producing bacterial isolates of wheat were identified and characterized biochemically as well as on the basis of 16S rRNA gene sequences. Along these, 10 identified sugarcane isolates were also screened for zinc solubilizing ability on five different insoluble zinc sources. Out of 24, five strains, i.e., EPS 1 ( Pseudomonas fragi) , EPS 6 ( Pantoea dispersa) , EPS 13 ( Pantoea agglomerans) , PBS 2 ( E. cloacae) and LHRW1 ( Rhizobium sp.) were selected (based on their zinc solubilizing and PGP activities) for pot scale plant experiments. ZnCO 3 was used as zinc source and wheat seedlings were inoculated with these five strains, individually, to assess their effect on plant growth and development. The effect on plants was analyzed based on growth parameters and quantifying zinc content of shoot, root and grains using atomic absorption spectroscopy. Plant experiment was performed in two sets. For first set of plant experiments (harvested after 1 month), maximum shoot and root dry weights and shoot lengths were noted for the plants inoculated with Rhizobium sp. (LHRW1) while E. cloacae (PBS 2) increased both shoot and root lengths. Highest zinc content was found in shoots of E. cloacae (PBS 2) and in roots of P. agglomerans (EPS 13) followed by zinc supplemented control. For second set of plant experiment, when plants were harvested after three months, Pantoea dispersa (EPS 6), P. agglomerans (EPS 13) and E. cloacae (PBS 2) significantly increased shoot dry weights. However, significant increase in root dry weights and maximum zinc content was recorded for Pseudomonas fragi (EPS 1) inoculated plants, isolated from wheat rhizosphere. While maximum zinc content for roots was quantified in the control plants indicating the plant's inability to transport zinc to grains, supporting accelerated bioavailability of zinc to plant grains with zinc solubilizing rhizobacteria.

FIELD TEST OF CYCLODEXTRIN FOR ENHANCED IN-SITU FLUSHING OF MULTIPLE-COMPONENT IMMISCIBLE ORGANIC LIQUID CONTAMINATION: COMPARISON TO WATER FLUSHING

EPA Science Inventory

A pilot-scale field experiment was conducted to compare the remediation effectiveness of an enhanced-solubilization technique to that of water flushing for removal of multicomponent nonaqueous-phase organic liquid (NAPL) contaminants form a phreatic aquifer. This innovative remed...

SURFACTANT ENHANCED RECOVERY OF TETRACHLOROETHYLENE FROM A POROUS MEDIUM CONTAINING LOW PERMEABILITY LENSES. 2. NUMERICAL SIMULATION. (R825409)

EPA Science Inventory

Abstract

A numerical model of surfactant enhanced solubilization was developed and applied to the simulation of nonaqueous phase liquid recovery in two-dimensional heterogeneous laboratory sand tank systems. Model parameters were derived from independent, small-scale, ...

Cocrystal Transition Points: Role of Cocrystal Solubility, Drug Solubility, and Solubilizing Agents.

PubMed

Lipert, Maya P; Rodríguez-Hornedo, Naír

2015-10-05

In this manuscript we bring together concepts that are relevant to the solubilization and thermodynamic stability of cocrystals in the presence of drug solubilizing agents. Simple equations are derived that allow calculation of cocrystal solubilization and transition point solubility. Analysis of 10 cocrystals in 6 different solubilizing agents shows that cocrystal solubilization is quantitatively predicted from drug solubilization. Drug solubilizing agents such as surfactants and lipid-based media are also shown to induce cocrystal transition points, where drug and cocrystal solubilities are equal, and above which the cocrystal solubility advantage over drug is eliminated. We have discovered that cocrystal solubility at the transition point (S*) is independent of solubilizing agent, and can be predicted from knowledge of only the aqueous solubilities of drug and cocrystal. For 1:1 cocrystals, S* = (Scocrystal,aq)(2)/Sdrug,aq. S* is a key indicator of cocrystal thermodynamic stability and establishes the upper solubility limit below which cocrystal is more soluble than the constituent drug. These findings have important implications to tailor cocrystal solubility and stability in pharmaceutical formulations from commonly available drug solubility descriptors.

Accelerated oral nanomedicine discovery from miniaturized screening to clinical production exemplified by paediatric HIV nanotherapies

PubMed Central

Giardiello, Marco; Liptrott, Neill J.; McDonald, Tom O.; Moss, Darren; Siccardi, Marco; Martin, Phil; Smith, Darren; Gurjar, Rohan; Rannard, Steve P.; Owen, Andrew

2016-01-01

Considerable scope exists to vary the physical and chemical properties of nanoparticles, with subsequent impact on biological interactions; however, no accelerated process to access large nanoparticle material space is currently available, hampering the development of new nanomedicines. In particular, no clinically available nanotherapies exist for HIV populations and conventional paediatric HIV medicines are poorly available; one current paediatric formulation utilizes high ethanol concentrations to solubilize lopinavir, a poorly soluble antiretroviral. Here we apply accelerated nanomedicine discovery to generate a potential aqueous paediatric HIV nanotherapy, with clinical translation and regulatory approval for human evaluation. Our rapid small-scale screening approach yields large libraries of solid drug nanoparticles (160 individual components) targeting oral dose. Screening uses 1 mg of drug compound per library member and iterative pharmacological and chemical evaluation establishes potential candidates for progression through to clinical manufacture. The wide applicability of our strategy has implications for multiple therapy development programmes. PMID:27767027

Inhibition of Aspergillus niger Phosphate Solubilization by Fluoride Released from Rock Phosphate

PubMed Central

Mendes, Gilberto de Oliveira; Vassilev, Nikolay Bojkov; Bonduki, Victor Hugo Araújo; da Silva, Ivo Ribeiro; Ribeiro, José Ivo

2013-01-01

The simultaneous release of various chemical elements with inhibitory potential for phosphate solubilization from rock phosphate (RP) was studied in this work. Al, B, Ba, Ca, F, Fe, Mn, Mo, Na, Ni, Pb, Rb, Si, Sr, V, Zn, and Zr were released concomitantly with P during the solubilization of Araxá RP (Brazil), but only F showed inhibitory effects on the process at the concentrations detected in the growth medium. Besides P solubilization, fluoride decreased fungal growth, citric acid production, and medium acidification by Aspergillus niger. At the maximum concentration found during Araxá RP solubilization (22.9 mg F− per liter), fluoride decreased P solubilization by 55%. These findings show that fluoride negatively affects RP solubilization by A. niger through its inhibitory action on the fungal metabolism. Given that fluoride is a common component of RPs, the data presented here suggest that most of the microbial RP solubilization systems studied so far were probably operated under suboptimal conditions. PMID:23770895

An innovative sewage sludge reduction by using a combined mesophilic anaerobic and thermophilic aerobic process with thermal-alkaline treatment and sludge recirculation.

PubMed

Cho, Hyun Uk; Park, Sang Kyu; Ha, Jeong Hyub; Park, Jong Moon

2013-11-15

Lab-scale High Efficiency Digestion (HED) systems containing a Mesophilic Anaerobic Reactor (MAR), Thermophilic Aerobic Reactor (TAR), liquid/solid separation unit, and thermal-alkaline treatment were developed to evaluate the efficiencies of sludge reduction and methane production. The HED process was divided into three phases to examine the influence of sludge pretreatment and pretreated sludge recirculation using TCOD and VSS reduction, COD solubilization, and methane production. The VSS removal with a solid/liquid separation unit, sludge recirculation, and thermal-alkaline treatment drastically increased up to 95% compared to the feed concentration. In addition, the results of COD solubilization and VSS/TSS showed that the solubilization of cells and organic matters by the thermal-alkaline treatment was highly increased, which was also consistent with the SEM images. In particular, the methane production rate increased 24-fold when the feed sludge and recirculated sludge were pretreated together. Collectively, the HED experiments performed with sludge recirculation and thermal-alkaline treatment demonstrated that the HED systems can be successfully employed for highly efficient sewage sludge reduction and methane gas production. Copyright © 2013 Elsevier Ltd. All rights reserved.

Improved glucose-neopentyl glycol (GNG) amphiphiles for membrane protein solubilization and stabilization.

PubMed

Cho, Kyung Ho; Bae, Hyoung Eun; Das, Manabendra; Gellman, Samuel H; Chae, Pil Seok

2014-02-01

Membrane proteins are inherently amphipathic and undergo dynamic conformational changes for proper function within native membranes. Maintaining the functional structures of these biomacromolecules in aqueous media is necessary for structural studies but difficult to achieve with currently available tools, thus necessitating the development of novel agents with favorable properties. This study introduces several new glucose-neopentyl glycol (GNG) amphiphiles and reveals some agents that display favorable behaviors for the solubilization and stabilization of a large, multi-subunit membrane protein assembly. Furthermore, a detergent structure-property relationship that could serve as a useful guideline for the design of novel amphiphiles is discussed. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Thyroid peroxidase of the pig, dog, rat, and mouse. Solubilization and identification of isozymes by isoelectric focusing

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gonzalez-Lama, Z.; Feinstein, R.N.

Dog and pig thyroid peroxidase, which exist naturally in a largely insoluble form, can be solubilized by the use of 4 M urea, or of chlorhexidine, with small losses of total activity. In the mouse and the rat, the thyroid peroxidase occurs in a soluble form. The demonstration of these rodent thyroid peroxidases is therefore complicated by unavoidable contamination with peroxidatically acting hemoglobin and catalase; the demonstration of the presence of true peroxidase was achieved by isoelectric focusing on polyacrylamide gel slabs, which separates the various factors, and by the use of the catalase and peroxidase inhibitor 3-amino-1,2,4-triazole.

Protein recovery from rainbow trout (Oncorhynchus mykiss) processing byproducts via isoelectric solubilization/precipitation and its gelation properties as affected by functional additives.

PubMed

Chen, Yi-Chen; Jaczynski, Jacek

2007-10-31

Solubility of rainbow trout proteins was determined between pH 1.5 and 13.0 and various ionic strengths (IS). Minimum solubility occurred at pH 5.5; however, when IS = 0.2, the minimum solubility shifted toward more acidic pH. Isoelectric solubilization/precipitation was applied to trout processing byproducts (fish meat left over on bones, head, skin, etc.), resulting in protein recovery yields (Kjeldahl, dry basis) between 77.7% and 89.0%, depending of the pH used for solubilization and precipitation. The recovered protein contained 1.4-2.1% ash (dry basis), while the trout processing byproducts (i.e., starting material) 13.9%. Typical boneless and skinless trout fillets contain 5.5% ash, and therefore, the isoelectric solubilization/precipitation effectively removed impurities such as bones, scales, skin, etc., from the trout processing byproducts. The recovered proteins retained gel-forming ability as assessed with dynamic rheology, torsion test, and texture profile analysis (TPA). However, the recovered proteins failed to gel unless beef plasma protein (BPP) was added. Even with BPP, the recovered protein showed some proteolysis between 40 and 55 degrees C. Addition of potato starch, transglutaminase, and phosphate to the recovered proteins resulted in good texture of trout gels as confirmed by torsion test and TPA. Higher ( P < 0.05) shear stress and strain were measured for gels developed from basic pH treatments than the acidic counterparts. However, proteins recovered from acidic treatments had higher ( P < 0.05) lipid content than the basic treatments. This is probably why the gels from acidic treatments were whiter ( L* - 3 b*) ( P < 0.05) than those from the basic ones. Our study demonstrates that functional proteins can be efficiently recovered from low-value fish processing byproducts using isoelectric solubilization/precipitation and subsequently be used in value-added human foods.

Cocrystal Solubilization in Biorelevant Media and its Prediction from Drug Solubilization.

PubMed

Lipert, Maya P; Roy, Lilly; Childs, Scott L; Rodríguez-Hornedo, Naír

2015-12-01

This work examines cocrystal solubility in biorelevant media (FeSSIF, fed-state simulated intestinal fluid), and develops a theoretical framework that allows for the simple and quantitative prediction of cocrystal solubilization from drug solubilization. The solubilities of four hydrophobic drugs and seven cocrystals containing these drugs were measured in FeSSIF and in acetate buffer at pH 5.00. In all cases, the cocrystal solubility (Scocrystal ) was higher than the drug solubility (Sdrug ) in both buffer and FeSSIF; however, the solubilization ratio of drug, SRdrug = (SFeSSIF /Sbuffer )drug , was not the same as the solubilization ratio of cocrystal, SRcocrystal = (SFeSSIF /Sbuffer )cocrystal , meaning drug and cocrystal were not solubilized to the same extent in FeSSIF. This highlights the potential risk of anticipating cocrystal behavior in biorelevant media based on solubility studies in water. Predictions of SRcocrystal from simple equations based only on SRdrug were in excellent agreement with measured values. For 1:1 cocrystals, the cocrystal solubilization ratio (SR) can be obtained from the square root of the drug SR. For 2:1 cocrystals, SRcocrystal is found from (SRdrug )(2/3) . The findings in FeSSIF can be generalized to describe cocrystal behavior in other systems involving preferential solubilization of a drug such as surfactants, lipids, and other drug solubilizing media. © 2015 Wiley Periodicals, Inc. and the American Pharmacists Association.

High pH solubilization and chromatography-based renaturation and purification of recombinant human granulocyte colony-stimulating factor from inclusion bodies.

PubMed

Li, Ming; Fan, Hua; Liu, Jiahua; Wang, Minhong; Wang, Lili; Wang, Chaozhan

2012-03-01

Recombinant human granulocyte colony-stimulating factor (rhG-CSF) is a very efficient therapeutic protein drug which has been widely used in human clinics to treat cancer patients suffering from chemotherapy-induced neutropenia. In this study, rhG-CSF was solubilized from inclusion bodies by using a high-pH solution containing low concentration of urea. It was found that solubilization of the rhG-CSF inclusion bodies greatly depended on the buffer pH employed; alkalic pH significantly favored the solubilization. In addition, when small amount of urea was added to the solution at high pH, the solubilization was further enhanced. After solubilization, the rhG-CSF was renatured with simultaneous purification by using weak anion exchange, strong anion exchange, and hydrophobic interaction chromatography, separately. The results indicated that the rhG-CSF solubilized by the high-pH solution containing low concentration of urea had much higher mass recovery than the one solubilized by 8 M urea when using anyone of the three refolding methods employed in this work. In the case of weak anion exchange chromatography, the high pH solubilized rhG-CSF could get a mass recovery of 73%. The strategy of combining solubilization of inclusion bodies at high pH with refolding of protein using liquid chromatography may become a routine method for protein production from inclusion bodies.

Regulation of coal polymer degradation by fungi. Tenth Quartery report, October 1996--December 1996

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvine, R.L.; Bumpus, J.A.

1997-01-28

It has long been known that low rank coal such as leonardite can be solubilized by strong base (>pH 12). Recent discoveries have also shown that leonardite is solubilized by Lewis bases at considerably lower pH values and by fungi that secrete certain Lewis bases (i.e., oxalate ion). During the current reporting period we have studied the ability of a strong base (sodium hydroxide, pH 12), and two fungi, Phanerochaete chrysosporium and Trametes versicolor, to solubilize Argonne Premium Coals. In general, Argonne Premium Coals were relatively resistant to base mediated solubilization. However, when these coals were preoxidized (150{degrees}C for sevenmore » days), substantial amounts of several coals were solubilized. Most affected were the Lewiston-Stockton bituminous coal, the Beulah-Zap lignite, the Wyodak-Anderson subbituminous coal and the Blind Canyon bituminous coal. Argonne Premium Coals were previously shown by us to be relatively resistant to solubilization by sodium oxalate. When preoxidized coals were treated with sodium oxalate, only the Beulah-Zap lignite was substantially solubilized. Although very small amounts of the other preoxidized coals were solubilized by treatment with oxalate, the small amount of solubilization that did take place was generally increased relative to that observed for coals that were not preoxidized. None of the Argonne Premium Coals were solubilized by P. chrysosporium or T. versicolor. Of considerable interest, however, is the observation that P. chrysosporium and T. versicolor mediated extensive solubilization of Lewiston-Stockton bituminous coal, the Beulah-Zap lignite and the Wyodak-Anderson subbituminous coal.« less

Batch and semi-continuous anaerobic co-digestion of goose manure with alkali solubilized wheat straw: A case of carbon to nitrogen ratio and organic loading rate regression optimization.

PubMed

Hassan, Muhammad; Ding, Weimin; Umar, Muhammad; Rasool, Ghulam

2017-04-01

The present study focused on carbon to nitrogen ratio (C/N) and organic loading rate (OLR) optimization of goose manure (GM) and wheat straw (WS). Dealing the anaerobic digestion of poultry manure on industrial scale; the question of optimum C/N (mixing ratio) and OLR (daily feeding concentration) have significant importance still lack in literature. Therefore, Batch and CSTR co-digestion experiments of the GM and WS were carried out at mesophilic condition. The alkali (NaOH) solubilization pretreatment for the WS had greatly enhanced its anaerobic digestibility. The highest methane production was evaluated between the C/N of 20-30 during Batch experimentation while for CSTRs; the second applied OLR of (3g.VS/L.d) was proved as the optimum with maximum methane production capability of 254.65ml/g.VS for reactor B at C/N of 25. The C/N and OLR regression optimization models were developed for their commercial scale usefulness. Copyright © 2017 Elsevier Ltd. All rights reserved.

Microwave treatment of dairy manure for resource recovery: Reaction kinetics and energy analysis.

PubMed

Srinivasan, Asha; Liao, Ping H; Lo, Kwang V

2016-12-01

A newly designed continuous-flow 915 MHz microwave wastewater treatment system was used to demonstrate the effectiveness of the microwave enhanced advanced oxidation process (MW/H 2 O 2 -AOP) for treating dairy manure. After the treatment, about 84% of total phosphorus and 45% of total chemical oxygen demand were solubilized with the highest H 2 O 2 dosage (0.4% H 2 O 2 per %TS). The reaction kinetics of soluble chemical oxygen demand revealed activation energy to be in the range of 5-22 kJ mole -1 . The energy required by the processes was approximately 0.16 kWh per liter of dairy manure heated. A higher H 2 O 2 dosage used in the system had a better process performance in terms of solids solubilization, reaction kinetics, and energy consumption. Cost-benefit analysis for a farm-scale MW/H 2 O 2 -AOP treatment system was also presented. The results obtained from this study would provide the basic knowledge for designing an effective farm-scale dairy manure treatment system.

Biological solubilization of low-rank coal

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cohen, M.S.

1991-07-01

Low-ranked coals have been solubilized using cell-free extracts derived from liquid cultures of the white-rot fungus Trametes versicolor. The coal solubilizing agent (CSA) has been separated from the broth components and purified by several analytical techniques including rotary evaporation, reverse osmosis, and solvent extraction. The recrystallized CSA retains coal solubilizing activity. Results from polarography, FTIR, and x-ray crystallography confirm that the purified CSA crystals responsible for coal-solubilization are ammonium oxalate monohydrate. The mechanism of solubilization has been deduced to involve removal of divalent cations (particularly iron FE(III)) from low-rank coals. This is followed by dissolution of the macromolecular coal structure.more » 38 figs., 9 tabs.« less

Physical characterization and antioxidant activity of thymol solubilized Tween 80 micelles

PubMed Central

Deng, Ling-Li; Taxipalati, Maierhaba; Que, Fei; Zhang, Hui

2016-01-01

Attempts were made to solubilize thymol in Tween 80 micelle to study the solubilization mechanism of thymol and the effect of solubilization on its antioxidant activity. The maximum solubilized concentration of thymol in a 2.0% (w/v) Tween 80 micelle solution is 0.2 wt%. There was no significant difference in Z-average diameter between the empty micelles and thymol solubilized micelles. 1H NMR spectra indicated that 3-H and 4-H on the benzene ring of thymol interacted with the ester group between the hydrophilic head group and the hydrophobic tail group of Tween 80 by Van der Waals’ force. Ferric reducing antioxidant potential (FRAP) and cupric ion reducing antioxidant capacity (CUPRAC) assays showed that the reducing antioxidant activity of free thymol did not change after solubilized in Tween 80 micelles. Compared to free thymol, the solubilized thymol showed higher activities to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) and hydroxyl radicals. The present study suggested a possible preparation of thymol-carrying micelles with enhanced antioxidant activities that could be applied in food beverages. PMID:27905567

Solubilization of octane in cationic surfactant-anionic polymer complexes: Effect of ionic strength.

PubMed

Zhang, Hui; Deng, Lingli; Sun, Ping; Que, Fei; Weiss, Jochen

2016-01-01

Polymers may alter the ability of oppositely charged surfactant micelles to solubilize hydrophobic molecules depending on surfactant-polymer interactions. This study was conducted to investigate the effect of ionic strength on the solubilization thermodynamics of an octane oil-in-water emulsion in mixtures of an anionic polymer (carboxymethyl cellulose) and cationic cetyltrimethylammonium bromide (CTAB) surfactant micelles using isothermal titration calorimetry (ITC). Results indicated that the CTAB binding capacity of carboxymethyl cellulose increased with increasing NaCl concentrations up to 100 mM, and the thermodynamic behavior of octane solubilization in CTAB micelles, either in the absence or presence of polymer, was found to have a strong dependence on ionic strength. The increasing ionic strength caused the solubilization in CTAB micelles to be less endothermic or even exothermic, but increased the solubilization capacity. Based on the phase separation model, the solubilization was suggested to be driven by enthalpy. It is indicated that increasing ionic strength gave rise to a larger Gibbs energy decrease but a smaller unfavorable entropy increase for octane solubilization in cationic surfactant micelles. Copyright © 2015 Elsevier Inc. All rights reserved.

Physical characterization and antioxidant activity of thymol solubilized Tween 80 micelles.

PubMed

Deng, Ling-Li; Taxipalati, Maierhaba; Que, Fei; Zhang, Hui

2016-12-01

Attempts were made to solubilize thymol in Tween 80 micelle to study the solubilization mechanism of thymol and the effect of solubilization on its antioxidant activity. The maximum solubilized concentration of thymol in a 2.0% (w/v) Tween 80 micelle solution is 0.2 wt%. There was no significant difference in Z-average diameter between the empty micelles and thymol solubilized micelles. 1 H NMR spectra indicated that 3-H and 4-H on the benzene ring of thymol interacted with the ester group between the hydrophilic head group and the hydrophobic tail group of Tween 80 by Van der Waals' force. Ferric reducing antioxidant potential (FRAP) and cupric ion reducing antioxidant capacity (CUPRAC) assays showed that the reducing antioxidant activity of free thymol did not change after solubilized in Tween 80 micelles. Compared to free thymol, the solubilized thymol showed higher activities to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) and hydroxyl radicals. The present study suggested a possible preparation of thymol-carrying micelles with enhanced antioxidant activities that could be applied in food beverages.

Physical characterization and antioxidant activity of thymol solubilized Tween 80 micelles

NASA Astrophysics Data System (ADS)

Deng, Ling-Li; Taxipalati, Maierhaba; Que, Fei; Zhang, Hui

2016-12-01

Attempts were made to solubilize thymol in Tween 80 micelle to study the solubilization mechanism of thymol and the effect of solubilization on its antioxidant activity. The maximum solubilized concentration of thymol in a 2.0% (w/v) Tween 80 micelle solution is 0.2 wt%. There was no significant difference in Z-average diameter between the empty micelles and thymol solubilized micelles. 1H NMR spectra indicated that 3-H and 4-H on the benzene ring of thymol interacted with the ester group between the hydrophilic head group and the hydrophobic tail group of Tween 80 by Van der Waals’ force. Ferric reducing antioxidant potential (FRAP) and cupric ion reducing antioxidant capacity (CUPRAC) assays showed that the reducing antioxidant activity of free thymol did not change after solubilized in Tween 80 micelles. Compared to free thymol, the solubilized thymol showed higher activities to scavenge DPPH (2,2-diphenyl-1-picrylhydrazyl) and hydroxyl radicals. The present study suggested a possible preparation of thymol-carrying micelles with enhanced antioxidant activities that could be applied in food beverages.

Hydrolysis of biomass material

DOEpatents

Schmidt, Andrew J.; Orth, Rick J.; Franz, James A.; Alnajjar, Mikhail

2004-02-17

A method for selective hydrolysis of the hemicellulose component of a biomass material. The selective hydrolysis produces water-soluble small molecules, particularly monosaccharides. One embodiment includes solubilizing at least a portion of the hemicellulose and subsequently hydrolyzing the solubilized hemicellulose to produce at least one monosaccharide. A second embodiment includes solubilizing at least a portion of the hemicellulose and subsequently enzymatically hydrolyzing the solubilized hemicellulose to produce at least one monosaccharide. A third embodiment includes solubilizing at least a portion of the hemicellulose by heating the biomass material to greater than 110.degree. C. resulting in an aqueous portion that includes the solubilized hemicellulose and a water insoluble solids portion and subsequently separating the aqueous portion from the water insoluble solids portion. A fourth embodiment is a method for making a composition that includes cellulose, at least one protein and less than about 30 weight % hemicellulose, the method including solubilizing at least a portion of hemicellulose present in a biomass material that also includes cellulose and at least one protein and subsequently separating the solubilized hemicellulose from the cellulose and at least one protein.

Visualization of Detergent Solubilization of Membranes: Implications for the Isolation of Rafts

PubMed Central

Garner, Ashley E.; Smith, D. Alastair; Hooper, Nigel M.

2008-01-01

Although different detergents can give rise to detergent-resistant membranes of different composition, it is unclear whether this represents domain heterogeneity in the original membrane. We compared the mechanism of action of five detergents on supported lipid bilayers composed of equimolar sphingomyelin, cholesterol, and dioleoylphosphatidylcholine imaged by atomic force microscopy, and on raft and nonraft marker proteins in live cells imaged by confocal microscopy. There was a marked correlation between the detergent solubilization of the cell membrane and that of the supported lipid bilayers. In both systems Triton X-100 and CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) distinguished between the nonraft liquid-disordered (ld) and raft liquid ordered (lo) lipid phases by selectively solubilizing the ld phase. A higher concentration of Lubrol was required, and not all the ld phase was solubilized. The solubilization by Brij 96 occurred by a two-stage mechanism that initially resulted in the solubilization of some ld phase and then progressed to the solubilization of both ld and lo phases simultaneously. Octyl glucoside simultaneously solubilized both lo and ld phases. These data show that the mechanism of membrane solubilization is unique to an individual detergent. Our observations have significant implications for using different detergents to isolate membrane rafts from biological systems. PMID:17933878

Visualization of detergent solubilization of membranes: implications for the isolation of rafts.

PubMed

Garner, Ashley E; Smith, D Alastair; Hooper, Nigel M

2008-02-15

Although different detergents can give rise to detergent-resistant membranes of different composition, it is unclear whether this represents domain heterogeneity in the original membrane. We compared the mechanism of action of five detergents on supported lipid bilayers composed of equimolar sphingomyelin, cholesterol, and dioleoylphosphatidylcholine imaged by atomic force microscopy, and on raft and nonraft marker proteins in live cells imaged by confocal microscopy. There was a marked correlation between the detergent solubilization of the cell membrane and that of the supported lipid bilayers. In both systems Triton X-100 and CHAPS (3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate) distinguished between the nonraft liquid-disordered (l(d)) and raft liquid ordered (l(o)) lipid phases by selectively solubilizing the l(d) phase. A higher concentration of Lubrol was required, and not all the l(d) phase was solubilized. The solubilization by Brij 96 occurred by a two-stage mechanism that initially resulted in the solubilization of some l(d) phase and then progressed to the solubilization of both l(d) and l(o) phases simultaneously. Octyl glucoside simultaneously solubilized both l(o) and l(d) phases. These data show that the mechanism of membrane solubilization is unique to an individual detergent. Our observations have significant implications for using different detergents to isolate membrane rafts from biological systems.

Solubilization of bovine corpus-luteum adenylate cyclase in lubrol-PX, triton X-100 or digitonin and the stabilizing effect of sodium fluoride present in the solubilization medium.

PubMed

Young, J L; Stansfield, D A

1978-09-01

1. Adenylate cyclase activity of the washed 600g sediment of bovine corpus-luteum homogenate was solubilized by Lubrol-PX, Triton X-100 and digitonin. Digitonin was the least destructive of NaF-stimulated activity. 2. NaF, present in the solubilization medium together with MgSO4, increased the percentage yields of soluble activity from untreated 600g sediment and 600g sediment which had been preincubated with p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate). The stabilizing influence of NaF was most marked with digitonin. However, the highest specific activities of soluble enzyme were obtained with Lubrol-PX as solubilizing agent, since digitonin solubilized more membrane protein than does Lubrol-PX, and less of the activity of the digitonin-dispersed 600g sediment was recovered in the 105000g supernatant. 3. p[NH]ppG also has a stabilizing effect when present during the solubilization, but less so than NaF. 4. Both NaF and MgSO4 alone have a stabilizing effect during solubilization. The greatest amounts of soluble activity were obtained with both agents present in the solubilization medium, there being a synergistic effect.

Solubilization of bovine corpus-luteum adenylate cyclase in lubrol-PX, triton X-100 or digitonin and the stabilizing effect of sodium fluoride present in the solubilization medium.

PubMed Central

Young, J L; Stansfield, D A

1978-01-01

1. Adenylate cyclase activity of the washed 600g sediment of bovine corpus-luteum homogenate was solubilized by Lubrol-PX, Triton X-100 and digitonin. Digitonin was the least destructive of NaF-stimulated activity. 2. NaF, present in the solubilization medium together with MgSO4, increased the percentage yields of soluble activity from untreated 600g sediment and 600g sediment which had been preincubated with p[NH]ppG (guanosine 5'-[betagamma-imido]triphosphate). The stabilizing influence of NaF was most marked with digitonin. However, the highest specific activities of soluble enzyme were obtained with Lubrol-PX as solubilizing agent, since digitonin solubilized more membrane protein than does Lubrol-PX, and less of the activity of the digitonin-dispersed 600g sediment was recovered in the 105000g supernatant. 3. p[NH]ppG also has a stabilizing effect when present during the solubilization, but less so than NaF. 4. Both NaF and MgSO4 alone have a stabilizing effect during solubilization. The greatest amounts of soluble activity were obtained with both agents present in the solubilization medium, there being a synergistic effect. PMID:568467

Haloarchaea Endowed with Phosphorus Solubilization Attribute Implicated in Phosphorus Cycle

PubMed Central

Yadav, Ajar Nath; Sharma, Divya; Gulati, Sneha; Singh, Surender; Dey, Rinku; Pal, Kamal Krishna; Kaushik, Rajeev; Saxena, Anil Kumar

2015-01-01

Archaea are unique microorganisms that are present in ecological niches of high temperature, pH and salinity. A total of 157 archaea were obtained from thirteen sediment, water and rhizospheric soil samples collected from Rann of Kutch, Gujarat, India. With an aim to screen phosphate solubilizing archaea, a new medium was designed as Haloarchaea P Solubilization (HPS) medium. The medium supported the growth and P solubilization activity of archaea. Employing the HPS medium, twenty isolates showed the P-solubilization. Phosphate solubilizing archaea were identified as seventeen distinct species of eleven genera namely Haloarcula, Halobacterium, Halococcus, Haloferax, Halolamina, Halosarcina, Halostagnicola, Haloterrigena, Natrialba, Natrinema and Natronoarchaeum. Natrinema sp. strain IARI-WRAB2 was identified as the most efficient P-solubilizer (134.61 mg/L) followed by Halococcus hamelinensis strain IARI-SNS2 (112.56 mg/L). HPLC analysis detected seven different kinds of organic acids, namely: gluconic acid, citric acid, formic acid, fumaric acid succinic acid, propionic acid and tartaric acid from the cultures of these isolates. These phosphate solubilizing halophilic archaea may play a role in P nutrition to vegetation growing in these hypersaline soils. This is the first report for these haloarchaea to solubilize considerable amount of P by production of organic acids and lowering of pH. PMID:26216440

Regulation of coal polymer degradation by fungi. Fourth quarterly progress report, May 1995--June 1995

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvine, R.L.

1995-07-24

To test the hypothesis that coal (leonardite) Solubilization and the subsequent depolymerization of the solubilized coal macromolecules are distinct events in lignin degrading fungi. In addition to T versicolor, Phanerochaete chrysosporium, another lignin degrading fungus that also has the ability to solubilize coal, will be studied. To test the hypothesis that the processes of coal (leonardite) solubilization and coal macro molecule depolymerization in lignin degrading fungi can be regulated by altering the nutritional status of the microorganism. Coal solubilization is expected to occur in nutrient rich media whereas depolymerization of solubilized coal macromolecules is expected to occur in nutrient limitedmore » media. To determine the role of extracellular enzymes (laccases, lignin peroxidases and Mn peroxidases) that are secreted by lignin degrading fungi during coal solubilization or coal macro molecule depolymerization. To assess the role of enzymatically generated oxygen radicals, non-radical active oxygen species, veratryl alcohol radicals and Mn{sup +++} complexes in coal macro molecule depolymerization. To characterize products of coal solubilization and coal macro molecule depolymerization that are formed by T. versicolor and P. chrysosporium and their respective extracellular enzymes. Solubilization products formed using oxalic acid and other metal chelators will also be characterized and compared.« less

REGULATION OF COAL POLYMER DEGRADATION BY FUNGI

DOE Office of Scientific and Technical Information (OSTI.GOV)

John A. Bumpus

1998-11-30

A variety of lignin degrading fungi mediate solubilization and subsequent biodegradation of coal macromolecules (a.k.a. coal polymer) from highly oxidized low rank coals such as leonardites. It appears that oxalate or possibly other metal chelators (i.e., certain Krebs Cycle intermediates) mediate solubilization of low rank coals while extracellular oxidases have a role in subsequent oxidation of solubilized coal macromolecule. These processes are under nutritional control. For example, in the case of P. chrysosporium, solubilization of leonardite occurred when the fungi were cultured on most but not all nutrient agars tested and subsequent biodegradation occurred only in nutrient nitrogen limited cultures.more » Lignin peroxidases mediate oxidation of coal macromolecule in a reaction that is dependent on the presence of veratryl alcohol and hydrogen peroxide. Kinetic evidence suggests that veratryl alcohol is oxidized to the veratryl alcohol cation radical which then mediates oxidation of the coal macromolecule. Results by others suggest that Mn peroxidases mediate formation of reactive Mn{sup 3+} complexes which also mediate oxidation of coal macromolecule. A biomimetic approach was used to study solubilization of a North Dakota leonardite. It was found that a concentration {approximately}75 mM sodium oxalate was optimal for solubilization of this low rank coal. This is important because this is well above the concentration of oxalate produced by fungi in liquid culture. Higher local concentrations probably occur in solid agar cultures and thus may account for the observation that greater solubilization occurs in agar media relative to liquid media. The characteristics of biomimetically solubilized leonardite were similar to those of biologically solubilized leonardite. Perhaps our most interesting observation was that in addition to oxalate, other common Lewis bases (phosphate/hydrogen phosphate/dihydrogen phosphate and bicarbonate/carbonate ions) are able to mediate substantial solubilization of leonardite at physiological pH values. Lastly, we present evidence that some fungi appear to possess coal solubilization ability in which the initial events of solubilization is not mediated by oxalate ion.« less

Sub-CMC solubilization of dodecane by rhamnolipid in saturated porous media

NASA Astrophysics Data System (ADS)

Yang, X.; Zhong, H.; Zhang, H.; Brusseau, M. L.

2016-12-01

Sub-CMC solubilization of dodecane by rhamnolipid in saturated porous mediaXin Yang1,Hua Zhong1, 2, 3 *, Hui Zhang1, Mark L Brusseau31 College of Environmental Science and Engineering, Hunan University, Changsha 410082, China;2 School of Water Resources and Hydropower Engineering, Wuhan University, Wuhan 430072, China;3 Department of Soil, Water and Environmental Science, University of Arizona, Tucson, Arizona 85721;*Corresponding author, E-mail: zhonghua@hnu.edu.cn, Tel: +86-731-88664182Purpose: Investigate solubilization of dodecane by monorhamnolipid at sub-CMC concentrations in porous media under dynamic flow conditions. Testify aggregate formation mechanism for the solubilization. Methods:One-dimension column experiment was implemented to test dodecane solubilization in glass beads by rhamnolipid at sub-CMC concentrations, and the effect of solubilization on the residual NAPL morphology was examined using X-ray tomography. A two-dimension flow cell was used to examine mobilization and solubilization of dodecane in quartz sand by sub-CMC rhamnolipid. The result of solubilization was compared to that of two synthetic surfactants, SDBS and Triton X-100, and a solvent, ethanol. Size, zeta potential and the morphology of particles in the effluent were also examined. Results: Results of the column and 2-D flow cell studies show enhancement of dodecane solubility by sub-CMC monorhamnolipid in the porous medium. Retention of rhamnolipid and detection of nano-size aggregates show that the solubilization is based on a sub-CMC aggregate-formation mechanism. The rhamnolipid is more efficient for the solubilization compared to the synthetic surfactants and ethanol, and significant solubilization could occur at a rhamnolipid concentration that did not cause mobilization. Conclusions:Results of the study demonstrate the aggregate-based solubilization of dodecane in porous media by rhamnolipid at sub-CMC concentrations. These results indicate a strategy of employing low concentrations of rhamnolipid for surfactant-enhanced aquifer remediation (SEAR), which may promote the cost-effectiveness of rhamnolipid application and overcome the drawbacks of using surfactants at hyper-CMC concentrations.

Recovery of bioactive protein from bacterial inclusion bodies using trifluoroethanol as solubilization agent.

PubMed

Upadhyay, Vaibhav; Singh, Anupam; Jha, Divya; Singh, Akansha; Panda, Amulya K

2016-06-08

Formation of inclusion bodies poses a major hurdle in recovery of bioactive recombinant protein from Escherichia coli. Urea and guanidine hydrochloride have routinely been used to solubilize inclusion body proteins, but many times result in poor recovery of bioactive protein. High pH buffers, detergents and organic solvents like n-propanol have been successfully used as mild solubilization agents for high throughput recovery of bioactive protein from bacterial inclusion bodies. These mild solubilization agents preserve native-like secondary structures of proteins in inclusion body aggregates and result in improved recovery of bioactive protein as compared to conventional solubilization agents. Here we demonstrate solubilization of human growth hormone inclusion body aggregates using 30% trifluoroethanol in presence of 3 M urea and its refolding into bioactive form. Human growth hormone was expressed in E. coli M15 (pREP) cells in the form of inclusion bodies. Different concentrations of trifluoroethanol with or without addition of low concentration (3 M) of urea were used for solubilization of inclusion body aggregates. Thirty percent trifluoroethanol in combination with 3 M urea was found to be suitable for efficient solubilization of human growth hormone inclusion bodies. Solubilized protein was refolded by dilution and purified by anion exchange and size exclusion chromatography. Purified protein was analyzed for secondary and tertiary structure using different spectroscopic tools and was found to be bioactive by cell proliferation assay. To understand the mechanism of action of trifluoroethanol, secondary and tertiary structure of human growth hormone in trifluoroethanol was compared to that in presence of other denaturants like urea and guanidine hydrochloride. Trifluoroethanol was found to be stabilizing the secondary structure and destabilizing the tertiary structure of protein. Finally, it was observed that trifluoroethanol can be used to solubilize inclusion bodies of a number of proteins. Trifluoroethanol was found to be a suitable mild solubilization agent for bacterial inclusion bodies. Fully functional, bioactive human growth hormone was recovered in high yield from inclusion bodies using trifluoroethanol based solubilization buffer. It was also observed that trifluoroethanol has potential to solubilize inclusion bodies of different proteins.

Solubilization of poorly water-soluble drug carbamezapine in pluronic micelles: effect of molecular characteristics, temperature and added salt on the solubilizing capacity.

PubMed

Kadam, Yogesh; Yerramilli, Usha; Bahadur, Anita

2009-08-01

The solubilization of a poorly water-soluble antiepileptic drug, carbamazepine (CBZ), in a series of micelle-forming PEO-PPO-PEO block copolymers with combinations of blocks having different molecular weight was studied. The drug solubility and micelle-water partition coefficient (P) were determined using UV-vis spectroscopy. Dynamic light scattering on copolymer solutions was used to measure size and polydispersity of nanoaggregates. Solubilization of carbamezapine increased with the rise in temperature and concentration of block copolymers, but no significant increase was observed with added salt (NaCl). The solubilization is also discussed from a thermodynamics viewpoint, by considering the standard free energy of solubilization (DeltaG degrees ).

Characterization of Protein Detergent Complexes by NMR, Light Scattering, and Analytical Ultracentrifugation

PubMed Central

Maslennikov, Innokentiy; Krupa, Martin; Dickson, Christopher; Esquivies, Luis; Blain, Katherine; Kefala, Georgia; Choe, Senyon; Kwiatkowski, Witek

2009-01-01

Bottlenecks in expression, solubilization, purification and crystallization hamper the structural study of integral membrane proteins (IMPs). Successful crystallization is critically dependent on the purity, stability and oligomeric homogeneity of an IMP sample. These characteristics are in turn strongly influenced by the type and concentration of the detergents used in IMP preparation. By utilizing the techniques and analytical tools we earlier developed for the characterization of protein-detergent complexes (PDCs) (Maslennikov et al., 2007), we demonstrate that for successful protein extraction from E. coli membrane fractions, the solubilizing detergent associates preferentially to IMPs rather than to membrane lipids. Notably, this result is contrary to the generally accepted mechanism of detergent-mediated IMP solubilization. We find that for one particular member of the family of proteins studied (E. coli receptor kinases, which is purified in mixed multimeric states and oligomerizes through its transmembrane region), the protein oligomeric composition is largely unaffected by a 10-fold increase in protein concentration, by alteration of micelle properties through addition of other detergents to the PDC sample, or by a 20-fold variation in the detergent concentration used for solubilization of the IMP from the membrane. We observed that the conditions used for expression of the IMP, which impact protein density in the membrane, has the greatest influence on the IMP oligomeric structure. Finally, we argue that for concentrating PDCs smaller than 30 kDa, stirred concentration cells are less prone to over-concentration of detergent and are therefore more effective than centrifugal ultrafiltration devices. PMID:19214777

Microbial solubilization of coal

DOEpatents

Strandberg, Gerald W.; Lewis, Susan N.

1990-01-01

This invention deals with the solubilization of coal using species of Streptomyces. Also disclosed is an extracellular component from a species of Streptomyces, said component being able to solubilize coal.

Tuning polarity and improving charge transport in organic semiconductors

NASA Astrophysics Data System (ADS)

Oh, Joon Hak; Han, A.-Reum; Yu, Hojeong; Lee, Eun Kwang; Jang, Moon Jeong

2013-09-01

Although state-of-the-art ambipolar polymer semiconductors have been extensively reported in recent years, highperformance ambipolar polymers with tunable dominant polarity are still required to realize on-demand, target-specific, high-performance organic circuitry. Herein, dithienyl-diketopyrrolopyrrole (TDPP)-based polymer semiconductors with engineered side-chains have been synthesized, characterized and employed in ambipolar organic field-effect transistors, in order to achieve controllable and improved electrical properties. Thermally removable tert-butoxycarbonyl (t-BOC) groups and hybrid siloxane-solubilizing groups are introduced as the solubilizing groups, and they are found to enable the tunable dominant polarity and the enhanced ambipolar performance, respectively. Such outstanding performance based on our molecular design strategies makes these ambipolar polymer semiconductors highly promising for low-cost, large-area, and flexible electronics.

Advanced oxidation process using hydrogen peroxide/microwave system for solubilization of phosphate.

PubMed

Liao, Ping Huang; Wong, Wayne T; Lo, Kwang Victor

2005-01-01

An advanced oxidation process (AOP) combining hydrogen peroxide and microwave heating was used for the solubilization of phosphate from secondary municipal sludge from an enhanced biological phosphorus removal process. The microwave irradiation is used as a generator agent of oxidizing radicals as well as a heating source in the process. This AOP process could facilitate the release of a large amount of the sludge-bound phosphorus from the sewage sludge. More than 84% of the total phosphorous could be released at a microwave heating time of 5 min at 170 degrees C. This innovative process has the potential of being applied to simple sludge treatment processes in domestic wastewater treatment and to the recovery of phosphorus from the wastewater.

The solubilization of low-ranked coals by microorganisms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Strandberg, G.W.

1987-07-09

Late in 1984, our Laboratory was funded by the Pittsburgh Energy Technology Center, US Department of Energy, to investigate the potential utility of microorganisms for the solubilization of low-ranked coals. Our approach has been multifacited, including studies of the types of microorganisms involved, appropriate conditions for their growth and coal-solubilization, the suceptibility of different coals to microbial action, the chemical and physical nature of the product, and potential bioprocess designs. A substantial number of fungal species have been shown to be able to solubilize coal. Cohen and Gabrielle reported that two lignin-degrading fungi, Polyporous (Trametes) versicolor and Poria monticola couldmore » solubilize lignite. Ward has isolated several diverse fungi from nature which are capable of degrading different lignites, and our Laboratory has isolated three coal-solubilizing fungi which were found growing on a sample of Texas lignite. The organisms we studied are shown in Table 1. The perceived significance of lignin degradation led us to examine two lignin-degrading strains of the genus Streptomyces. As discussed later, these bacteria were capable of solubilizing coal; but, in the case of at least one, the mechanism was non-enzymatic. The coal-solubilizing ability of other strains of Streptomyces was recently reported. Fakoussa and Trueper found evidence that a strain of Pseudomonas was capble of solubizing coal. It would thus appear that a diverse array of microorganisms possess the ability to solubilize coal. 16 refs.« less

Sewage sludge solubilization by high-pressure homogenization.

PubMed

Zhang, Yuxuan; Zhang, Panyue; Guo, Jianbin; Ma, Weifang; Fang, Wei; Ma, Boqiang; Xu, Xiangzhe

2013-01-01

The behavior of sludge solubilization using high-pressure homogenization (HPH) treatment was examined by investigating the sludge solid reduction and organics solubilization. The sludge volatile suspended solids (VSS) decreased from 10.58 to 6.67 g/L for the sludge sample with a total solids content (TS) of 1.49% after HPH treatment at a homogenization pressure of 80 MPa with four homogenization cycles; total suspended solids (TSS) correspondingly decreased from 14.26 to 9.91 g/L. About 86.15% of the TSS reduction was attributed to the VSS reduction. The increase of homogenization pressure from 20 to 80 MPa or homogenization cycle number from 1 to 4 was favorable to the sludge organics solubilization, and the protein and polysaccharide solubilization linearly increased with the soluble chemical oxygen demand (SCOD) solubilization. More proteins were solubilized than polysaccharides. The linear relationship between SCOD solubilization and VSS reduction had no significant change under different homogenization pressures, homogenization cycles and sludge solid contents. The SCOD of 1.65 g/L was solubilized for the VSS reduction of 1.00 g/L for the three experimental sludge samples with a TS of 1.00, 1.49 and 2.48% under all HPH operating conditions. The energy efficiency results showed that the HPH treatment at a homogenization pressure of 30 MPa with a single homogenization cycle for the sludge sample with a TS of 2.48% was the most energy efficient.

Evaluation of the effect of temperature, NaOH concentration and time on solubilization of palm oil mill effluent (POME) using response surface methodology (RSM).

PubMed

Chou, K W; Norli, I; Anees, A

2010-11-01

In this study, palm oil mill effluent (POME) was solubilized by batch thermo-alkaline pre-treatments. A three-factor central composite design (CCD) was applied to identify the optimum COD solubilization condition. The individual and interactive effects of three factors, temperature, NaOH concentration and reaction time, on solubilization of POME were evaluated by employing response surface methodology (RSM). The experimental results showed that temperature, NaOH concentration and reaction time all had an individual significant effect on the solubilization of POME. But these three factors were independent, or there was insignificant interaction on the response. The maximum COD solubilization of 82.63% was estimated under the optimum condition at 32.5 degrees C, 8.83g/L of NaOH and 41.23h reaction time. The confirmation experiment of the predicted optimum conditions verified that the RSM with the central composite design was useful for optimizing the solubilization of POME.

Food-grade microemulsions based on nonionic emulsifiers: media to enhance lycopene solubilization.

PubMed

Spernath, Aviram; Yaghmur, Anan; Aserin, Abraham; Hoffman, Roy E; Garti, Nissim

2002-11-06

Water-dilutable food-grade microemulsions consisting of ethoxylated sorbitan esters, and in some cases blended with other emulsifiers, water, (R)-(+)-limonene, ethanol, and propylene glycol, have been prepared. These microemulsions are of growing interest to the food industry as vehicles for delivering and enhancing solubilization of natural food supplements with nutritional and health benefits. Lycopene, an active natural lipophilic antioxidant from tomato, has solubilized in water-in-oil, bicontinuous, and oil-in-water types of microemulsions up to 10 times the oil [(R)-(+)-limonene] dissolution capacity. The effects of aqueous-phase dilution, nature of surfactant (hydrophilic-lypophilic balance), and mixed surfactant on solubilization capacity and solubilization efficiency were studied. Structural aspects studied by self-diffusion NMR were correlated to the solubilization capacity, and transformational structural changes were identified.

Recent progress in cell-free solubilization of coal

DOE Office of Scientific and Technical Information (OSTI.GOV)

Cohen, M.S.; Aronson, H.; Feldman, K.

1988-01-01

Low rank coal has been solubilized using cell-free filtrates separated from cultures of Polyporus versicolor. Solubilization has been obtained with neat filtrates and with fractions collected from the neat filtrates after gel permeation chromatography. The coal solubilizing enzymes have been collected in enriched fractions with gpc. This increased relative purity has allowed the determination of the average molecular weight of this enzyme by gel permeation chromatography and by polyacrylamide gel electrophoresis. Rates of coal solubilization are dependent on the size of coal particles, mass of coal, temperature, pH, concentration of the cell-free filtrate, and the concentration of several inorganic ions.

Identification of Heterotrophic Zinc Mobilization Processes among Bacterial Strains Isolated from Wheat Rhizosphere (Triticum aestivum L.).

PubMed

Costerousse, Benjamin; Schönholzer-Mauclaire, Laurie; Frossard, Emmanuel; Thonar, Cécile

2018-01-01

Soil and plant inoculation with heterotrophic zinc-solubilizing bacteria (ZSB) is considered a promising approach for increasing zinc (Zn) phytoavailability and enhancing crop growth and nutritional quality. Nevertheless, it is necessary to understand the underlying bacterial solubilization processes to predict their repeatability in inoculation strategies. Acidification via gluconic acid production remains the most reported process. In this study, wheat rhizosphere soil serial dilutions were plated on several solid microbiological media supplemented with scarcely soluble Zn oxide (ZnO), and 115 putative Zn-solubilizing isolates were directly detected based on the formation of solubilization halos around the colonies. Eight strains were selected based on their Zn solubilization efficiency and siderophore production capacity. These included one strain of Curtobacterium , two of Plantibacter , three strains of Pseudomonas , one of Stenotrophomonas , and one strain of Streptomyces In ZnO liquid solubilization assays, the presence of glucose clearly stimulated organic acid production, leading to medium acidification and ZnO solubilization. While solubilization by Streptomyces and Curtobacterium was attributed to the accumulated production of six and seven different organic acids, respectively, the other strains solubilized Zn via gluconic, malonic, and oxalic acids exclusively. In contrast, in the absence of glucose, ZnO dissolution resulted from proton extrusion (e.g., via ammonia consumption by Plantibacter strains) and complexation processes (i.e., complexation with glutamic acid in cultures of Curtobacterium ). Therefore, while gluconic acid production was described as a major Zn solubilization mechanism in the literature, this study goes beyond and shows that solubilization mechanisms vary among ZSB and are strongly affected by growth conditions. IMPORTANCE Barriers toward a better understanding of the mechanisms underlying zinc (Zn) solubilization by bacteria include the lack of methodological tools for isolation, discrimination, and identification of such organisms. Our study proposes a direct bacterial isolation procedure, which prevents the need to screen numerous bacterial candidates (for which the ability to solubilize Zn is unknown) for recovering Zn-solubilizing bacteria (ZSB). Moreover, we confirm the potential of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) as a quick and accurate tool for the identification and discrimination of environmental bacterial isolates. This work also describes various Zn solubilization processes used by wheat rhizosphere bacteria, including proton extrusion and the production of different organic acids among bacterial strains. These processes were also clearly affected by growth conditions (i.e., solid versus liquid cultures and the presence and absence of glucose). Although highlighted mechanisms may have significant effects at the soil-plant interface, these should only be transposed cautiously to real ecological situations. Copyright © 2017 American Society for Microbiology.

NMR structural and dynamical investigation of the isolated voltage-sensing domain of the potassium channel KvAP: implications for voltage gating.

PubMed

Shenkarev, Zakhar O; Paramonov, Alexander S; Lyukmanova, Ekaterina N; Shingarova, Lyudmila N; Yakimov, Sergei A; Dubinnyi, Maxim A; Chupin, Vladimir V; Kirpichnikov, Mikhail P; Blommers, Marcel J J; Arseniev, Alexander S

2010-04-28

The structure and dynamics of the isolated voltage-sensing domain (VSD) of the archaeal potassium channel KvAP was studied by high-resolution NMR. The almost complete backbone resonance assignment and partial side-chain assignment of the (2)H,(13)C,(15)N-labeled VSD were obtained for the protein domain solubilized in DPC/LDAO (2:1) mixed micelles. Secondary and tertiary structures of the VSD were characterized using secondary chemical shifts and NOE contacts. These data indicate that the spatial structure of the VSD solubilized in micelles corresponds to the structure of the domain in an open state of the channel. NOE contacts and secondary chemical shifts of amide protons indicate the presence of tightly bound water molecule as well as hydrogen bond formation involving an interhelical salt bridge (Asp62-R133) that stabilizes the overall structure of the domain. The backbone dynamics of the VSD was studied using (15)N relaxation measurements. The loop regions S1-S2 and S2-S3 were found mobile, while the S3-S4 loop (voltage-sensor paddle) was found stable at the ps-ns time scale. The moieties of S1, S2, S3, and S4 helices sharing interhelical contacts (at the level of the Asp62-R133 salt bridge) were observed in conformational exchange on the micros-ms time scale. Similar exchange-induced broadening of characteristic resonances was observed for the VSD solubilized in the membrane of lipid-protein nanodiscs composed of DMPC, DMPG, and POPC/DOPG lipids. Apparently, the observed interhelical motions represent an inherent property of the VSD of the KvAP channel and can play an important role in the voltage gating.

Preliminary characterization of Thy-1.1 and Ag-B antigens from rat tissues solubilized in detergents

PubMed Central

Letarte-Muirhead, Michelle; Acton, Ronald T.; Williams, Alan F.

1974-01-01

1. A radioactive binding assay for Thy-1.1 alloantigen which functions in the presence of detergents was established by using glutaraldehyde-fixed thymocytes as target cells. Thy-1.1 activity in detergent extracts was then assayed by measuring inhibition of the binding assay. 2. Solubilization of Thy-1.1 from whole thymocytes, and their membranes by a large number of non-ionic detergents and deoxycholate was studied. In the same extracts Ag-B(4) histocompatibility antigenic activities were measured. With the exception of Nonidet P-40, the detergents did not affect the antigenicity of Thy-1.1, but only Lubrol-PX and deoxycholate gave effective solubilization as measured by activity remaining in the supernatant after centrifugation at 200000g for 40min. With Ag-B(4) antigen, Triton X-100, Triton X-67 and Nonidet P-40 gave effective solubilization as well as Lubrol-PX and deoxycholate. Solubilization of Thy-1.1 activity from leukaemia cells and a brain homogenate was also studied, but none of the non-ionic detergents gave satisfactory results with these tissues. 3. Extracts from thymocyte membranes were further examined by gel filtration and sucrose gradient centrifugation. The Thy-1.1 activity behaved as a single component in deoxycholate with a density similar to that of a globular protein, but in Lubrol-PX the antigen was contained in a low-density complex. In Lubrol-PX extracts Ag-B(4) was also found in aggregates not observed in deoxycholate. 4. The s20,w values for Thy-1.1 and Ag-B(4) antigens in deoxycholate were 2.4 and 4.4, and v̄ values were 0.70 and 0.75 respectively. The Stokes radius observed for Thy-1.1 was 3.1nm and for Ag-B(4) 5.3nm. By using these values the molecular weights for the antigen–detergent complexes were calculated to be 28000 for Thy-1.1 and 100000 for Ag-B(4). PMID:4219284

Phosphate Solubilization Potentials of Rhizosphere Isolates from Central Anatolia (Turkey)

NASA Astrophysics Data System (ADS)

Ogut, M.; Er, F.

2009-04-01

Plant available-phosphorus (P) is usually low in Anatolian soils due mainly to the precipitation as calcium (Ca) and magnesium (Mg) phosphates in alkaline conditions. Phosphate solubilizing microorganisms (PSM) can enhance plant P-availability by dissolving the hardly soluble-P within the rhizosphere, which is the zone that surrounds the plant roots. PSM's can be used as seed- or soil-inocula to increase plant P-uptake and the overall growth. A total of 162 PSM's were isolated from the rhizosphere of wheat plants excavated from different fields located along a 75 km part of a highway in Turkey. The mean, the standart deviation, and the median for solubilized-P (ppm) in a 24 h culture in a tricalcium phosphate broth were 681, 427, and 400 for glucose; 358, 266, and 236 for sucrose; and 102, 117, and 50 for starch, respectively. There was not a linear relationship between the phosphate solubilized in the liquid cultures and the solubilization index obtained in the Pikovskaya's agar. Nine isolates representing both weak and strong solubilizers [Bacillus megaterium (5), Bacillus pumilis (1), Pseudomonas syringae pv. phaseolica (1), Pseudomonas fluorescens (1), Arthrobacter aurescens (1) as determined by the 16S rRNA gene sequence analysis] were further studied in a five day incubation. Pseudomonas syringae pv. phaseolica solubilized statistically (P<0.05) higher phosphate (409 ppm) than all the other strains did. There was not a statistically significant (P<0.05) difference in solubilized-P among the Bacillus strains. The pH of the medium fell to the levels between 4 and 5 from the initial neutrality. The phosphate solubilizing strains variably produced gluconic, 2-keto-D-gluconic, glycolic, acetic and butyric acids. The organic acids produced by these microorganisms seem to be the major source of phosphate solubilization in vitro.

Major factors influencing bacterial leaching of heavy metals (Cu and Zn) from anaerobic sludge.

PubMed

Couillard, D; Chartier, M; Mercier, G

1994-01-01

Anaerobically digested sewage sludges were treated for heavy metal removal through a biological solubilization process called bacterial leaching (bioleaching). The solubilization of copper and zinc from these sludges is described in this study: using continuously stirred tank reactors with and without sludge recycling at different mean hydraulic residence times (1, 2, 3 and 4 days). Significant linear equations were established for the solubilization of zinc and copper according to relevant parameters: oxygen reduction potential (ORP), pH and residence time (t). Zinc solubilization was related to the residence time with a r2 (explained variance) of 0.82. Considering only t=2 and 3 days explained variance of 0.31 and 0.24 were found between zinc solubilization as a function of ORP and pH indicating a minor importance of those two factors for this metal in the range of pH and ORP experimented. Cu solubilization was weakly correlated to mean hydraulic residence time (r2=0.48), while it was highly correlated to ORP (r2=0.80) and pH (r2=0.62) considering only t of 2 and 3 days in the case of pH and ORP. The ORP dependence of Cu solubilization has been clearly demonstrated in this study. In addition to this, the importance of the substrate concentration for Cu solubilization has been confirmed. The hypothesis of a biological solubilization of Cu by the indirect mechanism has been supported. The results permit, under optimum conditions, the drawing of linear equations which will allow prediction of metal solubilization efficiencies from the parameters pH (Cu), ORP (Cu) and residence time (Cu and Zn), during the treatment. The linear regressions will be a useful tool for routine operation of the process.

Isolation of phosphate solubilizing bacteria and their potential for lead immobilization in soil.

PubMed

Park, Jin Hee; Bolan, Nanthi; Megharaj, Mallavarapu; Naidu, Ravi

2011-01-30

Lead (Pb), a highly toxic heavy metal forms stable compounds with phosphate (P). The potential of phosphate solubilizing bacteria (PSB) to immobilize Pb by enhancing solubilization of insoluble P compounds was tested in this research. Eighteen different PSB strains isolated from P amended and Pb contaminated soils were screened for their efficiency in P solubilization. The PSB isolated from P amended soils solubilized 217-479 mg/L of P while the PSB from Pb contaminated soil solubilized 31-293 mg/L of P. Stepwise multiple regression analysis and P solubility kinetics indicated that the major mechanism of P solubilization by PSB is the pH reduction through the release of organic acids. From the isolated bacteria, two PSB were chosen for Pb immobilization and these bacteria were identified as Pantoea sp. and Enterobacter sp., respectively. The PSB significantly increased P solubilization by 25.0% and 49.9% in the case of Pantoea sp., and 63.3% and 88.6% in the case of Enterobacter sp. for 200 and 800 mg/kg of rock phosphate (RP) addition, respectively, thereby enhancing the immobilization of Pb by 8.25-13.7% in the case of Pantoea sp. and 14.7-26.4% in the case of Enterobacter sp. The ability of PSB to solubilize P, promote plant growth, and immobilize Pb can be used for phytostabilization of Pb contaminated soils. Copyright © 2010 Elsevier B.V. All rights reserved.

Modeling the Influence of Different Processes on Iron Solubilization in Mineral Aerosols: From the Gobi Desert to the North Pacific Ocean

NASA Astrophysics Data System (ADS)

Meskhidze, N.; Chameides, W. L.; Chen, G.

2002-12-01

Atmospheric transport is the only known means to deliver dissolved iron from the continents to remote oceanic areas. Dissolved iron is one of the necessary nutrients for photosynthesis of microscopic, single-celled marine organisms (phytoplankton) that grow abundantly in oceans around the world. Alteration of dissolved iron fluxes may substantially affect ocean ecosystem productivity and even exert a global-scale influence on climate by affecting the rate at which atmospheric CO2 is fixed by oceanic biota. On continents, iron mainly exists in forms of highly insoluble minerals (iron-oxides and iron-aluminosilicates) and the processes that can solubilize iron in mineral aerosols during their long-range transport remain poorly understood. In this work we attempt to elucidate the key processes that control the solubilization of iron in mineral aerosols using a simple Lagrangian box model to simulate the transport and chemical alteration of iron-containing mineral aerosols as they are transported from the east coast of China to the remote western North Pacific Ocean. Model parameters and initial conditions are set using a combination of soil and aerosol data from the Gobi Desert, as well as from measurements made during specific PEM-West B flights that encountered dust storm plumes over the Pacific Ocean that had originated in China. Our preliminary results indicate that the amount of acidic pollutants in the air along the dust transport pathways can have a significant effect on the amount of iron that is solubilized in advecting mineral aerosols. This suggests that there may be a link between the flux of the dissolved iron to the remote North Pacific Ocean and the rate at which pollutants such as sulfur dioxide, nitrogen oxides, and ammonia are emitted in East Asia.

Fluidized-bed bioreactor system for the microbial solubilization of coal

DOEpatents

Scott, C.D.; Strandberg, G.W.

1987-09-14

A fluidized-bed bioreactor system for the conversion of coal into microbially solubilized coal products. The fluidized-bed bioreactor continuously or periodically receives coal and bio-reactants and provides for the production of microbially solubilized coal products in an economical and efficient manner. An oxidation pretreatment process for rendering coal uniformly and more readily susceptible to microbial solubilization may be employed with the fluidized-bed bioreactor. 2 figs.

Multifunctional properties of phosphate-solubilizing microorganisms grown on agro-industrial wastes in fermentation and soil conditions.

PubMed

Vassileva, Maria; Serrano, Mercedes; Bravo, Vicente; Jurado, Encarnación; Nikolaeva, Iana; Martos, Vanessa; Vassilev, Nikolay

2010-02-01

One of the most studied approaches in solubilization of insoluble phosphates is the biological treatment of rock phosphates. In recent years, various techniques for rock phosphate solubilization have been proposed, with increasing emphasis on application of P-solubilizing microorganisms. The P-solubilizing activity is determined by the microbial biochemical ability to produce and release metabolites with metal-chelating functions. In a number of studies, we have shown that agro-industrial wastes can be efficiently used as substrates in solubilization of phosphate rocks. These processes were carried out employing various technologies including solid-state and submerged fermentations including immobilized cells. The review paper deals critically with several novel trends in exploring various properties of the above microbial/agro-wastes/rock phosphate systems. The major idea is to describe how a single P-solubilizing microorganism manifests wide range of metabolic abilities in different environments. In fermentation conditions, P-solubilizing microorganisms were found to produce various enzymes, siderophores, and plant hormones. Further introduction of the resulting biotechnological products into soil-plant systems resulted in significantly higher plant growth, enhanced soil properties, and biological (including biocontrol) activity. Application of these bio-products in bioremediation of disturbed (heavy metal contaminated and desertified) soils is based on another important part of their multifunctional properties.

Effects of L-arginine on solubilization and purification of plant membrane proteins.

PubMed

Arakawa, Junji; Uegaki, Masamichi; Ishimizu, Takeshi

2011-11-01

Biochemical analysis of membrane proteins is problematic at the level of solubilization and/or purification because of their hydrophobic nature. Here, we developed methods for efficient solubilization and purification of membrane proteins using L-arginine. The addition of 100 mM of basic amino acids (L-arginine, L-lysine, and L-ornithine) to a detergent-containing solubilization buffer enhanced solubilization (by 2.6-4.3 fold) of a model membrane protein-polygalacturonic acid synthase. Of all the amino acids, arginine was the most effective additive for solubilization of this membrane protein. Arginine addition also resulted in the best solubilization of other plant membrane proteins. Next, we examined the effects of arginine on purification of a model membrane protein. In anion-exchange chromatography, the addition of arginine to the loading and elution buffers resulted in a greater recovery of a membrane protein. In ultrafiltration, the addition of arginine to a protein solution significantly improved the recovery of a membrane protein. These results were thought to be due to the properties of arginine that prevent aggregation of hydrophobic proteins. Taken together, the results of our study showed that arginine is useful for solubilization and purification of aggregate-prone membrane proteins. Copyright © 2011 Elsevier Inc. All rights reserved.

Solubilization of lipids and lipid phases by the styrene-maleic acid copolymer.

PubMed

Dominguez Pardo, Juan J; Dörr, Jonas M; Iyer, Aditya; Cox, Ruud C; Scheidelaar, Stefan; Koorengevel, Martijn C; Subramaniam, Vinod; Killian, J Antoinette

2017-01-01

A promising tool in membrane research is the use of the styrene-maleic acid (SMA) copolymer to solubilize membranes in the form of nanodiscs. Since membranes are heterogeneous in composition, it is important to know whether SMA thereby has a preference for solubilization of either specific types of lipids or specific bilayer phases. Here, we investigated this by performing partial solubilization of model membranes and analyzing the lipid composition of the solubilized fraction. We found that SMA displays no significant lipid preference in homogeneous binary lipid mixtures in the fluid phase, even when using lipids that by themselves show very different solubilization kinetics. By contrast, in heterogeneous phase-separated bilayers, SMA was found to have a strong preference for solubilization of lipids in the fluid phase as compared to those in either a gel phase or a liquid-ordered phase. Together the results suggest that (1) SMA is a reliable tool to characterize native interactions between membrane constituents, (2) any solubilization preference of SMA is not due to properties of individual lipids but rather due to properties of the membrane or membrane domains in which these lipids reside and (3) exploiting SMA resistance rather than detergent resistance may be an attractive approach for the isolation of ordered domains from biological membranes.

The effect of steroids and nucleotides on solubilized bilirubin uridine diphosphate glucuronyltransferase

PubMed Central

Adlard, B. P. F.; Lathe, G. H.

1970-01-01

1. It was confirmed that bilirubin glucuronyltransferase can be obtained in solubilized form from rat liver microsomes. 2. Michaelis–Menten kinetics were not followed by the enzyme with bilirubin as substrate when the bilirubin/albumin ratio was varied. High concentrations of bilirubin were inhibitory. 3. The Km for UDP-glucuronic acid at the optimum bilirubin concentration was 0.46mm. 4. Low concentrations of Ca2+ were inhibitory in the absence of Mg2+ but stimulatory in its presence; the converse applied for EDTA. 5. UDP-N-acetylglucosamine and UDP-glucose enhanced conjugation by untreated, but not by solubilized microsomes. 6. The apparent 9.5-fold increase in activity after solubilization was probably due to the absence of UDP-glucuronic acid pyrophosphatase activity in the solubilized preparation. 7. The activation of solubilized enzyme activity by ATP was considered to be a result of chelation of inhibitory metal ions. 8. The solubilized enzyme activity was inhibited by UMP and UDP. The effect of UMP was not competitive with respect to UDP-glucuronic acid. 9. A number of steroids inhibited the solubilized enzyme activity. The competitive effects of stilboestrol, oestrone sulphate and 3β-hydroxyandrost-5-en-17-one, with respect to UDP-glucuronic acid, may be explained on an allosteric basis. PMID:4251180

Determining orientation and direction of DNA sequences

DOEpatents

Goodwin, Edwin H.; Meyne, Julianne

2000-01-01

Determining orientation and direction of DNA sequences. A method by which fluorescence in situ hybridization can be made strand specific is described. Cell cultures are grown in a medium containing a halogenated nucleotide. The analog is partially incorporated in one DNA strand of each chromatid. This substitution takes place in opposite strands of the two sister chromatids. After staining with the fluorescent DNA-binding dye Hoechst 33258, cells are exposed to long-wavelength ultraviolet light which results in numerous strand nicks. These nicks enable the substituted strand to be denatured and solubilized by heat, treatment with high or low pH aqueous solutions, or by immersing the strands in 2.times.SSC (0.3M NaCl+0.03M sodium citrate), to name three procedures. It is unnecessary to enzymatically digest the strands using Exo III or another exonuclease in order to excise and solubilize nucleotides starting at the sites of the nicks. The denaturing/solubilizing process removes most of the substituted strand while leaving the prereplication strand largely intact. Hybridization of a single-stranded probe of a tandem repeat arranged in a head-to-tail orientation will result in hybridization only to the chromatid with the complementary strand present.

Multimeric species in equilibrium in detergent-solubilized Na,K-ATPase.

PubMed

Yoneda, Juliana Sakamoto; Scanavachi, Gustavo; Sebinelli, Heitor Gobbi; Borges, Júlio Cesar; Barbosa, Leandro R S; Ciancaglini, Pietro; Itri, Rosangela

2016-08-01

In this work, we find an equilibrium between different Na,K-ATPase (NKA) oligomeric species solubilized in a non-ionic detergent C12E8 by means of Dynamic Light Scattering (DLS), Analytical Ultracentrifugation (AUC), Small Angle X-ray Scattering (SAXS), Spectrophotometry (absorption at 280/350nm) and enzymatic activity assay. The NKA sample after chromatography purification presented seven different populations as identified by AUC, with monomers and tetramers amounting to ∼55% of the total protein mass in solution. These two species constituted less than 40% of the total protein mass after increasing the NKA concentration. Removal of higher-order oligomer/aggregate species from the NKA solution using 220nm-pore filter resulted in an increase of the specific enzymatic activity. Nevertheless, the enzyme forms new large aggregates over an elapsed time of 20h. The results thus point out that C12E8-solubilized NKA is in a dynamic equilibrium of monomers, tetramers and high-order oligomers/subunit aggregates. These latter have low or null activity. High amount of detergent leads to the dissociation of NKA into smaller aggregates with no enzymatic activity. Copyright © 2016. Published by Elsevier B.V.

Digestive kinetics determines bioavailability of pollutants. Final report, 1 July 1993--30 September 1998

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mayer, L.M.

1998-09-30

The bioavailability of sedimentary contaminants to animals in harbor sediments was addressed by studying the mechanisms by which animals solubilize contaminants during feeding and digestion. Digestive physiology work on many different animal species revealed patterns of enzymes, surfactants and dissolved organic matter that correlate with feeding mode, phyletic position, and diet. Incubation of digestive fluids to dissolve contaminants from polluted sediments was developed to provide numerical estimates of bioavailability, and showed that much higher fractions of total contaminant loading are available than predicted by currently established, aqueous equilibrium approaches. The kinetics of reactions are slow enough that variations in feedingmore » rates will influence overall bioavailability. Experimental manipulations showed mechanisms of bioavailability. Dissolved amino acids, in the form of enzyme proteins and hydrolyzed food, are responsible for solubilization of metals such as copper. At high levels, copper can inactivate digestive enzymes. Metals in sedimentary sulfide minerals were largely impervious to digestive fluid attack. Surfactants are responsible for most solubilization of polycyclic aromatic hydrocarbons (PAH), though other agents also appear to play a role. Bioavailability of both metals and PAH can be limited by saturating the digestive agents responsible for their dissolution.« less

Micro-scale displacement of NAPL by surfactant and microemulsion in heterogeneous porous media

NASA Astrophysics Data System (ADS)

Javanbakht, Gina; Arshadi, Maziar; Qin, Tianzhu; Goual, Lamia

2017-07-01

Industrial processes such as remediation of oil-contaminated aquifers and enhanced oil recovery (EOR) often utilize chemical additives to increase the removal of non-aqueous phase liquids (NAPLs) from subsurface formations. Although the majority of crude oils are classified as LNAPLs, they often contain heavy molecules (DNAPLs) such as asphaltenes that tend to adsorb on minerals and alter their wettability. Effective additives are therefore those that can reduce the threshold capillary pressure, thus mobilizing LNAPL inside pore spaces and solubilizing DNAPL from rock surfaces. Nonionic surfactants in brine have often been injected to oil or contaminated aquifer formations in order to enhance NAPL displacement through IFT reduction. Recent studies revealed that surfactant-based microemulsions have a higher tendency to alter the wettability of surfaces, compared to surfactants alone, leading to more effective NAPL removal. However, the impact of these additives on pore-scale displacement mechanisms and multi-phase fluid occupancy in porous media is, to date, still unclear. In this study, x-ray microtomography experiments were performed to investigate the impact of surfactants and microemulsions on the mobilization and solubilization of NAPL in heterogeneous rocks. Saturation profiles indicated that an incremental NAPL removal was attained by addition of microemulsion to brine, compared with surfactant. Residual cluster size distributions revealed that microemulsions could break up large clusters into smaller disconnected ones, improving their mobilization in the rock. In-situ contact angle measurements showed that microemulsions could reverse the wettability of rough contaminated surfaces to a higher extent than surfactants. Unlike surfactant alone, the surfactant-solvent blend in the carrier fluid of microemulsions was able to penetrate rough grain surfaces, particularly those of dolomite cement, and desorb asphaltenes in the form of small-emulsified NAPL droplets, which were eventually washed away by the continuous flow process. The greater wettability alteration caused by microemulsions resulted in a lower threshold capillary pressure, which in turn promoted the mobilization of NAPL ganglia more than surfactant alone.

Research and engineering assessment of biological solubilization of phosphate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rogers, R.D.; McIlwain, M.E.; Losinski, S.J.

This research and engineering assessment examined a microbial phosphate solubilization process as a method of recovering phosphate from phosphorus containing ore compared to the existing wet acid and electric arc methods. A total of 860 microbial isolates, collected from a range of natural environments were tested for their ability to solubilize phosphate from rock phosphate. A bacterium (Pseudomonas cepacia) was selected for extensive characterization and evaluation of the mechanism of phosphate solubilization and of process engineering parameters necessary to recover phosphate from rock phosphate. These studies found that concentration of hydrogen ion and production of organic acids arising from oxidationmore » of the carbon source facilitated microbial solubilization of both pure chemical insoluble phosphate compounds and phosphate rock. Genetic studies found that phosphate solubilization was linked to an enzyme system (glucose dehydrogenase). Process-related studies found that a critical solids density of 1% by weight (ore to liquid) was necessary for optimal solubilization. An engineering analysis evaluated the cost and energy requirements for a 2 million ton per year sized plant, whose size was selected to be comparable to existing wet acid plants.« less

Solubilization of human cells by the styrene-maleic acid copolymer: Insights from fluorescence microscopy.

PubMed

Dörr, Jonas M; van Coevorden-Hameete, Marleen H; Hoogenraad, Casper C; Killian, J Antoinette

2017-11-01

Extracting membrane proteins from biological membranes by styrene-maleic acid copolymers (SMAs) in the form of nanodiscs has developed into a powerful tool in membrane research. However, the mode of action of membrane (protein) solubilization in a cellular context is still poorly understood and potential specificity for cellular compartments has not been investigated. Here, we use fluorescence microscopy to visualize the process of SMA solubilization of human cells, exemplified by the immortalized human HeLa cell line. Using fluorescent protein fusion constructs that mark distinct subcellular compartments, we found that SMA solubilizes membranes in a concentration-dependent multi-stage process. While all major intracellular compartments were affected without a strong preference, plasma membrane solubilization was found to be generally slower than the solubilization of organelle membranes. Interestingly, some plasma membrane-localized proteins were more resistant against solubilization than others, which might be explained by their presence in specific membrane domains with differing properties. Our results support the general applicability of SMA for the isolation of membrane proteins from different types of (sub)cellular membranes. Copyright © 2017 Elsevier B.V. All rights reserved.

Solubilization of cyclohexane in aqueous solutions of sodium. cap alpha. -alkyl alkanoates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sagitani, H.; Suzuki, T.; Nagai, M.

1982-01-01

The effect of branched alkyl chain length and the position of the COONa group on the solubilizing power of n-alkane sodium carboxylates was studied. The lipophilic property and the amount of solubilized cyclohexane increased with the branched chain length of branched soaps, and with the change of the position of the -COONa group from 3 to 7 in the alkyl chain of pentadecane -3, -5, and -7 sodium carboxylates. Alpha-branched soaps having proper branched alkyl chains were better solubilizers for cyclohexane than straight chain compounds. The amount of cyclohexane solublized by C/sub 10/ H/sub 21/ CH(C/sub 6/H/sub 13/) COONa wasmore » about three times greater than the amount solubilized by C/sub 17/ H/sub 35/ COONa. There was a marked increase in the solubilization of cyclohexane replacing ..cap alpha..-branched fatty acid soaps with optimum amount of cosurfactants such as C/sub 8/H/sub 17/ (OCH/sub 2/CH/sub 2/)/sub 2/OH. Namely, solubilization increased markedly at the optimum hydrophile-lipophile balance of mixed surfactant. 21 references.« less

The solubilization of bone and dentin collagens by pepsin. Effect of cross-linkages and non-collagen components.

PubMed

Carmichael, D J; Dodd, C M; Veis, A

1977-03-28

Bone and dentin collagen are less susceptible to solubilization by pepsin digestion then is skin collagen. Digestion at 4 degrees C for 72 h solubilized only 35.3% of bovine cortical bone and 5.6% of bovine dentin compared with nearly 100% dissolution of bovine skin. Sodium dodecyl sulfate-acrylamide gel electrophoresis and molecular sieve chromatography showed that, for bone and dentin, intact alpha chains and cross-linked aggregates of beta, gamma and higher weight remained intact after pepsin solubilization but lower molecular weight fragments also were prevalent indicating chain scission in helical regions. Electron microscopic examination of segment long spacing precipitates of the soluble collagens confirmed the presence of solubilized polymerized collagen. The principal reducible cross-link in both bone and dentin was the precursor of dihydroxylsinonorleucine and this cross-link was also present in the solubilized collagens. Small amounts of non-collagenous proteins and glycosaminoglycans of different compositions in dentin and bone resisted extraction before pepsin digestion. However, the differences in solubilization of the collagens have been related to differences in cross-linkage placement.

Thermo-tolerant phosphate-solubilizing microbes for multi-functional biofertilizer preparation.

PubMed

Chang, Cheng-Hsiung; Yang, Shang-Shyng

2009-02-01

In order to prepare the multi-functional biofertilizer, thermo-tolerant phosphate-solubilizing microbes including bacteria, actinomycetes, and fungi were isolated from different compost plants and biofertilizers. Except Streptomycesthermophilus J57 which lacked pectinase, all isolates possessed amylase, CMCase, chitinase, pectinase, protease, lipase, and nitrogenase activities. All isolates could solubilize calcium phosphate and Israel rock phosphate; various isolates could solubilize aluminum phosphate, iron phosphate, and hydroxyapatite. During composting, biofertilizers inoculated with the tested microbes had a significantly higher temperature, ash content, pH, total nitrogen, soluble phosphorus content, and germination rate than non-inoculated biofertilizer; total organic carbon and carbon-to-nitrogen ratio showed the opposite pattern. Adding these microbes can shorten the period of maturity, improve the quality, increase the soluble phosphorus content, and enhance the populations of phosphate-solubilizing and proteolytic microbes in biofertilizers. Therefore, inoculating thermo-tolerant phosphate-solubilizing microbes into agricultural and animal wastes represents a practical strategy for preparing multi-functional biofertilizer.

Solubilization of adenylyl cyclase from human myometrium in a alphas-coupled form.

PubMed

Bajo, Ana M; Prieto, Juan C; Valenzuela, Pedro; Martinez, Pilar; Guijarro, Luis G

2003-08-01

Adenylyl cyclase (AC) was extracted from human myometrium with either non-ionic (Lubrol-PX or Triton X-100) or zwitterionic (3-[3-cholamidopropyl)dimethylammonio]-1-propanesulfonate, CHAPS) detergents. The soluble enzyme was stimulated by forskolin, a hydrophobic activator, in the presence of Mg2+ indicating that the catalytic subunit had not been damaged after solubilization. The enzyme was also activated by 5'-guanylyl imidodiphosphate (Gpp(NH)p) showing that the catalytic unit was not separated from stimulatory guanine nucleotide binding protein (Gs) during the extraction. Both activators showed different effects on the stimulatory efficacy and potency of AC activity solobulized with detergents. Gel filtration of Lubrol-PX and CHAPS extracts over a Sepharose CL-2B column partially resolved AC and its complexes. The chromatographic profile for Lubrol-solubilized AC presented a main peak of about 200 kDa whereas CHAPS-solubilized AC showed a dominant peak of about 1100 kDa. The heterodisperse peaks obtained revealed that the catalytic AC subunit was not separated from Gs proteins after gel filtration, and that AC could be associated with other cellular proteins. When Lubrol extract was submitted to anionic-exchange chromatography, the enzyme was purified about 7.5 fold (enzymatic activity of 48.1 pmol/min/mg of protein). The catalytic subunit was co-eluted with both AC-activating proteins Galphas large (52.2 kDa) and Galphas small (48.7 kDa). This is the first demonstration of the stable physical association of AC with both alphas subunits of G proteins in human myometrium.

Biochar enhances Aspergillus niger rock phosphate solubilization by increasing organic acid production and alleviating fluoride toxicity.

PubMed

Mendes, Gilberto de Oliveira; Zafra, David Lopez; Vassilev, Nikolay Bojkov; Silva, Ivo Ribeiro; Ribeiro, José Ivo; Costa, Maurício Dutra

2014-05-01

During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F(-)) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F(-) adsorbents [activated alumina (Al2O3) and biochar] on RP solubilization by Aspergillus niger was examined. Al2O3 adsorbed part of the F(-) released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F(-) while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F(-) measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F(-) per liter can be removed from solution by biochar when added at 3 g liter(-1) to the culture medium. Thus, biochar acted as an F(-) sink during RP solubilization and led to an F(-) concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F(-) and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F(-), the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP.

Biochar Enhances Aspergillus niger Rock Phosphate Solubilization by Increasing Organic Acid Production and Alleviating Fluoride Toxicity

PubMed Central

Mendes, Gilberto de Oliveira; Zafra, David Lopez; Vassilev, Nikolay Bojkov; Silva, Ivo Ribeiro; Ribeiro, José Ivo

2014-01-01

During fungal rock phosphate (RP) solubilization, a significant quantity of fluoride (F−) is released together with phosphorus (P), strongly inhibiting the process. In the present study, the effect of two F− adsorbents [activated alumina (Al2O3) and biochar] on RP solubilization by Aspergillus niger was examined. Al2O3 adsorbed part of the F− released but also adsorbed soluble P, which makes it inappropriate for microbial RP solubilization systems. In contrast, biochar adsorbed only F− while enhancing phosphate solubilization 3-fold, leading to the accumulation of up to 160 mg of P per liter. By comparing the values of F− measured in solution at the end of incubation and those from a predictive model, it was estimated that up to 19 mg of F− per liter can be removed from solution by biochar when added at 3 g liter−1 to the culture medium. Thus, biochar acted as an F− sink during RP solubilization and led to an F− concentration in solution that was less inhibitory to the process. In the presence of biochar, A. niger produced larger amounts of citric, gluconic, and oxalic acids, whether RP was present or not. Our results show that biochar enhances RP solubilization through two interrelated processes: partial removal of the released F− and increased organic acid production. Given the importance of organic acids for P solubilization and that most of the RPs contain high concentrations of F−, the proposed solubilization system offers an important technological improvement for the microbial production of soluble P fertilizers from RP. PMID:24610849

Blocking monocyte transmigration in in vitro system by a human antibody scFv anti-CD99. Efficient large scale purification from periplasmic inclusion bodies in E. coli expression system.

PubMed

Moricoli, Diego; Muller, William Anthony; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Watson, Richard; Fiori, Valentina; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

2014-06-01

Migration of leukocytes into site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells, inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies and the absence of toxic reagents utilized for solubilization and refolding step of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting, we herein describe an efficient and large scale production of the antibody fragments expressed in E. coli as periplasmic insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signaling. This protocol can be useful for the successful purification of other monomeric scFvs which are expressed as periplasmic inclusion bodies in bacterial systems. Copyright © 2014 Elsevier B.V. All rights reserved.

Biochemical solubilization of toxic salts from residual geothermal brines and waste waters

DOEpatents

Premuzic, Eugene T.; Lin, Mow S.

1994-11-22

A method of solubilizing metal salts such as metal sulfides in a geothermal sludge using mutant Thiobacilli selected for their ability to metabolize metal salts at high temperature is disclosed, The method includes the introduction of mutated Thiobacillus ferrooxidans and Thiobacillus thiooxidans to a geothermal sludge or brine. The microorganisms catalyze the solubilization of metal salts, For instance, in the case of metal sulfides, the microorganisms catalyze the solubilization to form soluble metal sulfates.

Rock inhabiting potassium solubilizing bacteria from Kerala, India: characterization and possibility in chemical K fertilizer substitution.

PubMed

Anjanadevi, Indira Parameswaran; John, Neetha Soma; John, Kuzhivilayil Susan; Jeeva, Muthulekshmi Lajapathy; Misra, Raj Shekhar

2016-01-01

The role of rock inhabiting bacteria in potassium (K) solubilization from feldspar and their application in crop nutrition through substitution of fertilizer K was explored through the isolation of 36 different bacteria from rocks of a major hill station at Ponmudi in Thiruvananthapuram, Kerala, India. A comprehensive characterization of K solubilization from feldspar was achieved with these isolates which indicated that the K solubilizing efficiency increases with decrease in pH and increase in viscosity and viable cell count. Based on the level of K solubilization, two potent isolates were selected and identified as Bacillus subtilis ANctcri3 and Bacillus megaterium ANctcri7. Exopolysaccharide production, scanning electron microscopic and fourier transform infrared spectroscopic studies with these efficient strains conclusively depicted the role of low pH, increase in viscosity, and bacterial attachment in K solubilization. They were also found to be efficient in phosphorus (P) solubilization, indole acetic acid production as well as tolerant to wide range of physiological conditions. Moreover, the applicability of K containing rock powder as a carrier for K solubilizing bacteria was demonstrated. A field level evaluation on the yield of a high K demanding tuberous vegetable crop, elephant foot yam (Amorphophallus paeoniifolius (dennst.) nicolson) established the possibility of substituting chemical K fertilizer with these biofertilizer candidates successfully. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Growth promotion of peanut (Arachis hypogaea L.) and maize (Zea mays L.) plants by single and mixed cultures of efficient phosphate solubilizing bacteria that are tolerant to abiotic stress and pesticides.

PubMed

Anzuay, María Soledad; Ciancio, María Gabriela Ruiz; Ludueña, Liliana Mercedes; Angelini, Jorge Guillermo; Barros, Germán; Pastor, Nicolás; Taurian, Tania

2017-06-01

The aims of this study were, to analyze in vitro phosphate solubilization activity of six native peanut bacteria and to determine the effect of single and mixed inoculation of these bacteria on peanut and maize plants. Ability to produce organic acids and cofactor PQQ, to solubilize FePO 4 and AlPO 4 and phosphatase activity were analyzed. Also, the ability to solubilize phosphate under abiotic stress and in the presence of pesticides of the selected bacteria was determined. The effect of single and mixed bacterial inocula was analyzed on seed germination, maize plant growth and in a crop rotation plant assay with peanut and maize. The six strains produced gluconic acid and five released cofactor PQQ into the medium. All bacteria showed ability to solubilize phosphate from FePO 4 and AlPO 4 and phosphatase activity. The ability of the bacteria to solubilize tricalcium phosphate under abiotic stress and in presence of pesticides indicated encouraging results. Bacterial inoculation on peanut and maize increased seed germination, plant́s growth and P content. Phosphate solubilizing bacteria used in this study showed efficient phosphate mineralizing and solubilization ability and would be potential P-biofertilizers for peanut and maize. Copyright © 2017 Elsevier GmbH. All rights reserved.

Thermochemical pretreatment of lignocellulose residues: assessment of the effect on operational conditions and their interactions on the characteristics of leachable fraction.

PubMed

Vásquez, Denisse; Contreras, Elsa; Palma, Carolyn; Carvajal, Andrea

2015-01-01

Annually, large amounts of agricultural residues are produced in Chile, which can be turned into a good opportunity to diversify the energy matrix. These residues have a slow hydrolysis stage during anaerobic digestion; therefore, the application of a pretreatment seems to be an alternative to improve the process. This work focused on applying a thermochemical pretreatment with NaOH on two lignocellulosic residues. The experiments were performed according to a 2(4) factorial design. The factors studied in a 2(4) factorial design were: temperature (60 and 120 °C), pretreatment time (10 and 30 minutes), NaOH dose (2 and 4%), and residue size (<1 and 1-3 mm for wheat straw; 1-5 and 5-10 mm for corn stover). The analyzed response variables were the solubilization of organic matter, and the biodegradability of the lignocellulose hydrolysate. The statistical analysis of the data allowed the identification of the experimental conditions that maximized solubilization of organic matter and biodegradability. The main results showed that more aggressive experimental conditions could increase the breaking down of the structure; in addition, the time of pretreatment was not significant. Conversely, the less aggressive experimental conditions, regarding regent dosage and downsizing, favored the release of biodegradable organic matter. The main conclusion of this study was the identification of the operational conditions of the thermochemical pretreatment that promote maximum biogas production, which was caused due to the solubilization of a large amount of organic matter, but not because of the increase in biodegradability of the released organic matter.

Solubilization and characterization of haloperidol-sensitive (+)-( sup 3 H)SKF-10,047 binding sites (sigma sites) from rat liver membranes

DOE Office of Scientific and Technical Information (OSTI.GOV)

McCann, D.J.; Su, T.P.

1991-05-01

The zwitterionic detergent 3-((3-cholamidopropyl)dimethylamino)-1-propanesulfonate (CHAPS) produced optimal solubilization of (+)-({sup 3}H)SKF-10,047 binding sites from rat liver membranes at a concentration of 0.2%, well below the critical micellular concentration of the detergent. The pharmacological selectivity of the liver (+)-({sup 3}H)SKF-10,047 binding sites corresponds to that of sigma sites from rat and guinea pig brain. When the affinities of 18 different drugs at (+)-({sup 3}H)SKF-10,047 binding sites in membranes and solubilized preparations were compared, a correlation coefficient of 0.99 and a slope of 1.03 were obtained, indicating that the pharmacological selectivity of rat liver sigma sites is retained after solubilization. In addition,more » the binding of 20 nM ({sup 3}H)progesterone to solubilized rat liver preparations was found to exhibit a pharmacological selectivity appropriate for sigma sites. A stimulatory effect of phenytoin on (+)-({sup 3}H)SKF-10,047 binding to sigma sites persisted after solubilization. When the solubilized preparation was subjected to molecular sizing chromatography, a single peak exhibiting specific (+)-({sup 3}H)SKF-10,047 binding was obtained. The binding activity of this peak was stimulated symmetrically when assays were performed in the presence of 300 microM phenytoin. The molecular weight of the CHAPS-solubilized sigma site complex was estimated to be 450,000 daltons. After solubilization with CHAPS, rat liver sigma sites were enriched to 12 pmol/mg of protein. The present results demonstrate a successful solubilization of sigma sites from rat liver membranes and provide direct evidence that the gonadal steroid progesterone binds to sigma sites. The results also suggest that the anticonvulsant phenytoin binds to an associated allosteric site on the sigma site complex.« less

Medium pH, carbon and nitrogen concentrations modulate the phosphate solubilization efficiency of Penicillium purpurogenum through organic acid production.

PubMed

Scervino, J M; Papinutti, V L; Godoy, M S; Rodriguez, M A; Della Monica, I; Recchi, M; Pettinari, M J; Godeas, A M

2011-05-01

To study phosphate solubilization in Penicillium purpurogenum as function of medium pH, and carbon and nitrogen concentrations. Tricalcium phosphate (CP) solubilization efficiency of P. purpurogenum was evaluated at acid or alkaline pH using different C and N sources. Glucose- and (NH(4) )(2) SO(4) -based media showed the highest P solubilization values followed by fructose. P. purpurogenum solubilizing ability was higher in cultures grown at pH 6·5 than cultures at pH 8·5. Organic acids were detected in both alkaline and neutral media, but the relative percentages of each organic acid differed. Highest P release coincided with the highest organic acids production peak, especially gluconic acid. When P. purpurogenum grew in alkaline media, the nature and concentration of organic acids changed at different N and C concentrations. A factorial categorical experimental design showed that the highest P-solubilizing activity, coinciding with the highest organic acid production, corresponded to the highest C concentration and lowest N concentration. The results described in the present study show that medium pH and carbon and nitrogen concentrations modulate the P solubilization efficiency of P. purpurogenum through the production of organic acids and particularly that of gluconic acid. In the P solubilization optimization studies, glucose and (NH(4) )(2) SO(4) as C and N sources allowed a higher solubilization efficiency at high pH. This organism is a potentially proficient soil inoculant, especially in P-poor alkaline soils where other P solubilizers fail to release soluble P. Further work is necessary to elucidate whether these results can be extrapolated to natural soil ecosystems, where different pH values are present. Penicillium purpurogenum could be used to develop a bioprocess for the manufacture of phosphatic fertilizer with phosphate calcium minerals. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

Solubility of amphotericin B in water-lecithin-dispersions and lecithin-based submicron emulsions.

PubMed

Salerno, Claudia; Perez, Sebastian; Monteagudo, Ezequiel; Carlucci, Adriana; Bregni, Carlos

2013-01-01

The aim of this work was to evaluate water-lecithin-dispersions (WLDs) as carriers for amphotericin B (AmB) and to compare the drug solubility in WLDs and O/W lecithin-based submicron emulsions (SMEs) in order to evaluate the influence of lecithin content on the dosage form solubilization of the active compound. WLDs and different SMEs with either 1.2 or 2.4% of lecithin were prepared. WLD with 2.4% lecithin show a 10-fold increase in solubilization of AmB compared with 1.2% lecithin WLD. SMEs with 1.2% lecithin show an increase of over 400 times in solubilization compared with WLD containing the same concentration of lecithin, whereas SMEs with 2.4% lecithin show an increase of over 40 times compared with the corresponding WLD. Drug solubilization in SMEs with 2.4% lecithin is not significantly greater than in those containing 1.2% lecithin. The content of surfactant Brij 97 ® had a significant influence on drug solubilization in SMEs (P < 0.05). Results indicate that indicate that SMEs are proper systems to solubilize AmB. It can be assumed that solubilization is due to the formulation microstructure and not to the separate components themselves.

Effects of solubilization on the inhibition of the p-type ATPase from maize roots by N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline.

PubMed

Brauer, D K; Gurriel, M; Tu, S I

1992-12-01

The biochemical events utilized by transport proteins to convert the chemical energy from the hydrolysis of ATP into an electro-chemical gradient are poorly understood. The inhibition of the plasma membrane ATPase from corn (Zea mays L.) roots by N-(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline (EEDQ) was compared to that of ATPase solubilized with N-tetradecyl-N,N-dimethyl-3-ammonio-1-propane-sulfonate (3-14) to provide insight into the minimal functional unit. The chromatographic behavior of the 3-14-solubilized ATPase activity during size exclusion chromatography and glycerol gradient centrifugation indicated that the solubilized enzyme was in a monomeric form. Both plasma membrane-bound and solubilized ATPase were inhibited by EEDQ in a time- and concentration-dependent manner consistent with a first-order reaction. When the log of the reciprocal of the half-time for inhibition was plotted as a function of the log of the EEDQ concentration, straight lines were obtained with slopes of approximately 0.5 and 1.0 for membrane-bound and 3-14-solubilized ATPase, respectively, indicating a change in the number of polypeptides per functional ATPase complex induced by solubilization with 3-14.

Biochemical solubilization of toxic salts from residual geothermal brines and waste waters

DOEpatents

Premuzic, E.T.; Lin, M.S.

1994-11-22

A method of solubilizing metal salts such as metal sulfides in a geothermal sludge using mutant Thiobacilli selected for their ability to metabolize metal salts at high temperature is disclosed. The method includes the introduction of mutated Thiobacillus ferrooxidans and Thiobacillus thiooxidans to a geothermal sludge or brine. The microorganisms catalyze the solubilization of metal salts. For instance, in the case of metal sulfides, the microorganisms catalyze the solubilization to form soluble metal sulfates. 54 figs.

Microbial solubilization of phosphate

DOEpatents

Rogers, R.D.; Wolfram, J.H.

1993-10-26

A process is provided for solubilizing phosphate from phosphate containing ore by treatment with microorganisms which comprises forming an aqueous mixture of phosphate ore, microorganisms operable for solubilizing phosphate from the phosphate ore and maintaining the aqueous mixture for a period of time and under conditions operable to effect the microbial solubilization process. An aqueous solution containing soluble phosphorus can be separated from the reacted mixture by precipitation, solvent extraction, selective membrane, exchange resin or gravity methods to recover phosphate from the aqueous solution. 6 figures.

Microbial solubilization of phosphate

DOEpatents

Rogers, Robert D.; Wolfram, James H.

1993-01-01

A process is provided for solubilizing phosphate from phosphate containing ore by treatment with microorganisms which comprises forming an aqueous mixture of phosphate ore, microorganisms operable for solubilizing phosphate from the phosphate ore and maintaining the aqueous mixture for a period of time and under conditions operable to effect the microbial solubilization process. An aqueous solution containing soluble phosphorous can be separated from the reacted mixture by precipitation, solvent extraction, selective membrane, exchange resin or gravity methods to recover phosphate from the aqueous solution.

Regulation of coal polymer degradation by fungi. Eighth quarterly report, [January--March 1996

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvine, R.L.; Bumpus, J.A.

1996-07-28

Progress is reported on solubilization of low-rank coal by enzyme activity derived from Trametes versicolor or P. chrysosporium. Specifically during the reporting period efforts were directed towards the determining the effect of pH on solubilization of leonardite, the role of laccase in low coal solubilization and metabolism, the decolorization of soluble coal macromolecule by P. chrysosprium and T. versicolor in solid agar gel, and the solubilization of low rank coal in slurry cultures and solid phase reactors.

Biochemical bond breaking in coal: Third quarterly report, (April through June 1987)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Not Available

1987-07-01

Major research efforts are presently being conducted in three principal areas of focus: (1) optimization of coal biosolubilization; (2) characterization of the solubilized products resulting from microbial coal depolymerization; and (3) degradation of model compounds to assess potential interunit linkages which may be attacked by whole culture or cell-free culture supernatants containing extracellular enzymes. Initial evaluations of the various combinations of microbes, coals, and coal pretreatments indicated that CP1 and CP1 + 2 solubilized all of the coals selected for this project at substantially higher rates than S. setonii or T. versicolor. The ARC CP1 + 2 consortium was chosenmore » as the primary culture for detailed evaluation of coal biosolubilization and model compound degradation. Studies were conducted to determine if solubilization of coal by CP1 + 2 supernatants could be enhanced by elevating the temperature. Solubilization of both untreated Leonardite and HNO3 treated Wyodak (Smith-Roland) subbituminous coal was increased when elevating the temperature from ambient to 35C. The initial solubilization rate (T0 - 1 hour) of Leonardite at 22C was 16 OD units/hour and at 35C was 18 OD units/hour. Thus, an elevation of 13C enhanced solubilization of this coal by 12.5%. The effect of temperature on solubilization of Wyodak coal appeared to be more pronounced. Solubilization of HNO3 treated coals by the CP organisms is not only relatively rapid, but is also extensive. The relatively rapid and extensive coal solubilization attainable by CP1 + 2 has enabled us to produce quantities of product sufficient for analytical methods development and for characterization of the coal products. Initial attempts have been made to characterize the depolymerized products using HPLC and GC/MS. 9 figs., 3 tabs.« less

Isolation and characterization of phosphate-solubilizing bacteria from seagrass rhizosphere soil

NASA Astrophysics Data System (ADS)

Ghosh, Upasana; Subhashini, Ponnambalam; Dilipan, Elangovan; Raja, Subramanian; Thangaradjou, Thirunavukarassu; Kannan, Lakshmanan

2012-03-01

Phosphate-solubilizing bacterial strains (6 Nos.) were isolated from the rhizosphere soils of two seagrasses ( Halophila ovalis (R. Br.) Hook and Halodule pinifolia (Miki) Hartog) in the Vellar estuary. Experimental studies found that the strain PSSG6 was effective in phosphate solubilization with Phosphate Solubilization efficiency index E = 375 ± 8.54, followed by the strain PSSG5 with Phosphate Solubilization efficiency index E = 275 ± 27.3. Of the 6 strains isolated, the strains PSSG4 and PSSG5 belonged to the genus Bacillus, and PSSG1, PSSG2 and PSSG3 were identified as Citrobacter sp., Shigella sp., and Klebsiella sp., respectively, by conventional method, and PSSG6 was identified as Bacillus circulans using conventional and molecular methods.

Effect of organic acids production and bacterial community on the possible mechanism of phosphorus solubilization during composting with enriched phosphate-solubilizing bacteria inoculation.

PubMed

Wei, Yuquan; Zhao, Yue; Shi, Mingzi; Cao, Zhenyu; Lu, Qian; Yang, Tianxue; Fan, Yuying; Wei, Zimin

2018-01-01

Enriched phosphate-solubilizing bacteria (PSB) agent were acquired by domesticated cultivation, and inoculated into kitchen waste composting in different stages. The effect of different treatments on organic acids production, tricalcium phosphate (TCP) solubilization and their relationship with bacterial community were investigated during composting. Our results pointed out that inoculation affected pH, total acidity and the production of oxalic, lactic, citric, succinic, acetic and formic acids. We also found a strong advantage in the solubilization of TCP and phosphorus (P) availability for PSB inoculation especially in the cooling stage. Redundancy analysis and structural equation models demonstrated inoculation by different methods changed the correlation of the bacterial community composition with P fractions as well as organic acids, and strengthened the cooperative function related to P transformation among species during composting. Finally, we proposed a possible mechanism of P solubilization with enriched PSB inoculation, which was induced by bacterial community and organic acids production. Copyright © 2017 Elsevier Ltd. All rights reserved.

Maltose-neopentyl glycol (MNG) amphiphiles for solubilization, stabilization and crystallization of membrane proteins.

PubMed

Chae, Pil Seok; Rasmussen, Søren G F; Rana, Rohini R; Gotfryd, Kamil; Chandra, Richa; Goren, Michael A; Kruse, Andrew C; Nurva, Shailika; Loland, Claus J; Pierre, Yves; Drew, David; Popot, Jean-Luc; Picot, Daniel; Fox, Brian G; Guan, Lan; Gether, Ulrik; Byrne, Bernadette; Kobilka, Brian; Gellman, Samuel H

2010-12-01

The understanding of integral membrane protein (IMP) structure and function is hampered by the difficulty of handling these proteins. Aqueous solubilization, necessary for many types of biophysical analysis, generally requires a detergent to shield the large lipophilic surfaces of native IMPs. Many proteins remain difficult to study owing to a lack of suitable detergents. We introduce a class of amphiphiles, each built around a central quaternary carbon atom derived from neopentyl glycol, with hydrophilic groups derived from maltose. Representatives of this maltose-neopentyl glycol (MNG) amphiphile family show favorable behavior relative to conventional detergents, as manifested in multiple membrane protein systems, leading to enhanced structural stability and successful crystallization. MNG amphiphiles are promising tools for membrane protein science because of the ease with which they may be prepared and the facility with which their structures may be varied.

Regulation of coal polymer degradation by fungi. Eighth quarterly report, [April--June 1996

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvine, R.L.; Bumpus, J.A.

1996-07-28

This project addresses the solubilization of low-rank coal (leonardite) by lignin degrading fungi. During this reporting period efforts were focused on determining the effect of pH on coal solubilization by oxalate ion and other biologically important compounds that might function as metal chelators, on the role of laccase in coal solubilization and metabolism, on decolorization of soluble coal macromolecule by Phanerochaete chrysosporium and T. versicolor in solid agar media, and on solubilization of coal in slurry cultures and solid phase reactors.

Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nelson, C.A.; Seamon, K.B.

1988-01-01

The binding of (/sup 3/H)forskolin to proteins solubilized from bovine brain membranes was studied by precipitating proteins with polyethylene glycol and separating (/sup 3/H)forskolin bound to protein from free (/sup 3/H)forskolin by rapid filtration. The K/sub d/ for (/sup 3/H)forskolin binding to solubilized proteins was 14 nM which was similar to that for (/sup 3/H)forskolin binding sites in membranes from rat brain and human platelets. Forskolin analogs competed for (/sup 3/H)forskolin binding sites with the same rank potency in both brain membranes and in proteins solubilized from brain membranes. (/sup 3/H)forskolin bound to proteins solubilized from membranes with a Bmaxmore » of 38 fmolmg protein which increased to 94 fmolmg protein when GppNHp was included in the binding assay. In contrast, GppNHp had no effect on (/sup 3/H)forskolin binding to proteins solubilized from membranes preactivated with GppNHp. Solubilized adenylate cyclase from non-preactivated membranes had a basal activity of 130 pmolmgmin which was increased 7-fold by GppNHp. In contrast, adenylate cyclase from preactivated membranes had a basal activity of 850 pmolmgmin which was not stimulated by GppNHp or forskolin« less

Solubilization conditions for bovine heart mitochondrial membranes allow selective purification of large quantities of respiratory complexes I, III, and V.

PubMed

Shimada, Satoru; Maeda, Shintaro; Hikita, Masahide; Mieda-Higa, Kaoru; Uene, Shigefumi; Nariai, Yukiko; Shinzawa-Itoh, Kyoko

2018-04-24

Ascertaining the structure and functions of mitochondrial respiratory chain complexes is essential to understanding the biological mechanisms of energy conversion; therefore, numerous studies have examined these complexes. A fundamental part of that research involves devising a method for purifying samples with good reproducibility; the samples obtained need to be stable and their constituents need to retain the same structure and functions they possess when in mitochondrial membranes. Submitochondrial bovine heart particles were isolated using differential centrifugation to adjust to a membrane concentration of 46.0% (w/v) or 31.5% (w/v) based on weight. After 0.7% (w/v) deoxycholic acid, 0.4% (w/v) decyl maltoside, and 7.2% (w/v) potassium chloride were added to the mitochondrial membranes, those membranes were solubilized. At a membrane concentration of 46%, complex V was selectively solubilized, whereas at a concentration of 31.5% (w/v), complexes I and III were solubilized. Two steps-sucrose density gradient centrifugation and anion-exchange chromatography on a POROS HQ 20 μm column-enabled selective purification of samples that retained their structure and functions. These two steps enabled complexes I, III, and V to be purified in two days with a high yield. Complexes I, III, and V were stabilized with n-decyl-β-D-maltoside. A total of 200 mg-300 mg of those complexes from one bovine heart (1.1 kg muscle) was purified with good reproducibility, and the complexes retained the same functions they possessed while in mitochondrial membranes. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

Improved solubilization of activated sludge by ozonation in pressure cycles.

PubMed

Cheng, Chia-Jung; Hong, P K Andy; Lin, Cheng-Fang

2012-05-01

The generation of a large volume of activated sludge (AS) from wastewater treatment has increasingly become a great burden on the environment. Anaerobic digestion is routinely practiced for excess waste sludge; however, the process retention time is long because of kinetic limitation in the hydrolysis step. We tested the feasibility of applying ozone in pressure cycles to enhance the disintegration and solubilization of AS with the goal to prepare them for digestion using reduced ozone dose and contact time. The AS was subjected to repetitive pressure cycles in a closed vessel in which an ozone gas mixture was compressed into the slurry to reach 1040 kPa in the headspace to be followed by rapid venting. For a returned AS with total COD (tCOD) of 8200 mg L(-1), a dose of 0.01 gO(3)g(-1) total suspended solids (TSS) delivered via 20 pressure cycles within 16 min resulted in a 37-fold increase of the sCOD/tCOD ratio (due to increased soluble COD, i.e. sCOD) and a 25% reduction of TSS, in comparison to a dose of 0.08 gO(3)g(-1) TSS via bubbling contact over 15 min that resulted in a 15-fold increase of the sCOD/tCOD ratio and a 12% reduction of TSS. Sludge solubilization was evidenced by increased dissolved contents of total phosphorous (from 10 to 64 mg L(-1)), total nitrogen (from 14 to 120 mg L(-1)), and protein (from <15 to 39 mg L(-1)) in the sludge suspension after treatment, indicating significant solubilization of AS. Copyright © 2012 Elsevier Ltd. All rights reserved.

Solubilization of poorly soluble photosensitizer hypericin by polymeric micelles and polyethylene glycol.

PubMed

Búzová, Diana; Kasák, Peter; Miškovský, Pavol; Jancura, Daniel

2013-06-01

Hypericin (Hyp) is a promising photosensitizer for photodiagnostic and photodynamic therapy of cancer. However, Hyp has a large conjugated system and in aqueous solutions forms insoluble aggregates which do not possess biological activity. This makes intravenous injection of Hyp problematic and restricts its medical applications. To overcome this problem, Hyp is incorporated into drug delivery systems which can increase its solubility and bioavailability. One of the possibilities is utilization of polymeric micelles. The most used hydrophilic block for preparation of polymeric micelles is polyethylen glycol (PEG). PEG is a polymer which for its lack of immunogenicity, antigenicity and toxicity obtained approval for use in human medicine. In this work we have studied the solubilization of Hyp aggregates in the presence of PEG-PE and PEG-cholesterol micelles. The concentration of polymeric micelles which allows total monomerization of Hyp corresponds to the critical micellar concentration of these micelles (~10(-6) M). We have also investigated the effect of the molecular weight and concentration of PEG on the transition of aggregated Hyp to its monomeric form. PEGs with low molecular weight (< 1000 g/mol) do not significantly contribute to the solubilization of Hyp. However, PEGs with molecular weight > 2000 g/mol efficiently transform Hyp aggregates to the monomeric state of this photosensitizer.

Properties of bovine erythrocyte acetylcholinesterase solubilized by phosphatidylinositol-specific phospholipase C1.

PubMed

Taguchi, R; Ikezawa, H

1987-10-01

The properties of acetylcholinesterase solubilized from bovine erythrocyte membrane by phosphatidylinositol (PI)-specific phospholipase C of Bacillus thuringiensis or with a detergent, Lubrol-PX, were studied. The activity of Lubrol-PX-solubilized acetylcholinesterase was broadly distributed in the fractions having Ve/Vo = 1.0-2.0 in gel filtration on a Sepharose 6B column. The intermediary fractions (Ve/Vo = 1.3-1.7) were collected as "the middle active Sepharose 6B eluate" and characterized on the basis of enzymology and protein chemistry. When this eluate was treated with PI-specific phospholipase C, the major activity peak was obtained in the later fractions with Ve/Vo = 1.75-2.0 on the same column chromatography. Lubrol-solubilized and phospholipase C-treated acetylcholinesterase preparations were different in the thermostability, the elution profiles of chromatography on Mono Q, butyl-Toyopearl and phenyl-Sepharose columns, and the affinity to phospholipid micelles. On treatment with PI-specific phospholipase C, Lubrol-solubilized acetylcholinesterase became more thermostable. The phospholipase C-treated enzyme was eluted at lower NaCl concentration from the Mono Q column than the Lubrol-solubilized enzyme. The most important difference was observed in the hydrophobicity of these two enzyme preparations. The Lubrol-solubilized enzyme shows high affinity to phospholipid micelles and hydrophobic adsorbents such as butyl-Toyopearl and phenyl-Sepharose. However, this hydrophobicity was lost when acetylcholinesterase was solubilized from bovine erythrocyte membrane by PI-specific phospholipase C. The presence of myo-inositol was confirmed in the purified preparation of acetylcholinesterase by gas chromatography (GC)-mass spectrometry (MS).(ABSTRACT TRUNCATED AT 250 WORDS)

The selective solubilization of different murine splenocyte membrane fractions with lubrol WX and triton X-100 distinguishes two forms of Ia antigens. A cell surface (alpha, beta) and an intracellular (alpha, Ii, beta).

PubMed

Moosic, J P; Sung, E; Nilson, A; Jones, P P; McKean, D J

1982-08-25

The selective solubilization of different murine lymphocyte membrane compartments with several nonionic detergents was used to study the subcellular distribution of two distinct forms of lymphocyte cell recognition structures (Ia antigens). Ia antigens were isolated with a monoclonal anti-Ia immunoadsorbent from murine splenocytes that had been solubilized with four different nonionic detergents. Analyses of the immunoprecipitates indicated that Lubrol WX was selectively solubilizing a subpopulation of Ia consisting of mature highly glycosylated alpha and beta polypeptides which were not associated with Ii polypeptide. A second Ia subpopulation consisting of less glycosylated cytoplasmic precursor alpha and beta polypeptides associated with Ii polypeptide was immunoprecipitated from the Lubrol WX-insoluble material after solubilizing this material with Triton X-100. Comparable results were obtained when HLA-DR antigens were similarly isolated from cultured human lymphoblastoid cells. This selective solubilization phenomenon was not unique to Ia antigens. Only mature highly glycosylated H-2K molecules were immunoprecipitated from the Lubrol WX-soluble material while the less glycosylated precursor H-2K molecules were immunoprecipitated from the Triton X-100-solubilized Lubrol-insoluble material. These data directly demonstrate that the Ii polypeptide is exclusively associated with the intracellular Ia antigen cytoplasmic precursor molecules. These data also indicate that, under the conditions used in these experiments, Lubrol WX does not completely solubilize integral membrane proteins that have previously been shown to be associated with the rough endoplasmic reticulum.

Combined application of bio-organic phosphate and phosphorus solubilizing bacteria (Bacillus strain MWT 14) improve the performance of bread wheat with low fertilizer input under an arid climate.

PubMed

Tahir, Muhammad; Khalid, Umaira; Ijaz, Muhammad; Shah, Ghulam Mustafa; Naeem, Muhammad Asif; Shahid, Muhammad; Mahmood, Khalid; Ahmad, Naveed; Kareem, Fazal

2018-04-24

This study was aimed to investigate the effect of bio-organic phosphate either alone or in combination with phosphorus solubilizing bacteria strain (Bacillus MWT-14) on the growth and productivity of two wheat cultivars (Galaxy-2013 and Punjab-2011) along with recommended (150-100NPkgha -1 ) and half dose (75-50NPkgha -1 ) of fertilizers. The combined application of bio-organic phosphate and the phosphorous solubilizing bacteria strain at either fertilizer level significantly improved the growth, yield parameters and productivity of both wheat cultivars compared to non-inoculated control treatments. The cultivar Punjab-2011 produced the higher chlorophyll contents, crop growth rate, and the straw yield at half dose of NP fertilizer; while Galaxy-2013, with the combined application of bio-organic phosphate and phosphorous solubilizing bacteria under recommended NP fertilizer dose. Combined over both NP fertilizer levels, the combined use of bio-organic phosphate and phosphorous solubilizing bacteria enhanced the grain yield of cultivar Galaxy-2013 by 54.3% and that of cultivar Punjab-2011 by 83.3%. The combined application of bio-organic phosphate and phosphorous solubilizing bacteria also increased the population of phosphorous solubilizing bacteria, the soil organic matter and phosphorous contents in the soil. In conclusion, the combined application of bio-organic phosphate and phosphorous solubilizing bacteria offers an eco-friendly option to harvest the better wheat yield with low fertilizer input under arid climate. Copyright © 2018 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Mild-temperature thermochemical pretreatment of green macroalgal biomass: Effects on solubilization, methanation, and microbial community structure.

PubMed

Jung, Heejung; Baek, Gahyun; Kim, Jaai; Shin, Seung Gu; Lee, Changsoo

2016-01-01

The effects of mild-temperature thermochemical pretreatments with HCl or NaOH on the solubilization and biomethanation of Ulva biomass were assessed. Within the explored region (0-0.2M HCl/NaOH, 60-90°C), both methods were effective for solubilization (about 2-fold increase in the proportion of soluble organics), particularly under high-temperature and high-chemical-dose conditions. However, increased solubilization was not translated into enhanced biogas production for both methods. Response surface analysis statistically revealed that HCl or NaOH addition enhances the solubilization degree while adversely affects the methanation. The thermal-only treatment at the upper-limit temperature (90°C) was estimated to maximize the biogas production for both methods, suggesting limited potential of HCl/NaOH treatment for enhanced Ulva biomethanation. Compared to HCl, NaOH had much stronger positive and negative effects on the solubilization and methanation, respectively. Methanosaeta was likely the dominant methanogen group in all trials. Bacterial community structure varied among the trials according primarily to HCl/NaOH addition. Copyright © 2015 Elsevier Ltd. All rights reserved.

Solubilization of insoluble zinc compounds by Gluconacetobacter diazotrophicus and the detrimental action of zinc ion (Zn2+) and zinc chelates on root knot nematode Meloidogyne incognita.

PubMed

Saravanan, V S; Kalaiarasan, P; Madhaiyan, M; Thangaraju, M

2007-03-01

To examine the zinc (Zn) solubilization potential and nematicidal properties of Gluconacetobacter diazotrophicus. Atomic Absorption Spectrophotometer, Differential Pulse Polarography and Gas Chromatography Coupled Mass Spectrometry were used to estimate the total Zn and Zn(2+) ions and identify the organic acids present in the culture supernatants. The effect of culture filtrate of Zn-amended G. diazotrophicus PAl5 on Meloidogyne incognita in tomato was examined under gnotobiotic conditions. Gluconacetobacter diazotrophicus PAl5 effectively solubilized the Zn compounds tested and 5-ketogluconic acid was identified as the major organic acid aiding the solubilization of zinc oxide. The presence of Zn compounds in the culture filtrates of G. diazotrophicus enhanced the mortality and reduced the root penetration of M. incognita under in vitro conditions. 5-ketogluconic acid produced by G. diazotrophicus mediated the solubilization process and the available Zn(2+) ions enhanced the nematicidal activity of G. diazotrophicus against M. incognita. Zn solubilization and enhanced nematicidal activity of Zn-amended G. diazotrophicus provides the possibility of exploiting it as a plant growth promoting bacteria.

Endocytosis of GPI-linked membrane folate receptor-alpha

PubMed Central

1996-01-01

GPI-linked membrane folate receptors (MFRs) have been implicated in the receptor-mediated uptake of reduced folate cofactors and folate-based chemotherapeutic drugs. We have studied the biosynthetic transport to and internalization of MFR isoform alpha in KB-cells. MFR-alpha was synthesized as a 32-kD protein and converted in a maturely glycosylated 36-38-kD protein 1 h after synthesis. 32-kD MFR-alpha was completely soluble in Triton X-100 at 0 degree C. In contrast, only 33% of the 36- 38-kD species could be solubilized at these conditions whereas complete solubilization was obtained in Triton X-100 at 37 degrees C or in the presence of saponin at 0 degree C. Similar solubilization characteristics were found when MFR-alpha at the plasma membrane was labeled with a crosslinkable 125I-labeled photoaffinity-analog of folic acid as a ligand. Triton X-100-insoluble membrane domains containing MFR-alpha could be separated from soluble MFR-alpha on sucrose flotation gradients. Only Triton X-100 soluble MFR-alpha was internalized from the plasma membrane. The reduced-folate-carrier, an integral membrane protein capable of translocating (anti-)folates across membranes, was completely excluded from the Triton X-100- resistant membrane domains. Internalized MFR-alpha recycled slowly to the cell surface during which it remained soluble in Triton X-100 at 0 degree C. Using immunoelectron microscopy, we found MFR-alpha along the entire endocytic pathway: in clathrin-coated buds and vesicles, and in small and large endosomal vacuoles. In conclusion, our data indicate that a large fraction, if not all, of internalizing MFR-alpha bypasses caveolae. PMID:8567728

Endocytosis of GPI-linked membrane folate receptor-alpha.

PubMed

Rijnboutt, S; Jansen, G; Posthuma, G; Hynes, J B; Schornagel, J H; Strous, G J

1996-01-01

GPI-linked membrane folate receptors (MFRs) have been implicated in the receptor-mediated uptake of reduced folate cofactors and folate-based chemotherapeutic drugs. We have studied the biosynthetic transport to and internalization of MFR isoform alpha in KB-cells. MFR-alpha was synthesized as a 32-kD protein and converted in a maturely glycosylated 36-38-kD protein 1 h after synthesis. 32-kD MFR-alpha was completely soluble in Triton X-100 at 0 degree C. In contrast, only 33% of the 36-38-kD species could be solubilized at these conditions whereas complete solubilization was obtained in Triton X-100 at 37 degrees C or in the presence of saponin at 0 degree C. Similar solubilization characteristics were found when MFR-alpha at the plasma membrane was labeled with a crosslinkable 125I-labeled photoaffinity-analog of folic acid as a ligand. Triton X-100-insoluble membrane domains containing MFR-alpha could be separated from soluble MFR-alpha on sucrose flotation gradients. Only Triton X-100 soluble MFR-alpha was internalized from the plasma membrane. The reduced-folate-carrier, an integral membrane protein capable of translocating (anti-)folates across membranes, was completely excluded from the Triton X-100-resistant membrane domains. Internalized MFR-alpha recycled slowly to the cell surface during which it remained soluble in Triton X-100 at 0 degree C. Using immunoelectron microscopy, we found MFR-alpha along the entire endocytic pathway: in clathrin-coated buds and vesicles, and in small and large endosomal vacuoles. In conclusion, our data indicate that a large fraction, if not all, of internalizing MFR-alpha bypasses caveolae.

Sodium-potassium-activated adenosine triphosphatase of electrophorus electric organ. X. Immunochemical properties of the Lubrol-solubilized enzume and its constituent polypeptides.

PubMed

Jean, D H; Albers, R W; Koval, G J

1975-02-10

Detergent (Lubrol WX)-solubilized sodium-potassium-activated adenosine triphosphatase ((Na+ + K+)-ATPase) of electrophorus electric organ contains two major constituent polypeptides with molecular weights of 96,000 and 58,000 which can be readily demonstrated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. These two polypeptides can be clearly separated and can be obtained in milligram quantities by preparative sodium dodecyl sulfate gel electrophoresis. The separated polypeptides, after removal of sodium dodecyl sulfate, and Lubrol-solubilized (Na+ + K+)-ATPase activity to some degree. Moreover, the degree of inhibition is directly proportional to the increasing amounts of antisera. The inhibition is maximal 4 weeks after the first injection. Immunodiffusion in 1% agar gel indicated that only Lubrol-solubilized enzyme antiserum, but not 58,000-dalton or 96,00-dalton polypeptide antiserum, gives one major precipitin band. However, specific complex formation between each polypeptide antiserum and Lubrol-solubilized enzyme occurs. This was demonstrated indirectly. After incubating Lubrol-solubilized enzyme with increasing amounts of polypeptide antisera at 37 degrees for 15 min, they were placed in the side wells of an immunodiffusion plate with antiserum against Lubrol-solubilized enzyme in the central well. The intensity of the precipitin band decreased with increasing amounts of polypeptide antisera. Thus, the results indicate that both 96,000-dalton and 58,000-dalton polypeptides are integral subunits of (Na+ + K+)-ATPase.

The performance and microbial diversity of temperature-phased hyperthermophilic and thermophilic anaerobic digestion system fed with organic waste.

PubMed

Lee, M Y; Cheon, J H; Hidaka, T; Tsuno, H

2008-01-01

The objective of this study was to evaluate the performances and microbial diversities for development of the effective hyperthermophilic digester system that consists of a hyperthermophilic reactor and hyperthermophilic or thermophilic reactor in series. Lab-scale reactors were operated continuously fed with artificial kitchen garbage. The effect of temperature on the acidification step was firstly investigated. Results indicated that 20.8% of COD solubilization was achieved at 70 degrees C, with 12.6% at 80 degrees C. The average protein solubilization reached 31% at 80 degrees C. Methane conversion efficiency following the acidification was around 85% on average at 55 degrees C, but decreased with increasing temperature and methane gas was not produced over 73 degrees C. As well, bacteria affiliated with the methanogens dominated the population below 65 degrees C, while those affiliated with acidogens were predominant over 73 degrees C. These results indicated that the hyperthermophilic process has considerable benefits to treat wastewater or waste containing high concentration of protein.

Maltose-neopentyl glycol (MNG) amphiphiles for solubilization, stabilization and crystallization of membrane proteins

PubMed Central

Chae, Pil Seok; Rasmussen, Søren G. F.; Rana, Rohini; Gotfryd, Kamil; Chandra, Richa; Goren, Michael A.; Kruse, Andrew C.; Nurva, Shailika; Loland, Claus J.; Pierre, Yves; Drew, David; Popot, Jean-Luc; Picot, Daniel; Fox, Brian G.; Guan, Lan; Gether, Ulrik; Byrne, Bernadette; Kobilka, Brian; Gellman, Samuel H.

2011-01-01

The understanding of integral membrane protein (IMP) structure and function is hampered by the difficulty of handling these proteins. Aqueous solubilization, necessary for many types of biophysical analysis, generally requires a detergent to shield the large lipophilic surfaces displayed by native IMPs. Many proteins remain difficult to study owing to a lack of suitable detergents. We introduce a class of amphiphiles, each of which is built around a central quaternary carbon atom derived from neopentyl glycol, with hydrophilic groups derived from maltose. Representatives of this maltose-neopentyl glycol (MNG) amphiphile family display favorable behavior relative to conventional detergents, as tested on multiple membrane protein systems, leading to enhanced structural stability and successful crystallization. MNG amphiphiles are promising tools for membrane protein science because of the ease with which they may be prepared and the facility with which their structures may be varied. PMID:21037590

Lipid-protein nanodiscs promote in vitro folding of transmembrane domains of multi-helical and multimeric membrane proteins.

PubMed

Shenkarev, Zakhar O; Lyukmanova, Ekaterina N; Butenko, Ivan O; Petrovskaya, Lada E; Paramonov, Alexander S; Shulepko, Mikhail A; Nekrasova, Oksana V; Kirpichnikov, Mikhail P; Arseniev, Alexander S

2013-02-01

Production of helical integral membrane proteins (IMPs) in a folded state is a necessary prerequisite for their functional and structural studies. In many cases large-scale expression of IMPs in cell-based and cell-free systems results in misfolded proteins, which should be refolded in vitro. Here using examples of the bacteriorhodopsin ESR from Exiguobacterium sibiricum and full-length homotetrameric K(+) channel KcsA from Streptomyces lividans we found that the efficient in vitro folding of the transmembrane domains of the polytopic and multimeric IMPs could be achieved during the protein encapsulation into the reconstructed high-density lipoprotein particles, also known as lipid-protein nanodiscs. In this case the self-assembly of the IMP/nanodisc complexes from a mixture containing apolipoprotein, lipids and the partially denatured protein solubilized in a harsh detergent induces the folding of the transmembrane domains. The obtained folding yields showed significant dependence on the properties of lipids used for nanodisc formation. The largest recovery of the spectroscopically active ESR (~60%) from the sodium dodecyl sulfate (SDS) was achieved in the nanodiscs containing anionic saturated lipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPG) and was approximately twice lower in the zwitterionic DMPC lipid. The reassembly of tetrameric KcsA from the acid-dissociated monomer solubilized in SDS was the most efficient (~80%) in the nanodiscs containing zwitterionic unsaturated lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC). The charged and saturated lipids provided lower tetramer quantities, and the lowest yield (<20%) was observed in DMPC. The overall yield of the ESR and KcsA folding was mainly restricted by the efficiency of the protein encapsulation into the nanodiscs. Copyright © 2012 Elsevier B.V. All rights reserved.

Recombinant production of biologically active giant grouper (Epinephelus lanceolatus) growth hormone from inclusion bodies of Escherichia coli by fed-batch culture.

PubMed

Chung, Wen-Jen; Huang, Chi-Lung; Gong, Hong-Yi; Ou, Tsung-Yin; Hsu, Jue-Liang; Hu, Shao-Yang

2015-06-01

Growth hormone (GH) performs important roles in regulating somatic growth, reproduction, osmoregulation, metabolism and immunity in teleosts, and thus, it has attracted substantial attention in the field of aquaculture application. Herein, giant grouper GH (ggGH) cDNA was cloned into the pET28a vector and expressed in Shuffle® T7 Competent Escherichia coli. Recombinant N-terminal 6× His-tagged ggGH was produced mainly in insoluble inclusion bodies; the recombinant ggGH content reached 20% of total protein. For large-scale ggGH production, high-cell density E. coli culture was achieved via fed-batch culture with pH-stat. After 30h of cultivation, a cell concentration of 41.1g/l dry cell weight with over 95% plasmid stability was reached. Maximal ggGH production (4.0g/l; 22% total protein) was achieved via mid-log phase induction. Various centrifugal forces, buffer pHs and urea concentrations were optimized for isolation and solubilization of ggGH from inclusion bodies. Hydrophobic interactions and ionic interactions were the major forces in ggGH inclusion body formation. Complete ggGH inclusion body solubilization was obtained in PBS buffer at pH 12 containing 3M urea. Through a simple purification process including Ni-NTA affinity chromatography and refolding, 5.7mg of ggGH was obtained from 10ml of fed-batch culture (45% recovery). The sequence and secondary structure of the purified ggGH were confirmed by LC-MS/MS mass spectrometry and circular dichroism analysis. The cell proliferation-promoting activity was confirmed in HepG2, ZFL and GF-1 cells with the WST-1 colorimetric bioassay. Copyright © 2015 Elsevier Inc. All rights reserved.

Direct Evidence of Solution-Mediated Superoxide Transport and Organic Radical Formation in Sodium-Oxygen Batteries.

PubMed

Xia, Chun; Fernandes, Russel; Cho, Franklin H; Sudhakar, Niranjan; Buonacorsi, Brandon; Walker, Sean; Xu, Meng; Baugh, Jonathan; Nazar, Linda F

2016-09-07

Advanced large-scale electrochemical energy storage requires cost-effective battery systems with high energy densities. Aprotic sodium-oxygen (Na-O2) batteries offer advantages, being comprised of low-cost elements and possessing much lower charge overpotential and higher reversibility compared to their lithium-oxygen battery cousins. Although such differences have been explained by solution-mediated superoxide transport, the underlying nature of this mechanism is not fully understood. Water has been suggested to solubilize superoxide via formation of hydroperoxyl (HO2), but direct evidence of these HO2 radical species in cells has proven elusive. Here, we use ESR spectroscopy at 210 K to identify and quantify soluble HO2 radicals in the electrolyte-cold-trapped in situ to prolong their lifetime-in a Na-O2 cell. These investigations are coupled to parallel SEM studies that image crystalline sodium superoxide (NaO2) on the carbon cathode. The superoxide radicals were spin-trapped via reaction with 5,5-dimethyl-pyrroline N-oxide at different electrochemical stages, allowing monitoring of their production and consumption during cycling. Our results conclusively demonstrate that transport of superoxide from cathode to electrolyte leads to the nucleation and growth of NaO2, which follows classical mechanisms based on the variation of superoxide content in the electrolyte and its correlation with the crystallization of cubic NaO2. The changes in superoxide content upon charge show that charge proceeds through the reverse solution process. Furthermore, we identify the carbon-centered/oxygen-centered alkyl radicals arising from attack of these solubilized HO2 species on the diglyme solvent. This is the first direct evidence of such species, which are likely responsible for electrolyte degradation.

Limiting solubilizing capacity of some nonionic surfactants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wan, L.S.C.

1980-12-01

This report gives an account of the attempts to solubilize corn oil. A fixed quantity of corn oil or oily dispersion containing corn oil and a sorbitan ester was added to a series of 25 ml of polysorbate solutions of increasing concentration. This investigation showed that corn oil is not solubilized by either aqueous solutions of polyoxyethylene sorbitan esters or by a combination of these surfactants with sorbitan esters. The findings suggest that nonionic surfactants of the polyoxyethylene sorbitan ester type as well as the sorbitan esters have limiting capacities to solubilize extremely hydrophobic substances such as corn oil. 19more » references.« less

Solubilization and other studies on adenylate cyclase of baker's yeast.

PubMed Central

Varimo, K; Londesborough, J

1976-01-01

1. Adenylate cyclase of Saccharomyces cerevisiae was sedimented from mechanically disintegrated preparations of yeast over an unusually wide range of centrifugal forces. 2. The enzyme was readily solubilized by Ficoll and by Lubrol PX. Lubrol caused a 2-fold activation. 3. Both particle-bound and Lubrol-solubilized enzyme had an apparent Km for ATP of 1.6 mM in the presence of 0.4 mM-cyclic AMP and 5 mM-MnCl2 at pH 6.2 and 30 degrees C. 4. The Lubrol-solubilized enzyme behaved on gel filtration as a monodisperse protein with an apparent mol.wt. of about 450000. PMID:793584

Biomediated continuous release phosphate fertilizer

DOEpatents

Goldstein, Alan H.; Rogers, Robert D.

1999-01-01

A composition is disclosed for providing phosphate fertilizer to the root zone of plants. The composition comprises a microorganism capable of producing and secreting a solubilization agent, a carbon source for providing raw material for the microorganism to convert into the solubilization agent, and rock phosphate ore for providing a source of insoluble phosphate that is solubilized by the solubilization agent and released as soluble phosphate. The composition is provided in a physical form, such as a granule, that retains the microorganism, carbon source, and rock phosphate ore, but permits water and soluble phosphate to diffuse into the soil. A method of using the composition for providing phosphate fertilizer to plants is also disclosed.

Solubilization of menthol by platycodin D in aqueous solution: an integrated study of classical experiments and dissipative particle dynamics simulation.

PubMed

Ding, Haiou; Yin, Qianqian; Wan, Guang; Dai, Xingxing; Shi, Xinyuan; Qiao, Yanjiang

2015-03-01

Menthol (M) and platycodin D (PD) are the main active ingredients in Mentha haplocalyx and Platycodon grandiflorum A. DC., respectively. They are commonly used in combination in traditional Chinese medicine. In this study, laboratory experiments and computer simulations were used to investigate the solubilization of M by PD, which was believed to be one of the main causes of the synergistic effect of M. haplocalyx and P. grandiflorum A. DC. Results showed that both the method by which M was added and the concentration of PD had significant effects on the solubilization efficiency of M, and these influences were closely associated with each other. Temperature, an important environmental condition, was also found to have a significant effect on the solubilization effect of PD. These findings not only clarify the molecular basis of the solubilization effect, including amount solubilized at the macroscale and the structures of the micelles, and the drug loading mechanisms and processing at the mesoscale. This work may provide some guidance for the further development of saponins and fundamental research in the drug delivery system. Copyright © 2015 Elsevier B.V. All rights reserved.

HemX is required for production of 2-ketogluconate, the predominant organic anion required for inorganic phosphate solubilization by Burkholderia sp. Ha185.

PubMed

Hsu, Pei-Chun Lisa; Condron, Leo; O'Callaghan, Maureen; Hurst, Mark R H

2015-12-01

The bacterium Burkholderia sp. Ha185 readily solubilizes inorganic phosphate by releasing the low molecular weight organic anion, 2-ketogluconate. Using random transposon mutagenesis and in silico analysis, a mutation that caused almost complete abolition of phosphate solubilization was located within hemX, which is part of the hem operon. Burkholderia sp. Ha185 HemX is a multidomain protein, predicted to encode a bifunctional uroporphyrinogen-III synthetase/uroporphyrin-III C-methyltransferase, which has not previously been implicated in phosphate solubilization. Complementation of hemX restored the ability of the mutant to solubilize phosphate in both plate and liquid cultures. Based on a combination of organic-anion profiling, quantitative polymerase chain reaction and in silico analyses, hemX was confirmed to be solely responsible for hydroxyapatite solubilization in Burkholderia sp. Ha185. It is proposed that the biosynthesis of a yet to be determined redox cofactor by HemX is the main pathway for generating 2-ketogluconate via a haem-dependent gluconate 2-dehydrogenase in Burkholderia sp. Ha185. © 2015 Society for Applied Microbiology and John Wiley & Sons Ltd.

Remediation of soil-bound polynuclear aromatic hydrocarbons using nonionic surfactants

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yeom, IckTae; Ghosh, Mriganka; Cox, C.

1996-12-31

The solubilization and biodegradation of soil-bound PAHs from a manufactured gas plant (MGP) site soil was investigated using surfactants. Three nonionic polyoxyethylene (POE) surfactants, Triton X-100, Tween 80, and Brij 35, were used. The fate of four PAHs, phenanthrene, anthracene, pyrene, and benzo(a)pyrene were monitored during the remediation process. The measured concentrations of solubilized PAHs agreed well with those estimated using micelle-water partitioning coefficient, K{sub m}, and Raoult`s law. The solubilization of soil-bound PAHs by surfactants is a slow, nonequilibrium process. Diffusion of PAH molecules within the weathered soil-tar matrix is proposed as the rate-limiting step in solubilizing PAHs frommore » such soils. A radial diffusion model is used to describe solubilization of PAHs by surfactant washing. The model predicts experimental results fairly well at low surfactant dosages while at high dosages it somewhat overestimates the extent of solubilization. Biodegradation studies were performed using a natural consortium of microorganisms enriched from PAH-contaminated soils. Surfactants enhanced biodegradation of PAHs except for Tween 80. However, biodegradation of surfactants themselves appear to attenuate the beneficial effects of surfactant-mediated bioremediation.« less

PCE solubilization and mobilization by commercial humic acid

NASA Astrophysics Data System (ADS)

Johnson, William P.; John, W. Wynn

1999-01-01

In this paper, comparison is made of terms describing solubilization of hydrophobic organic compounds (HOC) by dissolved humic substances (DHS) and commercial non-ionic surfactants. This paper examines the ability of a commercial humic acid (Aldrich humic acid) to solubilize and mobilize tetrachlorothene (PCE) residual in porous media. The constant for solubilization of PCE by Aldrich humic acid is shown to be a factor of two to thirty times less than that published for dodecyl alcohol ethoxylate surfactants, showing that Aldrich humic acid is less capable than some non-ionic surfactants at solubilizing residual PCE. The depression of PCE-water interfacial tension in the presence of DHS is shown to be significantly less than published values for a non-ionic surfactant, and surfactant mixtures, indicating that the DHS used in this study is less prone to cause mobilization of non-aqueous phase liquids relative to surfactants. Several possible advantages of DHS use in the remediation of subsurface media contaminated with HOC are described, including the ability of DHS to solubilize HOC irrespective of the DHS concentration, and potential lesser tendency of DHS to depress the interfacial tension between non-aqueous phases and water relative to surfactants (an advantage when mobilization is undesired).

An ability of endophytic bacteria from nutgrass (cyperus rotundus) from lafau beach of north nias in producing indole acetic acid and in solubilizing phosphate

NASA Astrophysics Data System (ADS)

Zega, Atriani; Suryanto, Dwi; Yurnaliza

2018-03-01

Endophytic bacteria have taken much attention for their potency to promote plant growth. This study was aimed to isolate endophytic bacteria from nutgrass (Cyperus rotundus) and to examine their potency in producing indole acetic acid (IAA) and in solubilizing phosphate. Isolation of endophytic bacteria was done by slicing and sterilizing root, stem, and leaf sample surface with alcohol 70% and sodium hypochlorite 2%, followed by incubation of the sliced samples in nutrient agar medium. Morphological characterization and simple biochemical tests were performed on bacterial isolates. All bacterial isolates were examined for their ability to produce indole acetic acid and to solubilize phosphate. Three isolates (AZ5, AZ12 and AZ6) out of fifteen indicated the ability to produce indole acetic acid and to solubilize phosphate. IAA producing test using spectrophotometry method showed that AZ5, AZ12,and AZ6 produce more IAA with concentration of 49,91, 48,18, and 44,45 ppm, respectively. Phosphate solubilizing test using Pikovskaya agar medium showed that the three isolates were able to solubilize phosphate with index of 6.27, 3,31, and 3.41 respectively.

Recovery of active N-acetyl-D-glucosamine 2-epimerase from inclusion bodies by solubilization with non-denaturing buffers.

PubMed

Lu, Shih-Chin; Lin, Sung-Chyr

2012-01-05

Overexpression of recombinant N-acetyl-D-glucosamine 2-epimerase, one of the key enzymes for the synthesis of N-acetylneuraminic acid, in E. coli led to the formation of protein inclusion bodies. In this study we report the recovery of active epimerase from inclusion bodies by direct solubilization with Tris buffer. At pH 7.0, 25% of the inclusion bodies were solubilized with Tris buffer. The specific activity of the solubilized proteins, 2.08±0.02 U/mg, was similar to that of the native protein, 2.13±0.01 U/mg. The result of circular dichroism spectroscopy analysis indicated that the structure of the solubilized epimerase obtained with pH 7.0 Tris buffer was similar to that of the native epimerase purified from the clarified cell lysate. As expected, the extent of deviation in CD spectra increased with buffer pH. The total enzyme activity recovered by solubilization from inclusion bodies, 170.41±10.06 U/l, was more than 2.5 times higher than that from the clarified cell lysate, 67.32±5.53 U/l. The results reported in this study confirm the hypothesis that the aggregation of proteins into inclusion bodies is reversible and suggest that direct solubilization with non-denaturing buffers is a promising approach for the recovery of active proteins from inclusion bodies, especially for aggregation-prone multisubunit proteins. Copyright © 2011 Elsevier Inc. All rights reserved.

The removal characteristics of natural organic matter in the recycling of drinking water treatment sludge: Role of solubilized organics.

PubMed

Zhou, Zhiwei; Yang, Yanling; Li, Xing; Ji, Siyang; Zhang, Hao; Wang, Shuai; Zeng, Qingping; Han, Xinghang

2016-01-01

To clarify the role of solubilized organics derived from drinking water treatment sludge (DWTS) in the elimination of natural organic matter (NOM) in the DWTS recycling process, a probe sonoreactor at a frequency of 25 kHz was used to solubilize the organics at varied specific energies. The coagulation behavior related to NOM removal in recycling the sonicated DWTS with and without solubilized organics was evaluated, and the effect on organic fractionations in coagulated water was determined. The study results could provide useful implications in designing DWTS recycling processes that avoid the enrichment of organic matter. Our results indicate that DWTS was disrupted through a low release of soluble chemical oxygen demand (SCOD) and proteins, which could deteriorate the coagulated water quality under the specific energy of 37.87-1212.1 kW h/kg TS. The optimal coagulation behavior for NOM removal was achieved by recycling the sonicated DWTS without solubilized organics at 151.5 kW h/kg TS specific energy. Recycling the sonicated DWTS could increase the enrichment potential of weakly hydrophobic acid, hydrophilic matter, and <3 kDa fractions; the enrichment risks could be reduced by discharging the solubilized organics. Fluorescent characteristic analysis indicated that when recycling the sonicated DWTS without solubilized organics, the removal of humic-like substances was limited, whereas removal of protein-like substances was enhanced, lowering the enrichment potential of protein-like substances. Copyright © 2015. Published by Elsevier B.V.

Proteomic analysis of the renal effects of simulated occupational jet fuel exposure.

PubMed

Witzmann, F A; Bauer, M D; Fieno, A M; Grant, R A; Keough, T W; Lacey, M P; Sun, Y; Witten, M L; Young, R S

2000-03-01

We analyzed protein expression in the cytosolic fraction prepared from whole kidneys in male Swiss-Webster mice exposed 1 h/day for five days to aerosolized JP-8 jet fuel at a concentration of 1000 mg/m3, simulating military occupational exposure. Kidney cytosol samples were solubilized and separated via large-scale, high-resolution two-dimensional electrophoresis (2-DE) and gel patterns scanned, digitized and processed for statistical analysis. Significant changes in soluble kidney proteins resulted from jet fuel exposure. Several of the altered proteins were identified by peptide mass finger-printing and related to ultrastructural abnormalities, altered protein processing, metabolic effects, and paradoxical stress protein/detoxification system responses. These results demonstrate a significant but comparatively moderate JP-8 effect on protein expression in the kidney and provide novel molecular evidence of JP-8 nephrotoxicity. Human risk is suggested by these data but conclusive assessment awaits a noninvasive search for biomarkers in JP-8 exposed humans.

Substrate-modulated unwinding of transmembrane helices in the NSS transporter LeuT.

PubMed

Merkle, Patrick S; Gotfryd, Kamil; Cuendet, Michel A; Leth-Espensen, Katrine Z; Gether, Ulrik; Loland, Claus J; Rand, Kasper D

2018-05-01

LeuT, a prokaryotic member of the neurotransmitter:sodium symporter (NSS) family, is an established structural model for mammalian NSS counterparts. We investigate the substrate translocation mechanism of LeuT by measuring the solution-phase structural dynamics of the transporter in distinct functional states by hydrogen/deuterium exchange mass spectrometry (HDX-MS). Our HDX-MS data pinpoint LeuT segments involved in substrate transport and reveal for the first time a comprehensive and detailed view of the dynamics associated with transition of the transporter between outward- and inward-facing configurations in a Na + - and K + -dependent manner. The results suggest that partial unwinding of transmembrane helices 1/5/6/7 drives LeuT from a substrate-bound, outward-facing occluded conformation toward an inward-facing open state. These hitherto unknown, large-scale conformational changes in functionally important transmembrane segments, observed for LeuT in detergent-solubilized form and when embedded in a native-like phospholipid bilayer, could be of physiological relevance for the translocation process.

Microemulsions containing long-chain oil ethyl oleate improve the oral bioavailability of piroxicam by increasing drug solubility and lymphatic transportation simultaneously.

PubMed

Xing, Qiao; Song, Jia; You, Xiuhua; Xu, Dongling; Wang, Kexin; Song, Jiaqi; Guo, Qin; Li, Pengyu; Wu, Chuanbin; Hu, Haiyan

2016-09-25

Drug solubility and lymphatic transport enhancements are two main pathways to improve drug oral bioavailability for microemulsions. However, it is not easy to have both achieved simultaneously because excipients used for improving lymphatic transport were usually insufficient in forming microemulsions and solubilizing drugs. Our research is to explore whether ethyl oleate, an oil effective in developing microemulsions with desired solubilizing capability, could increase bioavailability to a higher extent by enhancing lymphatic transport. As a long-chain oil, ethyl oleate won larger microemulsion area than short-chain tributyrin and medium-chain GTCC. In contrast, long-chain soybean oil failed to prepare microemulsions. The solubility of piroxicam in ethyl oleate microemulsions (ME-C) increased by about 30 times than in water. ME-C also won significantly higher AUC0-t compared with tributyrin microemulsions (ME-A) and GTCC microemulsions (ME-B). Oral bioavailability in ME-C decreased by 38% after lymphatic transport was blocked by cycloheximide, severer than those in ME-A and ME-B (8% and 34%). These results suggest that improving lymphatic transport and solubility simultaneously might be a novel strategy to increase drug oral bioavailability to a higher extent than increasing solubility only. Ethyl oleate is a preferred oil candidate due to its integrated advantages of high solubilizing capability, large microemulsion area and effective lymphatic transport. Copyright © 2016 Elsevier B.V. All rights reserved.

Oxygen radical absorbance capacity (ORAC) of cyclodextrin-solubilized flavonoids, resveratrol and astaxanthin as measured with the ORAC-EPR method

PubMed Central

Sueishi, Yoshimi; Ishikawa, Misa; Yoshioka, Daisuke; Endoh, Nobuyuki; Oowada, Shigeru; Shimmei, Masashi; Fujii, Hirotada; Kotake, Yashige

2012-01-01

Recently, we proposed an oxygen radical absorbance capacity method that directly quantifies the antioxidant’s scavenging capacity against free radicals and evaluated the radical scavenging abilities for water soluble antioxidant compounds. In this study, we determined the radical scavenging abilities of lipophilic antioxidants which were solubilized by cyclodextrin in water. Commonly employed fluorescence-based method measures the antioxidant’s protection capability for the fluorescent probe, while we directly quantify free-radical level using electron paramagnetic resonance spin trapping technique. In addition, the spin trapping-based method adopted controlled UV-photolysis of azo-initiator for free radical generation, but in fluorescence-based method, thermal decomposition of azo-initiator was utilized. We determined the radical scavenging abilities of seven well-known lipophilic antioxidants (five flavonoids, resveratrol and astaxanthin), using methylated β-cyclodextrin as a solubilizer. The results indicated that the agreement between spin trapping-based and fluorescence-based values was only fair partly because of a large variation in the previous fluorescence-based data. Typical radical scavenging abilities in trolox equivalent unit are: catechin 0.96; epicatechin 0.94; epigallocatechin gallate 1.3; kaempferol 0.37; myricetin 3.2; resveratrol 0.64; and astaxanthin 0.28, indicating that myricetin possesses the highest antioxidant capacity among the compounds tested. We sorted out the possible causes of the deviation between the two methods. PMID:22448093

Oxygen radical absorbance capacity (ORAC) of cyclodextrin-solubilized flavonoids, resveratrol and astaxanthin as measured with the ORAC-EPR method.

PubMed

Sueishi, Yoshimi; Ishikawa, Misa; Yoshioka, Daisuke; Endoh, Nobuyuki; Oowada, Shigeru; Shimmei, Masashi; Fujii, Hirotada; Kotake, Yashige

2012-03-01

Recently, we proposed an oxygen radical absorbance capacity method that directly quantifies the antioxidant's scavenging capacity against free radicals and evaluated the radical scavenging abilities for water soluble antioxidant compounds. In this study, we determined the radical scavenging abilities of lipophilic antioxidants which were solubilized by cyclodextrin in water. Commonly employed fluorescence-based method measures the antioxidant's protection capability for the fluorescent probe, while we directly quantify free-radical level using electron paramagnetic resonance spin trapping technique. In addition, the spin trapping-based method adopted controlled UV-photolysis of azo-initiator for free radical generation, but in fluorescence-based method, thermal decomposition of azo-initiator was utilized. We determined the radical scavenging abilities of seven well-known lipophilic antioxidants (five flavonoids, resveratrol and astaxanthin), using methylated β-cyclodextrin as a solubilizer. The results indicated that the agreement between spin trapping-based and fluorescence-based values was only fair partly because of a large variation in the previous fluorescence-based data. Typical radical scavenging abilities in trolox equivalent unit are: catechin 0.96; epicatechin 0.94; epigallocatechin gallate 1.3; kaempferol 0.37; myricetin 3.2; resveratrol 0.64; and astaxanthin 0.28, indicating that myricetin possesses the highest antioxidant capacity among the compounds tested. We sorted out the possible causes of the deviation between the two methods.

Soluble arabinoxylan enhances large intestinal microbial health biomarkers in pigs fed a red meat-containing diet.

PubMed

Williams, Barbara A; Zhang, Dagong; Lisle, Allan T; Mikkelsen, Deirdre; McSweeney, Christopher S; Kang, Seungha; Bryden, Wayne L; Gidley, Michael J

2016-04-01

The aim of this study was to investigate how moderately increased dietary red meat combined with a soluble fiber (wheat arabinoxylan [AX]) alters the large intestinal microbiota in terms of fermentative end products and microbial community profiles in pigs. Four groups of 10 pigs were fed Western-type diets containing two amounts of red meat, with or without a solubilized wheat AX-rich fraction for 4 wk. After euthanasia, fermentative end products (short-chain fatty acids, ammonia) of digesta from four sections of large intestine were measured. Di-amino-pimelic acid was a measure of total microbial biomass, and bacterial profiles were determined using a phylogenetic microarray. A factorial model determined effects of AX and meat content. Arabinoxylan was highly fermentable in the cecum, as indicated by increased concentrations of short-chain fatty acids (particularly propionate). Protein fermentation end products were decreased, as indicated by the reduced ammonia and branched-chain ratio although this effect was less prominent distally. Microbial profiles in the distal large intestine differed in the presence of AX (including promotion of Faecalibacterium prausnitzii), consistent with an increase in carbohydrate versus protein fermentation. Increased di-amino-pimelic acid (P < 0.0001) suggested increased microbial biomass for animals fed AX. Solubilized wheat AX has the potential to counteract the effects of dietary red meat by reducing protein fermentation and its resultant toxic end products such as ammonia, as well as leading to a positive shift in fermentation end products and microbial profiles in the large intestine. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

Human Intestinal Fluid Layer Separation: The Effect On Colloidal Structures & Solubility Of Lipophilic Compounds.

PubMed

Danny, Riethorst; Amitava, Mitra; Filippos, Kesisoglou; Wei, Xu; Jan, Tack; Joachim, Brouwers; Patrick, Augustijns

2018-05-23

In addition to individual intestinal fluid components, colloidal structures are responsible for enhancing the solubility of lipophilic compounds. The present study investigated the link between as well as the variability in the ultrastructure of fed state human intestinal fluids (FeHIF) and their solubilizing capacity for lipophilic compounds. For this purpose, FeHIF samples from 10 healthy volunteers with known composition and ultrastructure were used to determine the solubility of four lipophilic compounds. In light of the focus on solubility and ultrastructure, the study carefully considered the methodology of solubility determination in relation to colloid composition and solubilizing capacity of FeHIF. To determine the solubilizing capacity of human and simulated intestinal fluids, the samples were saturated with the compound of interest, shaken for 24 h, and centrifuged. When using FeHIF, solubilities were determined in the micellar layer of FeHIF, i.e. after removing the upper (lipid) layer (standard procedure), as well as in 'full' FeHIF (without removal of the upper layer). Compound concentrations were determined using HPLC-UV/fluorescence. To link the solubilizing capacity with the ultrastructure, all human and simulated fluids were imaged using transmission electron microscopy (TEM) before and after centrifugation and top layer (lipid) removal. Comparing the ultrastructure and solubilizing capacity of individual FeHIF samples demonstrated a high intersubject variability in postprandial intestinal conditions. Imaging of FeHIF after removal of the upper layer clearly showed that only micellar structures remain in the lower layer. This observation suggests that larger colloids such as vesicles and lipid droplets are contained in the upper, lipid layer. The solubilizing capacity of most FeHIF samples substantially increased with inclusion of this lipid layer. The relative increase in solubilizing capacity upon inclusion of the lipid layer was most pronounced in samples that contained mainly vesicles alongside the micelles. Current fed state simulated intestinal fluids do not contain the larger colloids observed in the lipid layer of FeHIF and can only simulate the solubilizing capacity of the micellar layer of FeHIF. While the importance of drug molecules solubilized in the micellar layer of postprandial intestinal fluids for absorption has been extensively demonstrated previously, the in-vivo relevance of drug solubilization in the lipid layer is currently unclear. In the dynamic environment of the human gastrointestinal tract, drug initially entrapped in larger postprandial colloids may become available for absorption upon lipid digestion and uptake. The current study, demonstrating the substantial solubilization of lipophilic compounds in the larger colloids of postprandial intestinal fluids, warrants further research in this field. Copyright © 2018. Published by Elsevier B.V.

Biomediated continuous release phosphate fertilizer

DOEpatents

Goldstein, A.H.; Rogers, R.D.

1999-06-15

A composition is disclosed for providing phosphate fertilizer to the root zone of plants. The composition comprises a microorganism capable of producing and secreting a solubilization agent, a carbon source for providing raw material for the microorganism to convert into the solubilization agent, and rock phosphate ore for providing a source of insoluble phosphate that is solubilized by the solubilization agent and released as soluble phosphate. The composition is provided in a physical form, such as a granule, that retains the microorganism, carbon source, and rock phosphate ore, but permits water and soluble phosphate to diffuse into the soil. A method of using the composition for providing phosphate fertilizer to plants is also disclosed. 13 figs.

Effects of temperature and glucose limitation on coal solubilization by Candida ML13

DOE Office of Scientific and Technical Information (OSTI.GOV)

Evans, B.

1991-04-01

Biological processing has received considerable attention in recent years as a technology for the utilization of low-ranked coals. Several fungi and actinomycetes have been shown to liquefy highly oxidized coal in pure culture under aerobic conditions. This report describes the optimization of cultural conditions for coal solubilization by Candida sp. ML13, an organism originally isolated from a naturally weathered coal seam. Coal solubilization by surface cultures of Candida sp. has previously been demonstrated. The author describes here the elicitation of the activity in submerged cultures as well as the effect of carbohydrate concentration, carbon source, temperature, and agitation rate onmore » coal solubilization by this organism.« less

Exploring the Arabidopsis proteome: influence of protein solubilization buffers on proteome coverage.

PubMed

Marondedze, Claudius; Wong, Aloysius; Groen, Arnoud; Serrano, Natalia; Jankovic, Boris; Lilley, Kathryn; Gehring, Christoph; Thomas, Ludivine

2014-12-31

The study of proteomes provides new insights into stimulus-specific responses of protein synthesis and turnover, and the role of post-translational modifications at the systems level. Due to the diverse chemical nature of proteins and shortcomings in the analytical techniques used in their study, only a partial display of the proteome is achieved in any study, and this holds particularly true for plant proteomes. Here we show that different solubilization and separation methods have profound effects on the resulting proteome. In particular, we observed that the type of detergents employed in the solubilization buffer preferentially enriches proteins in different functional categories. These include proteins with a role in signaling, transport, response to temperature stimuli and metabolism. This data may offer a functional bias on comparative analysis studies. In order to obtain a broader coverage, we propose a two-step solubilization protocol with first a detergent-free buffer and then a second step utilizing a combination of two detergents to solubilize proteins.

Exploring the Arabidopsis Proteome: Influence of Protein Solubilization Buffers on Proteome Coverage

PubMed Central

Marondedze, Claudius; Wong, Aloysius; Groen, Arnoud; Serrano, Natalia; Jankovic, Boris; Lilley, Kathryn; Gehring, Christoph; Thomas, Ludivine

2014-01-01

The study of proteomes provides new insights into stimulus-specific responses of protein synthesis and turnover, and the role of post-translational modifications at the systems level. Due to the diverse chemical nature of proteins and shortcomings in the analytical techniques used in their study, only a partial display of the proteome is achieved in any study, and this holds particularly true for plant proteomes. Here we show that different solubilization and separation methods have profound effects on the resulting proteome. In particular, we observed that the type of detergents employed in the solubilization buffer preferentially enriches proteins in different functional categories. These include proteins with a role in signaling, transport, response to temperature stimuli and metabolism. This data may offer a functional bias on comparative analysis studies. In order to obtain a broader coverage, we propose a two-step solubilization protocol with first a detergent-free buffer and then a second step utilizing a combination of two detergents to solubilize proteins. PMID:25561235

Fluidized-bed bioreactor process for the microbial solubiliztion of coal

DOEpatents

Scott, Charles D.; Strandberg, Gerald W.

1989-01-01

A fluidized-bed bioreactor system for the conversion of coal into microbially solubilized coal products. The fluidized-bed bioreactor continuously or periodically receives coal and bio-reactants and provides for the production of microbially solubilized coal products in an economical and efficient manner. An oxidation pretreatment process for rendering coal uniformly and more readily susceptible to microbial solubilization may be employed with the fluidized-bed bioreactor.

A novel solubilization technique for poorly soluble drugs through the integration of nanocrystal and cocrystal technologies.

PubMed

Karashima, Masatoshi; Kimoto, Kouya; Yamamoto, Katsuhiko; Kojima, Takashi; Ikeda, Yukihiro

2016-10-01

The aim of the present study was to develop a novel solubilization technique consisting of a nano-cocrystal suspension by integrating cocrystal and nanocrystal formulation technologies to maximize solubilization over current solubilizing technologies. Monodisperse carbamazepine-saccharin, indomethacin-saccharin, and furosemide-caffeine nano-cocrystal suspensions, as well as a furosemide-cytosine nano-salt suspension, were successfully prepared with particle sizes of less than 300nm by wet milling with the stabilizers hydroxypropyl methylcellulose and sodium dodecyl sulfate. Interestingly, the properties of resultant nano-cocrystal suspensions were dramatically changed depending on the physicochemical and structural properties of the cocrystals. In the formulation optimization, the concentration and ratio of the stabilizers also influenced the zeta potentials and particles sizes of the resultant nano-cocrystal suspensions. Raman spectroscopic analysis revealed that the crystalline structures of the cocrystals were maintained in the nanosuspensions, and were physically stable for at least one month. Furthermore, their dissolution profiles were significantly improved over current solubilization-enabling technologies, nanocrystals, and cocrystals. In the present study, we demonstrated that nano-cocrystal formulations can be a new promising option for solubilization techniques to improve the absorption of poorly soluble drugs, and can expand the development potential of poorly soluble candidates in the pharmaceutical industry. Copyright © 2016 Elsevier B.V. All rights reserved.

Aggregate-based sub-CMC Solubilization of n-Alkanes by Monorhamnolipid Biosurfactant.

PubMed

Zhong, Hua; Yang, Xin; Tan, Fei; Brusseau, Mark L; Yang, Lei; Liu, Zhifeng; Zeng, Guangming; Yuan, Xingzhong

2016-03-01

Solubilization of n -decane, dodecane, tetradecane and hexadecane by monorhamnolipid biosurfactant (monoRL) at concentrations near the critical micelle concentration (CMC) was investigated. The apparent solubility of all the four alkanes increases linearly with increasing monoRL concentration either below or above CMC. The capacity of solubilization presented by the molar solubilization ratio (MSR), however, is stronger at monoRL concentrations below CMC than above CMC. The MSR decreases following the order dodecane > decane > tetradecane > hexadecane at monoRL concentration below CMC. Formation of aggregates at sub-CMC monoRL concentrations was demonstrated by dynamic light scattering (DLS) and cryo-transmission electron microscopy examination. DLS-based size ( d ) and zeta potential of the aggregates decrease with increasing monoRL concentration. The surface excess ( Γ ) of monoRL calculated based on alkane solubility and aggregate size data increases rapidly with increasing bulk monoRL concentration, and then asymptotically approaches the maximum surface excess ( Γ max ). Relation between Γ and d indicates that the excess of monoRL molecules at the aggregate surface greatly impacts the surface curvature. The results demonstrate formation of aggregates for alkane solubilization at monoRL concentrations below CMC, indicating the potential of employing low-concentration rhamnolipid for enhanced solubilization of hydrophobic organic compounds.

Fungal extracellular phosphatases: their role in P cycling under different pH and P sources availability.

PubMed

Della Mónica, I F; Godoy, M S; Godeas, A M; Scervino, J M

2018-01-01

The aim of this work is to analyse the effect of pH, fungal identity and P chemical nature on microbial development and phosphatase release, discussing solubilization and mineralization processes in P cycling. P solubilizing fungi (Talaromyces flavus, T. helicus L, T. helicus N, T. diversus and Penicillium purpurogenum) were grown under three pH conditions (6, 6·5 and 8·5) and with different inorganic (calcium, iron, aluminium and rock) and organic (lecithin and phytate) P sources. P solubilization, mineralization, growth and phosphatase production were recorded. Acid and neutral environments maximized fungal development and P recycling. P chemical nature changed the phosphatases release pattern depending on the fungal identity. Acid phosphatase activity was higher than alkaline phosphatases, regardless of pH or sample times. Alkaline phosphatases were affected by a combination of those factors. P chemical nature and pH modify fungal growth, P mineralization and solubilization processes. The underlying fungal identity-dependent metabolism governs the capacity and efficiency of P solubilization and mineralization. P solubilization and mineralization processes are interrelated and simultaneously present in soil fungi. This study constitutes a reference work to improve the selection of fungal bioinoculants in different environmental conditions, highlighting their role in P cycling. © 2017 The Society for Applied Microbiology.

The potential of fluorinated surfactants in membrane biochemistry.

PubMed

Shepherd, F H; Holzenburg, A

1995-01-01

Detergents are important reagents in membrane biochemistry. Since each membrane system studied places different demands on the detergent in terms of desirous physicochemical properties, detergents new to biochemistry must continuously be sought. Ammonium perfluorooctanoate (APFO) was investigated, as representative of fluorinated surfactants, in terms of its suitability as a "biological detergent." It did not interfere with the Markwell modification of the Lowry procedure at detergent concentrations of up to 2% (w/v). Critical micellization concentration (cmc) values (0.013-0.0275 M) for this detergent were determined in a number of buffers of biological interest. It was demonstrated that the detergent can be removed by dialysis, albeit slowly. This slow removal may be particularly useful for reconstitution/crystallization studies. Solubilization studies on several membrane systems containing the proteins listed (the major protein of the membrane sector of the vacuolar H(+)-ATPase (16 kDa protein); photosystem II; equine herpes virus (EHV) envelope proteins) indicate that it is a potent solubilizing agent, likely to enhance the yield in cases where solubilization has already been demonstrated, and, in other cases, to solubilize proteins formerly recalcitrant to solubilization. The removal of APFO from solubilized 16-kDa protein by means of Extracti-Gel D resin as a means of exchanging detergents quickly and with a minimum requirement for second detergent was investigated.

Phosphate Solubilization and Gene Expression of Phosphate-Solubilizing Bacterium Burkholderia multivorans WS-FJ9 under Different Levels of Soluble Phosphate.

PubMed

Zeng, Qingwei; Wu, Xiaoqin; Wang, Jiangchuan; Ding, Xiaolei

2017-04-28

Phosphate-solubilizing bacteria (PSB) have the ability to dissolve insoluble phosphate and enhance soil fertility. However, the growth and mineral phosphate solubilization of PSB could be affected by exogenous soluble phosphate and the mechanism has not been fully understood. In the present study, the growth and mineral phosphate-solubilizing characteristics of PSB strain Burkholderia multivorans WS-FJ9 were investigated at six levels of exogenous soluble phosphate (0, 0.5, 1, 5, 10, and 20 mM). The WS-FJ9 strain showed better growth at high levels of soluble phosphate. The phosphate-solubilizing activity of WS-FJ9 was reduced as the soluble phosphate concentration increased, as well as the production of pyruvic acid. Transcriptome profiling of WS-FJ9 at three levels of exogenous soluble phosphate (0, 5, and 20 mM) identified 446 differentially expressed genes, among which 44 genes were continuously up-regulated when soluble phosphate concentration was increased and 81 genes were continuously down-regulated. Some genes related to cell growth were continuously up-regulated, which would account for the better growth of WS-FJ9 at high levels of soluble phosphate. Genes involved in glucose metabolism, including glycerate kinase, 2-oxoglutarate dehydrogenase, and sugar ABC-type transporter, were continuously down-regulated, which indicates that metabolic channeling of glucose towards the phosphorylative pathway was negatively regulated by soluble phosphate. These findings represent an important first step in understanding the molecular mechanisms of soluble phosphate effects on the growth and mineral phosphate solubilization of PSB.

Regulation of Coal Polymer Degradation by Fungi

DOE Office of Scientific and Technical Information (OSTI.GOV)

NONE

1998-09-01

During this reporting period we have further studied the oxidation of soluble coal macromolecules by lignin peroxidase from Phanerochaete chrysosporium . Previous studies by others have suggested that a soluble fraction (coal macromolecule B-111) from a nitric acid solubilized North Dakota Lignite is depolymerized by this enzyme. Our investigations indicate that fraction B-111 is a substrate for lignin peroxidase as this material is decolorized in the presence of lignin peroxidase H8 and hydrogen peroxide. Of interest, however, is the observation that little, if any, depolymerization of this material occurs. Instead, it appears that lignin peroxidase and coal macromolecule B-111 formmore » a precipitate. These results are similar to those observed in our investigations of lignin peroxidase mediated oxidation of oxalate solubilize coal macromolecule. Previous studies in our laboratory using a spectrophotometric assay suggested that, in addition to oxalate, several other fungal metabolites are able to solubilize leonardite. We have reinvestigated this phenomenon using a more reliable gravimetric procedure for assessing solubilization. Our results confirm our earlier findings that malate, oxaloacetate and citrate are effective solubilizing agents whereas succinate, fumarate and x-ketoglutarate solubilize relatively small amounts of leonardite. Finally, we have studied the composition of the insoluble material remaining following extensive solubilization by sodium oxalate. The ratio of hydrogen to carbon is increased in the insoluble material relative to the parent leonardite. However, the ratio of oxygen to carbon is also increased in the insoluble material. Thus, the insoluble material does not appear to be more highly reduced that the parent leonardite and is not likely to be a better fuel that the parent material.« less

Characterization of (/sup 3/H)pirenzepine binding to muscarinic cholinergic receptors solubilized from rat brain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Luthin, G.R.; Wolfe, B.B.

Membranes prepared from rat cerebral cortex were solubilized in buffer containing 1% digitonin. Material present in the supernatant after centrifugation at 147,000 X g was shown to contain binding sites for both (/sup 3/H)quinuclidinyl benzilate ((/sup 3/H)QNB) and (/sup 3/H)pirenzepine ((/sup 3/H)PZ). Recovery of binding sites was approximately 25% of the initial membrane-bound (/sup 3/H)QNB binding sites. The Kd values for (/sup 3/H)QNB and (/sup 3/H)PZ binding to solubilized receptors were 0.3 nM and 0.1 microM, respectively. As has been observed previously in membrane preparations, (/sup 3/H)PZ appeared to label fewer solubilized binding sites than did (/sup 3/H)QNB. Maximum bindingmore » values for (/sup 3/H)PZ and (/sup 3/H)QNB binding to solubilized receptors were approximately 400 and 950 fmol/mg of protein, respectively. Competition curves for PZ inhibiting the binding of (/sup 3/H)QNB, however, had Hill slopes of 1, with a Ki value of 0.24 microM. The k1 and k-1 for (/sup 3/H)PZ binding were 3.5 X 10(6) M-1 min-1 and 0.13 min-1, respectively. The muscarinic receptor antagonists atropine, scopolamine and PZ inhibited the binding of (/sup 3/H)QNB and (/sup 3/H)PZ to solubilized receptors with Hill slopes of 1, as did the muscarinic receptor agonist oxotremorine. The muscarinic receptor agonist carbachol competed for (/sup 3/H)QNB and (/sup 3/H)PZ binding with a Hill slope of less than 1 in cerebral cortex, but not in cerebellum. GTP did not alter the interactions of carbachol or oxotremorine with the solubilized receptor. Together, these data suggest that muscarinic receptor sites solubilized from rat brain retain their abilities to interact selectively with muscarinic receptor agonists and antagonists.« less

[Screening, identification and phosphate-solubilizing characteristics of phosphate-solubilizing bacteria strain D2 (Pantoea sp.)in rhizosphere of Pinus tabuliformis in iron tailings yard.

PubMed

Wang, Jun Juan; Yan, Ai Hua; Wang, Wei; Li, Ji Quan; Li, Yu Ling

2016-11-18

Two strains of phosphate-solubilizing bacteria were isolated from the rhizosphere of Pinus tabuliformis in iron tailings vegetation restoration areas in Malan Town, Qianan City, Hebei Pro-vince. The bacterial strain D2 with strong phosphate-solubilizing capacity was obtained via screening with plate and shake flask. Based on the morphology, physiology and biochemistry, and the sequence analysis of 16S rDNA, the D2 was identified as a member of Pantoea sp. A fermentation experiment was conducted to investigate the effect of carbon and nitrogen sources on the phosphate-solubilizing capacity of the strain D2; under different nitrogen sources, the organic acids in liquid culture, as well as their types and contents were determined by high performance liquid chromatography. The results showed that the strain D2 was capable of efficiently solubilizing tricalcium phosphate, and the highest value of available phosphorus was up to 392.13 mg·L -1 in liquid culture. The strain D2 displayed the strongest phosphate-solubilizing capability when glucose and ammonium sulfate were used as carbon and nitrogen sources in the culture media, respectively. Under varied nitrogen sources, the resulting organic acids and their types and contents were different. When the nitrogen source in culture media was ammonium sulfate, ammonium chloride, potassium nitrate, sodium nitrate or ammonium nitrate, all four organic acids, including oxalic acid, formic acid, acetic acid and citric acid, were produced. In addition, malic acid was uniquely produced when ammonium sulfate, ammonium chloride or ammonium nitrate was used as the nitrogen source. By Pearson's correlation analysis, a significant positive correlation between the acetic acid content and the available phosphorus content was found (r=0.886, P<0.05), suggesting that acetic acid produced by strain D2 played an important role in promoting inorganic phosphorus dissolution, which was most likely to be one of the important phosphate-solubilizing mechanisms of the strain.

Blocking monocyte transmigration in in vitro system by an anti-CD99 human antibody in single chain fragment variable (scFv) format. Efficient large scale purification of biological active scFv from inclusion bodies in E. coli expression system

PubMed Central

Moricoli, Diego; Muller, William A.; Carbonella, Damiano Cosimo; Balducci, Maria Cristina; Dominici, Sabrina; Fiori, Valentina; Watson, Richard; Weber, Evan; Cianfriglia, Maurizio; Scotlandi, Katia; Magnani, Mauro

2015-01-01

Migration of leukocytes into a site of inflammation involves several steps mediated by various families of adhesion molecules. CD99 play a significant role in transendothelial migration (TEM) of leukocytes. Inhibition of TEM by specific monoclonal antibody (mAb) can provide a potent therapeutic approach to treating inflammatory conditions. However, the therapeutic utilization of whole IgG can lead to an inappropriate activation of Fc receptor-expressing cells inducing serious adverse side effects due to cytokine release. In this regard, specific recombinant antibody in single chain variable fragments (scFvs) originated by phage library may offer a solution by affecting TEM function in a safe clinical context. However, this consideration requires large scale production of functional scFv antibodies under GMP conditions and hence, the absence of toxic reagents utilized for the solubilization and refolding steps of inclusion bodies that may discourage industrial application of these antibody fragments. In order to apply the scFv anti-CD99 named C7A in a clinical setting we herein describe an efficient and large scale production of the antibody fragments expressed in E.coli as insoluble protein avoiding gel filtration chromatography approach, and laborious refolding step pre- and post-purification. Using differential salt elution which is a simple, reproducible and effective procedure we are able to separate scFv in monomer format from aggregates. The purified scFv antibody C7A exhibits inhibitory activity comparable to an antagonistic conventional mAb, thus providing an excellent agent for blocking CD99 signalling. Thanks to the original purification protocol that can be extended to other scFvs that are expressed as inclusion bodies in bacterial systems, the scFv anti-CD99 C7A herein described represents the first step towards the construction of new antibody therapeutic. PMID:24798881

Screening of detergents for solubilization, purification and crystallization of membrane proteins: a case study on succinate:ubiquinone oxidoreductase from Escherichia coli

PubMed Central

Shimizu, Hironari; Nihei, Coh-ichi; Inaoka, Daniel Ken; Mogi, Tatushi; Kita, Kiyoshi; Harada, Shigeharu

2008-01-01

Succinate:ubiquinone oxidoreductase (SQR) was solubilized and purified from Escherichia coli inner membranes using several different detergents. The number of phospholipid molecules bound to the SQR molecule varied greatly depending on the detergent combination that was used for the solubilization and purification. Crystallization conditions were screened for SQR that had been solubilized and purified using 2.5%(w/v) sucrose monolaurate and 0.5%(w/v) Lubrol PX, respectively, and two different crystal forms were obtained in the presence of detergent mixtures composed of n-alkyl-oligoethylene glycol monoether and n-alkyl-maltoside. Crystallization took place before detergent phase separation occurred and the type of detergent mixture affected the crystal form. PMID:18765923

Screening of detergents for solubilization, purification and crystallization of membrane proteins: a case study on succinate:ubiquinone oxidoreductase from Escherichia coli.

PubMed

Shimizu, Hironari; Nihei, Coh-ichi; Inaoka, Daniel Ken; Mogi, Tatushi; Kita, Kiyoshi; Harada, Shigeharu

2008-09-01

Succinate:ubiquinone oxidoreductase (SQR) was solubilized and purified from Escherichia coli inner membranes using several different detergents. The number of phospholipid molecules bound to the SQR molecule varied greatly depending on the detergent combination that was used for the solubilization and purification. Crystallization conditions were screened for SQR that had been solubilized and purified using 2.5%(w/v) sucrose monolaurate and 0.5%(w/v) Lubrol PX, respectively, and two different crystal forms were obtained in the presence of detergent mixtures composed of n-alkyl-oligoethylene glycol monoether and n-alkyl-maltoside. Crystallization took place before detergent phase separation occurred and the type of detergent mixture affected the crystal form.

Effect of phospholipid, detergent and protein-protein interaction on stability and phosphoenzyme isomerization of soluble sarcoplasmic reticulum Ca-ATPase.

PubMed

Vilsen, B; Andersen, J P

1987-12-30

The purpose of the present study was to elucidate the separate roles of lipid, detergent and protein-protein interaction for stability and catalytic properties of sarcoplasmic reticulum Ca-ATPase solubilized in the non-ionic detergent octa(ethylene glycol) monododecyl ether (C12E8). The use of large-zone high-performance liquid chromatography permitted us to define the self-association state of Ca-ATPase peptide at various detergent, phospholipid and protein concentrations, and also during enzymatic turnover with ATP. Conditions were established for monomerization of Ca-ATPase in the presence of a high concentration of phospholipid relative to detergent. The lipid-saturated monomeric preparation was relatively resistant to inactivation in the absence of Ca2+, whereas delipidated enzyme in monomeric or in oligomeric form was prone to inactivation. Kinetics of phosphoenzyme turnover were examined in the presence and absence of Mg2+. Dephosphorylation rates were sensitive to Mg2+, irrespective of whether the peptide was present in soluble monomeric form or was membrane-bound. C12E8-solubilized monomer without added phospholipid was, however, characterized by a fast initial phase of dephosphorylation in the absence of Mg2+. This was not observed with monomer saturated with phospholipid or with monomer solubilized in myristoylglycerophosphocholine or deoxycholate. The mechanism underlying this difference was shown to be a C12E8-induced acceleration of conversion of ADP-sensitive phosphoenzyme (E1P) to ADP-insensitive phosphoenzyme (E2P). The phosphoenzyme isomerization rate was also found to be enhanced by low-affinity binding of ATP. This was demonstrated both in membrane-bound and in soluble monomeric Ca-ATPase. Our results indicate that a single peptide chain constitutes the target for modulation of phosphoenzyme turnover by Mg2+ and ATP, and that detergent effects, distinct from those arising from disruption of protein-protein contacts, are the major determinants of kinetic differences between C12E8-solubilized and membrane-bound enzyme preparations.

Impact of Solubilizing Additives on Supersaturation and Membrane Transport of Drugs.

PubMed

Raina, Shweta A; Zhang, Geoff G Z; Alonzo, David E; Wu, Jianwei; Zhu, Donghua; Catron, Nathaniel D; Gao, Yi; Taylor, Lynne S

2015-10-01

Many enabling formulations give rise to supersaturated solutions wherein the solute possesses higher thermodynamic activity gradients than the solute in a saturated solution. Since flux across a membrane is driven by solute activity rather than concentration, understanding how solute thermodynamic activity varies with solution composition, particularly in the presence of solubilizing additives, is important in the context of passive absorption. In this study, a side-by-side diffusion cell was used to evaluate solute flux for solutions of nifedipine and felodipine in the absence and presence of different solubilizing additives at various solute concentrations. At a given solute concentration above the equilibrium solubility, it was observed that the solubilizing additives could reduce the membrane flux, indicating that the extent of supersaturation can be reduced. However, the flux could be increased back to the same maximum value (which was determined by the concentration where liquid-liquid phase separation (LLPS) occurred) by increasing the total solute concentration. Qualitatively, the shape of the curves of solute flux through membrane as a function of total solute concentration is the same in the absence and presence of solubilizing additives. Quantitatively, however, LLPS occurs at higher solute concentrations in the presence of solubilizing additives. Moreover, the ratios of the LLPS onset concentration and equilibrium solubility vary significantly in the absence and presence of additives. These findings clearly point out the flaws in using solute concentration in estimating solute activity or supersaturation, and reaffirm the use of flux measurements to understand supersaturated systems. Clear differentiation between solubilization and supersaturation, as well as thorough understanding of their respective impacts on membrane transport kinetics is important for the rational design of enabling formulations for poorly soluble compounds.

Solubilization of protein aggregates by the acid stress chaperones HdeA and HdeB.

PubMed

Malki, Abderrahim; Le, Hai-Tuong; Milles, Sigrid; Kern, Renée; Caldas, Teresa; Abdallah, Jad; Richarme, Gilbert

2008-05-16

The acid stress chaperones HdeA and HdeB of Escherichia coli prevent the aggregation of periplasmic proteins at acidic pH. We show in this report that they also form mixed aggregates with proteins that have failed to be solubilized at acidic pH and allow their subsequent solubilization at neutral pH. HdeA, HdeB, and HdeA and HdeB together display an increasing efficiency for the solubilization of protein aggregates at pH 3. They are less efficient for the solubilization of aggregates at pH 2, whereas HdeB is the most efficient. Increasing amounts of periplasmic proteins draw increasing amounts of chaperone into pellets, suggesting that chaperones co-aggregate with their substrate proteins. We observed a decrease in the size of protein aggregates in the presence of HdeA and HdeB, from very high molecular mass aggregates to 100-5000-kDa species. Moreover, a marked decrease in the exposed hydrophobicity of aggregated proteins in the presence of HdeA and HdeB was revealed by 1,1'-bis(4-anilino)naphtalene-5,5'-disulfonic acid binding experiments. In vivo, during the recovery at neutral pH of acid stressed bacterial cells, HdeA and HdeB allow the solubilization and renaturation of protein aggregates, including those formed by the maltose receptor MalE, the oligopeptide receptor OppA, and the histidine receptor HisJ. Thus, HdeA and HdeB not only help to maintain proteins in a soluble state during acid treatment, as previously reported, but also assist, both in vitro and in vivo, in the solubilization at neutral pH of mixed protein-chaperone aggregates formed at acidic pH, by decreasing the size of protein aggregates and the exposed hydrophobicity of aggregated proteins.

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates toC. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(–) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15%more » (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance.« less

Regulation of the plasma membrane type III phosphatidylinositol 4-kinase by positively charged compounds.

PubMed

Yang, W; Boss, W F

1994-08-15

The effects of positively charged compounds on a plasma membrane, type III phosphatidylinositol 4-kinase were studied. To determine whether the enzyme would respond differently to the compounds in a membrane-associated versus a soluble state, both the plasma membrane and solubilized (released by 0.01% (v/v) Triton X-100) PI 4-kinase were used. Spermidine, spermine, polylysine, cardiotoxin, melittin, and histone stimulated the solubilized PI 4-kinase but had little effect on or weakly stimulated the membrane-associated PI 4-kinase. Polyarginine inhibited membrane-associated PI 4-kinase 75% and inhibited the solubilized PI 4-kinase 30%, indicating that charge alone was not sufficient for activation. Polyarginine also eliminated the activation of the solubilized PI 4-kinase by a PI 4-kinase activator protein, PIK-A49. Calmodulin, a common calcium-binding protein, at micromolar levels strongly inhibited solubilized PI 4-kinase activity but did not inhibit membrane-associated PI 4-kinase activity. The inhibition of the solubilized PI 4-kinase by calmodulin was calcium independent. Calcium alone (1 microM-0.1 mM) inhibited PI 4-kinase activity only slightly (< 30%). The differential effects of the positively charged compounds on the solubilized and membrane-associated PI 4-kinase were not due to substrate availability because both enzymes were assayed in the presence of excess PI (0.6 mM) and 0.3% (v/v) Triton X-100. The data suggest that positively charged compounds affected the enzyme activity not only by interacting with the substrates or products of the reaction but also by interacting with the PI 4-kinase or regulatory components in the plasma membrane.

Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation

PubMed Central

Nandimath, Arusha P.; Karad, Dilip D.; Gupta, Shantikumar G.; Kharat, Arun S.

2017-01-01

Background and Objectives: Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. Materials and Methods: In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Results: Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml−1. Conclusion: As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting. PMID:29296275

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

DOE PAGES

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.; ...

2017-06-16

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates toC. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(–) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15%more » (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance.« less

Consortium inoculum of five thermo-tolerant phosphate solubilizing Actinomycetes for multipurpose biofertilizer preparation.

PubMed

Nandimath, Arusha P; Karad, Dilip D; Gupta, Shantikumar G; Kharat, Arun S

2017-10-01

Alkaline pH of the soil facilitates the conversion of phosphate present in phosphate fertilizer applied in the field to insoluble phosphate which is not available to plants. Problem of soluble phosphate deficiency arises, primarily due to needless use of phosphate fertilizer. We sought to biofertilizer with the thermo-tolerant phosphate solubilizing actinomycetes consortium that could convert insoluble phosphate to soluble phosphate at wider temperature range. In the present investigation consortium of five thermo-tolerant phosphate solubilizing actinomycetes was applied for preparation of inoculum to produce multipurpose bio-fertilizer. Phosphates solubilizing thermo-tolerant 32 actinomycetes strains were processed for identification with the use of PIBWIN software and were screened for phosphate solubilizing activity. Amongst these five actinomycetes were selected on the basis of their ability to produce cellulase, chitinase, pectinase, protease, lipase, amylase and phosphate solubilizing enzymes. Ability to produce these enzymes at 28°C and 50°C were examined. Biofertilizer was prepared by using agricultural waste as a raw material. While preparation of bio-fertilizer the pH decreased from 7.5 to 4.3 and temperature increased up to 74°C maximum at the end of 4 th week and in subsequent week it started to decline gradually till it reached around 50°C, which was found to be stable up to eighth week. This thermo-tolerant actinomycetes consortium released soluble phosphate of up to 46.7 μg ml -1 . As the mesophilic organisms die out at high temperature of composting hence thormo-tolerant actinomycetes would be the better substitute for preparation of phosphate solubilizing bio-fertilizer with added potential to degrade complex macromolecules in composting.

UTSI/CFFF MHD Program Completion and Related Activities.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvin, R.L.; Bumpus, J.A.

1997-10-31

During this reporting period we have further studied the oxidation of soluble coal macromolecules by lignin peroxidase from Phanerochaete chrysosporium. Previous studies by others have suggested that a soluble fraction (coal macromolecule B-111) from a nitric acid solubilized North Dakota Lignite is depolymerized by this enzyme. Our investigations indicate that fraction B-111 is a substrate for lignin peroxidase as this material is decolorized in the presence of lignin peroxidase H{sub 8} and hydrogen peroxide. Of interest, however, is the observation that little, if any, depolymerization of this material occurs. Instead, it appears that lignin peroxidase and coal macromolecule B-111 formmore » a precipitate. These results are similar to those observed in our investigations of lignin peroxidase mediated oxidation of oxalate solubilize coal macromolecule. Previous studies in our laboratory using a spectrophotometric assay suggested that, in addition to oxalate, several other fungal metabolites are able to solubilize leonardite. We have reinvestigated this phenomenon using a more reliable gravimetric procedure for assessing solubilization. Our results confirm our earlier findings that malate, oxaloacetate and citrate are effective solubilizing agents whereas succinate, fumarate and {alpha}-ketoglutarate solubilize relatively small amounts of leonardite. Finally, we have studied the composition of the insoluble material remaining following extensive solubilization by sodium oxalate. The ratio of hydrogen to carbon is increased in the insoluble material relative to the parent leonardite. However, the ratio of oxygen to carbon is also increased in the insoluble material. Thus, the insoluble material does not appear to be more highly reduced that the parent leonardite and is not likely to be a better fuel that the parent material.« less

Detergent effects on enzyme activity and solubilization of lipid bilayer membranes.

PubMed

Womack, M D; Kendall, D A; MacDonald, R C

1983-09-07

Over 50 detergents were tested to establish which would be most effective in releasing proteins from membrane-bounded compartments without denaturing them. Various concentrations each of detergent were tested for two activities: (1) solubilization of egg phospholipid liposomes as measured by reduction of turbidity and (2) effect of detergent concentration on the activities of soluble, hydrolytic enzymes. Those detergents most effective in solubilizing 0.2% lipid and least detrimental to enzymes were five pure, synthetic compounds recently introduced: CHAPS, CHAPSO, Zwittergents 310 and 312, and octylglucoside. Industrial detergents were generally much inferior, insofar as they solubilized membranes inefficiently and/or inactivated certain hydrolytic enzymes readily. The five detergents were characterized by (a) an unusually high critical micelle concentration and (b) a preference for forming mixed micelles with lipids instead of forming pure micelles, as indicated by an ability to solubilize lipid at concentrations of detergent significantly below the critical micelle concentration. This characteristic permits solubilization of high concentrations of membrane below the critical micelle concentration of the detergent so that protein denaturation is minimized. A generally applicable guideline that emerged from this study is that detergents should be used at approximately their critical micelle concentration which should not be exceeded by the concentration of membrane. Similar considerations should apply to the use of detergents in purifying and reconstituting intrinsic membrane proteins.

Root Associated Bacillus sp. Improves Growth, Yield and Zinc Translocation for Basmati Rice (Oryza sativa) Varieties

PubMed Central

Shakeel, Muhammad; Rais, Afroz; Hassan, Muhammad Nadeem; Hafeez, Fauzia Yusuf

2015-01-01

Plant associated rhizobacteria prevailing in different agro-ecosystems exhibit multiple traits which could be utilized in various aspect of sustainable agriculture. Two hundred thirty four isolates were obtained from the roots of basmati-385 and basmati super rice varieties growing in clay loam and saline soil at different locations of Punjab (Pakistan). Out of 234 isolates, 27 were able to solubilize zinc (Zn) from different Zn ores like zinc phosphate [Zn3 (PO4)2], zinc carbonate (ZnCO3) and zinc oxide (ZnO). The strain SH-10 with maximum Zn solubilization zone of 24 mm on Zn3 (PO4)2ore and strain SH-17 with maximum Zn solubilization zone of 14–15 mm on ZnO and ZnCO3ores were selected for further studies. These two strains solubilized phosphorous (P) and potassium (K) in vitro with a solubilization zone of 38–46 mm and 47–55 mm respectively. The strains also suppressed economically important rice pathogens Pyricularia oryzae and Fusarium moniliforme by 22–29% and produced various biocontrol determinants in vitro. The strains enhanced Zn translocation toward grains and increased yield of basmati-385 and super basmati rice varieties by 22–49% and 18–47% respectively. The Zn solubilizing strains were identified as Bacillus sp. and Bacillus cereus by 16S rRNA gene analysis. PMID:26635754

Application of Potential Phosphate-Solubilizing Bacteria and Organic Acids on Phosphate Solubilization from Phosphate Rock in Aerobic Rice

PubMed Central

Jusop, Shamshuddin; Naher, Umme Aminun; Othman, Radziah; Razi, Mohd Ismail

2013-01-01

A study was conducted at Universiti Putra Malaysia to determine the effect of phosphate-solubilizing bacteria (PSB) and organic acids (oxalic & malic) on phosphate (P) solubilization from phosphate rock (PR) and growth of aerobic rice. Four rates of each organic acid (0, 10, 20, and 30 mM), and PSB strain (Bacillus sp.) were applied to aerobic rice. Total bacterial populations, amount of P solubilization, P uptake, soil pH, and root morphology were determined. The results of the study showed significantly high P solubilization in PSB with organic acid treatments. Among the two organic acids, oxalic acid was found more effective compared to malic acid. Application of oxalic acid at 20 mM along with PSB16 significantly increased soluble soil P (28.39 mg kg−1), plant P uptake (0.78 P pot−1), and plant biomass (33.26 mg). Addition of organic acids with PSB and PR had no influence on soil pH during the planting period. A higher bacterial population was found in rhizosphere (8.78 log10 cfu g−1) compared to the nonrhizosphere and endosphere regions. The application of organic acids along with PSB enhanced soluble P in the soil solution, improved root growth, and increased plant biomass of aerobic rice seedlings without affecting soil pH. PMID:24288473

Comparison of the Solubilization Properties of Polysorbate 80 and Isopropanol/Water Solvent Systems for Organic Compounds Extracted from Three Pharmaceutical Packaging Configurations.

PubMed

Zdravkovic, Steven A

2016-10-10

It has been reported that the presence of polysorbate 80 in a pharmaceutical product's formulation may increase the number and/or amount of impurities leached from materials used during its manufacture, storage, and/or administration. However, it is uncertain if/how the solubilization properties of this surfactant compare to non-surfactant solvent systems. The goal of this study is to provide insight into this area of uncertainty by comparing the solubilization properties of polysorbate 80 to those of isopropanol/water solutions while in contact with a plasticized polyvinylchloride parenteral delivery bag, a single-use type manufacturing bag, and a polypropylene bottle. These properties were determined via a binding experiment, in which a set of model compounds was introduced into the solutions, and via an extraction experiment, in which compounds were extracted from the packaging material by the solutions. In both experiments, the amount of each compound present at equilibrium was assayed to determine the extent they were solubilized by the solution from the packaging material. Results from these experiments illustrate differences in the magnitude of solubilization obtained from solutions containing polysorbate 80 as compared to those composed of isopropanol/water. However, it was also demonstrated that their solubilization properties can be linked via a mathematical model. Copyright © 2016 Elsevier B.V. All rights reserved.

Sub-CMC solubilization of dodecane by rhamnolipid in saturated porous media.

PubMed

Zhong, Hua; Zhang, Hui; Liu, Zhifeng; Yang, Xin; Brusseau, Mark L; Zeng, Guangming

2016-09-13

Experiments were conducted with a two-dimensional flow cell to examine the effect of monorhamnolipid surfactant at sub-CMC concentrations on solubilization of dodecane in porous media under dynamic flow conditions. Quartz sand was used as the porous medium and artificial groundwater was used as the background solution. The effectiveness of the monorhamnolipid was compared to that of SDBS, Triton X-100, and ethanol. The results demonstrated the enhancement of dodecane solubility by monorhamnolipid surfactant at concentrations lower than CMC. The concentrations (50-210 μM) are sufficiently low that they do not cause mobilization of the dodecane. Retention of rhamnolipid in the porous medium and detection of nano-size aggregates in the effluent show that the solubilization is based on a sub-CMC aggregate-formation mechanism, which is significantly stronger than the solubilization caused by the co-solvent effect. The rhamnolipid biosurfactant is more efficient for the solubilization compared to the synthetic surfactants. These results indicate a strategy of employing low concentrations of rhamnolipid for surfactant-enhanced aquifer remediation (SEAR), which may overcome the drawbacks of using surfactants at hyper-CMC concentrations.

Potential Application of Biohydrogen Production Liquid Waste as Phosphate Solubilizing Agent-A Study Using Soybean Plants.

PubMed

Sarma, Saurabh Jyoti; Brar, Satinder Kaur; LeBihan, Yann; Buelna, Gerardo

2016-03-01

With CO2 free emission and a gravimetric energy density higher than gasoline, diesel, biodiesel, and bioethanol, biohydrogen is a promising green renewable energy carrier. During fermentative hydrogen production, 60-70 % of the feedstock is converted to different by-products, dominated by organic acids. In the present investigation, a simple approach for value addition of hydrogen production liquid waste (HPLW) containing these compounds has been demonstrated. In soil, organic acids produced by phosphate solubilizing bacteria chelate the cations of insoluble inorganic phosphates (e.g., Ca3 (PO4)2) and make the phosphorus available to the plants. Organic acid-rich HPLW, therefore, has been evaluated as soil phosphate solubilizer. Application of HPLW as soil phosphate solubilizer was found to improve the phosphorus uptake of soybean plants by 2.18- to 2.74-folds. Additionally, 33-100 % increase in seed germination rate was also observed. Therefore, HPLW has the potential to be an alternative for phosphate solubilizing biofertilizers available in the market. Moreover, the strategy can be useful for phytoremediation of phosphorus-rich soil.

Regulation of Coal Polymer Degradation by Fungi

DOE Office of Scientific and Technical Information (OSTI.GOV)

Irvin, R.L.; Bumpus, J.A.

1997-04-30

Previous studies in our laboratory used a spectrophotometric assay to study biomimetic solubilization of leonardite by sodium oxalate. It was found, however, that in extended incubations of several days, this assay resulted in overestimation of the percent of leonardite that was solubilized. This problem did not appear to be significant for short term incubations (ie., up to -24 h) and was circumvented in long term incubations by using a gravimetric assay to assay for solubilization. In other studies during this reporting period we examined oxalate production by P. chrysosporium and T. versicolor grown in Fahreus-Reinhammar medium in agitated pelleted culture.more » It was found that in this system concentrations of oxalate are produced that are much lower than those that would be optimal for leonardite solubilization.« less

Homogeneous time-resolved G protein-coupled receptor-ligand binding assay based on fluorescence cross-correlation spectroscopy.

PubMed

Antoine, Thomas; Ott, David; Ebell, Katharina; Hansen, Kerrin; Henry, Luc; Becker, Frank; Hannus, Stefan

2016-06-01

G protein-coupled receptors (GPCRs) mediate many important physiological functions and are considered as one of the most successful therapeutic target classes for a wide spectrum of diseases. Drug discovery projects generally benefit from a broad range of experimental approaches for screening compound libraries and for the characterization of binding modes of drug candidates. Owing to the difficulties in solubilizing and purifying GPCRs, assay formats have been so far mainly limited to cell-based functional assays and radioligand binding assays. In this study, we used fluorescence cross-correlation spectroscopy (FCCS) to analyze the interaction of detergent-solubilized receptors to various types of GPCR ligands: endogenous peptides, small molecules, and a large surrogate antagonist represented by a blocking monoclonal antibody. Our work demonstrates the suitability of the homogeneous and time-resolved FCCS assay format for a robust, high-throughput determination of receptor-ligand binding affinities and kinetic rate constants for various therapeutically relevant GPCRs. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

[Solubilization Specificities Interferon beta-1b from Inclusion Bodies].

PubMed

Zhuravko, A S; Kononova, N V; Bobruskin, A I

2015-01-01

A new solubilization method of recombinant interferon beta-1b (IFNβ-1b) from the inclusion bodies was developed. This method allows to extract the target protein selectively in the solutions of different alcohols, such as ethanol, propanol and isopropanol. It was shown that the more effective IFNβ-1b solubilization was achieved in the 55% propanol solution. This method allowed to extract the target protein from inclusion bodies around 85-90%, and significantly reduced Escherichia coli content in the solubilizate, in comparison with standard methods.

Flow-through pretreatment of lignocellulosic biomass with inorganic nanoporous membranes

DOEpatents

Bhave, Ramesh R.; Lynd, Lee; Shao, Xiongjun

2018-04-03

A process for the pretreatment of lignocellulosic biomass is provided. The process generally includes flowing water through a pretreatment reactor containing a bed of particulate ligno-cellulosic biomass to produce a pressurized, high-temperature hydrolyzate exit stream, separating solubilized compounds from the hydrolyzate exit stream using an inorganic nanoporous membrane element, fractionating the retentate enriched in solubilized organic components and recycling the permeate to the pretreatment reactor. The pretreatment process provides solubilized organics in concentrated form for the subsequent conversion into biofuels and other chemicals.

Design of microemulsion system suitable for the oral delivery of poorly aqueous soluble beta-carotene.

PubMed

Peng, Cheng; Svirskis, Darren; Lee, Sung Je; Oey, Indrawati; Kwak, Hae-Soo; Chen, Guanyu; Bunt, Craig; Wen, Jingyuan

2017-02-14

Beta-carotene is a potent antioxidant for maintaining human health. However, its oral absorption is low due to poor aqueous solubility of less than 1 μg/ml. A microemulsion delivery system was designed to solubilize beta-carotene toward enhancing its oral bioavailability. From seven pseudoternary diagrams constructed, three systems were selected with large microemulsion areas suitable for oral administration and dilution in the predominately aqueous gastrointestinal fluids. Conductivity and rheology characterization were conducted along four dilution lines within the selected systems. Three pseudoternary-phase diagrams were selected with large microemulsion regions, >60% of the total phase diagram area, which provide microemulsions with higher drug-loading capacity. A phenomenon was observed by which both propylene glycol and Capmul MCM EP stabilize the microstructure of the microemulsions has been proposed based on the characterization studies. An optimal bicontinuous microemulsion formulation was selected comprising 12% orange oil, 24% Capmul MCM, 18% Tween 20, 6% Labrasol, 20% propylene glycol and 20% water, with a high beta-carotene loading capacity of 140.8 μg/ml and droplet size of 117.4 nm. In conclusion, the developed novel microemulsion formulation allows solubilizing beta-carotene and is a promising basis for further development as a functional beverage.

New mechanistically based model for predicting reduction of biosolids waste by ozonation of return activated sludge.

PubMed

Isazadeh, Siavash; Feng, Min; Urbina Rivas, Luis Enrique; Frigon, Dominic

2014-04-15

Two pilot-scale activated sludge reactors were operated for 98 days to provide the necessary data to develop and validate a new mathematical model predicting the reduction of biosolids production by ozonation of the return activated sludge (RAS). Three ozone doses were tested during the study. In addition to the pilot-scale study, laboratory-scale experiments were conducted with mixed liquor suspended solids and with pure cultures to parameterize the biomass inactivation process during exposure to ozone. The experiments revealed that biomass inactivation occurred even at the lowest doses, but that it was not associated with extensive COD solubilization. For validation, the model was used to simulate the temporal dynamics of the pilot-scale operational data. Increasing the description accuracy of the inactivation process improved the precision of the model in predicting the operational data. Copyright © 2014 Elsevier B.V. All rights reserved.

Solubilization of a membrane protein by combinatorial supercharging.

PubMed

Hajduczki, Agnes; Majumdar, Sudipta; Fricke, Marie; Brown, Isola A M; Weiss, Gregory A

2011-04-15

Hydrophobic and aggregation-prone, membrane proteins often prove too insoluble for conventional in vitro biochemical studies. To engineer soluble variants of human caveolin-1, a phage-displayed library of caveolin variants targeted the hydrophobic intramembrane domain with substitutions to charged residues. Anti-selections for insolubility removed hydrophobic variants, and positive selections for binding to the known caveolin ligand HIV gp41 isolated functional, folded variants. Assays with several caveolin binding partners demonstrated the successful folding and functionality by a solubilized, full-length caveolin variant selected from the library. This caveolin variant allowed assay of the direct interaction between caveolin and cavin. Clustered along one face of a putative helix, the solubilizing mutations suggest a structural model for the intramembrane domain of caveolin. The approach provides a potentially general method for solubilization and engineering of membrane-associated proteins by phage display.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Dahlberg, M.D.; Bockrath, B.C.; Speers, V.A.

Oxidized coals, including a naturally oxidized lignite identified as leonardite, are solubilized and sometimes degraded further by a variety of fungi and bacteria. Evidence for biosolubilization of coal was first presented by Fakoussa, and Cohen and Gabriele. Subsequent studies concentrated on screening organisms, characterization of the product, and determination of the biochemical mechanisms. Mechanisms of biosolubilization are poorly known and may vary with the species used and the media. There is evidence for both enzymatic degradation and alkaline solubilization. The objective of this study was to discover critical factors in solubilization and biosolubilization mechanisms by testing a variety of growthmore » media, growth conditions, and fungi. Lignin-degrading species were emphasized because of similarities between the structures in lignin and in low-rank coals. The results indicate that during idiophase (secondary metabolism), the fungi produce alkaline materials that solubilize leonardite.« less

Influence of alkyl chain length compatibility on microemulsion structure and solubilization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bansal, V.K.; O'Connell, J.P.; Shah, D.O.

1980-06-01

The water solubilization capacity of water/oil microemulsions is studied as a function of alkyl chain length of oil (C/sub 8/ to C/sub 16/), surfactant (C/sub 14/ and C/sub 18/ fatty acid soaps), and alcohol (C/sub 4/ to C/sub 7/). Sodium stearate and sodium myristate were used as surfactants. For n-butanol microemulsions the maximum amount of water solubilized in the microemulsion decreased continuously with increasing oil chain length; for n-heptanol it increased continuously. For n-pentanol and n-hexanol systems, water solubilization reached a maximum when the oil chain length plus alcohol chain length was equal to that of the surfactant. The electricmore » resistance and dielectric constant of the microemulsions also are measured as a function of alkyl chain length of the oil. 48 references.« less

Method of remediation of contaminants in porous media through minimization of bouyancy effects

DOEpatents

Shook, G. Michael; Pope, Gary A.

1999-01-01

A method for controlling vertical migration of contaminants in an aquifer includes introduction of a solubilizing solution having a surfactant and an alcohol or other light co-solvent. The surfactant is selected to solubilize the contaminant. The alcohol or other solvent is selected to provide the microemulsion with a substantially neutral buoyancy with respect to groundwater. The neutral buoyancy of the microemulsion prevents the normal downward movement which is typical of the solubilized dense non-aqueous phase liquid in surfactant-enhanced aquifer remediation. Thus, the risk that any significant amount of the solubilized dense non-aqueous contaminants will migrate vertically can be controlled. The relative tendency for vertical migration may also be reduced by increasing the injection rate or injected fluid viscosity (by adding polymer), or by reducing the well spacing.

Heat-solubilized curry spice curcumin inhibits antibody-antigen interaction in in vitro studies: a possible therapy to alleviate autoimmune disorders.

PubMed

Kurien, Biji T; D'Souza, Anil; Scofield, R Hal

2010-08-01

Chronic and complex autoimmune diseases, currently treated palliatively with immunosuppressives, require multi-targeted therapy for greater effectiveness. The naturally occurring polyphenol curcumin has emerged as a powerful "nutraceutical" that interacts with multiple targets to regress diseases safely and inexpensively. Up to 8 g/day of curcumin for 18 months was non-toxic to humans. However, curcumin's utility is limited by its aqueous insolubility. We have demonstrated a heat-mediated 12-fold increase in curcumin's aqueous solubility. Here, we show by SDS-PAGE and surface plasmon resonance that heat-solubilized curcumin binds to proteins. Based on this binding we hypothesized that heat-solubilized curcumin or turmeric would prevent autoantibody targeting of cognate autoantigens. Heat-solubilized curcumin/turmeric significantly decreased binding of autoantibodies from Sjögren's syndrome (up to 43/70%, respectively) and systemic lupus erythematosus (up to 52/70%, respectively) patients as well as an animal model of Sjögren's syndrome (up to 50/60%, respectively) to their cognate antigens. However, inhibition was not specific to autoimmunity. Heat-solubilized curcumin/turmeric also inhibited binding of commercial polyclonal anti-spectrin to spectrin (50/56%, respectively). Thus, we suggest that the multifaceted heat-solubilized curcumin can ameliorate autoimmune disorders. In addition, the non-toxic curcumin could serve as a new protein stain in SDS-PAGE even though it is less sensitive than the Coomassie system which involves toxic chemicals.

Efficiency of plant growth-promoting P-solubilizing Bacillus circulans CB7 for enhancement of tomato growth under net house conditions.

PubMed

Mehta, Preeti; Walia, Abhishek; Kulshrestha, Saurabh; Chauhan, Anjali; Shirkot, Chand Karan

2015-01-01

P-solubilizing bacterial isolate CB7 isolated from apple rhizosphere soil of Himachal Pradesh, India was identified as Bacillus circulans on the basis of phenotypic characteristics, biochemical tests, fatty acid methyl esters analysis, and 16S rRNA gene sequence. The isolate exhibited plant growth-promoting traits of P-solubilization, auxin, 1-aminocyclopropane-1-carboxylate deaminase activity, siderophore, nitrogenase activity, and antagonistic activity against Dematophora necatrix. In vitro studies revealed that P-solubilization and other plant growth-promoting traits were dependent on the presence of glucose in PVK medium and removal of yeast extract had no significant effect on plant growth-promoting traits. Plant growth-promoting traits of isolate CB7 were repressed in the presence of KH2 PO4 . P-solubilization activity was associated with the release of organic acids and a drop in the pH of the Pikovskaya's medium. HPLC analysis detected gluconic and citric acid as major organic acids in the course of P-solubilization. Remarkable increase was observed in seed germination (22.32%), shoot length (15.91%), root length (25.10%), shoot dry weight (52.92%) and root dry weight (31.4%), nitrogen (18.75%), potassium (57.69%), and phosphorus (22.22%) content of shoot biomass over control. These results demonstrate that isolate CB7 has the promising PGPR attributes to be developed as a biofertilizer to enhance soil fertility and promote plant growth. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Microbial Phosphorus Solubilization and Its Potential for Use in Sustainable Agriculture

PubMed Central

Alori, Elizabeth T.; Glick, Bernard R.; Babalola, Olubukola O.

2017-01-01

The use of excess conventional Phosphorus (P) fertilizers to improve agricultural productivity, in order to meet constantly increasing global food demand, potentially causes surface and ground water pollution, waterway eutrophication, soil fertility depletion, and accumulation of toxic elements such as high concentration of selenium (Se), arsenic (As) in the soil. Quite a number of soil microorganisms are capable of solubilizing/mineralizing insoluble soil phosphate to release soluble P and making it available to plants. These microorganisms improve the growth and yield of a wide variety of crops. Thus, inoculating seeds/crops/soil with Phosphate Solubilizing Microorganisms (PSM) is a promising strategy to improve world food production without causing any environmental hazard. Despite their great significance in soil fertility improvement, phosphorus-solubilizing microorganisms have yet to replace conventional chemical fertilizers in commercial agriculture. A better understanding of recent developments in PSM functional diversity, colonizing ability, mode of actions and judicious application should facilitate their use as reliable components of sustainable agricultural systems. In this review, we discussed various soil microorganisms that have the ability to solubilize phosphorus and hence have the potential to be used as bio fertilizers. The mechanisms of inorganic phosphate solubilization by PSM and the mechanisms of organic phosphorus mineralization are highlighted together with some factors that determine the success of this technology. Finally we provide some indications that the use of PSM will promote sustainable agriculture and conclude that this technology is ready for commercial exploitation in various regions worldwide. PMID:28626450

Phosphate dissolving fungi: Mechanism and application in alleviation of salt stress in wheat.

PubMed

Gaind, Sunita

2016-12-01

The present investigation reveals the solubilization efficiency of tri-calcium phosphate (TCP), Udaipur rock phosphate (URP), aluminium phosphate (AP) and ferric phosphate (FP) by Aspergillus niger (ITCC 6719) and Trichoderma harzianum (ITCC 6721) as function of carbon concentrations. Increasing glucose concentration from 1 to 7% in the growth medium, though improved the phosphorus (P) solubilization significantly but each fungal strain preferred different optimum carbon concentrations for mediating solubilization of different P sources. The two fungi employed different mechanisms to reduce medium pH for release of P from TCP, AP and FP. However, URP was solubilized solely through fungal production of citric, succinic, propionic, malic and acetic acid. A linear increase in citric acid production with increasing carbon concentration was recorded during FP solubilization by T. harzianum. The cell free culture filtrate of A. niger detected high phytase and low acid phosphatase activity titre whereas results were vice versa for T. harzianum. Both the fungal strains possessed plant growth promoting attributes such as auxin and sidreophore production and could solubilize Zn. In hydroponic system (with 60mM of sodium chloride concentration), supplementation with culture filtrate from each fungal strain increased the shoot growth of wheat seedlings significantly compared to non culture filtrate control. Use of A.niger as bio-inoculant could be a sustainable approach to improve soil P availability, promote plant growth and alleviate adverse effect of salt stress. Copyright © 2016 Elsevier GmbH. All rights reserved.

Role of the CipA Scaffoldin Protein in Cellulose Solubilization, as Determined by Targeted Gene Deletion and Complementation in Clostridium thermocellum

PubMed Central

Olson, Daniel G.; Giannone, Richard J.; Hettich, Robert L.

2013-01-01

The CipA scaffoldin protein plays a key role in the Clostridium thermocellum cellulosome. Previous studies have revealed that mutants deficient in binding or solubilizing cellulose also exhibit reduced expression of CipA. To confirm that CipA is, in fact, necessary for rapid solubilization of crystalline cellulose, the gene was deleted from the chromosome using targeted gene deletion technologies. The CipA deletion mutant exhibited a 100-fold reduction in cellulose solubilization rate, although it was eventually able to solubilize 80% of the 5 g/liter cellulose initially present. The deletion mutant was complemented by a copy of cipA expressed from a replicating plasmid. In this strain, Avicelase activity was restored, although the rate was 2-fold lower than that in the wild type and the duration of the lag phase was increased. The cipA coding sequence is located at the beginning of a gene cluster containing several other genes thought to be responsible for the structural organization of the cellulosome, including olpB, orf2p, and olpA. Tandem mass spectrometry revealed a 10-fold reduction in the expression of olpB, which may explain the lower growth rate. This deletion experiment adds further evidence that CipA plays a key role in cellulose solubilization by C. thermocellum, and it raises interesting questions about the differential roles of the anchor scaffoldin proteins OlpB, Orf2p, and SdbA. PMID:23204466

Hydrolysis and fractionation of lignocellulosic biomass

DOEpatents

Torget, Robert W.; Padukone, Nandan; Hatzis, Christos; Wyman, Charles E.

2000-01-01

A multi-function process is described for the hydrolysis and fractionation of lignocellulosic biomass to separate hemicellulosic sugars from other biomass components such as extractives and proteins; a portion of the solubilized lignin; cellulose; glucose derived from cellulose; and insoluble lignin from said biomass comprising one or more of the following: optionally, as function 1, introducing a dilute acid of pH 1.0-5.0 into a continual shrinking bed reactor containing a lignocellulosic biomass material at a temperature of about 94 to about 160.degree. C. for a period of about 10 to about 120 minutes at a volumetric flow rate of about 1 to about 5 reactor volumes to effect solubilization of extractives, lignin, and protein by keeping the solid to liquid ratio constant throughout the solubilization process; as function 2, introducing a dilute acid of pH 1.0-5.0, either as virgin acid or an acidic stream from another function, into a continual shrinking bed reactor containing either fresh biomass or the partially fractionated lignocellulosic biomass material from function 1 at a temperature of about 94-220.degree. C. for a period of about 10 to about 60 minutes at a volumetric flow rate of about 1 to about 5 reactor volumes to effect solubilization of hemicellulosic sugars, semisoluble sugars and other compounds, and amorphous glucans by keeping the solid to liquid ratio constant throughout the solubilization process; as function 3, optionally, introducing a dilute acid of pH 1.0-5.0 either as virgin acid or an acidic stream from another function, into a continual shrinking bed reactor containing the partially fractionated lignocellulosic biomass material from function 2 at a temperature of about 180-280.degree. C. for a period of about 10 to about 60 minutes at a volumetric flow rate of 1 to about 5 reactor volumes to effect solubilization of cellulosic sugars by keeping the solid to liquid ratio constant throughout the solubilization process; and as function 4, optionally, introducing a dilute acid of pH 1.0-5.0 either as virgin acid or an acidic stream from another function, into a continual shrinking bed reactor containing the partially fractionated lignocellulosic biomass material from function 3 at a temperature of about 180-280.degree. C. for a period of about 10 to about 60 minutes at a volumetric flow rate of about 1 to about 5 reactor volumes to effect solubilization of cellulosic sugars by keeping the solid to liquid ratio constant throughout the solubilization process.

A non-toxic microbial surfactant from Marinobacter hydrocarbonoclasticus SdK644 for crude oil solubilization enhancement.

PubMed

Zenati, Billal; Chebbi, Alif; Badis, Abdelmalek; Eddouaouda, Kamel; Boutoumi, Hocine; El Hattab, Mohamed; Hentati, Dorra; Chelbi, Manel; Sayadi, Sami; Chamkha, Mohamed; Franzetti, Andrea

2018-06-15

This study aims to investigate the ability of a biosurfactant produced by Marinobacter hydrocarbonoclasticus strain SdK644 isolated from hydrocarbon contaminated sediment to enhance the solubilization rate of crude oil contaminated seawater. Phylogenetic analysis shows that strain SdK644 was very closely related to M. hydrocarbonoclasticus with 16S rRNA gene sequence similarity of 97.44%. Using waste frying oil as inducer carbon source, the producing biosurfactant by strain SdK644 was applied to improve crude oil solubilization in seawater. The preliminary characterization of the produced biosurfactant by FT-IR analysis indicates its possible classification in a glycolipids group. Results from crude oil solubilization assay showed that SdK644 strain biosurfactant was 2-fold greater than Tween 80 surfactant in crude oil solubilization and 12-fold higher than seawater control, as shown by GC-MS analysis of aliphatic compounds. Furthermore, this bioactive compound was shown to be nontoxic against Artemia larvae in short-term acute toxicity bioassay. Generally, the results showed the possible use of M. hydrocarbonoclasticus strain SdK644 biosurfactant in bioremediation processes of the marine environments. Copyright © 2018 Elsevier Inc. All rights reserved.

Microbial screening test for lignite degradation: Quarterly progress report No. 9 for the period January-March 1987

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yen, Teh Fu

1987-03-01

Anaerobic fermentation of water soluble fraction of modified lignite was attempted. Solubilized lignite formed bioprecipitate after biodegradation. Fermentation of water solubilized lignite in enrichment media produced gases and organic acids. FT-IR spectra of solubilized lignite after biodegradation showed that the concentration of organic oxygen have decreased and that the concentration of -CH/sub 3/ terminal group have increased. Solubilized lignite may serve as sole carbon source by using selective media. Bacteria was suspected of being able to utilize fulvic-like materials from solubilized lignite. Isolation of anaerobic bacteria was achieved by surface culture, and it indicated morphological differences among isolated colonies. Alginatemore » gel entrapment, an immobilization method, was applied to T. versicolor fungal cells. Active fungal growth was observed from the immobilized spheres on sodium-alginate gel. It seems that the immobilized biocatalysts may be used to enhance the production of bioextract from lignite in a reactor system. Hydroxylation of lignite was accomplished through Fenton reaction at pH 7.5. FT-IR analysis showed that lignite treated with Fenton's reagent exhibits weaker aromatic bending and ether linkage than untreated lignite. 13 refs., 8 figs.« less

Revisiting the Supramolecular Organization of Photosystem II in Chlamydomonas reinhardtii*

PubMed Central

Tokutsu, Ryutaro; Kato, Nobuyasu; Bui, Khanh Huy; Ishikawa, Takashi; Minagawa, Jun

2012-01-01

Photosystem II (PSII) is a multiprotein complex that splits water and initiates electron transfer in photosynthesis. The central part of PSII, the PSII core, is surrounded by light-harvesting complex II proteins (LHCIIs). In higher plants, two or three LHCII trimers are seen on each side of the PSII core whereas only one is seen in the corresponding positions in Chlamydomonas reinhardtii, probably due to the absence of CP24, a minor monomeric LHCII. Here, we re-examined the supramolecular organization of the C. reinhardtii PSII-LHCII supercomplex by determining the effect of different solubilizing detergents. When we solubilized the thylakoid membranes with n-dodecyl-β-d-maltoside (β-DM) or n-dodecyl-α-d-maltoside (α-DM) and subjected them to gel filtration, we observed a clear difference in molecular mass. The α-DM-solubilized PSII-LHCII supercomplex bound twice more LHCII than the β-DM-solubilized supercomplex and retained higher oxygen-evolving activity. Single-particle image analysis from electron micrographs of the α-DM-solubilized and negatively stained supercomplex revealed that the PSII-LHCII supercomplex had a novel supramolecular organization, with three LHCII trimers attached to each side of the core. PMID:22801422

Enzymatic hydrolysis of flavonoids and pectic oligosaccharides from bergamot (Citrus bergamia Risso) peel.

PubMed

Mandalari, Giuseppina; Bennett, Richard N; Kirby, Andrew R; Lo Curto, Rosario B; Bisignano, Giuseppe; Waldron, Keith W; Faulds, Craig B

2006-10-18

Pectinolytic and cellulolytic enzymes (Pectinase 62L, Pectinase 690L, and Cellulase CO13P) were used to evaluate the solubilization of carbohydrates and low molecular weight flavonoids from bergamot peel, a major byproduct of the essential oil industry. The enzymes were characterized for main-chain and side-chain polysaccharide hydrolyzing activities and also against pure samples of various flavonoids previously identified in bergamot peel to determine various glycosidase activities. The addition of Pectinase 62L or 690L alone, or the combination of Pectinase 62L and Cellulase CO13P, was capable of solubilizing between 70 and 80% of the bergamot peel, and up to 90% of the flavonoid glycosides present were cleaved to their aglycones. Cellulase CO13P alone solubilized 62% of the peel but had no deglycosylating effect on the flavonoid glycosides. Over a 24-h time course, a rapid release of cell wall carbohydrates was observed after treatment with Pectinase 62L, with a concurrent gradual hydrolysis of the flavonoid glycosides. Size-exclusion chromatography of the solubilized extract showed that after 24-h incubation, the majority of the solubilized carbohydrates were present as monosaccharides with a smaller proportion of oligosaccharides.

Solubilization of myofibrillar proteins in water or low ionic strength media: Classical techniques, basic principles, and novel functionalities.

PubMed

Chen, Xing; Tume, Ron K; Xu, Xinglian; Zhou, Guanghong

2017-10-13

The qualitative characteristics of meat products are closely related to the functionality of muscle proteins. Myofibrillar proteins (MPs), comprising approximately 50% of total muscle proteins, are generally considered to be insoluble in solutions of low ionic strength (< 0.2 M), requiring high concentrations of salt (> 0.3 M) for solubilization. These soluble proteins are the ones which determine many functional properties of meat products, including emulsification and thermal gelation. In order to increase the utilization of meat and meat products, many studies have investigated the solubilization of MPs in water or low ionic strength media and determining their functionality. However, there still remains a lack of systematic information on the functional properties of MPs solubilized in this manner. Hence, this review will explore some typical techniques that have been used. The main procedures used for their solubilization, the fundamental principles and their functionalities in water (low ionic strength medium) are comprehensively discussed. In addition, advantages and disadvantages of each technique are summarized. Finally, future considerations are presented to facilitate progress in this new area and to enable water soluble muscle MPs to be utilized as novel meat ingredients in the food industry.

Isolation and characterization of novel plant growth promoting Micrococcus sp NII-0909 and its interaction with cowpea.

PubMed

Dastager, Syed G; Deepa, C K; Pandey, Ashok

2010-12-01

A phosphate-solubilizing bacterial strain NII-0909 isolated from the Western ghat forest soil in India was identified as Micrococcus sp on the basis of phenotypic characteristics, carbon source utilization pattern, fatty acid methyl esters analysis, and 16S rRNA gene sequence. The strain exhibited the plant growth-promoting attributes of phosphate solubilization, auxin production, 1-aminocyclopropane-1-carboxylate deaminase activity, and siderophore production. It was able to solubilize (122.4μg of Ca(3)PO(4) ml(-1)), and produce IAA (109μgml(-1)) at 30°C. P-solubilizing activity of the strain NII-0909 was associated with the release of organic acids and a drop in the pH of the NBRIP medium. HPLC analysis detected two organic acids in the course of P-solubilization. A significant increase in the growth of cow pea was recorded for inoculations under controlled conditions. Scanning electron microscopic study revealed the root colonization of strain on cow pea seedlings. These results demonstrate that isolates NII-0909 has the promising PGPR attributes to be develop as a biofertilizer to enhance soil fertility and promote the plant growth. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

FIELD ASSESSMENT OF MULTIPLE DNAPL REMEDIATION TECHNIQUES

EPA Science Inventory

Five DNAPL remediation technologies were evaluated in constructed test cells at the Dover National Test Site, Dover AFB, Delaware. The technologies were cosolvent solubilization, cosolvent mobilization, surfactant solubilization, complex sugar flushing and air sparging/soil vapor...

FIELD EVALUATION OF DNAPL EXTRACTION TECHNOLOGIES: PROJECT OVERVIEW

EPA Science Inventory

Five DNAPL remediation technologies were evaluated at the Dover National Test Site, Dover AFB, Delaware. The technologies were cosolvent solubilization, cosolvent mobilization, surfactant solubilization, complex sugar flushing and air sparging/soil vapor extraction. The effectiv...

Effect of Hydrotropic Compounds on the Self-Organization and Solubilization Properties of Cationic Surfactants

NASA Astrophysics Data System (ADS)

Gaynanova, G. A.; Valeeva, F. G.; Kushnazarova, R. A.; Bekmukhametova, A. M.; Zakharov, S. V.; Mirgorodskaya, A. B.; Zakharova, L. Ya.

2018-07-01

The effect hydrotropic additives (salts of aromatic acids and choline chloride) have on the micelle-forming properties (the critical concentrations of micelle formation and the Krafft temperature) of cationic surfactants, and on the solubilization capability of mono- and dicationic surfactants toward such hydrophobic compounds as a Sudan I spectral probe and curcumin natural dye, is considered. The factors that govern solubilization capacity, e.g., the structure of the head group of surfactants, the nature of the solubilizate and hydrotropic additives, and the pH of the medium are determined.

Intestinal receptor for heat-stable enterotoxin of Escherichia coli is tightly coupled to a novel form of particulate guanylate cyclase.

PubMed Central

Waldman, S A; Kuno, T; Kamisaki, Y; Chang, L Y; Gariepy, J; O'Hanley, P; Schoolnik, G; Murad, F

1986-01-01

A novel form of particulate guanylate cyclase tightly coupled by cytoskeletal components to receptors for heat-stable enterotoxin (ST) produced by Escherichia coli can be found in membranes from rat intestinal mucosa. Intestinal particulate guanylate cyclase was resistant to solubilization with detergent alone, with only 30% of the total enzyme activity being extracted with Lubrol-PX. Under similar conditions, 70% of this enzyme was solubilized from rat lung membranes. The addition of high concentrations of sodium chloride to the extraction buffer resulted in greater solubilization of particulate guanylate cyclase from intestinal membranes. Although extraction of intestinal membranes with detergent and salt resulted in greater solubilization of guanylate cyclase, a small fraction of the enzyme activity remained associated with the particulate fraction. This activity was completely resistant to solubilization with a variety of detergents and chaotropes. Particulate guanylate cyclase and the ST receptor solubilized by detergent retained their abilities to produce cyclic GMP and bind ST, respectively. However, ST failed to activate particulate guanylate cyclase in detergent extracts. In contrast, guanylate cyclase resistant to solubilization remained functional and coupled to the ST receptor since enzyme activation by ST was unaffected by various extraction procedures. The possibility that the ST receptor and particulate guanylate cyclase were the same molecule was explored. ST binding and cyclic GMP production were separated by affinity chromatography on GTP-agarose. Similarly, guanylate cyclase migrated as a 300,000-dalton protein, while the ST receptor migrated as a 240,000-dalton protein on gel filtration chromatography. Also, thiol-reactive agents such as cystamine and N-ethylmaleimide inhibited guanylate cyclase activation by ST, with no effect on receptor binding of ST. These data suggest that guanylate cyclase and the ST receptor are independent proteins coupled by cytoskeletal components in membranes of intestinal mucosa. PMID:2867046

Solubilization of bovine gelatin using power ultrasound: gelation without heating.

PubMed

Farahnaky, Asgar; Zendeboodi, Fatemeh; Azizi, Rezvan; Mesbahi, Gholamreza; Majzoobi, Mahsa

2017-04-01

The aim of this study was to investigate the efficacy of power ultrasound without using any heating stage in solubilizeing gelatin dispersions, and to characterize the mechanical and microstructural properties of the resulting gels using texture analysis and scanning electron microscopy, respectively. Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. For solubilising gelatin, an ultrasound equipment with a frequency of 20 kHz, amplitude of 100% and power range of 50-150 W was used. Aqueous gelatin dispersions (4% w/v) were subjected to ultrasound for different times (40-240 s) at a constant temperature of 13C. Applying ultrasound to gelatin dispersions caused increases in water absorption and water solubility of the hydrocolloid. The textural parameters of the resulting gelatin gels, increased with increasing time and power of ultrasound. Moreover, a generalized Maxwell model with three elements was used for calculating relaxation times of the gels. The microstructural observations by SEM showed that the structural cohesiveness of the gels increased by increasing ultrasonication time. Ultrasound-assisted solubilization of gelatin can have emerging implications for industrial uses in pharmaceuticals, food and non-food systems. Usually to prepare a gel from gelatin, a primary heating stage of at about 40C or above is required to solubilize gelatin macromolecules. Therefore, the use of gelatin as a hydrocolloid in food processings or pharmaceutical formulations which lack a heating step has been a technological and practical challenge. In this study solubilizing gelatin dispersions using power ultrasound without any heating was successfully performed. Ultrasound-assisted solubilisation of gelatin can have emerging implications for industrial uses in pharmaceuticals, food, and non-food systems, for example, to conserve heat sensitive compounds. © 2016 Wiley Periodicals, Inc.

Reductive solubilization of arsenic in a mining-impacted river floodplain: Influence of soil properties and temperature.

PubMed

Simmler, Michael; Bommer, Jérôme; Frischknecht, Sarah; Christl, Iso; Kotsev, Tsvetan; Kretzschmar, Ruben

2017-12-01

Mining activities have contaminated many riverine floodplains with arsenic (As). When floodplain soils become anoxic under water-saturated conditions, As can be released from the solid phase. Several microbially-driven As solubilization processes and numerous influential factors were recognized in the past. However, the interplay and relative importance of soil properties and the influence of environmental factors such as temperature remain poorly understood, especially considering the (co)variation of soil properties in a floodplain. We conducted anoxic microcosm experiments at 10, 17.5, and 25 °C using 65 representative soils from the mining-impacted Ogosta River floodplain in Bulgaria. To investigate the processes of As solubilization and its quantitative variation we followed the As and Fe redox dynamics in the solid and the dissolved phase and monitored a range of other solution parameters including pH, Eh, dissolved organic C, and dissolved Mn. We related soil properties to dissolved As observed after 20 days of microcosm incubation to identify key soil properties for As solubilization. Our results evidenced reductive dissolution of As-bearing Fe(III)-oxyhydroxides as the main cause for high solubilization. The availability of nutrients, most likely organic C as the source of energy for microorganisms, was found to limit this process. Following the vertical nutrient gradient common in vegetated soil, we observed several hundred μM dissolved As after 1-2 weeks for some topsoils (0-20 cm), while for subsoils (20-40 cm) with comparable total As levels only minor solubilization was observed. While high Mn contents were found to inhibit As solubilization, the opposite applied for higher temperature (Q 10 2.3-6.1 for range 10-25 °C). Our results suggest that flooding of nutrient-rich surface layers might be more problematic than water-saturation of nutrient-poor subsoil layers, especially in summer floodings when soil temperature is higher than in winter or spring. Copyright © 2017 Elsevier Ltd. All rights reserved.

Preparation and Extraction of Insoluble (Inclusion-Body) Proteins from Escherichia coli

PubMed Central

Palmer, Ira; Wingfield, Paul T.

2013-01-01

High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies. Inclusion bodies are normally formed in the cytoplasm; however, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies can be recovered from cell lysates by low speed centrifugation. Following preextaction (or washing) protein is extracted from washed pellets using guanidine·HCl. The solubilized and unfolded protein is either directly folded as described in UNIT 6.1 or further purified by gel filtration in the presence of guanidine·HCl as described here. A support protocol describes the removal of guanidine·HCl from column fractions so they can be monitored by SDS-PAGE. High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies (UNITS 5.1 & 6.1). Inclusion bodies are normally formed in the cytoplasm; alternatively, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies are not restricted to E. coli; they can also form in yeast, mammalian, and insect cells. Inclusion bodies recovered from cell lysates by low-speed centrifugation are heavily contaminated with E. coli cell wall and outer membrane components. The latter are largely removed by selective extraction with detergents and low concentrations of either urea or guanidine·HCl to produce so-called washed pellets. These basic steps result in a significant purification of the recombinant protein, which usually makes up ~60% of the washed pellet protein. The challenge, therefore, is not to purify the recombinant-derived protein, but to solubilize it and then fold it into native and biologically active protein. Basic Protocol 1 describes preparation of washed pellets and solubilization of the protein using guanidine·HCl. The extracted protein, which is unfolded, is either directly folded as described in UNIT 6.5 or further purified by gel filtration in the presence of guanidine·HCl as in basic Protocol 2. A Support Protocol describes the removal of guanidine·HCl from column fractions so they can be monitored by SDS-PAGE (UNIT 10.1). Other methods discussed in the Commentary section of this unit include the direct purification of polyhistidine-tagged proteins solubilized in guanidine·HCl, preparative removal of guanidine·HCl by reversed-phase chromatography as a prelude to protein folding, and the solubilization of inclusion bodies with anionic detergents. PMID:23151747

Preparation and Extraction of Insoluble (Inclusion-Body) Proteins from Escherichia coli

PubMed Central

Palmer, Ira; Wingfield, Paul T.

2012-01-01

High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies. Inclusion bodies are normally formed in the cytoplasm; however, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies can be recovered from cell lysates by low speed centrifugation. Following preextaction (or washing) protein is extracted from washed pellets using guanidine·HCl. The solubilized and unfolded protein is either directly folded as described in UNIT 6.1 or further purified by gel filtration in the presence of guanidine·HCl as described here. A support protocol describes the removal of guanidine·HCl from column fractions so they can be monitored by SDS-PAGE. High-level expression of many recombinant proteins in Escherichia coli leads to the formation of highly aggregated protein commonly referred to as inclusion bodies (UNITS 5.1 & 6.1). Inclusion bodies are normally formed in the cytoplasm; alternatively, if a secretion vector is used, they can form in the periplasmic space. Inclusion bodies are not restricted to E. coli; they can also form in yeast, mammalian, and insect cells. Inclusion bodies recovered from cell lysates by low-speed centrifugation are heavily contaminated with E. coli cell wall and outer membrane components. The latter are largely removed by selective extraction with detergents and low concentrations of either urea or guanidine·HCl to produce so-called washed pellets. These basic steps result in a significant purification of the recombinant protein, which usually makes up ~60% of the washed pellet protein. The challenge, therefore, is not to purify the recombinant-derived protein, but to solubilize it and then fold it into native and biologically active protein. Basic Protocol 1 describes preparation of washed pellets and solubilization of the protein using guanidine·HCl. The extracted protein, which is unfolded, is either directly folded as described in UNIT 6.5 or further purified by gel filtration in the presence of guanidine·HCl as in basic Protocol 2. A Support Protocol describes the removal of guanidine·HCl from column fractions so they can be monitored by SDS-PAGE (UNIT 10.1). Other methods discussed in the Commentary section of this unit include the direct purification of polyhistidine-tagged proteins solubilized in guanidine·HCl, preparative removal of guanidine·HCl by reversed-phase chromatography as a prelude to protein folding, and the solubilization of inclusion bodies with anionic detergents. PMID:18429271

Thermodynamic and Spectroscopic Investigation of Interactions between Reactive Red 223 and Reactive Orange 122 Anionic Dyes and Cetyltrimethyl Ammonium Bromide (CTAB) Cationic Surfactant in Aqueous Solution

PubMed Central

Irfan, Muhammad; Usman, Muhammad; Mansha, Asim; Rasool, Nasir; Ibrahim, Muhammad; Rana, Usman Ali; Siddiq, Mohammad; Zia-Ul-Haq, Muhammad; Jaafar, Hawa Z. E.; Khan, Salah Ud-Din

2014-01-01

The present study describes the conductometric and spectroscopic study of the interaction of reactive anionic dyes, namely, reactive red 223 and reactive orange 122 with the cationic surfactant cetyltrimethyl ammonium bromide (CTAB). In a systematic investigation, the electrical conductivity data was used to calculate various thermodynamic parameters such as free energy (ΔG), enthalpy (ΔH), and the entropy (ΔS) of solubilization. The trend of change in these thermodynamic quantities indicates toward the entropy driven solubilization process. Moreover, the results from spectroscopic data reveal high degree of solubilization, with strong interactions observed in the cases of both dyes and the CTAB. The spontaneous nature of solubilization and binding was evident from the observed negative values of free energies (ΔG p and ΔG b). PMID:25243216

Effects of microwave and alkali induced pretreatment on sludge solubilization and subsequent aerobic digestion.

PubMed

Chang, Chia-Jung; Tyagi, Vinay Kumar; Lo, Shang-Lien

2011-09-01

Individual and combined effects of microwave (MW) and alkali pretreatments on sludge disintegration and subsequent aerobic digestion of waste activated sludge (WAS) were studied. Pretreatments with MW (600W-85°C-2 min), conventional heating (520 W-80°C-12 min) and alkali (1.5 g NaOH/L - pH 12-30 min) achieved 8.5%, 7% and 18% COD solubilization, respectively. However, combined MW-alkali pretreatment synergistically enhanced sludge solubilization and achieved 46% COD solubilization, 20% greater than the additive value of MW alone and alkali alone (8.5+18%=26.5%). Moreover, the results of the batch aerobic digestion study on MW-alkali pretreated sludge showed 93% and 63% reductions in SCOD and VSS concentrations, respectively, at 16 days of SRT. The VSS reduction was 20% higher than that of WAS without pretreatment. Copyright © 2011 Elsevier Ltd. All rights reserved.

Anaerobic microbial dissolution of lead and production of organic acids

DOEpatents

Francis, A.J.; Dodge, C.; Chendrayan, K.

1986-02-28

The present invention relates to a method of solubilizing lead, in the form of lead oxide, found in industrial wastes, before these wastes are dumped into the environment. The lead is solubilized by dissolving the lead oxide in the wastes through contact with an anaerobic bacterial culture containing the bacterium ATCC No. 53464. The solubilized lead can then be removed from the wastes by chemical separation. It could also be removed by extending the contact period with the bacterial culture. As the culture grows, the solubilized lead is removed from the wastes by bioaccumulation by the microorganism or by immobilization by a polymer-like material produced by the microorganism. At this point, the lead is then removed from the wastes when the waste material is separated from the bacterial culture. If desired, the bacterial culture could be digested at this point to yield relatively pure lead for further industrial use.

The styrene-maleic acid copolymer: a versatile tool in membrane research.

PubMed

Dörr, Jonas M; Scheidelaar, Stefan; Koorengevel, Martijn C; Dominguez, Juan J; Schäfer, Marre; van Walree, Cornelis A; Killian, J Antoinette

2016-01-01

A new and promising tool in membrane research is the detergent-free solubilization of membrane proteins by styrene-maleic acid copolymers (SMAs). These amphipathic molecules are able to solubilize lipid bilayers in the form of nanodiscs that are bounded by the polymer. Thus, membrane proteins can be directly extracted from cells in a water-soluble form while conserving a patch of native membrane around them. In this review article, we briefly discuss current methods of membrane protein solubilization and stabilization. We then zoom in on SMAs, describe their physico-chemical properties, and discuss their membrane-solubilizing effect. This is followed by an overview of studies in which SMA has been used to isolate and investigate membrane proteins. Finally, potential future applications of the methodology are discussed for structural and functional studies on membrane proteins in a near-native environment and for characterizing protein-lipid and protein-protein interactions.

New spectrophotometric estimation of indomethacin capsules with niacinamide as hydrotropic solubilizing agent.

PubMed

Maheshwari, R K; Rathore, Amit; Agrawal, Archana; Gupta, Megha A

2011-07-01

Hydrotropic solubilization process involves cooperative intermolecular interaction with several balancing molecular forces, rather than either a specific complexation event or a process dominated by a medium effect, such as co-solvency or salting-in. In the present investigation, hydrotropic solution of 2 M niacinamide was employed as the solubilizing agent to solubilize the poorly water-soluble drug, indomethacin, from the capsule dosage form for spectrophotometric determination in ultraviolet region. Hydrotropic agent used did not interfere in the spectrophotometric analysis. In preliminary solubility studies, it was found that there was more than fivefold enhancement in the aqueous solubility of indomethacin (poorly water-soluble drug) in 2 M niacinamide solution as compared to its aqueous solubility at 28 ± 1°C. The proposed method is new, simple, safe, environmentally friendly, economic, accurate and cost-effective and can be successfully employed in routine analysis.

Solubilization and Interaction Studies of Bile Salts with Surfactants and Drugs: a Review.

PubMed

Malik, Nisar Ahmad

2016-05-01

In this review, bile salt, bile salt-surfactant, and bile salt-drug interactions and their solubilization studies are mainly focused. Usefulness of bile salts in digestion, absorption, and excretion of various compounds and their rare properties in ordering the shape and size of the micelles owing to the presence of hydrophobic and hydrophilic faces are taken into consideration while compiling this review. Bile salts as potential bio-surfactants to solubilize drugs of interest are also highlighted. This review will give an insight into the selection of drugs in different applications as their properties get modified by interaction with bile salts, thus influencing their solution behavior which, in turn, modifies the phase-forming behavior, microemulsion, and clouding phenomenon, besides solubilization. Finally, their future perspectives are taken into consideration to assess their possible uses as bio-surfactants without side effects to human beings.

Sequencing biological acidification of waste-activated sludge aiming to optimize phosphorus dissolution and recovery.

PubMed

Guilayn, Felipe; Braak, Etienne; Piveteau, Simon; Daumer, Marie-Line

2017-06-01

Phosphorus (P) recovery in wastewater treatment plants (WWTP) as pure crystals such as struvite (MgNH 4 PO 4 .6H 2 O), potassium struvite (KMgPO 4 .6H 2 O) and calcium phosphates (e.g. Ca 3 (PO 4 ) 2 ) is an already feasible technique that permits the production of green and marketable fertilizers and the reduction of operational costs. Commercial crystallizers can recovery more than 90% of soluble P. However, most of the P in WWTP sludge is unavailable for the processes (not dissolved). P solubilization and separation are thus the limiting steps in P-crystallization. With an innovative two-step sequencing acidification strategy, the current study has aimed to improve biological P solubilization on waste-activated sludge (WAS) from a full-scale plant. In the first step (P-release), low charges of organic waste were used as co-substrates of WAS pre-fermentation, seeking to produce volatile fatty acids to feed the P-release by Polyphosphate-accumulating organisms, while keeping its optimal metabolic pH (6-7). In this phase, milk serum, WWTP grease, urban organic waste and collective restaurant waste were individually applied as co-substrates. In the second step (P-dissolution), pH 4 was aimed at as it allows the dissolution of the most common precipitated species of P. Biological acidification was performed by white sugar addition, as a carbohydrate-rich organic waste model, which was compared to chemical acidification by HCl (12M) addition. With short retention times (48-96 h) and without inoculum application, all experiences succeeded on P solubilization (37-55% of soluble P), principally when carbohydrate-rich co-substrates were applied. Concentrations from 270 to 450 mg [Formula: see text] were achieved. [Formula: see text].

Geochemical hosts of solubilized radionuclides in uranium mill tailings

USGS Publications Warehouse

Landa, E.R.; Bush, C.A.

1990-01-01

The solubilization and subsequent resorption of radionuclides by ore components or by reaction products during the milling of uranium ores may have both economic and environmental consequences. Particle-size redistribution of radium during milling has been demonstrated by previous investigators; however, the identification of sorbing components in the tailings has received little experimental attention. In this study, uranium-bearing sandstone ore was milled, on a laboratory scale, with sulfuric acid. At regular intervals, filtrate from this suspension was placed in contact with mixtures of quartz sand and various potential sorbents which occur as gangue in uranium ores; the potential sorbents included clay minerals, iron and aluminum oxides, feldspar, fluorspar, barite, jarosite, coal, and volcanic glass. After equilibration, the quartz sand-sorbent mixtures were separated from the filtrate and radioassayed by gamma-spectrometry to determine the quantities of 238U, 230Th, 226Ra, and 210Pb sorbed, and the radon emanation coefficients. Sorption of 238U was low in all cases, with maximal sorptions of 1-2% by the bentonite- and coal-bearing samples. 230Th sorption also was generally less than 1%; maximal sorption here was observed in the fluorspar-bearing sample and appears to be associated with the formation of gypsum during milling. 226Ra and 210 Pb generally showed higher sorption than the other nuclides - more than 60% of the 26Ra solubilized from the ore was sorbed on the barite-bearing sample. The mechanism (s) for this sorption by a wide variety of substrates is not yet understood. Radon emanation coefficients of the samples ranged from about 5 to 30%, with the coal-bearing samples clearly demonstrating an emanating power higher than any of the other materials. ?? 1990.

Simplified in vitro refolding and purification of recombinant human granulocyte colony stimulating factor using protein folding cation exchange chromatography.

PubMed

Vemula, Sandeep; Dedaniya, Akshay; Thunuguntla, Rahul; Mallu, Maheswara Reddy; Parupudi, Pavani; Ronda, Srinivasa Reddy

2015-01-30

Protein folding-strong cation exchange chromatography (PF-SCX) has been employed for efficient refolding with simultaneous purification of recombinant human granulocyte colony stimulating factor (rhG-CSF). To acquire a soluble form of renatured and purified rhG-CSF, various chromatographic conditions, including the mobile phase composition and pH was evaluated. Additionally, the effects of additives such as urea, amino acids, polyols, sugars, oxidizing agents and their amalgamations were also investigated. Under the optimal conditions, rhG-CSF was efficaciously solubilized, refolded and simultaneously purified by SCX in a single step. The experimental results using ribose (2.0M) and arginine (0.6M) combination were found to be satisfactory with mass yield, purity and specific activity of 71%, ≥99% and 2.6×10(8)IU/mg respectively. Through this investigation, we concluded that the SCX refolding method was more efficient than conventional methods which has immense potential for the large-scale production of purified rhG-CSF. Copyright © 2014 Elsevier B.V. All rights reserved.

Comparison of phosphorylated proteins in intact rat spermatozoa from caput and cauda epididymidis.

PubMed

Chulavatnatol, M; Panyim, S; Wititsuwannakul, D

1982-02-01

Spermatozoa from rat epididymis were incubated with [32P] orthophosphate and the radioactively labeled proteins were solubilized for analysis by electrophoresis in SDS-gels or in two-dimensional gels by isoelectric focusing and SDS electrophoresis. Three major phosphorylated protein bands of Mr 42,700, 56,200, and 76,200 were identified together with several minor phosphorylated proteins. The phosphorylated proteins of Mr 42,700 and 76,200 were more heterogeneous in charge than the one of Mr 56,000. The major phosphorylated proteins were not found in the isolated heads of cytosol derived from sperm sonicate. They were not solubilized by 1% Triton X-100 and 2 mM DTT, which removed the plasma membrane and mitochondria, but they were solubilized by 6 M urea and 5 mM DTT away from the insoluble fibrous sheath which contained no appreciable radioactivity. Most of the major phosphorylated bands were solubilized by 2% SDS and 4 mM DTT, leaving the insoluble outer dense fiber-connecting piece (ODF-CP) complex with some of the proteins. The ODF-CP complex of the spermatozoa from the cauda epididymis contained more of the major phosphorylated bands than did that of the spermatozoa from the caput region. Treatment with 1% SDS alone can solubilize about half of the major phosphorylated bands from the spermatozoa of the caput region and essentially none from the spermatozoa of the caudal part. The latter required 1% SDS and 13 mM DTT to achieve solubilization, suggesting the formation of disulfide bonds holding the three major phosphorylated proteins to some intracellular structure during sperm maturation.

Phospholipids-embedded fully dilutable liquid nanostructures. Part 2: The role of sodium diclofenac.

PubMed

Amsalem, Orit; Aserin, Abraham; Garti, Nissim

2010-12-01

Complex pseudo-ternary phase diagrams based on sucrose monolaurate (SE), propylene glycol (PG), and phosphatidylcholine (PC) as the "surfactant phase"; triacetin (TA) and decaglycerol ester (10G1CC) as the "oil phase"; and water as the aqueous phase were constructed, into which we solubilized the water-insoluble drug (sodium diclofenac, Na-DFC). In our previous study we demonstrated that the solubilization of Na-DFC in the oil+surfactant phases (prior to diluting it with water), was 90-fold greater than its dissolution in water, and that the system was pH-dependent. The greatest Na-DFC solubilization capacity was obtained at pH 7.2. In this study we examined the effect of the solubilization of Na-DFC in a phosphatidylcholine system using DLS, viscosity, electrical conductivity, SAXS, SD-NMR, and cryo-TEM measurements. It was found that: (1) the system remains micellar after aqueous dilution but with greater polydispersity and greater variety of shapes. We concluded that the structures in the absence of water (but in the presence of PG) were of direct spherical micelles (∼4 nm) mixed with elongated cylindrical micelles (12-140 nm); (2) the aqueous dilution causes fragmentation of the cylinders into smaller spherical micelles; (3) solubilization of Na-DFC behaving like a kosmotropic agent or "structure maker" yields mostly spherical swollen micelles and more ordered systems than in its absence; and (4) Na-DFC is solubilized at the interface of the micelles without swelling the droplets. Copyright © 2010 Elsevier B.V. All rights reserved.

Comparative study of the interaction of CHAPS and Triton X-100 with the erythrocyte membrane.

PubMed

Rodi, P M; Bocco Gianello, M D; Corregido, M C; Gennaro, A M

2014-03-01

The zwitterionic detergent CHAPS, a derivative of the bile salts, is widely used in membrane protein solubilization. It is a "facial" detergent, having a hydrophilic side and a hydrophobic back. The objective of this work is to characterize the interaction of CHAPS with a cell membrane. To this aim, erythrocytes were incubated with a wide range of detergent concentrations in order to determine CHAPS partition behavior, and its effects on membrane lipid order, hemolytic effects, and the solubilization of membrane phospholipids and cholesterol. The results were compared with those obtained with the nonionic detergent Triton X-100. It was found that CHAPS has a low affinity for the erythrocyte membrane (partition coefficient K=0.06mM(-1)), and at sub-hemolytic concentrations it causes little effect on membrane lipid order. CHAPS hemolysis and phospholipid solubilization are closely correlated. On the other side, binding of Triton X-100 disorders the membrane at all levels, and has independent mechanisms for hemolysis and solubilization. Differential behavior was observed in the solubilization of phospholipids and cholesterol. Thus, the detergent resistant membranes (DRM) obtained with the two detergents will have different composition. The behaviors of the two detergents are related to the differences in their molecular structures, suggesting that CHAPS does not penetrate the lipid bilayer but binds in a flat position on the erythrocyte surface, both in intact and cholesterol depleted erythrocytes. A relevant result for Triton X-100 is that hemolysis is not directly correlated with the solubilization of membrane lipids, as it is usually assumed. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation for rock phosphate solubilization in fermentation and soil-plant system using a stress-tolerant phosphate-solubilizing Aspergillus niger WHAK1.

PubMed

Xiao, Chunqiao; Zhang, Huaxiang; Fang, Yujuan; Chi, Ruan

2013-01-01

A strain WHAK1, identified as Aspergillus niger, was isolated from Yichang phosphate mines in Hubei province of China. The fungus developed a phosphate solubilization zone on modified National Botanical Research Institute's phosphate growth (NBRIP) agar medium, supplemented with tricalcium phosphate. The fungus was applied in a repeated-batch fermentation process in order to test its effect on solubilization of rock phosphate (RP). The results showed that A. niger WHAK1 could effectively solubilize RP in NBRIP liquid medium and released soluble phosphate in the broth, which can be illustrated by the observation of scanning electron microscope, energy-dispersive X-ray microanalysis, and Fourier transform infrared spectroscopy. Acidification of the broth seemed to be the major mechanism for RP solubilization by the fungus. Indeed, multiple organic acids (mainly gluconic acid) were detected in the broth by high-performance liquid chromatography analysis. These organic acids caused a significant drop of pH and an obvious rise of titratable acidity in the broth. The fungus also exhibited high levels of tolerance against temperature, pH, salinity, and desiccation stresses, although a significant decline in the fungal growth and release of soluble phosphate was marked under increasing intensity of stress parameters. Further, the fungus was introduced into the soil supplemented with RP to analyze its effect on plant growth and phosphate uptake of wheat plants. The result revealed that inoculation of A. niger WHAK1 significantly increased the growth and phosphate uptake of wheat plants in the RP-amended soil compared to the control soil.

Dietary fibers solubilized in water or an oil emulsion induce satiation through CCK-mediated vagal signaling in mice.

PubMed

Rasoamanana, Rojo; Chaumontet, Catherine; Nadkarni, Nachiket; Tomé, Daniel; Fromentin, Gilles; Darcel, Nicolas

2012-11-01

This study focused on the fate of the satiating potency of dietary fibers when solubilized in a fat-containing medium. Fourteen percent of either guar gum (GG) or fructo-oligosaccharide (FOS) or a mixture of the 2 (GG-FOS, 5% GG and 9% FOS) were solubilized in water or an oil emulsion (18-21% rapeseed oil in water, v:v) and administered by gavage to mice before their food intake was monitored. When compared with water (control), only GG-FOS solubilized in water or in the oil emulsion reduced daily energy intake by 21.1 and 14.1%, respectively. To further describe this effect, the meal pattern was characterized and showed that GG-FOS increased satiation without affecting satiety by diminishing the size and duration of meals for up to 9 h after administration independently of the solubilization medium. The peripheral blockade of gut peptide receptors showed that these effects were dependent on the peripheral signaling of cholecystokinin but not of glucagon-like peptide 1, suggesting that anorectic signals emerge from the upper intestine rather than from distal segments. Measurements of neuronal activation in the nucleus of solitary tract supported the hypothesis of vagal satiation signaling because a 3-fold increase in c-Fos protein expression was observed in that nucleus after the administration of GG-FOS, independently of the solubilization medium. Taken together, these data suggest that a mixture of GG and FOS can maintain its appetite suppressant effect in fatty media. Adding these dietary fibers to fat-containing foods might therefore be useful in managing food intake.

Application and evaluation of scale dissolver treatments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Fielder, G.D.

1994-12-31

In order to provide an improved basis for the design of barium sulfate scale dissolver treatments both laboratory testing and monitoring of field applications were carried out. The deleterious effects of mixing produced water with dissolver prior to contacting scale are shown. Increasing total dissolved solids (TDS) levels can reduce dissolution depending upon temperature. Precomplexation with divalent cations reduces the capacity of the dissolver to solubilize solid scales. Magnesium may adversely affect dissolver performance at elevated temperatures. Several oil and gas wells were closely monitored during initial flowback after treatment. Samples were collected on a frequent basis and analyzed formore » pH, dissolver content, chlorides and various cations. The resulting data were used to construct flowback profiles for evaluation of the treatments. Evidence of scale dissolution is presented. The presence of an incompatible flush brine was discovered in one case and possible reverse order of addition of preflush and dissolver in another. The importance of establishing and following treatment procedures is briefly discussed.« less

Improving the apo-state detergent stability of NTS1 with CHESS for pharmacological and structural studies.

PubMed

Scott, Daniel J; Kummer, Lutz; Egloff, Pascal; Bathgate, Ross A D; Plückthun, Andreas

2014-11-01

The largest single class of drug targets is the G protein-coupled receptor (GPCR) family. Modern high-throughput methods for drug discovery require working with pure protein, but this has been a challenge for GPCRs, and thus the success of screening campaigns targeting soluble, catalytic protein domains has not yet been realized for GPCRs. Therefore, most GPCR drug screening has been cell-based, whereas the strategy of choice for drug discovery against soluble proteins is HTS using purified proteins coupled to structure-based drug design. While recent developments are increasing the chances of obtaining GPCR crystal structures, the feasibility of screening directly against purified GPCRs in the unbound state (apo-state) remains low. GPCRs exhibit low stability in detergent micelles, especially in the apo-state, over the time periods required for performing large screens. Recent methods for generating detergent-stable GPCRs, however, offer the potential for researchers to manipulate GPCRs almost like soluble enzymes, opening up new avenues for drug discovery. Here we apply cellular high-throughput encapsulation, solubilization and screening (CHESS) to the neurotensin receptor 1 (NTS1) to generate a variant that is stable in the apo-state when solubilized in detergents. This high stability facilitated the crystal structure determination of this receptor and also allowed us to probe the pharmacology of detergent-solubilized, apo-state NTS1 using robotic ligand binding assays. NTS1 is a target for the development of novel antipsychotics, and thus CHESS-stabilized receptors represent exciting tools for drug discovery. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of phosphate-solubilizing bacteria on phosphorus dynamics and the bacterial community during composting of sugarcane industry waste.

PubMed

Estrada-Bonilla, German A; Lopes, Cintia M; Durrer, Ademir; Alves, Paulo R L; Passaglia, Nicolle; Cardoso, Elke J B N

2017-07-01

Sugarcane processing generates a large quantity of residues, such as filter cake and ashes, which are sometimes composted prior to their amendment in soil. However, important issues still have to be addressed on this subject, such as the description of bacterial succession that occurs throughout the composting process and the possibilities of using phosphate-solubilizing bacteria (PSB) during the process to improve phosphorus (P) availability in the compost end product. Consequently, this study evaluated the bacterial diversity and P dynamics during the composting process when inoculated with Pseudomonas aeruginosa PSBR12 and Bacillus sp. BACBR01. To characterize the bacterial community structure during composting, and to compare PSB-inoculated compost with non-inoculated compost, partial sequencing of the bacterial 16S rRNA gene and sequential P fractionation were used. The data indicated that members of the order Lactobacillales prevailed in the early stages of composting for up to 30 days, mostly due to initial changes in pH and the C/N ratio. This dominant bacterial group was then slowly replaced by Bacillales during a composting process of up to 60 days. In addition, inoculation of PSB reduced the levels of Ca-bound P by 21% and increased the labile organic P fraction. In PSB-inoculated compost, Ca-P compound solubilization occurred concomitantly with an increase of the genus Bacillus. The bacterial succession and the final community is described in compost from sugarcane residues and the possible use of these inoculants to improve P availability in the final compost is validated. Copyright © 2017 Elsevier GmbH. All rights reserved.

Detergent-free purification of ABC (ATP-binding-cassette) transporters.

PubMed

Gulati, Sonali; Jamshad, Mohammed; Knowles, Timothy J; Morrison, Kerrie A; Downing, Rebecca; Cant, Natasha; Collins, Richard; Koenderink, Jan B; Ford, Robert C; Overduin, Michael; Kerr, Ian D; Dafforn, Timothy R; Rothnie, Alice J

2014-07-15

ABC (ATP-binding-cassette) transporters carry out many vital functions and are involved in numerous diseases, but study of the structure and function of these proteins is often hampered by their large size and membrane location. Membrane protein purification usually utilizes detergents to solubilize the protein from the membrane, effectively removing it from its native lipid environment. Subsequently, lipids have to be added back and detergent removed to reconstitute the protein into a lipid bilayer. In the present study, we present the application of a new methodology for the extraction and purification of ABC transporters without the use of detergent, instead, using a copolymer, SMA (polystyrene-co-maleic acid). SMA inserts into a bilayer and assembles into discrete particles, essentially solubilizing the membrane into small discs of bilayer encircled by a polymer, termed SMALPs (SMA lipid particles). We show that this polymer can extract several eukaryotic ABC transporters, P-glycoprotein (ABCB1), MRP1 (multidrug-resistance protein 1; ABCC1), MRP4 (ABCC4), ABCG2 and CFTR (cystic fibrosis transmembrane conductance regulator; ABCC7), from a range of different expression systems. The SMALP-encapsulated ABC transporters can be purified by affinity chromatography, and are able to bind ligands comparably with those in native membranes or detergent micelles. A greater degree of purity and enhanced stability is seen compared with detergent solubilization. The present study demonstrates that eukaryotic ABC transporters can be extracted and purified without ever being removed from their lipid bilayer environment, opening up a wide range of possibilities for the future study of their structure and function.

Backbone dynamics of a model membrane protein: assignment of the carbonyl carbon /sup 13/C NMR resonances in detergent-solubilized M13 coat protein

DOE Office of Scientific and Technical Information (OSTI.GOV)

Henry, G.D.; Weiner, J.H.; Sykes, B.D.

The major coat protein of the filamentous bacteriophage M13 is a 50-residue amphiphilic polypeptide which is inserted, as an integral membrane-spanning protein, in the inner membrane of the Escherichia coli host during infection. /sup 13/C was incorporated biosynthetically into a total of 23 of the peptide carbonyls using labeled amino acids (alanine, glycine, lysine, phenylalanine, and proline). The structure and dynamics of carbonyl-labeled M13 coat protein were monitored by /sup 13/C nuclear magnetic resonance (NMR) spectroscopy. Assignment of many resonances was achieved by using protease digestion, pH titration, or labeling of the peptide bond with both /sup 13/C and /supmore » 15/N. The carbonyl region of the natural-abundance /sup 13/C NMR spectrum of M13 coat protein in sodium dodecyl sulfate solution shows approximately eight backbone carbonyl resonances with line widths much narrower than the rest. Three of these more mobile residues correspond to assigned peaks (glycine-3, lysine-48, and alanine-49) in the individual amino acid spectra, and another almost certainly arises from glutamic acid-2. A ninth residue, alanine-1, also gives rise to a very narrow carbonyl resonance if the pH is well above or below the pK/sub a/ of the terminal amino group. These data suggest that only about four residues at either end of the protein experience large-amplitude spatial fluctuations; the rest of the molecule is essentially rigid on the time scale of the overall rotational tumbling of the protein-detergent complex. The relative exposure of different regions of detergent-bound protein was monitored by limited digestion with proteinase K. Comparable spectra and digestion patterns were obtained when the protein was solubilized in sodium deoxycholate, suggesting that the coat protein binds both amphiphiles in a similar fashion.« less

SURFACTANT-ENHANCED SOLUBILIZATION OF RESIDUAL DODECANE IN SOIL COLUMNS - 1. EXPERIMANTAL INVESTIGATION

EPA Science Inventory

The solubilization of dodecane by polyoxyethylene (20) sorbitan monooleate, a nonionic surfactant, was investigated as a potential means of recoveringnonaqueous-phase liquids from contaminated aquifers. Residual saturations of dodecane were established by injecting ¹⁴C...

Development of a new biofertilizer with a high capacity for N2 fixation, phosphate and potassium solubilization and auxin production.

PubMed

Leaungvutiviroj, Chaveevan; Ruangphisarn, Pimtida; Hansanimitkul, Pikul; Shinkawa, Hidenori; Sasaki, Ken

2010-01-01

Biofertilizers that possess a high capacity for N(2) fixation (Azotobacter tropicalis), and consist of phosphate solubilizing bacteria (Burkhoderia unamae), and potassium solubilizing bacteria (Bacillus subtilis) and produce auxin (KJB9/2 strain), have a high potential for growth and yield enhancement of corn and vegetables (Chinese kale). For vegetables, the addition of biofertilizer alone enhanced growth 4 times. Moreover, an enhancement of growth by 7 times was observed due to the addition of rock phosphate and K-feldspar, natural mineral fertilizers, in combination with the biofertilizer.

The solubilization of fatty acids in systems based on block copolymers and nonionic surfactants

NASA Astrophysics Data System (ADS)

Mirgorodskaya, A. B.; Yatskevich, E. I.; Zakharova, L. Ya.

2010-12-01

The solubilizing action of micellar, microemulsion, and polymer-colloid systems formed on the basis of biologically compatible amphiphilic polymers and nonionic surfactants on capric, lauric, palmitic, and stearic acids was characterized quantitatively. Systems based on micelle forming oxyethyl compounds increased the solubility of fatty acids by more than an order of magnitude. Acid molecules incorporated into micelles increased their size and caused structural changes. Solubilization was accompanied by complete or partial destruction of intrinsic acid associates and an increase in their p K a by 1.5-2 units compared with water.

[Phosphate-solubilizing activity of aerobic methylobacteria].

PubMed

Agafonova, N V; Kaparullina, E N; Doronina, N V; Trotsenko, Iu A

2014-01-01

Phosphate-solubilizing activity was found in 14 strains of plant-associated aerobic methylobacteria belonging to the genera Methylophilus, Methylobacillus, Methylovorus, Methylopila, Methylobacterium, Delftia, and Ancyclobacter. The growth of methylobacteria on medium with methanol as the carbon and energy source and insoluble tricalcium phosphate as the phosphorus source was accompanied by a decrease in pH due to the accumulation of up to 7 mM formic acid as a methanol oxidation intermediate and by release of 120-280 μM phosphate ions, which can be used by both bacteria and plants. Phosphate-solubilizing activity is a newly revealed role of methylobacteria in phytosymbiosis.

Microbial solubilization of coals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Campbell, J.A.; Fredrickson, J.K.; Stewart, D.L.

1988-11-01

Microbial solubilization of coal may serve as a first step in a process to convert low-rank coals or coal-derived products to other fuels or products. For solubilization of coal to be an economically viable technology, a mechanistic understanding of the process is essential. Leonardite, a highly oxidized, low-rank coal, has been solubilized by the intact microorganism, cell-free filtrate, and cell-free enzyme of /ital Coriolus versicolor/. A spectrophotometric conversion assay was developed to quantify the amount of biosolubilized coal. In addition, a bituminous coal, Illinois No. 6, was solubilized by a species of /ital Penicillium/, but only after the coal hadmore » been preoxidized in air. Model compounds containing coal-related functionalities have been incubated with the leonardite-degrading fungus, its cell-free filtrate, and purified enzyme. The amount of degradation was determined by gas chromatography and the degradation products were identified by gas chromatography/mass spectrometry. We have also separated the cell-free filtrate of /ital C. versicolor/ into a <10,000 MW and >10,000 MW fraction by ultrafiltration techniques. Most of the coal biosolubilization activity is contained in the <10,000 MW fraction while the model compound degradation occurs in the >10,000 MW fraction. The >10,000 MW fraction appears to contain an enzyme with laccase-like activity. 10 refs., 8 figs., 5 tabs.« less

Solubilization of poorly water-soluble compounds using amphiphilic phospholipid polymers with different molecular architectures.

PubMed

Mu, Mingwei; Konno, Tomohiro; Inoue, Yuuki; Ishihara, Kazuhiko

2017-10-01

To achieve stable and effective solubilization of poorly water-soluble bioactive compounds, water-soluble and amphiphilic polymers composed of hydrophilic 2-methacryloyloxyethyl phosphorylcholine (MPC) units and hydrophobic n-butyl methacrylate (BMA) units were prepared. MPC polymers having different molecular architectures, such as random-type monomer unit sequences and block-type sequences, formed polymer aggregates when they were dissolved in aqueous media. The structure of the random-type polymer aggregate was loose and flexible. On the other hand, the block-type polymer formed polymeric micelles, which were composed of very stable hydrophobic poly(BMA) cores and hydrophilic poly(MPC) shells. The solubilization of a poorly water-soluble bioactive compound, paclitaxel (PTX), in the polymer aggregates was observed, however, solubilizing efficiency and stability were strongly depended on the polymer architecture; in other words, PTX stayed in the poly(BMA) core of the polymer micelle formed by the block-type polymer even when plasma protein was present in the aqueous medium. On the other hand, when the random-type polymer was used, PTX was transferred from the polymer aggregate to the protein. We conclude that water-soluble and amphiphilic MPC polymers are good candidates as solubilizers for poorly water-soluble bioactive compounds. Copyright © 2017 Elsevier B.V. All rights reserved.

Enhancement of clover growth by inoculation of P-solubilizing fungi and arbuscular mycorrhizal fungi.

PubMed

Souchie, Edson L; Azcón, Rosario; Barea, Jose M; Silva, Eliane M R; Saggin-Júnior, Orivaldo J

2010-09-01

This study evaluated the synergism between several P-solubilizing fungi isolates and arbuscular mycorrhizal fungi to improve clover ( Trifolium pratense) growth in the presence of Araxá apatite. Clover was sown directly in plastic pots with 300g of sterilized washed sand, vermiculite and sepiolite 1:1:1 (v:v:v) as substrate, and grown in a controlled environment chamber. The substrate was fertilized with 3 g L(-1) of Araxá apatite. A completely randomized design, in 8×2 factorial scheme (eight P-solubilizing fungi treatments with or without arbuscular mycorrhizal fungi)and four replicates were used. The P-solubilizing fungi treatments consisted of five Brazilian P-solubilizing fungi isolates (PSF 7, 9, 20, 21 and 22), two Spanish isolates ( Aspergillus niger and the yeast Yarowia lipolytica) and control (non-inoculated treatment). The greatest clover growth rate was recorded when Aspergillus niger and PSF 21 were co-inoculated with arbuscular mycorrhizal fungi. Aspergillus niger, PSF 7 and PSF 21 were the most effective isolates on increasing clover growth in the presence of arbuscular mycorrhizal fungi. Greater mycorrhizal colonization resulted in greater clover growth rate in most PSF treatments. PSF 7 was the best isolate to improve the establishment of mycorrhizal and rhizobia symbiosis.

Bacteria with Phosphate Solubilizing Capacity Alter Mycorrhizal Fungal Growth Both Inside and Outside the Root and in the Presence of Native Microbial Communities.

PubMed

Ordoñez, Yuli Marcela; Fernandez, Belen Rocio; Lara, Lidia Susana; Rodriguez, Alia; Uribe-Vélez, Daniel; Sanders, Ian R

2016-01-01

Arbuscular mycorrhizal fungi (AMF) and phosphate solubilizing Pseudomonas bacteria (PSB) could potentially interact synergistically because PSB solubilize phosphate into a form that AMF can absorb and transport to the plant. However, very little is known about the interactions between these two groups of microorganisms and how they influence the growth of each other. We tested whether different strains of bacteria, that have the capacity to solubilize phosphate, are able to grow along AMF hyphae and differentially influence the growth of AMF both outside the roots of carrot in in vitro conditions and inside the roots of potato in the presence of a microbial community. We found strong effects of AMF on the growth of the different bacterial strains. Different bacterial strains also had very strong effects on the growth of AMF extraradical hyphae outside the roots of carrot and on colonization of potato roots by AMF. The differential effects on colonization occurred in the presence of a microbial community. Our results show that these two important groups of rhizosphere microorganisms indeed interact with each other. Such interactions could potentially lead to synergistic effects between the two groups but this could depend on whether the bacteria truly solubilize phosphate in the rhizosphere in the presence of microbial communities.

In vitro digestion testing of lipid-based delivery systems: calcium ions combine with fatty acids liberated from triglyceride rich lipid solutions to form soaps and reduce the solubilization capacity of colloidal digestion products.

PubMed

Devraj, Ravi; Williams, Hywel D; Warren, Dallas B; Mullertz, Anette; Porter, Christopher J H; Pouton, Colin W

2013-01-30

In vitro digestion testing is of practical importance to predict the fate of drugs administered in lipid-based delivery systems. Calcium ions are often added to digestion media to increase the extent of digestion of long-chain triglycerides (LCTs), but the effects they have on phase behaviour of the products of digestion, and consequent drug solubilization, are not well understood. This study investigates the effect of calcium and bile salt concentrations on the rate and extent of in vitro digestion of soybean oil, as well as the solubilizing capacity of the digestion products for two poorly water-soluble drugs, fenofibrate and danazol. In the presence of higher concentrations of calcium ions, the solubilization capacities of the digests were reduced for both drugs. This effect is attributed to the formation of insoluble calcium soaps, visible as precipitates during the digestions. This reduces the availability of liberated fatty acids to form mixed micelles and vesicles, thereby reducing drug solubilization. The use of high calcium concentrations does indeed force in vitro digestion of LCTs but may overestimate the extent of drug precipitation that occurs within the intestinal lumen. Copyright © 2012 Elsevier B.V. All rights reserved.

Cyclodextrin-enhanced solubilization and removal of residual-phase chlorinated solvents from porous media

DOE Office of Scientific and Technical Information (OSTI.GOV)

Boving, T.B.; Wang, X.; Brusseau, M.L.

1999-03-01

The development of improved methods for remediation of contaminated aquifers has emerged as a significant environmental priority. One technology that appears to have considerable promise involves the use of solubilization agents such as surfactants and cosolvents for enhancing the removal of residual phase immiscible liquids. The authors examined the use of cyclodextrin, a glucose-based molecule, for solubilizing and removing residual-phase immiscible liquid from porous media. Batch experiments were conducted to measure the degree of trichloroethene (TCE) and tetrachloroethene (PCE) solubilization induced by hydroxypropyl-{beta}-cyclodextrin (HPCD) and methyl-{beta}-cyclodextrin (MCD). These studies revealed that the solubilities of TCE and PCE were enhanced bymore » up to 9.5 and 36.0 times, respectively. Column experiments were conducted to compare water and cyclodextrin-enhanced flushing of Borden sand containing residual saturations of TCE and PCE. The results indicate that solubilization and mass removal were enhanced substantially with the use of cyclodextrins. The effluent concentrations during the steady-state phase of the HPCD and MCD flushing experiments were close to the apparent solubilities measured with the batch experiments, indicating equilibrium concentrations were maintained during the initial phase of cyclodextrin flushing. Mobilization was observed for only the TCE-MCD and PCE-5%MCD experiments.« less

Solubilization of phencyclidine receptors from rat cerebral cortex in an active ligand binding site

DOE Office of Scientific and Technical Information (OSTI.GOV)

McVittie, L.D.; Sibley, D.R.

1989-01-01

A phencyclidine (PCP) receptor binding site has been solubilized in an active ligand-binding state from rat cerebral cortical membranes with sodium deoxycholate. Optimal receptor solubilization occurs at a detergent/protein ratio of 0.5 (w/w); for 5 mg protein/ml solubilized with 0.25% sodium deoxycholate, about 60% of the protein and 25% of the receptor is solubilized. Specific binding of either (/sup 3/H)-N-(1-(2-thienyl)cyclohexyl)piperidine ((/sup 3/H)TCP) or (/sup 3/H)MK-801 is measurable by filtration through Sephadex G-50 columns or glass fiber filters; more than 60% of the binding activity is stable after 48 h at 4/degrees/C. In the presence of detergent, (/sup 3/H)TCP binding exhibitsmore » a K/sub d/ of 250 nM, a B/sub max/ of 0.56 pmol/mg protein, and a pharmacological profile consistent with that of the membrane-bound PCP receptor, although most drugs bind with affinities 2 to 8 fold lower than in membranes. Upon reduction of detergent concentration, binding parameters approximate those for the membrane-bound receptor (/sup 3/H)TCP binding: K/sub d/ = 48 nM, M/sub max/ = 1.13 pmol/mg protein.« less

Photoacoustic analysis of the solubilization kinetics of pulmonary secretions from cystic fibrosis patients - secretor and non-secretor phenotypes

NASA Astrophysics Data System (ADS)

Barja, P. R.; Coelho, C. C.; Paiva, R. F.; Barboza, M. A.; Matos, L. C.; Matos, C. C. B.; Oliveira, L. V. F.

2010-03-01

Cystic fibrosis (CF) is an autosomal recessive inherited disease that increases viscoelasticity of pulmonary secretions. Affected patients are required to use therapeutic aerosols continuously. The expression of ABH glycoconjugates in exocrine secretions determines the nature of part of the carbohydrates present in these secretions, allowing the classification of individuals into the so-called "secretor" and "non secretor" phenotypes. The aim of this work was to employ photoacoustic (PA) measurements to monitor the solubilization kinetics of pulmonary secretions from CF patients, analyzing the influence of the secretor status in the solubilization kinetics of samples nebulized with different therapeutic aerosols. Sputum samples were obtained by spontaneous expectoration from positive and negative secretor CF patients. Each sample was nebulized with i) tobramycin, ii) alpha dornase, and iii) N-acetylcysteine in a PA cell; fitting of the data with the Boltzmann equation led to the determination of t0 (typical interaction time) and Δt (solubilization interval) for each curve. Differences between the secretor and non-secretor phenotypes were statistically significant in the groups for tobramycin and alpha dornase, but not for N-acetylcysteine. Results show that the secretor status influences the solubilization of pulmonary mucus of CF patients nebulized with tobramycin and alpha dornase.

Bioavailability of Carbohydrate Content in Natural and Transgenic Switchgrasses for the Extreme Thermophile Caldicellulosiruptor bescii

PubMed Central

Zurawski, Jeffrey V.; Khatibi, Piyum A.; Akinosho, Hannah O.; Straub, Christopher T.; Compton, Scott H.; Conway, Jonathan M.; Lee, Laura L.; Ragauskas, Arthur J.; Davison, Brian H.; Adams, Michael W. W.

2017-01-01

ABSTRACT Improving access to the carbohydrate content of lignocellulose is key to reducing recalcitrance for microbial deconstruction and conversion to fuels and chemicals. Caldicellulosiruptor bescii completely solubilizes naked microcrystalline cellulose, yet this transformation is impeded within the context of the plant cell wall by a network of lignin and hemicellulose. Here, the bioavailability of carbohydrates to C. bescii at 70°C was examined for reduced lignin transgenic switchgrass lines COMT3(+) and MYB Trans, their corresponding parental lines (cultivar Alamo) COMT3(−) and MYB wild type (WT), and the natural variant cultivar Cave-in-Rock (CR). Transgenic modification improved carbohydrate solubilization by C. bescii to 15% (2.3-fold) for MYB and to 36% (1.5-fold) for COMT, comparable to the levels achieved for the natural variant, CR (36%). Carbohydrate solubilization was nearly doubled after two consecutive microbial fermentations compared to one microbial step, but it never exceeded 50% overall. Hydrothermal treatment (180°C) prior to microbial steps improved solubilization 3.7-fold for the most recalcitrant line (MYB WT) and increased carbohydrate recovery to nearly 50% for the least recalcitrant lines [COMT3(+) and CR]. Alternating microbial and hydrothermal steps (T→M→T→M) further increased bioavailability, achieving carbohydrate solubilization ranging from 50% for MYB WT to above 70% for COMT3(+) and CR. Incomplete carbohydrate solubilization suggests that cellulose in the highly lignified residue was inaccessible; indeed, residue from the T→M→T→M treatment was primarily glucan and inert materials (lignin and ash). While C. bescii could significantly solubilize the transgenic switchgrass lines and natural variant tested here, additional or alternative strategies (physical, chemical, enzymatic, and/or genetic) are needed to eliminate recalcitrance. IMPORTANCE Key to a microbial process for solubilization of plant biomass is the organism's access to the carbohydrate content of lignocellulose. Economically viable routes will characteristically minimize physical, chemical, and biological pretreatment such that microbial steps contribute to the greatest extent possible. Recently, transgenic versions of plants and trees have been developed with the intention of lowering the barrier to lignocellulose conversion, with particular focus on lignin content and composition. Here, the extremely thermophilic bacterium Caldicellulosiruptor bescii was used to solubilize natural and genetically modified switchgrass lines, with and without the aid of hydrothermal treatment. For lignocellulose conversion, it is clear that the microorganism, plant biomass substrate, and processing steps must all be considered simultaneously to achieve optimal results. Whether switchgrass lines engineered for low lignin or natural variants with desirable properties are used, conversion will depend on microbial access to crystalline cellulose in the plant cell wall. PMID:28625990

Industrial-scale steam explosion pretreatment of sugarcane straw for enzymatic hydrolysis of cellulose for production of second generation ethanol and value-added products.

PubMed

Oliveira, Fernando M V; Pinheiro, Irapuan O; Souto-Maior, Ana M; Martin, Carlos; Gonçalves, Adilson R; Rocha, George J M

2013-02-01

Steam explosion at 180, 190 and 200°C for 15min was applied to sugarcane straw in an industrial sugar/ethanol reactor (2.5m(3)). The pretreated straw was delignificated by sodium hydroxide and hydrolyzed with cellulases, or submitted directly to enzymatic hydrolysis after the pretreatment. The pretreatments led to remarkable hemicellulose solubilization, with the maximum (92.7%) for pretreatment performed at 200°C. Alkaline treatment of the pretreated materials led to lignin solubilization of 86.7% at 180°C, and only to 81.3% in the material pretreated at 200°C. All pretreatment conditions led to high hydrolysis conversion of cellulose, with the maximum (80.0%) achieved at 200°C. Delignification increase the enzymatic conversion (from 58.8% in the cellulignin to 85.1% in the delignificated pulp) of the material pretreated at 180°C, but for the material pretreated at 190°C, the improvement was less remarkable, while for the pretreated at 200°C the hydrolysis conversion decreased after the alkaline treatment. Copyright © 2012 Elsevier Ltd. All rights reserved.

SOLUBILIZATION OF DODECANE, TETRACHLOROETHYLENE, AND 1,2-DICHLOROBENZENE IN MICELLAR SOLUTIONS OF ETHOXYLATED NONIONIC SURFACTANTS

EPA Science Inventory

Although surfactants have received considerable attention as a potential means for enhancing the recovery of organic compounds from the subsurface, only limited information is available regarding the micellar solubilization of common groundwater contaminants by nonionic surfactan...

Efficient Anaerobic Digestion of Microalgae Biomass: Proteins as a Key Macromolecule.

PubMed

Magdalena, Jose Antonio; Ballesteros, Mercedes; González-Fernandez, Cristina

2018-05-06

Biogas generation is the least complex technology to transform microalgae biomass into bioenergy. Since hydrolysis has been pointed out as the rate limiting stage of anaerobic digestion, the main challenge for an efficient biogas production is the optimization of cell wall disruption/hydrolysis. Among all tested pretreatments, enzymatic treatments were demonstrated not only very effective in disruption/hydrolysis but they also revealed the impact of microalgae macromolecular composition in the anaerobic process. Although carbohydrates have been traditionally recognized as the polymers responsible for the low microalgae digestibility, protease addition resulted in the highest organic matter solubilization and the highest methane production. However, protein solubilization during the pretreatment can result in anaerobic digestion inhibition due to the release of large amounts of ammonium nitrogen. The possible solutions to overcome these negative effects include the reduction of protein biomass levels by culturing the microalgae in low nitrogen media and the use of ammonia tolerant anaerobic inocula. Overall, this review is intended to evidence the relevance of microalgae proteins in different stages of anaerobic digestion, namely hydrolysis and methanogenesis.

Effective Application of Bicelles for Conformational Analysis of G Protein-Coupled Receptors by Hydrogen/Deuterium Exchange Mass Spectrometry

NASA Astrophysics Data System (ADS)

Duc, Nguyen Minh; Du, Yang; Thorsen, Thor S.; Lee, Su Youn; Zhang, Cheng; Kato, Hideaki; Kobilka, Brian K.; Chung, Ka Young

2015-05-01

G protein-coupled receptors (GPCRs) have important roles in physiology and pathology, and 40% of drugs currently on the market target GPCRs for the treatment of various diseases. Because of their therapeutic importance, the structural mechanism of GPCR signaling is of great interest in the field of drug discovery. Hydrogen/deuterium exchange mass spectrometry (HDX-MS) is a useful tool for analyzing ligand binding sites, the protein-protein interaction interface, and conformational changes of proteins. However, its application to GPCRs has been limited for various reasons, including the hydrophobic nature of GPCRs and the use of detergents in their preparation. In the present study, we tested the application of bicelles as a means of solubilizing GPCRs for HDX-MS studies. GPCRs (e.g., β2-adrenergic receptor [β2AR], μ-opioid receptor, and protease-activated receptor 1) solubilized in bicelles produced better sequence coverage (greater than 90%) than GPCRs solubilized in n-dodecyl-β-D-maltopyranoside (DDM), suggesting that bicelles are a more effective method of solubilization for HDX-MS studies. The HDX-MS profile of β2AR in bicelles showed that transmembrane domains (TMs) undergo lower deuterium uptake than intracellular or extracellular regions, which is consistent with the fact that the TMs are highly ordered and embedded in bicelles. The overall HDX-MS profiles of β2AR solubilized in bicelles and in DDM were similar except for intracellular loop 3. Interestingly, we detected EX1 kinetics, an important phenomenon in protein dynamics, at the C-terminus of TM6 in β2AR. In conclusion, we suggest the application of bicelles as a useful method for solubilizing GPCRs for conformational analysis by HDX-MS.

Sodium deoxycholate mediated enhanced solubilization and stability of hydrophobic drug Clozapine in pluronic micelles

NASA Astrophysics Data System (ADS)

Singla, Pankaj; Singh, Onkar; Chabba, Shruti; Aswal, V. K.; Mahajan, Rakesh Kumar

2018-02-01

In this report, the solubilization behaviour of a hydrophobic drug Clozapine (CLZ) in micellar suspensions of pluronics having different hydrophilic lipophilic balance (HLB) ratios viz. P84, F127 and F108 in the absence and presence of bile salt sodium deoxycholate (SDC) has been studied. UV-Vis spectroscopy has been exploited to determine the solubilization capacity of the investigated micellar systems in terms of drug loading efficiency, average number of drug molecules solubilized per micelle (ns), partition coefficient (P) and standard free energy of solubilization (Δ G°). The morphological and structural changes taking place in pluronics in different concentration regimes of SDC and with the addition of drug CLZ has been explored using dynamic light scattering (DLS) and small angle neutron scattering (SANS) measurements. The SANS results revealed that aggregation behaviour of pluronic-SDC mixed micelles gets improved in the presence of drug. The micropolarity measurements have been performed to shed light on the locus of solubilization of the drug in pure and mixed micellar systems. The compatibility between CLZ and drug carriers (pluronics and SDC) was confirmed using powder X-ray diffraction (PXRD) and Fourier transform infrared spectroscopy (FTIR) techniques. Among the investigated systems, P84-SDC mixed system was found to be highly efficient for CLZ loading. The long term stability data indicated that CLZ loaded P84-SDC mixed micellar formulation remained stable for 3 months at room temperature. Further, it was revealed that the CLZ loaded P84-SDC mixed micelles are converted into CLZ loaded pure P84 micelles at 30-fold dilutions which remain stable up to 48-fold dilutions. The results from the present studies suggest that P84-SDC mixed micelles can serve as suitable delivery vehicles for hydrophobic drug CLZ.

Physicochemical investigation of mixed surfactant microemulsions: water solubilization, thermodynamic properties, microstructure, and dynamics.

PubMed

Bardhan, Soumik; Kundu, Kaushik; Saha, Swapan K; Paul, Bidyut K

2013-12-01

In this contribution, we report on a systematic investigation of phase behavior and solubilization of water in water-in-heptane or decane aggregates stabilized by mixtures of polyoxyethylene (20) cetyl ether (Brij-58) and cetyltrimethylammonium bromide (CTAB) surfactants with varying compositions in conjugation with 1-pentanol (Pn) at fixed surfactant(s)/Pn ratio and temperature. Synergism in water solubilization was evidenced by the addition of CTAB to Brij-58 stabilized system in close proximity of equimolar composition in both oils. An attempt has been made to correlate composition dependent water solubilization and volume induced conductivity studies to provide insight into the solubilization mechanism of these mixed systems. Conductivity studies reveal the ascending curve in water solubilization capacity-(Brij-58:CTAB, w/w) profile as the interdroplet interaction branch indicating percolation of conductance and the descending curve is a curvature branch due to the rigidity of the interface in these systems. The microstructure of these systems as a function of surfactant composition has been determined by dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FTIR) measurements. FTIR study reveals increase and decrease in relative population of bound and bulk-like water, respectively, with increase in Brij-58:CTAB (w/w). DLS measurements showed that the droplet hydrodynamic diameter (Dh) decreases significantly with the increase in Brij-58:CTAB (w/w). Further, the interfacial composition and energetic parameters for the transfer of Pn from bulk oil to the interface were evaluated by the dilution method. Formation of temperature-insensitive microemulsions and temperature invariant droplet sizes are evidenced in the vicinity of the equimolar composition. The results are interpreted in terms of a proposed mechanism. Copyright © 2013 Elsevier Inc. All rights reserved.

Phosphate Solubilization Potential and Phosphatase Activity Of Rhizospheric Trichoderma Spp.

PubMed Central

Anil, Kapri; Lakshmi, Tewari

2010-01-01

Trichoderma sp., a well known biological control agent against several phytopathogens, was tested for its phosphate (P) solubilizing potential. Fourteen strains of Trichoderma sp. were isolated from the forest tree rhizospheres of pinus, deodar, bamboo, guava and oak on Trichoderma selective medium. The isolates were tested for their in-vitro P-solubilizing potential using National Botanical Research Institute Phosphate (NBRIP) broth containing tricalcium phosphate (TCP) as the sole P source, and compared with a standard culture of T. harzianum. All the cultures were found to solubilize TCP but with varying potential. The isolate DRT-1 showed maximum amount of soluble phosphate (404.07 εg.ml-1), followed by the standard culture of T. harzianum (386.42 εg.ml-1) after 96 h of incubation at 30±10C. Extra-cellular acid and alkaline phosphatases of the fungus were induced only in the presence of insoluble phosphorus source (TCP). High extra-cellular alkaline phosphatase activity was recorded for the isolate DRT-1 (14.50 U.ml-1) followed by the standard culture (13.41 U.ml-1) at 72h. The cultures showed much lesser acid phosphatase activities. Under glasshouse conditions, Trichoderma sp. inoculation increased chickpea (Cicer arietinum) growth parameters including shoot length, root length, fresh and dry weight of shoot as well as roots, in P-deficient soil containing only bound phosphate (TCP). Shoot weight was increased by 23% and 33% by inoculation with the isolate DRT-1 in the soil amended with 100 and 200 mg TCP kg-1 soil, respectively, after 60 d of sowing. The study explores high P-solubilizing potential of Trichoderma sp., which can be exploited for the solubilization of fixed phosphates present in the soil, thereby enhancing soil fertility and plant growth. PMID:24031556

Inhibition of liver fibrosis by solubilized coenzyme Q10: Role of Nrf2 activation in inhibiting transforming growth factor-beta1 expression

DOE Office of Scientific and Technical Information (OSTI.GOV)

Choi, Hoo-Kyun; Pokharel, Yuba Raj; Lim, Sung Chul

2009-11-01

Coenzyme Q10 (CoQ10), an endogenous antioxidant, is important in oxidative phosphorylation in mitochondria. It has anti-diabetic and anti-cardiovascular disease effects, but its ability to protect against liver fibrosis has not been studied. Here, we assessed the ability of solubilized CoQ10 to improve dimethylnitrosamine (DMN)-induced liver fibrogenesis in mice. DMN treatments for 3 weeks produced a marked liver fibrosis as assessed by histopathological examination and tissue 4-hydroxyproline content. Solubilized CoQ10 (10 and 30 mg/kg) significantly inhibited both the increases in fibrosis score and 4-hydroxyproline content induced by DMN. Reverse transcription-polymerase chain reaction and Western blot analyses revealed that solubilized CoQ10 inhibitedmore » increases in the transforming growth factor-beta1 (TGF-beta1) mRNA and alpha-smooth muscle actin (alpha-SMA) protein by DMN. Interestingly, hepatic glutamate-cysteine ligase (GCL) and glutathione S-transferase A2 (GSTA2) were up-regulated in mice treated with CoQ10. Solubilized CoQ10 also up-regulated antioxidant enzymes such as catalytic subunits of GCL and GSTA2 via activating NF-E2 related factor2 (Nrf2)/antioxidant response element (ARE) in H4IIE hepatoma cells. Moreover, CoQ10's inhibition of alpha-SMA and TGF-beta1 expressions disappeared in Nrf2-null MEF cells. In contrast, Nrf2 overexpression significantly decreased the basal expression levels of alpha-SMA and TGF-beta1 in Nrf2-null MEF cells. These results demonstrated that solubilized CoQ10 inhibited DMN-induced liver fibrosis through suppression of TGF-beta1 expression via Nrf2/ARE activation.« less

Physical characteristics of the gonadotropin receptor-hormone complexes formed in vivo and in vitro.

PubMed Central

Dufau, M L; Podesta, E J; Catt, K J

1975-01-01

The physical properties of detergent-solubilized gonadotropin receptor-hormone complexes, determined by density gradient centrifugation and gel filtration, were compared after in vivo and in vitro labeling of specific ovarian binding sites with radioiodinated human chorionic gonadotropin (hCG). Following intravenous administration of biologically active 125I-labeled hCG, up to 50% of the gonadotropin tracer was bound to the luteinized ovaries of immature female rats treated with pregnant mare serum/human chorionic gonadotropin. Comparable binding of 125I-labeled hCG was observed after equilibration of ovarian particles with the labeled hormone in vitro. The sedimentation properties of the solubilized receptor-hormone complexes formed in vivo were identical with those derived for the corresponding complexes formed in vitro and extracted with Triton X-100 and Lubrol PX, with sedimentation constants of 8.8 S for the Triton-solubilized complex and 7.0 S for the complex extracted with Lubrol PX. During analytical gel filtration of the Triton-solubilized receptor-hormone complex on Sepharose 6B in 0.1% Triton X-100, the partition coefficient (Kav) of the "in vivo" complex (0.32) was not significantly different from that of the complex formed in vitro (0.29). Gel filtration of the Lubrol-solubilized ovarian particles on Sepharose 6B in 0.5% Lubrol PX gave Kav values for the "in vivo" and "in vitro" labeled complexes of 0.36 and 0.32, respectively. These findings demonstrate that the physical properties of size and shape which determine the partition coefficient and sedimentation characteristics of detergent-solubilized gonadotropin receptor-hormone complexes formed in vitro are not distinguishable from those of the complexes extracted after specific interaction of the ovarian gonadotropin receptors with radioiodinated hCG in vivo. PMID:165502

Microbial liquefaction of peat for the production of synthetic fuels

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gunasekaran, M.

1988-01-01

Objectives of this study were: to evaluate the potential of using various microorganisms to hydrolyse and liquify peat; to determine the optimal conditions for peat hydrolysis and liquefaction; to study the co-metabolizable substances; to separate the compounds present in liquified peat by alumina and silica acid chromatography and capillary gas chromatography; and to identify the compounds in liquified peat by capillary GC-Mass spectrometry. Organisms used in the study include: Coprinus comatus, Coriolus hirsutus, Ganoderma lucidum, Lentinus edodes, Lenzites trabea, Phanerochaete chrysosporium, Pleurotus ostreatus, P. sapidus, Polyporus adjustus, Neurospora sitophila, Rhizophus arrhizus, Bacillus subtilis, Acinetobacter sp. and Alcaligenes sp. The fungimore » were maintained and cultivated in potato dextrose agar at 30 C. The bacteria were maintained in nutrient agar at 30 C. We have also initiated work on coal solubilization in addition to the studies on peat liquefaction. A relatively new substratum or semi-solid base for culture media called Pluronic F-127, or Polyol (BASF, New Jersey). Objectives of this study were: (1) to study the growth patterns of Candida ML 13 on pluronic as substratum; (2) to determine the rate of microbial coal solubilization on pluronic F-127 amended in different growth media; (3) to separate the mycelial mat of Candida ML 13 from unsolubilized coal particles and solubilized coal products from pluronic F-127; (4) to determine the effects of pH on microbial coal solubilization in pluronic F-127 media; (5) the effect of concentration of pluronic F-127 in media on coal solubilization; and, (6) to study the role of extracellular factors secreted by Candida ML 13 on coal solubilization in pluronic F-127 media. Results are discussed. 4 refs.« less

Zinc solubilizing Bacillus spp. potential candidates for biofortification in maize.

PubMed

Mumtaz, Muhammad Zahid; Ahmad, Maqshoof; Jamil, Moazzam; Hussain, Tanveer

2017-09-01

Bioaugmentation of Zn solubilizing rhizobacteria could be a sustainable intervention to increase bioavailability of Zn in soil which can be helpful in mitigation of yield loss and malnutrition of zinc. In present study, a number of pure rhizobacterial colonies were isolated from maize rhizosphere and screened for their ability to solubilize zinc oxide. These isolates were screened on the basis of zinc and phosphate solubilization, IAA production, protease production, catalase activity and starch hydrolysis. All the selected isolates were also positive for oxidase activity (except ZM22), HCN production (except ZM27) and utilization of citrate. More than 70% of isolates produces ammonia, hydrogen cyanide, siderophores, exopolysaccharides and cellulase. More than half of isolates also showed potential for urease activity and production of lipase. The ZM31 and S10 were the only isolates which showed the chitinase activity. All these isolates were evaluated in a jar trial for their ability to promote growth of maize under axenic conditions. Results revealed that inoculation of selected zinc solubilizing rhizobacterial isolates improved the growth of maize. In comparison, isolates ZM20, ZM31, ZM63 and S10 were best compared to other tested isolates in stimulating the growth attributes of maize like shoot length, root length, plant fresh and dry biomass. These strains were identified as Bacillus sp. (ZM20), Bacillus aryabhattai (ZM31 and S10) and Bacillus subtilis (ZM63) through 16S rRNA sequencing. This study indicated that inoculation of Zn solubilizing strains have potential to promote growth and can be the potential bio-inoculants for biofortification of maize to overcome the problems of malnutrition. Copyright © 2017 Elsevier GmbH. All rights reserved.

Low-temperature thermal pre-treatment of municipal wastewater sludge: Process optimization and effects on solubilization and anaerobic degradation.

PubMed

Nazari, Laleh; Yuan, Zhongshun; Santoro, Domenico; Sarathy, Siva; Ho, Dang; Batstone, Damien; Xu, Chunbao Charles; Ray, Madhumita B

2017-04-15

The present study examines the relationship between the degree of solubilization and biodegradability of wastewater sludge in anaerobic digestion as a result of low-temperature thermal pre-treatment. The main effect of thermal pre-treatment is the disintegration of cell membranes and thus solubilization of organic compounds. There is an established correlation between chemical oxygen demand (COD) solubilization and temperature of thermal pre-treatment, but results of thermal pre-treatment in terms of biodegradability are not well understood. Aiming to determine the impact of low temperature treatments on biogas production, the thermal pre-treatment process was first optimized based on an experimental design study on waste activated sludge in batch mode. The optimum temperature, reaction time and pH of the process were determined to be 80 °C, 5 h and pH 10, respectively. All three factors had a strong individual effect (p < 0.001), with a significant interaction effect for temp. pH 2 (p = 0.002). Thermal pre-treatments, carried out on seven different municipal wastewater sludges at the above optimum operating conditions, produced increased COD solubilization of 18.3 ± 7.5% and VSS reduction of 27.7 ± 12.3% compared to the untreated sludges. The solubilization of proteins was significantly higher than carbohydrates. Methane produced in biochemical methane potential (BMP) tests, indicated initial higher rates (p = 0.0013) for the thermally treated samples (k hyd up to 5 times higher), although the ultimate methane yields were not significantly affected by the treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of surfactants on apparent oxygen consumption of photosystem I isolated from Arthrospira platensis.

PubMed

Yu, Daoyong; Huang, Guihong; Xu, Fengxi; Ge, Baosheng; Liu, Shuang; Xu, Hai; Huang, Fang

2014-11-01

Surfactants play a significant role in solubilization of photosystem I (PSI) in vitro. Triton X-100 (TX), n-Dodecyl-β-D-maltoside (DDM), and sodium dodecyl sulfate (SDS) were employed to solubilize PSI particles in MES buffer to compare the effect of surfactant and its dosage on the apparent oxygen consumption rate of PSI. Through a combined assessment of sucrose density gradient centrifugation, Native PAGE and 77 K fluorescence with the apparent oxygen consumption, the nature of the enhancement of the apparent oxygen consumption activity of PSI by surfactants has been analyzed. Aggregated PSI particles can be dispersed by surfactant molecules into micelles, and the apparent oxygen consumption rate is higher for surfactant-solubilized PSI than for integral PSI particles. For DDM, PSI particles are solubilized mostly as the integral trimeric form. For TX, PSI particles are solubilized as incomplete trimeric and some monomeric forms. For the much harsher surfactant, SDS, PSI particles are completely solubilized as monomeric and its subunit forms. The enhancement of the oxygen consumption rate cannot be explained only by the effects of surfactant on the equilibrium between monomeric and trimeric forms of solubililized PSI. Care must be taken when the electron transfer activity of PSI is evaluated by methods based on oxygen consumption because the apparent oxygen consumption rate is influenced by uncoupled chlorophyll (Chl) from PSI, i.e., the larger the amount of uncoupled Chl, the higher the rate of apparent oxygen consumption. 77 K fluorescence spectra can be used to ensure that there is no uncoupled Chl present in the system. In order to eliminate the effect of trace uncoupled Chl, an efficient physical quencher of (1)O2, such as 1 mM NaN3, may be added into the mixture.

Macrophage Solubilization and Cytotoxicity of Indium-Containing Particles as in vitro Correlates to Pulmonary Toxicity in vivo

PubMed Central

Gwinn, William M.; Qu, Wei; Bousquet, Ronald W.; Price, Herman; Shines, Cassandra J.; Taylor, Genie J.; Waalkes, Michael P.; Morgan, Daniel L.

2015-01-01

Macrophage-solubilized indium-containing particles (ICPs) were previously shown in vitro to be cytotoxic. In this study, we compared macrophage solubilization and cytotoxicity of indium phosphide (InP) and indium-tin oxide (ITO) with similar particle diameters (∼1.5 µm) and then determined if relative differences in these in vitro parameters correlated with pulmonary toxicity in vivo. RAW 264.7 macrophages were treated with InP or ITO particles and cytotoxicity was assayed at 24 h. Ionic indium was measured in 24 h culture supernatants. Macrophage cytotoxicity and particle solubilization in vitro were much greater for InP compared with ITO. To correlate changes in vivo, B6C3F1 mice were treated with InP or ITO by oropharyngeal aspiration. On Days 14 and 28, bronchoalveolar lavage (BAL) and pleural lavage (PL) fluids were collected and assayed for total leukocytes. Cell differentials, lactate dehydrogenase activity, and protein levels were also measured in BAL. All lavage parameters were greatly increased in mice treated with InP compared with ITO. These data suggest that macrophage solubilization and cytotoxicity of some ICPs in vitro are capable of predicting pulmonary toxicity in vivo. In addition, these differences in toxicity were observed despite the two particulate compounds containing similar amounts of indium suggesting that solubilization, not total indium content, better reflects the toxic potential of some ICPs. Soluble InCl3 was shown to be more cytotoxic than InP to macrophages and lung epithelial cells in vitro further suggesting that ionic indium is the primary cytotoxic component of InP. PMID:25527823

Purification of inclusion bodies using PEG precipitation under denaturing conditions to produce recombinant therapeutic proteins from Escherichia coli.

PubMed

Chen, Huanhuan; Li, Ninghuan; Xie, Yueqing; Jiang, Hua; Yang, Xiaoyi; Cagliero, Cedric; Shi, Siwei; Zhu, Chencen; Luo, Han; Chen, Junsheng; Zhang, Lei; Zhao, Menglin; Feng, Lei; Lu, Huili; Zhu, Jianwei

2017-07-01

It has been documented that the purification of inclusion bodies from Escherichia coli by size exclusion chromatography (SEC) may benefit subsequent refolding and recovery of recombinant proteins. However, loading volume and the high cost of the column limits its application in large-scale manufacturing of biopharmaceutical proteins. We report a novel process using polyethylene glycol (PEG) precipitation under denaturing conditions to replace SEC for rapid purification of inclusion bodies containing recombinant therapeutic proteins. Using recombinant human interleukin 15 (rhIL-15) as an example, inclusion bodies of rhIL-15 were solubilized in 7 M guanidine hydrochloride, and rhIL-15 was precipitated by the addition of PEG 6000. A final concentration of 5% (w/v) PEG 6000 was found to be optimal to precipitate target proteins and enhance recovery and purity. Compared to the previously reported S-200 size exclusion purification method, PEG precipitation was easier to scale up and achieved the same protein yields and quality of the product. PEG precipitation also reduced manufacturing time by about 50 and 95% of material costs. After refolding and further purification, the rhIL-15 product was highly pure and demonstrated a comparable bioactivity with a rhIL-15 reference standard. Our studies demonstrated that PEG precipitation of inclusion bodies under denaturing conditions holds significant potential as a manufacturing process for biopharmaceuticals from E. coli protein expression systems.

Porous dendritic copper: an electrocatalyst for highly selective CO2 reduction to formate in water/ionic liquid electrolyte.

PubMed

Huan, Tran Ngoc; Simon, Philippe; Rousse, Gwenaëlle; Génois, Isabelle; Artero, Vincent; Fontecave, Marc

2017-01-01

Copper is currently extensively studied because it provides promising electrodes for carbon dioxide electroreduction. The original combination, reported here, of a nanostructured porous dendritic Cu-based material, characterized by electron microcopy (SEM, TEM) and X-ray diffraction methods, and a water/ionic liquid mixture as the solvent, contributing to CO 2 solubilization and activation, results in a remarkably efficient (large current densities at low overpotentials), stable and selective (large faradic yields) electrocatalytic system for the conversion of CO 2 into formic acid, a product with a variety of uses. These results provide new directions for the further improvement of Cu electrodes.

SURFACTANT-ENHANCED SOLUBILIZATION OF RESIDUAL DODECANE IN SOIL COLUMNS - 2. MATHEMATICAL MODELING

EPA Science Inventory

A mathematical model is developed to describe surfactant-enhanced solubilization of nonaqueous-phase liquids (NAPLs) in porous media. The model incorporates aqueous-phase transport equations for organic and surfactant components as well as a mass balance for the organic phase. Ra...

SURFACTANT ENHANCED SOLUBILIZATION OF RESIDUAL DODECANE IN SOIL COLUMNS 1. MATHEMATICAL MODELING

EPA Science Inventory

A mathematical model is developed to describe surfactant enhanced solubilization of nonaqueous phase liquids (NAPLS) in porous media. he model incorporates aqueous phase transport equations for organic and surfactant components as well as a mass balance on the organic phase. ate-...

DIRECT AND PHOTOACTIVATED TOXICITY OF A COMPLEX PETROLEUM MIXTURE: A COMPARISON OF SOLUBILIZATION METHODS

EPA Science Inventory

This work addresses several issues associated with the toxicity of a complex petroleum mixture (combined kerosene/diesel and crude oil), including developmental effects and early lifestage mortality, method of solubilization, and potential photo-activated and photo-modified toxic...

FIELD DEMONSTRATION STUDIES OF SURFACTANT-ENHANCED SOLUBILIZATION AND MOBILIZATION AT HILL AIR FORCE BASE, UTAH

EPA Science Inventory

Surfactant-enhanced subsurface remediation can dramatically improve contaminant removal rates compared to the traditional pump-and-treat technology. Surfactants can be used to significantly enhance the solubilization of non-aqueous phase liquids (NAPL) constituents, or they can b...

Gluconic acid production and phosphate solubilization by the plant growth-promoting bacterium Azospirillum spp.

NASA Astrophysics Data System (ADS)

Rodriguez, Hilda; Gonzalez, Tania; Goire, Isabel; Bashan, Yoav

2004-11-01

In vitro gluconic acid formation and phosphate solubilization from sparingly soluble phosphorus sources by two strains of the plant growth-promoting bacteria A. brasilense (Cd and 8-I) and one strain of A. lipoferum JA4 were studied. Strains of A. brasilense were capable of producing gluconic acid when grown in sparingly soluble calcium phosphate medium when their usual fructose carbon source is amended with glucose. At the same time, there is a reduction in pH of the medium and release of soluble phosphate. To a greater extent, gluconic acid production and pH reduction were observed for A. lipoferum JA4. For the three strains, clearing halos were detected on solid medium plates with calcium phosphate. This is the first report of in vitro gluconic acid production and direct phosphate solubilization by A. brasilense and the first report of P solubilization by A. lipoferum. This adds to the very broad spectrum of plant growth-promoting abilities of this genus.

Enhanced aqueous solubility of polycyclic aromatic hydrocarbons by green diester-linked cationic gemini surfactants and their binary solutions

NASA Astrophysics Data System (ADS)

Panda, Manorama; Fatma, Nazish; Kabir-ud-Din

2016-07-01

Three homologues of a novel biodegradable diester-linked cationic gemini surfactant series, CmH2m+1 (CH3)2N+(CH2COOCH2)2N+(CH3)2CmH2m+1.2Cl- (m-E2-m; m = 12, 14, 16), were used for investigation of the solubilization of polycyclic aromatic hydrocarbons (PAHs) such as naphthalene, anthracene and pyrene in single as well as binary surfactant solutions. Physicochemical parameters of the pure/mixed systems were derived by conductivity and surface tension measurements. Dissolution capacity of the equimolar binary surfactant solutions towards the PAHs was studied from the molar solubilization ratio (MSR), micelle-water partition coefficient (Km) and free energy of solubilization (ΔGs0) of the solubilizates. Influence of hydrophobic chain length of the dimeric surfactants on solubilization was characterized. Aqueous solubility of the PAHs was enhanced linearly with concentration of the surfactant in all the pure and mixed gemini-gemini surfactant systems.

Solubilization of phenanthrene above cloud point of Brij 30: a new application in biodegradation.

PubMed

Pantsyrnaya, T; Delaunay, S; Goergen, J L; Guseva, E; Boudrant, J

2013-06-01

In the present study a new application of solubilization of phenanthrene above cloud point of Brij 30 in biodegradation was developed. It was shown that a temporal solubilization of phenanthrene above cloud point of Brij 30 (5wt%) permitted to obtain a stable increase of the solubility of phenanthrene even when the temperature was decreased to culture conditions of used microorganism Pseudomonas putida (28°C). A higher initial concentration of soluble phenanthrene was obtained after the cloud point treatment: 200 against 120μM without treatment. All soluble phenanthrene was metabolized and a higher final concentration of its major metabolite - 1-hydroxy-2-naphthoic acid - (160 against 85μM) was measured in the culture medium in the case of a preliminary cloud point treatment. Therefore a temporary solubilization at cloud point might have a perspective application in the enhancement of biodegradation of polycyclic aromatic hydrocarbons. Copyright © 2013 Elsevier Ltd. All rights reserved.

Biotic and abiotic reduction and solubilization of Pu(IV)O₂•xH₂O(am) as affected by anthraquinone-2,6-disulfonate (AQDS) and ethylenediaminetetraacetate (EDTA).

PubMed

Plymale, Andrew E; Bailey, Vanessa L; Fredrickson, James K; Heald, Steve M; Buck, Edgar C; Shi, Liang; Wang, Zheming; Resch, Charles T; Moore, Dean A; Bolton, Harvey

2012-02-21

This study measured reductive solubilization of plutonium(IV) hydrous oxide (Pu(IV)O(2)·xH(2)O((am))) with hydrogen (H(2)) as electron donor, in the presence or absence of dissimilatory metal-reducing bacteria (DMRB), anthraquinone-2,6-disulfonate (AQDS), and ethylenediaminetetraacetate (EDTA). In PIPES buffer at pH 7 with excess H(2), Shewanella oneidensis and Geobacter sulfurreducens both solubilized <0.001% of 0.5 mM Pu(IV)O(2)·xH(2)O((am)) over 8 days, with or without AQDS. However, Pu((aq)) increased by an order of magnitude in some treatments, and increases in solubility were associated with production of Pu(III)((aq)). The solid phase of these treatments contained Pu(III)(OH)(3(am)), with more in the DMRB treatments compared with abiotic controls. In the presence of EDTA and AQDS, PuO(2)·xH(2)O((am)) was completely solubilized by S. oneidensis and G. sulfurreducens in ∼24 h. Without AQDS, bioreductive solubilization was slower (∼22 days) and less extensive (∼83-94%). In the absence of DMRB, EDTA facilitated reductive solubilization of 89% (without AQDS) to 98% (with AQDS) of the added PuO(2)·xH(2)O((am)) over 418 days. An in vitro assay demonstrated electron transfer to PuO(2)·xH(2)O((am)) from the S. oneidensis outer-membrane c-type cytochrome MtrC. Our results (1) suggest that PuO(2)·xH(2)O((am)) reductive solubilization may be important in reducing environments, especially in the presence of complexing ligands and electron shuttles, (2) highlight the environmental importance of polynuclear, colloidal Pu, (3) provide additional evidence that Pu(III)-EDTA is a more likely mobile form of Pu than Pu(IV)-EDTA, and (4) provide another example of outer-membrane cytochromes and electron-shuttling compounds facilitating bioreduction of insoluble electron acceptors in geologic environments.

Assessment of two carrier materials for phosphate solubilizing biofertilizers and their effect on growth of wheat (Triticum aestivum L.).

PubMed

Mukhtar, Salma; Shahid, Izzah; Mehnaz, Samina; Malik, Kauser A

2017-12-01

Biofertilizers are usually carrier-based inoculants containing beneficial microorganisms. Incorporation of microorganisms in carrier material enables easy-handling, long-term storage and high effectiveness of biofertilizers. Objective of the present study was to assess enriched biogas sludge and soil as biofertilizer carriers on growth and yield of wheat. Six phosphate solubilizing strains were used in this study. Three phosphate solubilizing strains, 77-NS2 (Bacillus endophyticus), 77-CS-S1 (Bacillus sphaericus) and 77-NS5 (Enterobacter aerogenes) were isolated from the rhizosphere of sugarcane, two strains, PSB5 (Bacillus safensis) and PSB12 (Bacillus megaterium) from the rhizosphere of wheat and one halophilic phosphate solubilizing strain AT2RP3 (Virgibacillus sp.) from the rhizosphere of Atriplex amnicola, were used as bioinoculants. Phosphate solubilization ability of these strains was checked in vitro in Pikovskaya medium, containing rock phosphate (RP) as insoluble P source, individually supplemented with three different carbon sources, i.e., glucose, sucrose and maltose. Maximum phosphate solubilization; 305.6μg/ml, 217.2μg/ml and 148.1μg/ml was observed in Bacillus strain PSB12 in Pikovskaya medium containing sucrose, maltose and glucose respectively. A field experiment and pot experiments in climate control room were conducted to study the effects of biogas sludge and enriched soil based phosphorous biofertilizers on growth of wheat. Bacillus strain PSB12 significantly increased root and shoot dry weights and lengths using biogas sludge as carrier material in climate control room experiments. While in field conditions, significant increase in root and shoot dry weights, lengths and seed weights was seen by PSB12 and PSB5 (Bacillus) and Enterobacter strain 77-NS5 using biogas sludge as carrier. PSB12 also significantly increased both root and shoot dry weights and lengths in field conditions when used as enriched soil based inoculum. These results indicated that bacterial isolates having plant beneficial traits such as P solubilization are more promising candidates as biofertilizer when used with carrier materials. Copyright © 2017 Elsevier GmbH. All rights reserved.

Amphiphiles for protein solubilization and stabilization

DOEpatents

Gellman, Samuel Helmer; Chae, Pil Seok; Laible, Philip D.; Wander, Marc J.

2012-09-11

The invention provides amphiphiles for manipulating membrane proteins. The amphiphiles can feature carbohydrate-derived hydrophilic groups and branchpoints in the hydrophilic moiety and/or in a lipophilic moiety. Such amphiphiles are useful as detergents for solubilization and stabilization of membrane proteins, including photosynthetic protein superassemblies obtained from bacterial membranes.

Amphiphiles for protein solubilization and stabilization

DOEpatents

Gellman, Samuel Helmer; Chae, Pil Seok; Laible, Phillip D; Wander, Marc J

2014-11-04

The invention provides amphiphiles for manipulating membrane proteins. The amphiphiles can feature carbohydrate-derived hydrophilic groups and branchpoints in the hydrophilic moiety and/or in a lipophilic moiety. Such amphiphiles are useful as detergents for solubilization and stabilization of membrane proteins, including photosynthetic protein superassemblies obtained from bacterial membranes.

Quantitation of buried contamination by use of solvents

NASA Technical Reports Server (NTRS)

Pappas, S. P.; Hsiao, P.; Hill, L. W.

1972-01-01

Solubilization studies were carried out on various cured silicone resins. A solvent spectrum was prepared. It was found that complete dissolution of cured silicone resins could be achieved without extensive physical degradation of samples. Based on the solubilization results, amine solvents were selected for spore viability studies.

Isolation and characterization of Chilembwe and Sinda Rock Phosphate solubilizing soil microorganisms

USDA-ARS?s Scientific Manuscript database

This study was conducted to isolate and characterize soil microorganisms capable of solubilizing Chilembwe and Sinda rock phosphates readily available in Zambia. Single isolates were obtained by direct plating and enrichment cultures with succinate, cellulose and glucose as the carbon sources. Isola...

Anaerobic microbial remobilization of coprecipitated metals

DOEpatents

Francis, Arokiasamy J.; Dodge, Cleveland J.

1994-10-11

A process is provided for solubilizing coprecipitated metals. Metals in wastestreams are concentrated by treatment with an iron oxide coprecipitating agent. The coprecipitated metals are solubilized by contacting the coprecipitate with a bacterial culture of a Clostridium species ATCC 53464. The remobilized metals can then be recovered and recycled.

Solubilization, partial purification, and properties of omega-conotoxin receptors associated with voltage-dependent calcium channels from rat brain synaptosomes.

PubMed

Rosenberg, R L; Isaacson, J S; Tsien, R W

1989-01-01

These experiments provide a starting point for biochemical characterization of Ca channels from neuronal membranes, using omega-CgTX as a specific marker. The purification of the omega-CgTX receptors is far from complete. Each of the purification steps described results in only a two- to fivefold enrichment of the receptor proteins, and is accompanied by a loss of receptor concentration and stability, so the maximal specific activity achieved by a combination of these steps falls several orders of magnitude short of that of a large, homogeneous, active protein. Nevertheless, these studies have yielded important information about the omega-CgTX receptor. The Stokes' radius, determined from gel exclusion chromatography, is approximately 87 A, and the sedimentation coefficient, determined from sucrose gradient sedimentation, is approximately 19 S. These values are similar to those found for the DHP receptors solubilized in digitonin. We have also found that at least some of the omega-CgTX receptors have complex carbohydrate moieties that are recognized by WGA, together with evidence of heterogeneity of receptor glycosylation. Additionally, we have been able to use the solubilized, partially purified receptors in cross-linking experiments to tentatively identify the molecular weights of the omega-CgTX targets from rat brain. A large peptide of approximately 300 kDa, similar to that identified in photoaffinity studies, is very clearly labeled by the chemical incorporation of [125I]omega-CgTX into partially purified receptor preparations, but some ambiguity remains because of the faint labeling of peptides in the 120-170-kDa range. The approximately 300-kDa peptide is much larger than any single peptide component of DHP receptors from skeletal muscle, and it may be related to a molecular combination of the 170-kDa and 135-kDa subunits of the DHP receptor. Because [125I]omega-CgTX presumably labels both N- and L-type neuronal Ca channels, both channel types will probably be found in the purified preparations. Thus, at some time, it will be necessary to separate DHP-sensitive L-type channels from preparations of L- and N-type channels identified by omega-CgTX binding.

Cytotoxic and anti-angiogenic paclitaxel solubilized and permeation-enhanced by natural product nanoparticles

PubMed Central

Liu, Zhijun; Zhang, Fang; Koh, Gar Yee; Dong, Xin; Hollingsworth, Javoris; Zhang, Jian; Russo, Paul S.; Yang, Peiying; Stout, Rhett W.

2014-01-01

Paclitaxel (PTX) is one of the most potent intravenous chemotherapeutic agents to date, yet an oral formulation has been problematic due to its low solubility and permeability. Using the recently discovered solubilizing properties of rubusoside (RUB), we investigated this unique PTX-RUB formulation. Paclitaxel was solubilized by RUB in water to levels of 1.6 to 6.3 mg/mL at 10 to 40% weight/volume. These, nanomicellar, PTX-RUB complexes were dried to a powder which was subsequently reconstituted in physiologic solutions. After 2.5 hrs in gastric fluid 85 to 99% of PTX-RUB remained soluble, while 79 to 96% remained soluble in intestinal fluid. The solubilization of PTX was mechanized by the formation of water-soluble spherical nanomicelles between PTX and RUB with an average diameter of 6.6 nm. Compared with Taxol®, PTX-RUB nanoparticles were nearly four times more permeable in Caco-2 cell monocultures. In a side-by-side comparison with DMSO-solubilized PTX, PTX-RUB maintained the same level of cytotoxicity against three human cancer cell lines with IC50 values ranging from 4 nM to 20 nM. Additionally, tubular formation and migration of HUVECs were inhibited at levels as low as 5 nM. These chemical and biological properties demonstrated by the PTX-RUB nanoparticles may improve oral bioavailability and enable further pharmacokinetic, toxicologic, and efficacy investigations. PMID:25243454

Effect of hydrothermal pre-treatment (HTP) on poultry slaughterhouse waste (PSW) sludge for the enhancement of the solubilization, physical properties, and biogas production through anaerobic digestion.

PubMed

Park, Seyong; Yoon, Young-Man; Han, Seong Kuk; Kim, Daegi; Kim, Ho

2017-06-01

This study is an assessment of the hydrothermal pre-treatment (HTP) of poultry slaughterhouse waste (PSW) sludge for the enhancement of the solubilization, physical properties, and biogas production through anaerobic digestion. This assessment was carried out to ascertain the optimal HTP temperature. The solubilization and physical properties efficacy was investigated by capillary suction time (CST), time to filter (TTF), and particle size. In addition, the anaerobic digestion was investigated through biochemical methane potential (BMP) tests and subsequent statistical analysis using the modified Gompertz model. HTP was found to have improved the solubilization of the PSW sludge with increasing HTP temperature. In addition, the results of the CST, TTF, and particle size decreased with increasing HTP temperature. These results of the assessment that was conducted in this study confirm that the HTP process indeed modifies the physical properties of PSWs to enhance the solubilization of organic solids. Nevertheless, the results of the BMP tests and the modified Gompertz model analysis show that the optimal HTP temperature of PSWs for anaerobic digestion is 190°C. These findings show that to achieve high conversion efficiency, an accurately designed pre-treatment step must be included in the overall anaerobic digestion process for wastewater treatment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Kinetics of degradation of surfactant-solubilized fluoranthene by a Sphingomonas paucimobilis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Willumsen, P.A.; Arvin, E.

To achieve a better quantitative understanding of the stimulating or inhibiting effect of surfactants on the metabolism of polycyclic aromatic hydrocarbons (PAHs), a biodegradation model describing solubilization, bioavailability, and biodegradation of crystalline fluoranthene is proposed and used to model experimental data. The degradation was investigated in batch systems containing the PAH-degrading bacterium Sphingomonas paucimobilis strain EPA505, the nonionic surfactant Triton X-100, and a fluoranthene-amended liquid mineral salts medium. Surfactant-enhanced biodegradation is complex; however, the biodegradation model predicted fluoranthene disappearance and the initial mineralization well. Surfactant-amendment did increase fluoranthene mineralization rates by strain EPA505; however, the increases were not proportional tomore » the rates of fluoranthene solubilization. The surfactant clearly influenced the microbial PAH metabolism as indicated by a rapid accumulation of colored products and by a surfactant -related decreased in the overall extent of fluoranthene mineralization. Model estimations of the bioavailability of micelle-solubilized fluoranthene, the relatively fast fluoranthene disappearance, and the accumulation of extracellular compounds in the degradation system suggest that low availability of micellar fluoranthene is not the only factor controlling surfactant-enhanced biodegradation. Also factors such as the extent of accumulation and bioavailability of the PAH metabolites and the crystalline solubilization rate in the presence of surfactants may determine the overall effect of surfactant-enhanced biodegradation of high molecular weight PAHs.« less

Solubilized liver extracellular matrix maintains primary rat hepatocyte phenotype in-vitro.

PubMed

Loneker, Abigail E; Faulk, Denver M; Hussey, George S; D'Amore, Antonio; Badylak, Stephen F

2016-04-01

Whole organ engineering and cell-based regenerative medicine approaches are being investigated as potential therapeutic options for end-stage liver failure. However, a major challenge of these strategies is the loss of hepatic specific function after hepatocytes are removed from their native microenvironment. The objective of the present study was to determine if solubilized liver extracellular matrix (ECM), when used as a media supplement, can better maintain hepatocyte phenotype compared to type I collagen alone or solubilized ECM harvested from a non-liver tissue source. Liver extracellular matrix (LECM) from four different species was isolated via liver tissue decellularization, solubilized, and then used as a media supplement for primary rat hepatocytes (PRH). The four species of LECM investigated were human, porcine, canine and rat. Cell morphology, albumin secretion, and ammonia metabolism were used to assess maintenance of hepatocyte phenotype. Biochemical and mechanical characterization of each LECM were also conducted. Results showed that PRH's supplemented with canine and porcine LECM maintained their phenotype to a greater extent compared to all other groups. PRH's supplemented with canine and porcine LECM showed increased bile production, increased albumin production, and the formation of multinucleate cells. The findings of the present study suggest that solubilized liver ECM can support in-vitro hepatocyte culture and should be considered for therapeutic and diagnostic techniques that utilize hepatocytes. © 2016 Wiley Periodicals, Inc.

Aggregate-based sub-CMC Solubilization of Hexadecane by Surfactants.

PubMed

Zhong, Hua; Yang, Lei; Zeng, Guangming; Brusseau, Mark L; Wang, Yake; Li, Yang; Liu, Zhifeng; Yuan, Xingzhong; Tan, Fei

Solubilization of hexadecane by two surfactants, SDBS and Triton X-100, at concentrations near the critical micelle concentration (CMC) and the related aggregation behavior was investigated in this study. Solubilization was observed at surfactant concentrations lower than CMC, and the apparent solubility of hexadecane increased linearly with surfactant concentration for both surfactants. The capacity of SDBS to solubilize hexadecane is stronger at concentrations below CMC than above CMC. In contrast, Triton X-100 shows no difference. The results of dynamic light scattering (DLS) and cryogenic TEM analysis show aggregate formation at surfactant concentrations lower than CMC. DLS-based size of the aggregates ( d ) decreases with increasing surfactant concentration. Zeta potential of the SDBS aggregates decreases with increasing SDBS concentration, whereas it increases for Triton X-100. The surface excess (Γ) of SDBS calculated based on hexadecane solubility and aggregate size data increases rapidly with increasing bulk concentration, and then asymptotically approaches the maximum surface excess (Γ max ). Conversely, there is only a minor increase in Γ for Triton X-100. Comparison of Γ and d indicates that excess of surfactant molecules at aggregate surface has great impact on surface curvature. The results of this study demonstrate formation of aggregates at surfactant concentrations below CMC for hexadecane solubilization, and indicate the potential of employing low-concentration strategy for surfactant application such as remediation of HOC contaminated sites.

Assessment of solubilization characteristics of different surfactants for carvedilol phosphate as a function of pH.

PubMed

Chakraborty, Subhashis; Shukla, Dali; Jain, Achint; Mishra, Brahmeshwar; Singh, Sanjay

2009-07-15

The effect of surfactants on the solubility of a new phosphate salt of carvedilol was investigated at different biorelevent pH to evaluate their solubilization capacity. Solutions of different classes of surfactants viz., anionic-sodium dodecyl sulfate (SDS) and sodium taurocholate (STC), cationic-cetyltrimethylammonium bromide (CTAB) and non-ionic-Tween 80 (T80) were prepared in the concentration range of 5-35 mmol dm(-3) in buffer solutions of pH 1.2, 3.0, 4.5, 5.8, 6.8 and 7.2. The solubility data were used to calculate the solubilization characteristics viz. molar solubilization capacity, water micelle partition coefficient, free energy of solubilization and binding constant. Solubility enhancement in basic pH was in following order: CTAB>T80>SDS>STC. CTAB and T80 showed remarkable solubility enhancement in acidic pH as well. Among the anionic surfactants, solubility in acidic medium was retarded except at pH 1.2 in case of SDS. Cationic and non-ionic surfactants were found to be suitable for enhancing the solubility of CP which can be employed for maintaining the in vitro sink condition in the basic dissolution medium. While anionic surfactants showed solubility retardant behavior which may be exploited in increasing the drug entrapment efficiency of a colloidal drug delivery system formulated by emulsification technique.

Effect of biosurfactants on the aqueous solubility of PCE and TCE.

PubMed

Albino, John D; Nambi, Indumathi M

2009-12-01

The effect of biosurfactants on the solubility of tetrachloroethylene (PCE) and trichloroethylene (TCE) was studied in batch experiments pertaining to their use for solubilization and mobilization of such contaminants in surfactant enhanced aquifer remediation. Biosurfactants, rhamnolipid and surfactin used in solubility studies were synthesized in our laboratory by Pseudomonas aeruginosa (MTCC 2297) and Bacillus subtilis (MTCC 2423), respectively. The efficiency of the biosurfactants in solubilizing the chlorinated solvents was compared to that of synthetic surfactants. The Weight Solubilization Ratio (WSR) values for solubilization of PCE and TCE by biosurfactants were very high compared to the values obtained for synthetic surfactants. Surfactin proved to be a better surfactant over rhamnolipid. The WSR of surfactin on solubilization of PCE and TCE were 3.83 and 12.5, respectively, whereas the values obtained for rhamnolipid were 2.06 and 8.36. The solubility of the chlorinated solvents by biosurfactants was considerably affected by the changes in pH. The aqueous solubility of PCE and TCE increased tremendously with decrease in pH. The solubility of biosurfactants was observed to decrease with the pH, favoring partitioning of surfactants into the chlorinated solvents in significant amounts at lower pH. The excessive accumulation of biosurfactants at the interface facilitated interfacial tension reductions resulting in higher solubility of the chlorinated solvents at pH less than 7.

Triterpenoids Amplify Anti-Tumoral Effects of Mistletoe Extracts on Murine B16.F10 Melanoma In Vivo

PubMed Central

Strüh, Christian M.; Jäger, Sebastian; Kersten, Astrid; Schempp, Christoph M.; Scheffler, Armin; Martin, Stefan F.

2013-01-01

Purpose Mistletoe extracts are often used in complementary cancer therapy although the efficacy of that therapy is controversially discussed. Approved mistletoe extracts contain mainly water soluble compounds of the mistletoe plant, i.e. mistletoe lectins. However, mistletoe also contains water-insoluble triterpenoids (mainly oleanolic acid) that have anti-tumorigenic effects. To overcome their loss in watery extracts we have solubilized mistletoe triterpenoids with cyclodextrins, thus making them available for in vivo cancer experiments. Experimental design B16.F10 subcutaneous melanoma bearing C57BL/6 mice were treated with new mistletoe extracts containing both water soluble compounds and solubilized triterpenoids. Tumor growth and survival was monitored. In addition, histological examinations of the tumor material and tumor surrounding tissue were performed. Results Addition of solubilized triterpenoids increased the anti-tumor effects of the mistletoe extracts, resulting in reduced tumor growth and prolonged survival of the mice. Histological examination of the treated tumors showed mainly tumor necrosis and some apoptotic cells with active caspase-3 and TUNEL staining. A significant decrease of CD31-positive tumor blood vessels was observed after treatment with solubilized triterpenoids and different mistletoe extracts. Conclusion We conclude that the addition of solubilized mistletoe triterpenoids to conventional mistletoe extracts improves the efficacy of mistletoe treatment and may represent a novel treatment option for malignant melanoma. PMID:23614029

Intestinal solubilization of particle-associated organic and inorganic mercury as a measure of bioavailability to benthic invertebrates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lawrence, A.L.; McAloon, K.M.; Mason, R.P.

1999-06-01

The bioavailability of particle-associated inorganic mercury (Hg{sub I}) and monomethylmercury (MMHg) was evaluated in vitro using digestive fluid of the deposit feeding lugworm, Arenicola marina. Digestive fluid, removed from the midgut of the polychaete, was incubated with contaminated sediment, and the proportion of Hg{sub I} or MMHg solubilized by the digestive fluid was determined. Digestive fluid was found to be a more effective solvent than seawater in solubilizing particle-associated Hg{sub I} or MMHg. A greater percentage of MMHg than Hg{sub I} was solubilized from most sediments, suggesting that sediment-associated MMHg is generally more readily available from sediment for biological uptake.more » The proportion of MMHg released from the sediment was inversely correlated with sediment organic matter content, decreasing exponentially with increasing organic matter content of the sediment. The results for Hg{sub I} were equivocal. MMHg bioaccumulation factors (BAFs) from previous studies showed a similar trend with organic content of sediment, suggesting that solubilization may be the process limiting the bioaccumulation of particle-bound MMHg. It is concluded that in vitro extraction with a deposit feeder`s digestive fluid provides a potential tool to study the process of Hg bioaccumulation via ingestion routes, although its application to various sediments and organisms needs further investigation.« less

Evaluation of pink-pigmented facultative methylotrophic bacteria for phosphate solubilization.

PubMed

Jayashree, Shanmugam; Vadivukkarasi, Ponnusamy; Anand, Kirupanithi; Kato, Yuko; Seshadri, Sundaram

2011-08-01

Thirteen pink-pigmented facultative methylotrophic (PPFM) strains isolated from Adyar and Cooum rivers in Chennai and forest soil samples in Tamil Nadu, India, along with Methylobacterium extorquens, M. organophilum, M. gregans, and M. komagatae were screened for phosphate solubilization in plates. P-solubilization index of the PPFMs grown on NBRIP-BPB plates for 7 days ranged from 1.1 to 2.7. The growth of PPFMs in tricalcium phosphate amended media was found directly proportional to the glucose concentration. Higher phosphate solubilization was observed in four strains MSF 32 (415 mg l(-l)), MDW 80 (301 mg l(-l)), M. komagatae (279 mg l(-l)), and MSF 34 (202 mg l(-l)), after 7 days of incubation. A drop in the media pH from 6.6 to 3.4 was associated with an increase in titratable acidity. Acid phosphatase activity was more pronounced in the culture filtrate than alkaline phosphatase activity. Adherence of phosphate to densely grown bacterial surface was observed under scanning electron microscope after 7-day-old cultures. Biochemical characterization and screening for methanol dehydrogenase gene (mxaF) confirmed the strains as methylotrophs. The mxaF gene sequence from MSF 32 clustered towards M. lusitanum sp. with 99% similarity. This study forms the first detailed report on phosphate solubilization by the PPFMs.

Development Of Nutrient And Water Recycling Capabilities In Algae Biofuels Production Systems. Final Summary Report

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lundquist, Tryg; Spierling, Ruth; Poole, Kyle

The objective of this project was to develop and demonstrate methods of recycling of water and nutrients for algal biofuels production. Recycling was accomplished both internal to the system and, in a broader sense, through import and reuse of municipal wastewater. Such an integrated system with wastewater input had not been demonstrated previously, and the performance was unknown, particularly in terms of influence of recycling on algal productivity and the practical extent of nutrient recovery from biomass residuals. Through long-term laboratory and pilot research, the project resulted in the following: 1. Bench-scale pretreatment of algal biomass did not sufficiently increasemore » methane yield of nutrient solubilization during anaerobic digestion to warrant incorporation of pre-treatment into the pilot plant. The trial pretreatments were high-pressure orifice homogenization, sonication, and two types of heat treatment. 2. Solubilization of biomass particulate nutrients by lab anaerobic digesters ranged from 20% to nearly 60% for N and 40-65% for P. Subsequent aerobic degradation of the anaerobically digested biomass simulated raceways receiving whole digestate and resulted in an additional 20-55% N solubilization and additional 20% P solubilization. 3. Comparisons of laboratory and pilot digesters showed that laboratory units were reasonable proxies for pilot-scale. 4. Pilot-scale anaerobic digesters were designed, installed, and operated to digest algal biomass. Nutrient re-solubilization by the digesters was monitored and whole digestate was successfully used as a fertilizer in pilot algae raceways. 5. Unheated, unmixed digesters achieved greater methane yield and nutrient solubilization than heated, mixed digesters, presumably due to longer the solids residence times in unmixed digesters. The unmixed, unheated pilot digesters yielded 0.16 L CH4/g volatile solids (VS) introduced with 0.15 g VS/L-d organic loading and 16oC average temperature. A conventional heated mixed lab digester yielded 0.22 L CH4/g VS with 0.25 g VS/L-d and 30oC. The highest yield (0.30 L CH4/g VS) was achieved by the unmixed lab digesters operated at a constant 20oC. All digesters were operated with a 40-d hydraulic residence time. 6. In general, 50-75% of initial particulate N and P could be solubilized during anaerobic digestion and available for subsequent rounds of algae cultivation. 7. Bench-scale experiments showed the recovery from hydrothermal liquefaction (HTL) wastewater of carbon via anaerobic digestion and of nutrients to grow algae. To satisfy the nitrogen demand of algae cultivation, HTL wastewater would be diluted 400-fold, which was found to eliminate inhibition of algae growth by HTL wastewater. 8. Anaerobic digestion methane yield was lower for algal biomass containing coagulants such as would be used to aid harvesting or dewatering. Depending on doses, starch-based coagulant decreased yield by 10-14% and aluminum chlorohydrate decreased it by 14-26%. The lowest yield was 0.28 L CH 4/g volatile solids introduced to the digesters. 9. Algae harvested from raceways operated on recycled water had methane yields 13% higher than algae from raceways operated on both recycled water and nutrients provided by algae digestate. The slightly lower yield was expected due to the presence of previously digested biomass from the digestate fertilizer. 10. Defined media was replenished with nutrients and recycled repeatedly in sequential batch growth of Chlorella sorokiniana (DOE 1412). This laboratory study tested for inhibition and accumulation of inhibiting compounds (allelopathic or auto-inhibitory substances), information that would help estimate the blowdown ratio needed for an integrated system. In laboratory experiments in which water was recycled a total of five times, each successive round of reuse resulted in an average 4±3% reduction in log-phase specific growth rates. However, linear-phase growth inhibition was only observed in the final fifth round of reuse. 11. No decline in productivity was detected after 15 rounds of water recycling with nutrients provided by whole digestate in lab cultivation. Lab tests allowed for steady light and temperature, increasing the ability to detect inhibition. 12. In initial pilot inhibition studies, wastewater growth media was reused once while productivity was monitored. Media reuse was accomplished with triplicate sets of 33-m2 raceways operated in series. First-round gross productivity (based on effluent biomass flow) averaged 23 g/m 2-day annually while second-round gross productivity averaged 19 g/m 2-day annually. In terms of net productivity (based on raceway effluent biomass minus influent biomass), the first-round productivity averaged 15 g/m2-d and second round averaged 13 g/m 2-d during June-September operation. The higher productivity in the first-round ponds was likely due to heterotrophic/mixotrophic growth on the wastewater organic matter. 13. In a culminating pilot experiment, coagulant was used to decrease the carry-over of unsettled algae into subsequent rounds of growth. Over nearly 8 months, 93% of the media (the equivalent of 14 rounds of water reuse) was recycled without significant productivity loss compared to controls. Ponds receiving both recycled water and nutrients had net productivities of 14-24 g/m 2-d during fall and mid-summer, respectively. 14. Techno-economic analysis of the proposed facility found minimum fuel selling price to range from $7.01/gallon gasoline equivalent without revenue other than fuel to $3.85/GGE with revenue from wastewater treatment fees and LCFS and RIN (Low Carbon Fuel Standard and Renewable Identification Numbers) credits. 15. Life cycle assessment indicated GHG emissions of 40.7 g CO 2/MJ fuel and a net energy ratio (energy required/energy produced) of 0.37.« less

Anaerobic microbial remobilization of coprecipitated metals

DOEpatents

Francis, A.J.; Dodge, C.J.

1994-10-11

A process is provided for solubilizing coprecipitated metals. Metals in waste streams are concentrated by treatment with an iron oxide coprecipitating agent. The coprecipitated metals are solubilized by contacting the coprecipitate with a bacterial culture of a Clostridium species ATCC 53464. The remobilized metals can then be recovered and recycled. 4 figs.

Detergent solubilization of the EGF receptor from A431 cells

NASA Technical Reports Server (NTRS)

Dayanidhi, R.; Rintoul, D. A.; Spooner, B. S. (Principal Investigator)

1993-01-01

Functional reconstitution of purified preparations of human epidermal growth factor receptor (EGFR) requires dissociation of the protein from its plasma membrane lipid environment. Solubilization of membrane proteins in this manner requires the use of detergents, which are known to disrupt plasma membrane lipid/protein interactions. We have investigated the ability of three nonionic detergents to solubilize the human EGFR selectively, and have also analyzed the effect of these various treatments on the intrinsic tyrosyl kinase activity of the receptor. The nonionic detergent known as n-octyl glucoside (n-octyl beta-D-glucopyranoside) was found to give the best combination of selectivity, yield, and maintenance of enzymatic activity of the human EGFR.

Biosolubilization of coal by Candida in glucose limited cultures

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mitter, J.; Guillory, L.; Bose, N.K.

1990-01-01

Coal biodegradation is attracting the attention of many workers because of its significance for efficient bioconversion of coal into useful chemicals. The authors work is based upon the beneficiation of a fungus (candida) on subbituminous coal. Candida was grown on both solid and liquid sabouraud medium and the coal solubilizing activity was studied at varying glucose concentration and temperature. Lower glucose concentration and higher temperature enhanced coal solubilizing activity by this fungus. Preliminary work has begun on analyzing organic extractions (alumina chromatography) of the liquid produced after microbial solubilization, including elemental analysis, solubility, molecular weights and chemical structure. This preliminarymore » work suggests that the candida could metabolize naturally occurring coal as substrate.« less

Enhancement of maize growth and alteration of the rhizosphere microbial community by phosphate-solubilizing fungus Aspergillus aculeatus P93

USDA-ARS?s Scientific Manuscript database

Over-application of phosphatic fertilizers has adversely impacted agricultural sustainability and the environment in China and other regions of the world resulting in the need for alternative strategies, such as the use of phosphate-solubilizing microbes. Aspergillus aculeatus P93, isolated from th...

A class-A GPCR solubilized under high hydrostatic pressure retains its ligand binding ability

USDA-ARS?s Scientific Manuscript database

The effect of high hydrostatic pressure (HHP) on the solubilization of a class-A G protein-coupled receptor, the silkmoth pheromone biosynthesis-activating neuropeptide receptor (PBANR), was investigated. PBANR was expressed in expresSF+ insect cells as a C-terminal fusion protein with EGFP. The mem...

Translocation and incorporation of strontium carbonate derived strontium into calcium oxalate crystals by the wood decay fungus Resinicium bicolor

Treesearch

Jon H. Connolly; Walter C. Shortle; Jody Jellison

1999-01-01

The white-rot wood decay fungus Resinicium bicolor (Abertini & Schwein.: Fr.) Parmasto was studied for its ability to solubilize and translocate ions from the naturally occurring mineral strontianite. Resinicium bicolor colonized a soil mixture culture medium containing strontianite sand, solubilized strontium ions from this...

Solubilization of coal in organic media

NASA Technical Reports Server (NTRS)

Lahaye, P.; Decroocq, D.

1977-01-01

The use of solvent extraction to solubilize coal is discussed. Simple extractions are described which are conducted at moderate temperatures to exclude extraneous chemical reactions which would lead to uncontrolled changes in the components of the treated coal. Sample preparation, extraction apparatus, and the determination of extract yield for different experiments are also described.

PH-sensitive dispersion of carbon nanotubes by myoglobin

NASA Astrophysics Data System (ADS)

Nie, Haiyu; Shen, Ganni; Sun, Junlin; Zhang, Tao

2017-03-01

A facile and effective method of dispersion of double-walled carbon nanotubes (DWNTs) was developed. At appropriate pH value and sonication, myoglobin helps the solubilization of DWNTs. The product is a pH-sensitive dispersion, which remains in a highly dispersed state at pH<3.0 and pH>10.0. This approach can be used to disperse DWNTs in scale. A reversible conversion of the highly dispersed state to the aggregated state could be observed by changing the pH value. This feature holds great promise for the development of pH sensors.

pH shift protein recovery with organic acids on texture and color of cooked gels.

PubMed

Paker, Ilgin; Beamer, Sarah; Jaczynski, Jacek; Matak, Kristen E

2015-01-01

Isoelectric solubilization and precipitation (ISP) processing uses pH shifts to separate protein from fish frames, which may increase commercial interest for silver carp. Texture and color properties of gels made from silver carp protein recovered at different pH strategies and organic acid types were compared. ISP was applied to headed gutted silver carp using 10 mol L(-1) sodium hydroxide (NaOH) and either glacial acetic acid (AA) or a (1:1) formic and lactic acid combination (F&L). Protein gels were made with recovered protein and standard functional additives. Texture profile analysis and the Kramer shear test showed that protein gels made from protein solubilized at basic pH values were firmer, harder, more cohesive, gummier and chewier (P < 0.05) than proteins solubilized under acidic conditions. Acidic solubilization led to whiter (P < 0.05) gels, and using F&L during ISP yielded whiter gels under all treatments (P < 0.05). Gels made from ISP-recovered silver carp protein using organic acids show potential for use as a functional ingredient in restructured foods. © 2014 Society of Chemical Industry.

Inosine-5'-monophosphate is a candidate agent to resolve rigor mortis of skeletal muscle.

PubMed

Matsuishi, Masanori; Tsuji, Mariko; Yamaguchi, Megumi; Kitamura, Natsumi; Tanaka, Sachi; Nakamura, Yukinobu; Okitani, Akihiro

2016-11-01

The object of the present study was to reveal the action of inosine-5'-monophosphate (IMP) toward myofibrils in postmortem muscles. IMP solubilized isolated actomyosin within a narrow range of KCl concentration, 0.19-0.20 mol/L, because of the dissociation of actomyosin into actin and myosin, but it did not solubilize the proteins in myofibrils with 0.2 mol/L KCl. However, IMP could solubilize both proteins in myofibrils with 0.2 mol/L KCl in the presence of 1 m mol/L pyrophosphate or 1.0-3.3 m mol/L adenosine-5'-diphosphate (ADP). Thus, we presumed that pyrophosphate and ADP released thin filaments composed of actin, and thick filaments composed of myosin from restraints of myofibrils, and then both filaments were solubilized through the IMP-induced dissociation of actomyosin. Thus, we concluded that IMP is a candidate agent to resolve rigor mortis because of its ability to break the association between thick and thin filaments. © 2016 Japanese Society of Animal Science.

Water solubilization capacity of pharmaceutical microemulsions based on Peceol®, lecithin and ethanol.

PubMed

Mouri, Abdelkader; Diat, Olivier; Lerner, Dan Alain; El Ghzaoui, Abdeslam; Ajovalasit, Alessia; Dorandeu, Christophe; Maurel, Jean-Claude; Devoisselle, Jean-Marie; Legrand, Philippe

2014-11-20

Biocompatible microemulsions composed of Peceol(®), lecithin, ethanol and water developed for encapsulation of hydrophilic drugs were investigated. The binary mixture Peceol(®)/ethanol was studied first. It was shown that the addition of ethanol to pure Peceol(®) has a significant fluidifying and disordering effect on the Peceol(®) supramolecular structure with an enhancement in water solubilization. The water solubilization capacity was improved by adding lecithin as a third component. It was then demonstrated that the ethanol/lecithin weight ratio played an important role in determining the optimal composition in term of water solubilization efficiency, a necessary property for a nutraceutical or pharmaceutical application. The optimal ethanol/lecithin weight ratio in the Peceol(®) rich region was found to be 40/60. Combination different techniques such as SAXS, fluorimetry, rheology and conductivity, we analyzed the water uptake within the microemulsion taking into account the partitioning of ethanol between polar and apolar domains. This ethanol distribution quantified along a water dilution line has a major effect on microemulsion properties. Copyright © 2014 Elsevier B.V. All rights reserved.

Engineering cocrystal solubility, stability, and pH(max) by micellar solubilization.

PubMed

Huang, Neal; Rodríguez-Hornedo, Naír

2011-12-01

Cocrystals offer great promise in enhancing drug aqueous solubilities, but face the challenge of conversion to a less soluble drug when in contact with solvent. This manuscript shows that differential solubilization of cocrystal components by micelles can impart thermodynamic stability to otherwise unstable cocrystals. The theoretical foundation for controlling cocrystal solubility and stability is presented by considering the contributions of micellar solubilization and ionization of cocrystal components. A surfactant critical stabilization concentration (CSC) and a solution pH (pH(max)) where cocrystal and drug are thermodynamically stable are shown to characterize cocrystal stability in micellar solutions. The solubility, CSC, and pH(max) of carbamazepine cocrystals in micellar solutions of sodium lauryl sulfate predicted by the models are in very good agreement with experimental measurements. The findings from this work demonstrate that cocrystal CSC and pH(max) can be tailored from the selection of coformer and solubilizing additives such as surfactants, thus providing an unprecedented level of control over cocrystal stability and solubility via solution phase chemistry. Copyright © 2011 Wiley-Liss, Inc.

Biosurfactant-producing strains in enhancing solubilization and biodegradation of petroleum hydrocarbons in groundwater.

PubMed

Liu, Hong; Wang, Hang; Chen, Xuehua; Liu, Na; Bao, Suriguge

2014-07-01

Three biosurfactant-producing strains designated as BS-1, BS-3, and BS-4 were screened out from crude oil-contaminated soil using a combination of surface tension measurement and oil spreading method. Thin layer chromatography and infrared analysis indicated that the biosurfactants produced by the three strains were lipopeptide, glycolipid, and phospholipid. The enhancement of solubilization and biodegradation of petroleum hydrocarbons in groundwater employing biosurfactant-producing strains was investigated. The three strain mixtures led to more solubilization of petroleum hydrocarbons in groundwater, and the solubilization rate was 10.5 mg l−1. The combination of biosurfactant-producing strains and petroleum-degrading strains exhibited a higher biodegradation efficiency of 85.4 % than the petroleum-degrading strains (71.2 %). Biodegradation was enhanced the greatest with biosurfactant-producing strains and petroleum-degrading strains in a ratio of 1:1. Fluorescence microscopy images illustrate that the oil dispersed into smaller droplets and emulsified in the presence of biosurfactant-producing strains, which attached to the oil. Thus, the biodegradation of petroleum hydrocarbons in groundwater was enhanced.

Solubilization of Tea Seed Oil in a Food-Grade Water-Dilutable Microemulsion

PubMed Central

Deng, Lingli; Que, Fei; Wei, Hewen; Xu, Guangwei; Dong, Xiaowei; Zhang, Hui

2015-01-01

Food-grade microemulsions containing oleic acid, ethanol, Tween 20, and water were formulated as a carrier system for tea seed oil (Camellia oleifera Abel.). The effect of ethanol on the phase behavior of the microemulsion system was clearly reflected in pseudo-ternary diagrams. The solubilization capacity and solubilization efficiency of tea seed oil dispersions were measured along the dilution line at a 70/30 surfactant/oil mass ratio with Tween 20 as the surfactant and oleic acid and ethanol (1:3, w/w) as the oil phase. The dispersed phase of the microemulsion (1.5% weight ratio of tea seed oil to the total amount of oil, surfactant, and tea seed oil) could be fully diluted with water without phase separation. Differential scanning calorimetry and viscosity measurements indicated that both the carrier and solubilized systems underwent a similar microstructure transition upon dilution. The dispersion phases gradually inverted from the water-in-oil phase (< 35% water) to the bicontinuous phase (40–45% water) and finally to the oil-in-water phase (> 45% water) along the dilution line. PMID:25996147

Solubilization of tea seed oil in a food-grade water-dilutable microemulsion.

PubMed

Deng, Lingli; Que, Fei; Wei, Hewen; Xu, Guangwei; Dong, Xiaowei; Zhang, Hui

2015-01-01

Food-grade microemulsions containing oleic acid, ethanol, Tween 20, and water were formulated as a carrier system for tea seed oil (Camellia oleifera Abel.). The effect of ethanol on the phase behavior of the microemulsion system was clearly reflected in pseudo-ternary diagrams. The solubilization capacity and solubilization efficiency of tea seed oil dispersions were measured along the dilution line at a 70/30 surfactant/oil mass ratio with Tween 20 as the surfactant and oleic acid and ethanol (1:3, w/w) as the oil phase. The dispersed phase of the microemulsion (1.5% weight ratio of tea seed oil to the total amount of oil, surfactant, and tea seed oil) could be fully diluted with water without phase separation. Differential scanning calorimetry and viscosity measurements indicated that both the carrier and solubilized systems underwent a similar microstructure transition upon dilution. The dispersion phases gradually inverted from the water-in-oil phase (< 35% water) to the bicontinuous phase (40-45% water) and finally to the oil-in-water phase (> 45% water) along the dilution line.

Dissolution and reconstitution of casein micelle containing dairy powders by high shear using ultrasonic and physical methods.

PubMed

Chandrapala, Jayani; Martin, Gregory J O; Kentish, Sandra E; Ashokkumar, Muthupandian

2014-09-01

The effect of shear on the solubilization of a range of dairy powders was investigated. The rate of solubilization of low solubility milk protein concentrate and micellar casein powders was examined during ultrasonication, high pressure homogenization and high-shear rotor-stator mixing and compared to low-shear overhead stirring. The high shear techniques were able to greatly accelerate the solubilization of these powders by physically breaking apart the powder agglomerates and accelerating the release of individual casein micelles into solution. This was achieved without affecting the structure of the solubilized proteins. The effect of high shear on the re-establishment of the mineral balance between the casein micelles and the serum was examined by monitoring the pH of the reconstituted skim milk powder after prior exposure to ultrasonication. Only minor differences in the re-equilibration of the pH were observed after sonication for up to 3 min, suggesting that the localized high shear forces exerted by sonication did not significantly affect the mass transfer of minerals from within the casein micelles. Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of solubilizing agents on mupirocin loading into and release from PEGylated nanoliposomes.

PubMed

Cern, Ahuva; Nativ-Roth, Einat; Goldblum, Amiram; Barenholz, Yechezkel

2014-07-01

Mupirocin was identified by quantitative structure property relationship models as a good candidate for remote liposomal loading. Mupirocin is an antibiotic that is currently restricted to topical administration because of rapid hydrolysis in vivo to its inactive metabolite. Formulating mupirocin in PEGylated nanoliposomes may potentially expand its use to parenteral administration by protecting it from degradation in the circulation and target it (by the enhanced permeability effect) to the infected tissue. Mupirocin is slightly soluble in aqueous medium and its solubility can be increased using solubilizing agents. The effect of the solubilizing agents on mupirocin remote loading was studied when the solubilizing agents were added to the drug loading solution. Propylene glycol was found to increase mupirocin loading, whereas polyethylene glycol 400 showed no effect. Hydroxypropyl-β-cyclodextrin (HPCD) showed a concentration-dependent effect on mupirocin loading; using the optimal HPCD concentration increased loading, but higher concentrations inhibited it. The inclusion of HPCD in the liposome aqueous phase while forming the liposomes resulted in increased drug loading and substantially inhibited drug release in serum. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Separation of Corn Fiber and Conversion to Fuels and Chemicals Phase II: Pilot-scale Operation

DOE Office of Scientific and Technical Information (OSTI.GOV)

Abbas, Charles; Beery, Kyle; Orth, Rick

2007-09-28

The purpose of the Department of Energy (DOE)-supported corn fiber conversion project, “Separation of Corn Fiber and Conversion to Fuels and Chemicals Phase II: Pilot-scale Operation” is to develop and demonstrate an integrated, economical process for the separation of corn fiber into its principal components to produce higher value-added fuel (ethanol and biodiesel), nutraceuticals (phytosterols), chemicals (polyols), and animal feed (corn fiber molasses). This project has successfully demonstrated the corn fiber conversion process on the pilot scale, and ensured that the process will integrate well into existing ADM corn wet-mills. This process involves hydrolyzing the corn fiber to solubilize 50%more » of the corn fiber as oligosaccharides and soluble protein. The solubilized fiber is removed and the remaining fiber residue is solvent extracted to remove the corn fiber oil, which contains valuable phytosterols. The extracted oil is refined to separate the phytosterols and the remaining oil is converted to biodiesel. The de-oiled fiber is enzymatically hydrolyzed and remixed with the soluble oligosaccharides in a fermentation vessel where it is fermented by a recombinant yeast, which is capable of fermenting the glucose and xylose to produce ethanol. The fermentation broth is distilled to remove the ethanol. The stillage is centrifuged to separate the yeast cell mass from the soluble components. The yeast cell mass is sold as a high-protein yeast cream and the remaining sugars in the stillage can be purified to produce a feedstock for catalytic conversion of the sugars to polyols (mainly ethylene glycol and propylene glycol) if desirable. The remaining materials from the purification step and any materials remaining after catalytic conversion are concentrated and sold as a corn fiber molasses. Additional high-value products are being investigated for the use of the corn fiber as a dietary fiber sources.« less

In Vitro Model Simulating Gastro-Intestinal Digestion in the Pediatric Population (Neonates and Young Infants).

PubMed

Kamstrup, Danna; Berthelsen, Ragna; Sassene, Philip Jonas; Selen, Arzu; Müllertz, Anette

2017-02-01

The focus on drug delivery for the pediatric population has been steadily increasing in the last decades. In terms of developing in vitro models simulating characteristics of the targeted pediatric population, with the purpose of predicting drug product performance after oral administration, it is important to simulate the gastro-intestinal conditions and processes the drug will encounter upon oral administration. When a drug is administered in the fed state, which is commonly the case for neonates, as they are typically fed every 3 h, the digestion of the milk will affect the composition of the fluid available for drug dissolution/solubilization. Therefore, in order to predict the solubilized amount of drug available for absorption, an in vitro model simulating digestion in the gastro-intestinal tract should be utilized. In order to simulate the digestion process and the drug solubilization taking place in vivo, the following aspects should be considered; physiologically relevant media, media volume, use of physiological enzymes in proper amounts, as well as correct pH and addition of relevant co-factors, e.g., bile salts and co-enzymes. Furthermore, physiological transit times and appropriate mixing should be considered and mimicked as close as possible. This paper presents a literature review on physiological factors relevant for digestion and drug solubilization in neonates. Based on the available literature data, a novel in vitro digestion model simulating digestion and drug solubilization in the neonate and young infant pediatric population (2 months old and younger) was designed.

Polymeric Curcumin Nanoparticle Pharmacokinetics and Metabolism in Bile Duct Cannulated Rats

PubMed Central

Zou, Peng; Helson, Lawrence; Maitra, Anirban; Stern, Stephan T.; McNeil, Scott E.

2013-01-01

The objective of this study was to compare the pharmacokinetics and metabolism of polymeric nanoparticle encapsulated (nanocurcumin), and solvent solubilized curcumin formulations in Sprague Dawley (SD) rats. Nanocurcumin is currently under development for cancer therapy. Since free, unencapsulated curcumin is rapidly metabolized and excreted in rats, upon i.v. administration of nanocurcumin only nanoparticle encapsulated curcumin can be detected in plasma samples. Hence, the second objective of this study was to utilize the metabolic instability of curcumin to assess in vivo drug release from nanocurcumin. Nanocurcumin and solvent solubilized curcumin were administered at 10 mg curcumin/kg by jugular vein to bile duct-cannulated male SD rats (n = 5). Nanocurcumin increased the plasma Cmax of curcumin 1749 fold relative to the solvent solubilized curcumin. Nanocurcumin also increased the relative abundance of curcumin and glucuronides in bile, but did not dramatically alter urine and tissue metabolite profiles. The observed increase in biliary and urinary excretion of both curcumin and metabolites for the nanocurcumin formulation suggested rapid, “burst” release of curcumin. Although the burst release observed in this study is a limitation for targeted tumor delivery, nanocurcumin still exhibits major advantages over solvent solubilized curcumin, as the nanoformulation does not result in the lung accumulation observed for the solvent solubilized curcumin and increases overall systemic curcumin exposure. Additionally, the remaining encapsulated curcumin fraction following burst release is available for tumor delivery via the enhanced permeation and retention effect commonly observed for nanoparticle formulations. PMID:23534919

Low-solubility particles and a Trojan-horse type mechanism of toxicity: the case of cobalt oxide on human lung cells

PubMed Central

2014-01-01

Background The mechanisms of toxicity of metal oxide particles towards lung cells are far from being understood. In particular, the relative contribution of intracellular particulate versus solubilized fractions is rarely considered as it is very challenging to assess, especially for low-solubility particles such as cobalt oxide (Co3O4). Methods This study was possible owing to two highly sensitive, independent, analytical techniques, based on single-cell analysis, using ion beam microanalysis, and on bulk analysis of cell lysates, using mass spectrometry. Results Our study shows that cobalt oxide particles, of very low solubility in the culture medium, are readily incorporated by BEAS-2B human lung cells through endocytosis via the clathrin-dependent pathway. They are partially solubilized at low pH within lysosomes, leading to cobalt ions release. Solubilized cobalt was detected within the cytoplasm and the nucleus. As expected from these low-solubility particles, the intracellular solubilized cobalt content is small compared with the intracellular particulate cobalt content, in the parts-per-thousand range or below. However, we were able to demonstrate that this minute fraction of intracellular solubilized cobalt is responsible for the overall toxicity. Conclusions Cobalt oxide particles are readily internalized by pulmonary cells via the endo-lysosomal pathway and can lead, through a Trojan-horse mechanism, to intracellular release of toxic metal ions over long periods of time, involving specific toxicity. PMID:24669904

Pesticide tolerant and phosphorus solubilizing Pseudomonas sp. strain SGRAJ09 isolated from pesticides treated Achillea clavennae rhizosphere soil.

PubMed

Rajasankar, R; Manju Gayathry, G; Sathiavelu, A; Ramalingam, C; Saravanan, V S

2013-05-01

In this study, an attempt was made to identify an effective phosphate solubilizing bacteria from pesticide polluted field soil. Based on the formation of solubilization halo on Pikovskaya's agar, six isolates were selected and screened for pesticide tolerance and phosphate (P) solubilization ability through liquid assay. The results showed that only one strain (SGRAJ09) obtained from Achillea clavennae was found to tolerate maximum level of the pesticides tested and it was phylogenetically identified as Pseudomonas sp. It possessed a wide range of pesticide tolerance, ranging from 117 μg mL(-1) for alphamethrin to 2,600 μg mL(-1) for endosulfan. The available P concentrations increased with the maximum and double the maximum dose of monocrotophos and imidacloprid, respectively. On subjected to FT-IR and HPLC analysis, the presence of organic acids functional group in the culture broth and the production of gluconic acid as dominant acid aiding the P solubilization were identified. On comparison with control broth, monocrotophos and imidacloprid added culture broth showed quantitatively high organic acids production. In addition to gluconic acid production, citric and acetic acids were also observed in the pesticide amended broth. Furthermore, the Pseudomonas sp. strain SGRAJ09 possessed all the plant growth promoting traits tested. In presence of monocrotophos and imidacloprid, its plant growth promoting activities were lower than that of the pesticides unamended treatment.

Effects of Lipid-Analog Detergent Solubilization on the Functionality and Lipidic Cubic Phase Mobility of the Torpedo californica Nicotinic Acetylcholine Receptor

PubMed Central

Padilla-Morales, Luis F.; Morales-Pérez, Claudio L.; De La Cruz-Rivera, Pamela C.; Asmar-Rovira, Guillermo; Báez-Pagán, Carlos A.

2011-01-01

Over the past three decades, the Torpedo californica nicotinic acetylcholine receptor (nAChR) has been one of the most extensively studied membrane protein systems. However, the effects of detergent solubilization on nAChR stability and function are poorly understood. The use of lipid-analog detergents for nAChR solubilization has been shown to preserve receptor stability and functionality. The present study used lipid-analog detergents from phospholipid-analog and cholesterol-analog detergent families for solubilization and affinity purification of the receptor and probed nAChR ion channel function using planar lipid bilayers (PLBs) and stability using analytical size exclusion chromatography (A-SEC) in the detergent-solubilized state. We also examined receptor mobility on the lipidic cubic phase (LCP) by measuring the nAChR mobile fraction and diffusion coefficient through fluorescence recovery after photobleaching (FRAP) experiments using lipid-analog and non-lipid-analog detergents. Our results show that it is possible to isolate stable and functional nAChRs using lipid-analog detergents, with characteristic ion channel currents in PLBs and minimal aggregation as observed in A-SEC. Furthermore, fractional mobility and diffusion coefficient values observed in FRAP experiments were similar to the values observed for these parameters in the recently LCP-crystallized β2-adrenergic receptor. The overall results show that phospholipid-analog detergents with 16 carbon acyl-chains support nAChR stability, functionality and LCP mobility. PMID:21922299

Acyclic cucurbit[n]uril-type molecular containers: influence of glycoluril oligomer length on their function as solubilizing agents.

PubMed

Gilberg, Laura; Zhang, Ben; Zavalij, Peter Y; Sindelar, Vladimir; Isaacs, Lyle

2015-04-07

We present the synthesis of a series of six new glycoluril derived molecular clips and acyclic CB[n]-type molecular containers (1–3) that all feature SO3(−) solubilizing groups but differ in the number of glycoluril rings between the two terminal dialkoxyaromatic sidewalls. We report the X-ray crystal structure of 3b which shows that its dialkoxynaphthalene sidewalls actively define a hydrophobic cavity with high potential to engage in π–π interactions with insoluble aromatic guests. Compounds 1–3 possess very good solubility characteristics (≥38 mM) and undergo only very weak self-association (Ks < 92 M(−1)) in water. The weak self-association is attributed to unfavorable SO3(−)···SO3(−) electrostatic interactions in the putative dimers 12–42. Accordingly, we created phase solubility diagrams to study their ability to act as solubilizing agents for four water insoluble drugs (PBS-1086, camptothecin, β-estradiol, and ziprasidone). We find that the containers 3a and 3b which feature three glycoluril rings between the terminal dialkoxy-o-xylylene and dialkoxynaphthalene sidewalls are less efficient solubilizing agents than 4a and 4b because of their smaller hydrophobic cavities. Containers 1 and 2 behave as molecular clip type receptors and therefore possess the ability to bind to and thereby solubilize aromatic drugs like camptothecin, ziprasidone, and PBS-1086.

Acyclic Cucurbit[n]uril-Type Molecular Containers: Influence of Glycoluril Oligomer Length on their Function as Solubilizing Agents

PubMed Central

Gilberg, Laura; Zhang, Ben; Zavalij, Peter Y.; Sindelar, Vladimir; Isaacs, Lyle

2015-01-01

We present the synthesis of a series of six new glycoluril derived molecular clips and acyclic CB[n]-type molecular containers (1 – 3) that all feature SO3− solubilizing groups but differ in the number of glycoluril rings between the two terminal dialkoxyaromatic sidewalls. We report the x-ray crystal structure of 3b which shows that its dialkoxynaphthalene sidewalls actively define a hydrophobic cavity with high potential to engage in π–π interactions with insoluble aromatic guests. Compounds 1 – 3 possess very good solubility characteristics (≥ 38 mM) and undergo only very weak self-association (Ks < 92 M−1) in water. The weak self-association is attributed to unfavorable SO3−•••SO3− electrostatic interactions in the putative dimers 12 – 42. Accordingly, we created phase solubility diagrams to study their ability to act as solubilizing agents for four water insoluble drugs (PBS-1086, camptothecin, β-estradiol, and ziprasidone). We find that the containers 3a and 3b which feature three glycoluril rings between the terminal dialkoxy-o-xylylene and dialkoxynaphthalene sidewalls are less efficient solubilizing agents than 4a and 4b because of their smaller hydrophobic cavities. Containers 1 and 2 behave as molecular clip type receptors and therefore possess the ability to bind to and thereby solubilize aromatic drugs like camptothecin and ziprasidone, and PBS-1086. PMID:25731639

Biotoxicity and bioavailability of hydrophobic organic compounds solubilized in nonionic surfactant micelle phase and cloud point system.

PubMed

Pan, Tao; Liu, Chunyan; Zeng, Xinying; Xin, Qiao; Xu, Meiying; Deng, Yangwu; Dong, Wei

2017-06-01

A recent work has shown that hydrophobic organic compounds solubilized in the micelle phase of some nonionic surfactants present substrate toxicity to microorganisms with increasing bioavailability. However, in cloud point systems, biotoxicity is prevented, because the compounds are solubilized into a coacervate phase, thereby leaving a fraction of compounds with cells in a dilute phase. This study extends the understanding of the relationship between substrate toxicity and bioavailability of hydrophobic organic compounds solubilized in nonionic surfactant micelle phase and cloud point system. Biotoxicity experiments were conducted with naphthalene and phenanthrene in the presence of mixed nonionic surfactants Brij30 and TMN-3, which formed a micelle phase or cloud point system at different concentrations. Saccharomyces cerevisiae, unable to degrade these compounds, was used for the biotoxicity experiments. Glucose in the cloud point system was consumed faster than in the nonionic surfactant micelle phase, indicating that the solubilized compounds had increased toxicity to cells in the nonionic surfactant micelle phase. The results were verified by subsequent biodegradation experiments. The compounds were degraded faster by PAH-degrading bacterium in the cloud point system than in the micelle phase. All these results showed that biotoxicity of the hydrophobic organic compounds increases with bioavailability in the surfactant micelle phase but remains at a low level in the cloud point system. These results provide a guideline for the application of cloud point systems as novel media for microbial transformation or biodegradation.

Phosphate solubilization and chromium (VI) remediation potential of Klebsiella sp. strain CPSB4 isolated from the chromium contaminated agricultural soil.

PubMed

Gupta, Pratishtha; Kumar, Vipin; Usmani, Zeba; Rani, Rupa; Chandra, Avantika

2018-02-01

In this study, an effort was made to identify an efficient phosphate solubilizing bacterial strain from chromium contaminated agricultural soils. Based on the formation of a solubilized halo around the colonies on Pikovskaya's agar amended with chromium (VI), 10 strains were initially screened out. Out of 10, strain CPSB4, which showed significantly high solubilization zone at different chromium concentrations, was selected for further study. The strain CPSB4 showed significant plant growth promotion traits with chromium (VI) stress under in-vitro conditions in broth. The plant growth promotion activities of the strain decreased regularly, but were not completely lost with the increase in concentration of chromium up to 200 mg L -1 . On subjected to FT-IR analysis, the presence of the functional group, indicating the organic acid aiding in phosphate solubilization was identified. At an optimal temperature of 30  ° C and pH 7.0, the strain showed around 93% chromium (VI) reduction under in-vitro conditions in broth study. In soil condition, the maximum chromium (VI) reduction obtained was 95% under in-vitro conditions. The strain CPSB4 was identified as Klebsiella sp. on the basis of morphological, biochemical and 16S rRNA gene sequencing. This study shows that the diverse role of the bacterial strain CPSB4 would be useful in the chromium contaminated soil as a good bioremediation and plant growth promoting agent as well. Copyright © 2017 Elsevier Ltd. All rights reserved.

Liquefaction of black thunder coal with counterflow reactor technology

DOE Office of Scientific and Technical Information (OSTI.GOV)

Parker, R.J.; Simpson, P.L.

There is currently a resurgence of interest in the use of carbon monoxide and water to promote the solubilization of low rank coals in liquefaction processes. The mechanism for the water shift gas reaction (WGSR) is well documented and proceeds via a formate ion intermediate at temperatures up to about 400{degrees}C. Coal solubilization is enhanced by CO/H{sub 2}O and by the solvent effect of the supercritical water. The WGSR is catalyzed by bases (alkali metal carbonates, hydroxides, acetates, aluminates). Many inorganic salts which promote catalytic hydrogenation are rendered inactive in CO/H{sub 2}O, although there is positive evidence for the benefitmore » of using pyrite for both the WGSR and as a hydrogenation catalyst. The temperatures at which coal solubilization occurs are insufficient to promote extensive cracking or upgrading of the solubilized coal. Therefore, a two step process might achieve these two reactions sequentially. Alberta Research Council (ARC) has developed a two-stage process for the coprocessing of low rank coals and petroleum resids/bitumens. This process was further advanced by utilizing the counterflow reactor (CFR) concept pioneered by Canadian Energy Developments (CED) and ARC. The technology is currently being applied to coal liquefaction. The two-stage process employs CO/H{sub 2}O at relatively mid temperature and pressure to solubilize the coal, followed by a more severe hydrocracking step. This paper describes the results of an autoclave study conducted to support a bench unit program on the direct liquefaction of coals.« less

Solubilization of phosphorus from phosphate rocks with Acidithiobacillus thiooxidans following a growing-then-recovery process.

PubMed

Calle-Castañeda, Susana M; Márquez-Godoy, Marco A; Hernández-Ortiz, Juan P

2017-12-29

Phosphorus is an essential nutrient for the synthesis of biomolecules and is particularly important in agriculture, as soils must be constantly supplemented with its inorganic form to ensure high yields and productivity. In this paper, we propose a process to solubilize phosphorus from phosphate rocks, where Acidithiobacillus thiooxidans cultures are pre-cultivated to foster the acidic conditions for bioleaching-two-step "growing-then-recovery"-. Our method solubilizes 100% of phosphorus, whereas the traditional process without pre-cultivation-single-step "growing-and-recovery"-results in a maximum of 56% solubilization. As a proof of principle, we demonstrate that even at low concentrations of the phosphate rock, 1% w/v, the bacterial culture is unviable and biological activity is not observed during the single-step process. On the other hand, in our method, the bacteria are grown without the rock, ensuring high acid production. Once pH levels are below 0.7, the mineral is added to the culture, resulting in high yields of biological solubilization. According to the Fourier Transform Infrared Spectroscopy spectrums, gypsum is the dominant phosphate phase after both the single- and two-step methods. However, calcite and fluorapatite, dominant in the un-treated rock, are still present after the single-step, highlighting the differences between the chemical and the biological methods. Our process opens new avenues for biotechnologies to recover phosphorus in tropical soils and in low-grade phosphate rock reservoirs.

β-Cyclodextrin-dextran polymers for the solubilization of poorly soluble drugs.

PubMed

di Cagno, Massimiliano; Terndrup Nielsen, Thorbjørn; Lambertsen Larsen, Kim; Kuntsche, Judith; Bauer-Brandl, Annette

2014-07-01

The aim of this study was to assess the potential of novel β-cyclodextrin (βCD)-dextran polymers for drug delivery. The size distribution of βCD-dextrans (for eventual parenteral administration), the influence of the dextran backbones on the stability of the βCD/drug complex, the solubilization efficiency of poorly soluble drugs and drug release properties were investigated. Size analysis of different βCD-dextrans was measured by size exclusion chromatography (SEC) and asymmetrical flow field-flow fractionation (AF4). Stability of drug/βCD-dextrans was assessed by isothermal titration calorimetry (ITC) and molar enthalpies of complexation and equilibrium constants compared to some commercially available βCD derivatives. For evaluation of the solubilization efficiency, phase-solubility diagrams were made employing hydrocortisone (HC) as a model of poorly soluble drugs, whereas reverse dialysis was used to detect potential drug supersaturation (increased molecularly dissolved drug concentration) as well as controlled release effects. Results indicate that all investigated βCD-polymers are of appropriate sizes for parenteral administration. Thermodynamic results demonstrate that the presence of the dextran backbone structure does not affect the stability of the βCD/drug complex, compared to native βCD and commercially available derivatives. Solubility studies evidence higher solubilizing abilities of these new polymers in comparison to commercially available βCDs, but no supersaturation states were induced. Moreover, drug release studies evidenced that diffusion of HC was influenced by the solubilization induced by the βCD-derivatives. Copyright © 2014 Elsevier B.V. All rights reserved.

Presence of a Large β(1-3)Glucan Linked to Chitin at the Saccharomyces cerevisiae Mother-Bud Neck Suggests Involvement in Localized Growth Control

PubMed Central

Blanco, Noelia; Arroyo, Javier

2012-01-01

Previous results suggested that the chitin ring present at the yeast mother-bud neck, which is linked specifically to the nonreducing ends of β(1-3)glucan, may help to suppress cell wall growth at the neck by competing with β(1-6)glucan and thereby with mannoproteins for their attachment to the same sites. Here we explored whether the linkage of chitin to β(1-3)glucan may also prevent the remodeling of this polysaccharide that would be necessary for cell wall growth. By a novel mild procedure, β(1-3)glucan was isolated from cell walls, solubilized by carboxymethylation, and fractionated by size exclusion chromatography, giving rise to a very high-molecular-weight peak and to highly polydisperse material. The latter material, soluble in alkali, may correspond to glucan being remodeled, whereas the large-size fraction would be the final cross-linked structural product. In fact, the β(1-3)glucan of buds, where growth occurs, is solubilized by alkali. A gas1 mutant with an expected defect in glucan elongation showed a large increase in the polydisperse fraction. By a procedure involving sodium hydroxide treatment, carboxymethylation, fractionation by affinity chromatography on wheat germ agglutinin-agarose, and fractionation by size chromatography on Sephacryl columns, it was shown that the β(1-3)glucan attached to chitin consists mostly of high-molecular-weight material. Therefore, it appears that linkage to chitin results in a polysaccharide that cannot be further remodeled and does not contribute to growth at the neck. In the course of these experiments, the new finding was made that part of the chitin forms a noncovalent complex with β(1-3)glucan. PMID:22366124

Presence of a large β(1-3)glucan linked to chitin at the Saccharomyces cerevisiae mother-bud neck suggests involvement in localized growth control.

PubMed

Cabib, Enrico; Blanco, Noelia; Arroyo, Javier

2012-04-01

Previous results suggested that the chitin ring present at the yeast mother-bud neck, which is linked specifically to the nonreducing ends of β(1-3)glucan, may help to suppress cell wall growth at the neck by competing with β(1-6)glucan and thereby with mannoproteins for their attachment to the same sites. Here we explored whether the linkage of chitin to β(1-3)glucan may also prevent the remodeling of this polysaccharide that would be necessary for cell wall growth. By a novel mild procedure, β(1-3)glucan was isolated from cell walls, solubilized by carboxymethylation, and fractionated by size exclusion chromatography, giving rise to a very high-molecular-weight peak and to highly polydisperse material. The latter material, soluble in alkali, may correspond to glucan being remodeled, whereas the large-size fraction would be the final cross-linked structural product. In fact, the β(1-3)glucan of buds, where growth occurs, is solubilized by alkali. A gas1 mutant with an expected defect in glucan elongation showed a large increase in the polydisperse fraction. By a procedure involving sodium hydroxide treatment, carboxymethylation, fractionation by affinity chromatography on wheat germ agglutinin-agarose, and fractionation by size chromatography on Sephacryl columns, it was shown that the β(1-3)glucan attached to chitin consists mostly of high-molecular-weight material. Therefore, it appears that linkage to chitin results in a polysaccharide that cannot be further remodeled and does not contribute to growth at the neck. In the course of these experiments, the new finding was made that part of the chitin forms a noncovalent complex with β(1-3)glucan.

Cell-wall polysaccharides and glycoproteins of parenchymatous tissues of runner bean (Phaseolus coccineus).

PubMed Central

Ryden, P; Selvendran, R R

1990-01-01

1. Polymers were solubilized from the cell walls of parenchyma from mature runner-bean pods with minimum degradation by successive extractions with cyclohexane-trans-1,2-diamine-NNN'N'-tetra-acetate (CDTA), Na2CO3 and KOH to leave the alpha-cellulose residue, which contained cross-linked pectic polysaccharides and Hyp-rich glycoproteins. These were solubilized with chlorite/acetic acid and cellulase. The polymers were fractionated by anion-exchange chromatography, and fractions were subjected to methylation analysis. 2. The pectic polysaccharides differed in their ease of extraction, and a small proportion were highly cross-linked. The bulk of the pectic polysaccharides solubilized by CDTA and Na2CO3 were less branched than those solubilized by KOH. There was good evidence that most of the pectic polysaccharides were not degraded during extraction. 3. The protein-containing fractions included Hyp-rich and Hyp-poor glycoproteins associated with easily extractable pectic polysaccharides, Hyp-rich glycoproteins solubilized with 4M-KOH+borate, the bulk of which were not associated with pectic polysaccharides, and highly cross-linked Hyp-rich glycoproteins. 4. Isodityrosine was not detected, suggesting that it does not have a (major) cross-linking role in these walls. Instead, it is suggested that phenolics, presumably linked to C-5 of 3,5-linked Araf residues of Hyp-rich glycoproteins, serve to cross-link some of the polymers. 5. There were two main types of xyloglucan, with different degrees of branching. The bulk of the less branched xyloglucans were solubilized by more-concentrated alkali. The anomeric configurations of the sugars in one of the highly branched xyloglucans were determined by 13C-n.m.r. spectroscopy. 6. The structural features of the cell-wall polymers and complexes are discussed in relation to the structure of the cell walls of parenchyma tissues. PMID:2167068

Solubilization and Stabilization of Isolated Photosystem I Complex with Lipopeptide Detergents

PubMed Central

Wang, Xiaoqiang; Huang, Guihong; Yu, Daoyong; Ge, Baosheng; Wang, Jiqian; Xu, Fengxi; Huang, Fang; Xu, Hai; Lu, Jian R.

2013-01-01

It is difficult to maintain a target membrane protein in a soluble and functional form in aqueous solution without biological membranes. Use of surfactants can improve solubility, but it remains challenging to identify adequate surfactants that can improve solubility without damaging their native structures and biological functions. Here we report the use of a new class of lipopeptides to solubilize photosystem I (PS-I), a well known membrane protein complex. Changes in the molecular structure of these surfactants affected their amphiphilicity and the goal of this work was to exploit a delicate balance between detergency and biomimetic performance in PS-I solubilization via their binding capacity. Meanwhile, the effects of these surfactants on the thermal and structural stability and functionality of PS-I in aqueous solution were investigated by circular dichroism, fluorescence spectroscopy, SDS-PAGE analysis and O2 uptake measurements, respectively. Our studies showed that the solubility of PS-I depended on both the polarity and charge in the hydrophilic head of the lipopeptides and the length of its hydrophobic tail. The best performing lipopeptides in favour of PS-I solubility turned out to be C14DK and C16DK, which were comparable to the optimal amphiphilicity of the conventional chemical surfactants tested. Lipopeptides showed obvious advantages in enhancing PS-I thermostability over sugar surfactant DDM and some full peptide amphiphiles reported previously. Fluorescence spectroscopy along with SDS-PAGE analysis demonstrated that lipopeptides did not undermine the polypeptide composition and conformation of PS-I after solubilization; instead they showed better performance in improving the structural stability and integrity of this multi-subunit membrane protein than conventional detergents. Furthermore, O2 uptake measurements indicated that PS-I solubilized with lipopeptides maintained its functionality. The underlying mechanism for the favorable actions of lipopeptide in PS-I solubilization and stabilization is discussed. PMID:24098786

THE MECHANISM OF ACTION OF COLCHICINE

PubMed Central

Wilson, Leslie; Meza, Isaura

1973-01-01

The thermal depolymerization procedure of Stephens (1970. J. Mol. Biol. 47:353) has been employed for solubilization of Strongylocentrotus purpuratus sperm tail outer doublet microtubules with the use of a buffer during solubilization which is of optimal pH and ionic strength for the preservation of colchicine binding activity of chick embryo brain tubulin. Colchicine binding values were corrected for first-order decay during heat solubilization at 50°C (t½ = 5.4 min) and incubation with colchicine at 37°C in the presence of vinblastine sulfate (t½ = 485 min). The colchicine binding properties of heat-solubilized outer doublet tubulin were qualitatively identical with those of other soluble forms of tubulin. The solubilized tubulin (mol wt, 115,000) bound 0.9 ± 0.2 mol of colchicine per mol of tubulin, with a binding constant of 6.3 x 105 liters/mol at 37°C. The colchicine binding reaction was both time and temperature dependent, and the binding of colchicine was prevented in a competitive manner by podophyllotoxin (Ki = 1.3 x 10-6 M). The first-order decay of colchicine binding activity was substantially decreased by the addition of the vinca alkaloids, vinblastine sulfate or vincristine sulfate, thus demonstrating the presence of a vinca alkaloid binding site(s) on the outer doublet tubulin. Tubulin contained within the assembled microtubules did not decay. Intact outer doublet microtubules bound less than 0.001 mol of colchicine per mol of tubulin contained in the microtubules, under conditions where soluble tubulin would have bound 1 mol of colchicine per mol of tubulin (saturating concentration of colchicine, no decay of colchicine binding activity). The presence of colchicine had no effect on the rate of solubilization of outer doublet microtubules during incubation at 37°C. Therefore, the colchicine binding site on tubulin is blocked (not available to bind colchicine) when the tubulin is in the assembled outer doublet microtubules. PMID:4747924

Ionic liquid-tolerant microorganisms and microbial communities for lignocellulose conversion to bioproducts

DOE PAGES

Yu, Chaowei; Simmons, Blake A.; Singer, Steven W.; ...

2016-11-12

Chemical and physical pretreatment of biomass is a critical step in the conversion of lignocellulose to biofuels and bioproducts. Ionic liquid (IL) pretreatment has attracted significant attention due to the unique ability of certain ILs to solubilize some or all components of the plant cell wall. However, these ILs inhibit not only the enzyme activities but also the growth and productivity of microorganisms used in downstream hydrolysis and fermentation processes. While pretreated biomass can be washed to remove residual IL and reduce inhibition, extensive washing is costly and not feasible in large-scale processes. IL-tolerant microorganisms and microbial communities have beenmore » discovered from environmental samples and studies begun to elucidate mechanisms of IL tolerance. The discovery of IL tolerance in environmental microbial communities and individual microbes has lead to the proposal of molecular mechanisms of resistance. Here, we review recent progress on discovering IL-tolerant microorganisms, identifying metabolic pathways and mechanisms of tolerance, and engineering microorganisms for IL tolerance. Research in these areas will yield new approaches to overcome inhibition in lignocellulosic biomass bioconversion processes and increase opportunities for the use of ILs in biomass pretreatment.« less

Ionic liquid-tolerant microorganisms and microbial communities for lignocellulose conversion to bioproducts.

PubMed

Yu, Chaowei; Simmons, Blake A; Singer, Steven W; Thelen, Michael P; VanderGheynst, Jean S

2016-12-01

Chemical and physical pretreatment of biomass is a critical step in the conversion of lignocellulose to biofuels and bioproducts. Ionic liquid (IL) pretreatment has attracted significant attention due to the unique ability of certain ILs to solubilize some or all components of the plant cell wall. However, these ILs inhibit not only the enzyme activities but also the growth and productivity of microorganisms used in downstream hydrolysis and fermentation processes. While pretreated biomass can be washed to remove residual IL and reduce inhibition, extensive washing is costly and not feasible in large-scale processes. IL-tolerant microorganisms and microbial communities have been discovered from environmental samples and studies begun to elucidate mechanisms of IL tolerance. The discovery of IL tolerance in environmental microbial communities and individual microbes has lead to the proposal of molecular mechanisms of resistance. In this article, we review recent progress on discovering IL-tolerant microorganisms, identifying metabolic pathways and mechanisms of tolerance, and engineering microorganisms for IL tolerance. Research in these areas will yield new approaches to overcome inhibition in lignocellulosic biomass bioconversion processes and increase opportunities for the use of ILs in biomass pretreatment.

Ionic liquid-tolerant microorganisms and microbial communities for lignocellulose conversion to bioproducts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yu, Chaowei; Simmons, Blake A.; Singer, Steven W.

Chemical and physical pretreatment of biomass is a critical step in the conversion of lignocellulose to biofuels and bioproducts. Ionic liquid (IL) pretreatment has attracted significant attention due to the unique ability of certain ILs to solubilize some or all components of the plant cell wall. However, these ILs inhibit not only the enzyme activities but also the growth and productivity of microorganisms used in downstream hydrolysis and fermentation processes. While pretreated biomass can be washed to remove residual IL and reduce inhibition, extensive washing is costly and not feasible in large-scale processes. IL-tolerant microorganisms and microbial communities have beenmore » discovered from environmental samples and studies begun to elucidate mechanisms of IL tolerance. The discovery of IL tolerance in environmental microbial communities and individual microbes has lead to the proposal of molecular mechanisms of resistance. Here, we review recent progress on discovering IL-tolerant microorganisms, identifying metabolic pathways and mechanisms of tolerance, and engineering microorganisms for IL tolerance. Research in these areas will yield new approaches to overcome inhibition in lignocellulosic biomass bioconversion processes and increase opportunities for the use of ILs in biomass pretreatment.« less

Application of preparative disk gel electrophoresis for antigen purification from inclusion bodies.

PubMed

Okegawa, Yuki; Koshino, Masanori; Okushima, Teruya; Motohashi, Ken

2016-02-01

Specific antibodies are a reliable tool to examine protein expression patterns and to determine the protein localizations within cells. Generally, recombinant proteins are used as antigens for specific antibody production. However, recombinant proteins from mammals and plants are often overexpressed as insoluble inclusion bodies in Escherichia coli. Solubilization of these inclusion bodies is desirable because soluble antigens are more suitable for injection into animals to be immunized. Furthermore, highly purified proteins are also required for specific antibody production. Plastidic acetyl-CoA carboxylase (ACCase: EC 6.4.1.2) from Arabidopsis thaliana, which catalyzes the formation of malonyl-CoA from acetyl-CoA in chloroplasts, formed inclusion bodies when the recombinant protein was overexpressed in E. coli. To obtain the purified protein to use as an antigen, we applied preparative disk gel electrophoresis for protein purification from inclusion bodies. This method is suitable for antigen preparation from inclusion bodies because the purified protein is recovered as a soluble fraction in electrode running buffer containing 0.1% sodium dodecyl sulfate that can be directly injected into immune animals, and it can be used for large-scale antigen preparation (several tens of milligrams). Copyright © 2015 Elsevier Inc. All rights reserved.

Pressure injection of solubilized benomyl for prevention and cure of oak wilt

Treesearch

Thomas W. Jones; Garold F. Gregory; Percy McWain

1973-01-01

A preliminary evaluation of the effectiveness of injecting solubilized benomyl into oaks for prevention or cure of oak wilt disease is presented. Symptom development was greatly reduced or prevented in trees injected with fungicide before inoculation. Symptom development was markedly arrested in diseased trees by fungicide injected before more than 10 percent of the...

Development of a water-soluble preparation of emamectin benzoate and its preventative effect against the wilting of pot-grown pine trees inoculated with the pine wood nematode, Bursaphelenchus xylophilus.

PubMed

Takai, K; Soejima, T; Suzuki, T; Kawazu, K

2001-05-01

Water-soluble preparations have been investigated to develop a trunk injection agent based on the poorly water-soluble anti-nematode emamectin benzoate. Following tests on the phytotoxicity of some solvents and solubilizers and demonstration of the ability of some solubilizers to dissolve emamectin benzoate in water, acetone + methanol was selected as the solvent and Polysorbate 80 as the solubilizer. This water-soluble preparation of emamectin benzoate prevented the wilting of pot-grown 4-year-old trees of the Japanese black pine, Pinus thunbergii, artificially inoculated with the pine wood nematode, Bursaphelenchus xylophilus, at a dose of 20 g emamectin benzoate per cubic metre of pine tree.

Method of recovering hazardous waste from phenolic resin filters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Meikrantz, D.H.; Bourne, G.L.; McFee, J.N.

1990-12-31

A method has been found for treating phenolic resin filter, whereby the filter is solubilized within the filter cartridge housing so the filter material can be removed from the cartridge housing in a remote manner. The invention consists of contacting the filter within the housing with an aqueous solution of about 8 to 12M nitric acid, at a temperature from about 110 to 190{degree}F, maintaining the contact for a period of time sufficient to solubilize the phenolic material within the housing, and removing the solubilized phenolic material from the housing, thereby removing the filter cartridge from the housing. Any hazardousmore » or other waste material can then be separated from the filter material by chemical or other means.« less

Geomycology. [fungal biosolubilization and accumulation of metals

NASA Technical Reports Server (NTRS)

Puerner, N. J.; Siegel, S. M.

1976-01-01

Fungi have long been known to have capabilities for reduction and alkylation of arsenate and selenate but their general capabilities for solubilizing and accumulating metallic substances have been given serious attention only in recent years. Common members of the Aspergillaceae cultured on boron, copper, lead and other metals or oxides can solubilize and concentrate the elements or their compounds. To account for biosolubilization of the metals, we have set up a model study, incubating selected metals, e.g., mercury, in solutions of various metabolites including L-lysine and citric acid. Results of 100-300 days incubation showed that many metals can in fact be readily solubilized, and in some cases more effectively at pH 6-7 than at pH 1.5-2.

Nutritional characteristics of moon dust for soil microorganisms

NASA Technical Reports Server (NTRS)

Ito, T.

1983-01-01

Approximately 46% of the lunar sample (10084,151), 125.42 mg, was solubilized in 680 ml 0.01 M salicylic acid. Atomic absorption spectroscopic analysis of the solubilized lunar sample showed the following amount of metal ions: Ca, 3.1; Mg, 4.0; K, 0.09; Na, 0.67; Fe, 7.3; Mn, 1.6; Cu, Ni, Cr, less than 0.1 each. All are in ppm. Salicylic acid used to solubilize the lunar sample was highly inhibitory to the growth of mixed soil microbes. However, the mineral part of the lunar extract stimulated the growth. For optimal growth of the soil microbes the following nutrients must be added to the moon extract; sources of carbon, nitrogen, sulfur, phosphorus, and magnesium in addition to water.

Response surface methodology for the optimization of sludge solubilization by ultrasonic pre-treatment

NASA Astrophysics Data System (ADS)

Zheng, Mingyue; Zhang, Xiaohui; Lu, Peng; Cao, Qiguang; Yuan, Yuan; Yue, Mingxing; Fu, Yiwei; Wu, Libin

2018-02-01

The present study examines the optimization of the ultrasonic pre-treatment conditions with response surface experimental design in terms of sludge disintegration efficiency (solubilisation of organic components). Ultrasonic pre-treatment for the maximum solubilization with residual sludge enhanced the SCOD release. Optimization of the ultrasonic pre-treatment was conducted through a Box-Behnken design (three variables, a total of 17 experiments) to determine the effects of three independent variables (power, residence time and TS) on COD solubilization of sludge. The optimal COD was obtained at 17349.4mg/L, when the power was 534.67W, the time was 10.77, and TS was 2%, while the SE of this condition was 28792J/kg TS.

Solubilization of Australian lignites by microorganisms

DOE Office of Scientific and Technical Information (OSTI.GOV)

Catcheside, D.E.A.; Mallett, K.J.; Cox, R.E.

1988-01-01

Australia has substantial lignite deposits, particularly in the Latrobe Valley in Victoria where 4.10/sup 10/ tons are accessible with available technologies. The authors have investigated the susceptibility of these coal to solubilization by microorganisms, including species additional to those already identified as active on North American lignites. The data presented here show that acid oxidized lignites from the Latrobe Valley are solubilized by each of seven species of microorganisms previously found to be active on Leonardite and oxidized North American lignites. These are the wood rot fungi: Trametes versicolor, Poria placenta and Phanerochaete chrysosporium, the lignin degrading prokaryote Streptomyces viridosporusmore » and three fungi isolated from lignite in Mississippi: Candida ML-13, Cunninghamelia YML-1 and Penicillium waksmanii.« less

Electrochemical detection of nitromethane vapors combined with a solubilization device.

PubMed

Delile, Sébastien; Aussage, Adeline; Maillou, Thierry; Palmas, Pascal; Lair, Virginie; Cassir, Michel

2015-01-01

During the past decade, the number of terrorism acts has increased and the need for efficient explosive detectors has become an urgent worldwide necessity. A prototype, Nebulex™, was recently developed in our laboratory. Basically, it couples the solubilization of an analyte from the atmosphere by a nebulization process and in-situ detection. This article presents the development and integration of an electrochemical sensor for the detection of nitromethane, a common chemical product that can be used to make an improvised explosive device. A gold screen-printed electrode was used in a flow-cell and a detection limit of 4.5 µM was achieved by square wave voltammetry. The detection method was also determined to be selective toward nitromethane over a large panel of interfering compounds. Detection tests with the Nebulex™ were thus carried out using a custom-made calibrated nitromethane vapor generator. Detection times of less than one minute were obtained for nitromethane contents of 8 and 90 ppmv. Further measurements were performed in a room-measurement configuration leading to detection times in the range of 1-2 min, clearly demonstrating the system's efficiency under quasi-real conditions. Copyright © 2014 Elsevier B.V. All rights reserved.

Structure and dynamics of a detergent-solubilized membrane protein: measurement of amide hydrogen exchange rates in M13 coat protein by /sub 1/H NMR spectroscopy

DOE Office of Scientific and Technical Information (OSTI.GOV)

O'Neil, J.D.J.; Sykes, B.D.

The coat protein of bacteriophage M13 is inserted into the inner membrane of Escherichia coli where it exists as an integral membrane protein during the reproductive cycle of the phage. The protein sequence consists of a highly hydrophobic 19-residue central segment flanked by an acidic 20-residue N-terminus and a basic 11-residue C-terminus. The authors have measured backbone amide hydrogen exchange of the protein solubilized in perdeuteriated sodium dodecyl sulfate using /sup 1/H nuclear magnetic resonance (NMR) spectroscopy. Direct proton exchange-out measurements in D/sub 2/O at 24 /sup 0/C were used to follow the exchange of the slowest amides in themore » protein. Multiple exponential fitting of the exchange data showed that these amides exchanged in two kinetic sets with exchange rates that differed by more than 100-fold. Steady-state saturation-transfer techniques were also used to measure exchange. These methods showed that 15-20 amides in the protein are very stable at 55/sup 0/C and that bout 30 amides have exchange rates retarded by at least 10/sup 5/-fold at 24/sup 0/C. Saturation-transfer studies also showed that the pH dependence of exchange in the hydrophilic termini was unusual. Relaxation and solid-state NMR experiments have previously shown that the majority of the protein backbone is rigid on the picosecond to microsecond time scale, except for the extreme ends of the molecule which are mobile. The hydrogen exchange results, which are sensitive to a much longer time scale, suggest a stable core with a progressive increase in amplitude or frequency of motions as the ends of the protein are approached.« less

Peptide stabilized amphotericin B nanodisks

PubMed Central

Tufteland, Megan; Pesavento, Joseph B.; Bermingham, Rachelle L.; Hoeprich, Paul D.; Ryan, Robert O.

2007-01-01

Nanometer scale apolipoprotein A-I stabilized phospholipid disk complexes (nanodisks; ND) have been formulated with the polyene antibiotic amphotericin B (AMB). The present studies were designed to evaluate if a peptide can substitute for the function of the apolipoprotein component of ND with respect to particle formation and stability. An 18-residue synthetic amphipathic α-helical peptide, termed 4F (Ac-D-W-F-K-A-F-Y-D-K-V-A-E-K-F-K-E-A-F-NH2), solubilized vesicles comprised of egg phosphatidylcholine (egg PC), dipentadecanoyl PC or dimyristoylphosphatidylcholine (DMPC) at rates greater than or equal to solubilization rates observed with human apolipoprotein A-I (apoA-I; 243 amino acids). Characterization studies revealed that interaction with DMPC induced a near doubling of 4F tryptophan fluorescence emission quantum yield (excitation 280 nm) and a ~7 nm blue shift in emission wavelength maximum. Inclusion of AMB in the vesicle substrate resulted in formation of 4F AMB-ND. Spectra of AMB containing particles revealed the antibiotic is a highly effective quencher of 4F tryptophan fluorescence emission, giving rise to a Ksv = 7.7 × 104. Negative stain electron microscopy revealed that AMB-ND prepared with 4F possessed a disk shaped morphology similar to ND prepared without AMB or prepared with apoA-I. In yeast and pathogenic fungi growth inhibition assays, 4F AMB-ND was as effective as apoA-I AMB-ND. The data indicate that AMB-ND generated using an amphipathic peptide in lieu of apoA-I form a discrete population of particles that possess potent biological activity. Given their intrinsic versatility, peptides may be preferred for scale up and clinical application of AMB-ND. PMID:17293004

Sequential hydrophile and lipophile solubilization as an efficient method for decellularization of porcine aortic valve leaflets: Structure, mechanical property and biocompatibility study.

PubMed

Qiao, Wei-Hua; Liu, Peng; Hu, Dan; Al Shirbini, Mahmoud; Zhou, Xian-Ming; Dong, Nian-Guo

2018-02-01

Antigenicity of xenogeneic tissues is the major obstacle to increased use of these materials in clinical medicine. Residual xenoantigens in decellularized tissue elicit the immune response after implantation, causing graft failure. With this in mind, the potential use is proposed of three protein solubilization-based protocols for porcine aortic valve leaflets decellularization. It was demonstrated that hydrophile solubilization alone achieved incomplete decellularization; lipophile solubilization alone (LSA) completely removed all cells and two most critical xenoantigens - galactose-α(1,3)-galactose (α-Gal) and major histocompatibility complex I (MHC I) - but caused severe alterations of the structure and mechanical properties; sequential hydrophile and lipophile solubilization (SHLS) resulted in a complete removal of cells, α-Gal and MHC I, and good preservation of the structure and mechanical properties. In contrast, a previously reported method using Triton X-100, sodium deoxycholate and IGEPAL CA-630 resulted in a complete removal of all cells and MHC I, but with remaining α-Gal epitope. LSA- and SHLS-treated leaflets showed significantly reduced leucocyte activation (polymorphonuclear elastase) upon interaction with human blood in vitro. When implanted subdermally in rats for 6 weeks, LSA- or SHLS-treated leaflets were presented with more biocompatible implants and all four decellularized leaflets were highly resistant to calcification. These findings illustrate that the SHLS protocol could be considered as a promising decellularization method for the decellularization of xenogeneic tissues in tissue engineering and regenerative medicine. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

Initial pH of medium affects organic acids production but do not affect phosphate solubilization.

PubMed

Marra, Leandro M; de Oliveira-Longatti, Silvia M; Soares, Cláudio R F S; de Lima, José M; Olivares, Fabio L; Moreira, Fatima M S

2015-06-01

The pH of the culture medium directly influences the growth of microorganisms and the chemical processes that they perform. The aim of this study was to assess the influence of the initial pH of the culture medium on the production of 11 low-molecular-weight organic acids and on the solubilization of calcium phosphate by bacteria in growth medium (NBRIP). The following strains isolated from cowpea nodules were studied: UFLA03-08 (Rhizobium tropici), UFLA03-09 (Acinetobacter sp.), UFLA03-10 (Paenibacillus kribbensis), UFLA03-106 (Paenibacillus kribbensis) and UFLA03-116 (Paenibacillus sp.). The strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 solubilized Ca3(PO4)2 in liquid medium regardless of the initial pH, although without a significant difference between the treatments. The production of organic acids by these strains was assessed for all of the initial pH values investigated, and differences between the treatments were observed. Strains UFLA03-09 and UFLA03-10 produced the same acids at different initial pH values in the culture medium. There was no correlation between phosphorus solubilized from Ca3(PO4)2 in NBRIP liquid medium and the concentration of total organic acids at the different initial pH values. Therefore, the initial pH of the culture medium influences the production of organic acids by the strains UFLA03-08, UFLA03-09, UFLA03-10 and UFLA03-106 but it does not affect calcium phosphate solubilization.

Preparation of trout liver microsomes for iron speciation in P-450 enzymes by AE-FPLC with ICP-(ORS)MS detection.

PubMed

Rodríguez-Cea, Andrés; de la Campa, María Rosario Fernández; Sanz-Medel, Alfredo

2005-01-01

Cytochromes P-450 are members of a superfamily of hemoproteins involved in the oxidative metabolism of various physiological and xenobiotic compounds in eukaryotes and prokaryotes. The multiplicity of this group of enzymes has been widely studied by chromatographic techniques, mainly high-performance liquid chromatography (HPLC). Because these enzymes are membrane-bound proteins, sample preparation for chromatographic separation of P-450 enzymes requires a solubilization step. The sample-preparation procedures are critical, because detergents affect not only the efficiency of protein solubilization but also their further chromatographic resolution. Trout liver microsomes have been taken here as a model sample to investigate iron speciation in cytochrome P-450. Trouts were treated intraperitoneally with beta-naphthoflavone, a potent inducer of some P-450 enzymes, and a microsomal suspension containing 7.4+/-0.1 nmol mL(-1) P-450 enzymes was obtained by ultracentrifugation. Lubrol PX was selected as detergent for solubilization, resulting in about 90% solubilization recovery. The solubilized cytochromes P-450 were further separated by AE-FPLC, with UV detection, or coupled to ICP-MS with an octapole reaction system, ICP-(ORS)MS (monitoring Fe signals at masses 54, 56, and 57). A sampling procedure and chromatographic conditions are developed and were successfully applied to iron speciation in trout liver P-450 enzymes. ICP-(ORS)MS detection of P-450 enzymes is Fe-specific and so will give accurate information on the prosthetic group of the protein, which can constitute an advantageous alternative to classical methods for detection of these hemoproteins.

Purification and Characterization of Two Voltage-Dependent Anion Channel Isoforms from Plant Seeds1

PubMed Central

Abrecht, Helge; Wattiez, Ruddy; Ruysschaert, Jean-Marie; Homblé, Fabrice

2000-01-01

Mitochondria were isolated from imbibed seeds of lentil (Lens culinaris) and Phaseolus vulgaris. We copurified two voltage-dependent anion channel from detergent solubilized mitochondria in a single purification step using hydroxyapatite. The two isoforms from P. vulgaris were separated by chromatofocusing chromatography in 4 m urea without any loss of channel activity. Channel activity of each isoform was characterized upon reconstitution into diphytanoyl phosphatidylcholine planar lipid bilayers. Both isoforms form large conductance channels that are slightly anion selective and display cation selective substates. PMID:11080295

FIELD IMPLEMENTATION OF A WINSOR TYPE I SURFACTANT/ALCOHOL MIXTURE FOR IN SITU SOLUBILIZATION OF A COMPLEX LNAPL AS A SINGLE-PHASE MICROEMULSION

EPA Science Inventory

A Winsor Type I surfactant/alcohol mixture was used as an in situ flushing agent to solubilize a muticomponent nonaqueous phase liquid (NAPL) as a single-phase microemulsion (SPME) in a hydraulically isolated test cell at Hill Air Force Base (AFB), Utah. The surfactant (polyoxye...

Chemical comminution and deashing of low-rank coals

DOEpatents

Quigley, David R.

1992-01-01

A method of chemically comminuting a low-rank coal while at the same time increasing the heating value of the coal. A strong alkali solution is added to a low-rank coal to solubilize the carbonaceous portion of the coal, leaving behind the noncarbonaceous mineral matter portion. The solubilized coal is precipitated from solution by a multivalent cation, preferably calcium.

Chemical comminution and deashing of low-rank coals

DOEpatents

Quigley, David R.

1992-12-01

A method of chemically comminuting a low-rank coal while at the same time increasing the heating value of the coal. A strong alkali solution is added to a low-rank coal to solubilize the carbonaceous portion of the coal, leaving behind the noncarbonaceous mineral matter portion. The solubilized coal is precipitated from solution by a multivalent cation, preferably calcium.

Refolding and purification of recombinant L-asparaginase from inclusion bodies of E. coli into active tetrameric protein

PubMed Central

Upadhyay, Arun K.; Singh, Anupam; Mukherjee, K. J.; Panda, Amulya K.

2014-01-01

A tetrameric protein of therapeutic importance, Escherichia coli L-asparaginase-II was expressed in Escherichia coli as inclusion bodies (IBs). Asparaginase IBs were solubilized using low concentration of urea and refolded into active tetrameric protein using pulsatile dilution method. Refolded asparaginase was purified in two steps by ion-exchange and gel filtration chromatographic techniques. The recovery of bioactive asparaginase from IBs was around 50%. The melting temperature (Tm) of the purified asparaginase was found to be 64°C. The specific activity of refolded, purified asparaginase was found to be comparable to the commercial asparaginase (190 IU/mg). Enzymatic activity of the refolded asparaginase was high even at four molar urea solutions, where the IB aggregates are completely solubilized. From the comparison of chemical denaturation data and activity at different concentrations of guanidine hydrochloride, it was observed that dissociation of monomeric units precedes the complete loss of helical secondary structures. Protection of the existing native-like protein structure during solubilization of IB aggregates with 4 M urea improved the propensity of monomer units to form oligomeric structure. Our mild solubilization technique retaining native-like structures, improved recovery of asparaginase in bioactive tetrameric form. PMID:25309524

Effect of hydrostatic pressure on gas solubilization in micelles.

PubMed

Meng, Bin; Ashbaugh, Henry S

2015-03-24

Molecular dynamics simulations of anionic sodium decylsulfate and nonionic pentaethylene glycol monodecyl ether micelles in water have been performed to examine the impact of hydrostatic pressure on argon solubilization as a function of pressure. The potential-of-mean force between the micelles and argon demonstrates that nonpolar gases are attracted to the interiors of both micelles. The affinity of argon for micelle interiors, however, decreases with increasing pressure as a result of the comparatively higher molar volume of argon inside assemblies. We evaluate solubility enhancement coefficients, which describe the drop in the solute chemical potential as a function of the micellized surfactant concentration, to quantify the impact of micellization on gas solubilization. While argon is similarly attracted to the hydrophobic cores of both micelles, the gas is more effectively sequestered within nonionic micelles compared with anionic micelles as a result of salting out by charged head groups and accompanying counterions. The solubility enhancement coefficients of both micelles decrease with increasing pressure, reflecting the changing forces observed in the potentials-of-mean force. An analytical liquid drop model is proposed to describe the pressure dependence of argon solubilization within micelles that captures the simulation solubility enhancement coefficients after fitting an effective micelle radius for each surfactant.

Solubilizing properties of new surface-active agents, products of catalytic oxyethylation of cholic acid.

PubMed

Kołodziejczyk, Michał Krzysztof; Nachajski, Michal Jakub; Lukosek, Marek; Zgoda, Marian Mikołaj

2013-01-01

Solubilizing properties of aqueous solutions of a series of surface-active agents, products of oxyethylation of cholic acid, were examined in the present study. The content of oxyethylated segments determined by means of the 1H NMR method enabled the verification of the molecular mass of surfactants along with the calculation of the structural hydrophilic-lipophilic balance (HLB), the solubility parameter delta1/2, and the required solubility level of balance HLB(R). Viscosimetric measurements enabled the calculation of the limiting viscosity number, the content-average molecular mass, the effective volume, the hydrodynamic radius of the surfactant micelle and their equilibrium adducts with rutin, diclofenac and loratadine (BCS Class II and III). By means of the spectrophotometric method (UV) the amount of the solubilized diclofenac, loratadine and rutin (rutoside) was determined in the equilibrium system (saturated solution) in the environment of aqueous solutions of cholic acid derivatives of n(TE) = 20-70. The obtained results serve as a basis for determining the solubilization mechanism of lipophilic therapeutic products and indirectly for estimating the influence of the above process on pharmaceutical as well as biological availability of a micellar adduct from model drug forms (Lindbladt lithogenolitic index).

Preparation and Cyclodextrin Solubilization of the Antibacterial Agent Benzoyl Metronidazole

PubMed Central

Yang, Shuo

2013-01-01

A one-pot method for the preparation of benzoyl metronidazole was achieved by using N,N′-carbonyldiimidazole as a coupling reagent. Moreover, it was found that the byproduct imidazole as the catalyst promoted the reaction. In addition, the β-cyclodextrin solubilization of benzoyl metronidazole was investigated by phase-solubility method. The phase-solubility studies indicated that the solubility of benzoyl metronidazole (S = 0.1435 g/L) was substantially increased 9.7-fold (S′ = 1.3881 g/L) by formation of 1 : 1 benzoyl metronidazole/β-cyclodextrin complexes in water, and the association constant K a value was determined to be 251 (±23) dm3/mol. Therefore, β-cyclodextrin can work as a pharmaceutical solubilizer for benzoyl metronidazole and may improve its oral bioavailability. PMID:23970831

Digestive kinetics determines bioavailability of pollutants. Final report, 1 June 1993--30 September 1998

DOE Office of Scientific and Technical Information (OSTI.GOV)

Jumars, P.A.; Mayer, L.M.

1999-04-19

The authors assayed digestive capabilities of marine deposit feeders (animals that eat sediments) by using fluorescently tagged substrates and contact-angle measurements of surfactancy. Polychaetes on average showed higher enzyme activities and surfactancy than echinoderms. They found that surfactants produced by deposit feeders substantially enhance their abilities to solubilize hydrophobic pollutants such as polycyclic aromatic hydrocarbons (PAHs). Amounts solubilized were consistent with incorporation into micelles of the surfactant. Kinetics of PAH uptake could be explained by passive diffusion. The authors also found that the digestive strategies of deposit feeders often produce concentrations of proteins (digestive enzymes plus products of protein digestion)more » that are sufficient to solubilize metals. Histidine residues in these proteins were found to be critical for copper binding.« less

Integrated chemical and multi-scale structural analyses for the processes of acid pretreatment and enzymatic hydrolysis of corn stover.

PubMed

Chen, Longjian; Li, Junbao; Lu, Minsheng; Guo, Xiaomiao; Zhang, Haiyan; Han, Lujia

2016-05-05

Corn stover was pretreated with acid under moderate conditions (1.5%, w/w, 121°C, 60min), and kinetic enzymolysis experiments were performed on the pretreated substrate using a mixture of Celluclast 1.5L (20FPU/g dry substrate) and Novozyme 188 (40CBU/g dry substrate). Integrated chemical and multi-scale structural methods were then used to characterize both processes. Chemical analysis showed that acid pretreatment removed considerable hemicellulose (from 19.7% in native substrate to 9.28% in acid-pretreated substrate) and achieved a reasonably high conversion efficiency (58.63% of glucose yield) in the subsequent enzymatic hydrolysis. Multi-scale structural analysis indicated that acid pretreatment caused structural changes via cleaving acetyl linkages, solubilizing hemicellulose, relocating cell wall surfaces and enlarging substrate porosity (pore volume increased from 0.0067cm(3)/g in native substrate to 0.019cm(3)/g in acid-pretreated substrate), thereby improving the polysaccharide digestibility. Copyright © 2016 Elsevier Ltd. All rights reserved.

Hydrodynamic properties of the gonadotropin receptor from a murine Leydig tumor cell line are altered by desensitization

DOE Office of Scientific and Technical Information (OSTI.GOV)

Rebois, R.V.; Bradley, R.M.; Titlow, C.C.

1987-10-06

The murine Leydig tumor cell line 1 (MLTC-1) contains gonadotropin receptors (GR) that are coupled to adenylate cyclase through the stimulatory guanine nucleotide binding protein (G/sub s/). The binding of human choriogonadotropin (hGC) causes MLTC-1 cells to accumulate cAMP. With time, the ability of MLTC-1 cells to respond to hCG is attenuated by a process called desensitization. The hydrodynamic properties of GR from control and desensitized MLTC-1 cells were studied. Sucrose density gradient sedimentation in H/sub 2/O and D/sub 2/O and gel filtration chromatography were used to estimate the Stokes radius (a), partial specific volume (v/sub c/), sedimentation coefficient (s/submore » 20,w/), and molecular weight (M/sub r/) of the detergent-solubilized hormone-receptor complex (hCG-GR). (/sup 125/I)hCG was bound to MLTC-1 cells under conditions that allow (37/sup 0/C) or prevent (0/sup 0/C) desensitization, and hCG-GR was solubilized in Triton X-100. In the absence of desensitization, control hCG-GR had a M/sub r/ of 213,000, whereas desensitized hCG-GR had a M/sub r/ of 158,000. Deglycosylated hCG (DG-HCG) is an antagonist that binds to GR with high affinity but fails to stimulate adenylate cyclase or cause desensitization. (/sup 125/I)DG-hCG was bound to MLTC-1 cells and DG-hCG-GR solubilized in Triton X-100. The hydrodynamic properties of DG-hCG-GR were the same as that for control hCG-GR. There was no evidence for the association of adenylate cyclase or G/sub s/ with GR in Triton X-100 solubilized preparations. When hCG was cross-linked to GR and solubilized with sodium dodecyl sulfate (SDS), the M/sub r/ was found to be 116,000, which was similar to that determined by SDS-polyacrylamide gel electrophoresis and less than that of the Triton X-100 solubilized control hCG-GR.« less

A solid-phase assay for the detection of anti-sperm antibodies.

PubMed

Okada, H; Kamidono, S; Owens, G R; Nagamatsu, G R; Addonizio, J C

1993-05-01

ELISA is an ideal assay method for a large-scale screening of anti-sperm antibodies among a large number of infertile males. However, conventional ELISA with whole spermatozoa needs time-consuming steps of centrifugation. A solid-phase assay used for detecting anti-sperm antibodies was established. This assay is suitable not only for detecting circulating anti-sperm antibodies of IgG, IgM, and IgA subclass simultaneously but also for screening hybridomas secreting anti-sperm monoclonal antibodies (mAbs). The microtiter plates, on which solubilized sperm antigens are fixed, can be stored at -80 degrees C for up to six months without losing reactivity with anti-sperm antibodies. Using this assay, 53 sera (13 were proven positive and 40 were proven negative for sperm agglutination antibody) were tested. Although the false-negative rate was 0%, the false-positive rate was 32%. One thousand one hundred sixty-five supernatants from hybridomas constructed with splenocytes of mice who were hyperimmunized with human sperm and nonsecreting myeloma cells were tested by this solid-phase assay and two anti-sperm mAb secreting clones were selected and established. It is recommended that for research work this assay could be used for the first screening of the hybridoma secreting anti-sperm mAb, and for clinical use this assay might be suitable for the first screening of sera of infertile patients. However, conventional bioassays should follow to confirm the biological meaning of the positivity.

Sucrose dependent mineral phosphate solubilization in Enterobacter asburiae PSI3 by heterologous overexpression of periplasmic invertases.

PubMed

Kumar, Chanchal; Wagh, Jitendra; Archana, G; Naresh Kumar, G

2016-12-01

Enterobacter asburiae PSI3 solubilizes mineral phosphates in the presence of glucose by the secretion of gluconic acid generated by the action of a periplasmic pyrroloquinoline quinone dependent glucose dehydrogenase. In order to achieve mineral phosphate solubilization phenotype in the presence of sucrose, plasmids pCNK4 and pCNK5 containing genes encoding the invertase enzyme of Zymomonas mobilis (invB) and of Saccharomyces cerevisiae (suc2) under constitutive promoters were constructed with malE signal sequence (in case of invB alone as the suc2 is secreted natively). When introduced into E. asburiae PSI3, E. a. (pCNK4) and E. a. (pCNK5) transformants secreted 21.65 ± 0.94 and 22 ± 1.3 mM gluconic acid, respectively, in the presence of 75 mM sucrose and they also solubilized 180 ± 4.3 and 438 ± 7.3 µM P from the rock phosphate. In the presence of a mixture of 50 mM sucrose and 25 mM glucose, E. a. (pCNK5) secreted 34 ± 2.3 mM gluconic acid and released 479 ± 8.1 µM P. Moreover, in the presence of a mixture of eight sugars (10 mM each) in the medium, E. a. (pCNK5) released 414 ± 5.3 µM P in the buffered medium. Thus, this study demonstrates incorporation of periplasmic invertase imparted P solubilization ability to E. asburiae PSI3 in the presence of sucrose and mixture of sugars.

Expression and solubilization of insect cell-based rabies virus glycoprotein and assessment of its immunogenicity and protective efficacy in mice.

PubMed

Ramya, R; Mohana Subramanian, B; Sivakumar, V; Senthilkumar, R L; Sambasiva Rao, K R S; Srinivasan, V A

2011-10-01

Rabies is a fatal zoonotic disease of serious public health and economic significance worldwide. The rabies virus glycoprotein (RVG) has been the major target for subunit vaccine development, since it harbors domains responsible for induction of virus-neutralizing antibodies, infectivity, and neurovirulence. The glycoprotein (G) was cloned using the baculovirus expression vector system (BEVS) and expressed in Spodoptera frugiperda (Sf-9) cells. In order to obtain a soluble form of G suitable for experimentation in mice, 18 different combinations of buffers and detergents were evaluated for their ability to solubilize the insect cell membrane-associated G. The combination that involved 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) detergent in lysis buffer 1, formulated with Tris, NaCl, 10% dimethyl sulfoxide (DMSO), and EDTA, gave the highest yield of soluble G, as evidenced by the experimental data. Subsequently, several other parameters, such as the concentration of CHAPS and the duration and temperature of the treatment for the effective solubilization of G, were optimized. The CHAPS detergent, buffered at a concentration of 0.4% to 0.7% (wt/vol) at room temperature (23 to 25°C) for 30 min to 1 h using buffer 1, containing 10% DMSO, resulted in consistently high yields. The G solubilized using CHAPS detergent was found to be immunogenic when tested in mice, as evidenced by high virus-neutralizing antibody titers in sera and 100% protection upon virulent intracerebral challenge with the challenge virus standard (CVS) strain of rabies virus. The results of the mice study indicated that G solubilized with CHAPS detergent retained the immunologically relevant domains in the native conformation, thereby paving the way for producing a cell-free and efficacious subunit vaccine.

Solubilization and Upgrading of High Polyethylene Terephthalate Loadings in a Low-Costing Bifunctional Ionic Liquid

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sun, Jian; Liu, Dajiang; Young, Robert P.

Polyethylene terephthalate (PET) is of significant commercial importance, but is difficult to recycle. Chemical inertness and resistance to biodegradation make the recycling of PET challenging and most solvents for PET are highly toxic. In this work, we demonstrate for the first time that a low cost (~$1.2/kg) and biocompatible ionic liquid (IL), cholinium phosphate ([Ch]3[PO4]) can play bifunctional roles in PET solubilization and glycolytic degradation. High loading of PET (10 wt%) is readily dissolved in [Ch]3[PO4] at relatively low temperatures (120 °C, 1h) and even in water-rich conditions. Tandem in situ confocal microscopy and Fourier Transform Infrared (FTIR) spectroscopy studiesmore » give detailed information on the solubilization mechanism in terms of morphological and chemical changes that occur. In depth analysis of PET-IL solution reveals that the high PET solubilization in [Ch]3[PO4] can be ascribed to significant PET depolymerization. Acid precipitation yields terephthalic acid as the dominant depolymerized monomer with a theoretical yield of ~95%. Further exploration shows that in the presence of ethylene glycol, [Ch]3[PO4] catalyzed glycolysis of PET can efficiently occur with ~100% PET conversion and ~60.6% bis(2-hydroxyethyl)terephthalate (BHET) yield under metal free conditions. The IL can be reused at least three times without an apparent decrease in activity. NMR analysis reveals that strong hydrogen bond interactions between EG and the IL play an important role for EG activation and promotion of the glycolysis reaction. This study opens up avenues for exploring environmentally benign and efficient technology of ILs for solubilizing and recycling postconsumer polyester plastics.« less

Rapid solubilization of insoluble phosphate by a novel environmental stress-tolerant Burkholderia vietnamiensis M6 isolated from ginseng rhizospheric soil.

PubMed

Park, Ki-Hyun; Lee, O-Mi; Jung, Ho-Il; Jeong, Jin-Ha; Jeon, Young-Dong; Hwang, Dae-Youn; Lee, Chung-Yeol; Son, Hong-Joo

2010-04-01

We isolated and characterized novel insoluble phosphate (P)-solubilizing bacteria tolerant to environmental factors like high salt, low and high pHs, and low temperature. A bacterium M6 was isolated from a ginseng rhizospheric soil and confirmed to belong to Burkholderia vietnamiensis by BIOLOG system and 16S rRNA gene analysis. The optimal cultural conditions for the solubilization of P were 2.5% (w/v) glucose, 0.015% (w/v) urea, and 0.4% (w/v) MgCl(2).6H(2)O along with initial pH 7.0 at 35 degrees C. High-performance liquid chromatography analysis showed that B. vietnamiensis M6 produced gluconic and 2-ketogluconic acids. During the culture, the pH was reduced with increase in gluconic acid concentration and was inversely correlated with P solubilization. Insoluble P solubilization in the optimal medium was about 902 mg l(-1), which was approximately 1.6-fold higher than the yield in NBRIP medium (580 mg l(-1)). B. vietnamiensis M6 showed resistance against different environmental stresses like 10-45 degrees C, 1-5% (w/v) salt, and 2-11 pH range. The maximal concentration of soluble P produced by B. vietnamiensis M6 from Ca(3)(PO(4))(2), CaHPO(4), and hydroxyapatite was 1,039, 2,132, and 1,754 mg l(-1), respectively. However, the strain M6 produced soluble P with 20 mg l(-1) from FePO(4) after 2 days and 100 mg l(-1) from AlPO(4) after 6 days, respectively. Our results indicate that B. vietnamiensis M6 could be a potential candidate for the development of biofertilizer applicable to environmentally stressed soil.

Tricalcium phosphate solubilization and nitrogen fixation by newly isolated Aneurinibacillus aneurinilyticus CKMV1 from rhizosphere of Valeriana jatamansi and its growth promotional effect.

PubMed

Chauhan, Anjali; Guleria, Shiwani; Balgir, Praveen P; Walia, Abhishek; Mahajan, Rishi; Mehta, Preeti; Shirkot, Chand Karan

Aneurinibacillus aneurinilyticus strain CKMV1 was isolated from rhizosphere of Valeriana jatamansi and possessed multiple plant growth promoting traits like production of phosphate solubilization (260mg/L), nitrogen fixation (202.91nmolethylenemL -1 h -1 ), indole-3-acetic acid (IAA) (8.1μg/mL), siderophores (61.60%), HCN (hydrogen cyanide) production and antifungal activity. We investigated the ability of isolate CKMV1 to solubilize insoluble P via mechanism of organic acid production. High-performance liquid chromatography (HPLC) study showed that isolate CKMV1 produced mainly gluconic (1.34%) and oxalic acids. However, genetic evidences for nitrogen fixation and phosphate solubilization by organic acid production have been reported first time for A. aneurinilyticus strain CKMV1. A unique combination of glucose dehydrogenase (gdh) gene and pyrroloquinoline quinone synthase (pqq) gene, a cofactor of gdh involved in phosphate solubilization has been elucidated. Nitrogenase (nif H) gene for nitrogen fixation was reported from A. aneurinilyticus. It was notable that isolate CKMV1 exhibited highest antifungal against Sclerotium rolfsii (93.58%) followed by Fusarium oxysporum (64.3%), Dematophora necatrix (52.71%), Rhizoctonia solani (91.58%), Alternaria sp. (71.08%) and Phytophthora sp. (71.37%). Remarkable increase was observed in seed germination (27.07%), shoot length (42.33%), root length (52.6%), shoot dry weight (62.01%) and root dry weight (45.7%) along with NPK (0.74, 0.36, 1.82%) content of tomato under net house condition. Isolate CKMV1 possessed traits related to plant growth promotion, therefore, could be a potential candidate for the development of biofertiliser or biocontrol agent and this is the first study to include the Aneurinibacillus as PGPR. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Lignocellulose fermentation and residual solids characterization for senescent switchgrass fermentation by Clostridium thermocellum in the presence and absence of continuous in situ ball-milling

DOE Office of Scientific and Technical Information (OSTI.GOV)

Balch, Michael L.; Holwerda, Evert K.; Davis, Mark F.

Milling during lignocellulosic fermentation, henceforth referred to as cotreatment, is investigated as an alternative to thermochemical pretreatment as a means of enhancing biological solubilization of lignocellulose. We investigate the impact of milling on soluble substrate fermentation by Clostridium thermocellum with comparison to yeast, document solubilization for fermentation of senescent switchgrass with and without ball milling, and characterize residual solids. Soluble substrate fermentation by C. thermocellum proceeded readily in the presence of continuous ball milling but was completely arrested for yeast. Total fractional carbohydrate solubilization achieved after fermentation of senescent switchgrass by C. thermocellum for 5 days was 0.45 without cotreatmentmore » or pretreatment, 0.81 with hydrothermal pretreatment (200 degrees C, 15 minutes, severity 4.2), and 0.88 with cotreatment. Acetate and ethanol were the main fermentation products, and were produced at similar ratios with and without cotreatment. Analysis of solid residues was undertaken using molecular beam mass spectrometry (PyMBMS) and solid-state nuclear magnetic resonance spectroscopy (NMR) in order to provide insight into changes in plant cell walls during processing via various modes. The structure of lignin present in residual solids remaining after fermentation with cotreatment appeared to change little, with substantially greater changes observed for hydrothermal pretreatment - particularly with respect to formation of C-C bonds. The observation of high solubilization with little apparent modification of the residue is consistent with cotreatment enhancing solubilization primarily by increasing the access of saccharolytic enzymes to the feedstock, and C. thermocellum being able to attack all the major linkages in cellulosic biomass provided that these linkages are accessible.« less

Comparison of in vitro digestive fluid extraction and traditional in vivo approaches as measures of polycyclic aromatic hydrocarbon bioavailability from sediments

DOE Office of Scientific and Technical Information (OSTI.GOV)

Weston, D.P.; Mayer, L.M.

1998-05-01

The bioavailability of particle-associated contaminants was measured by a new approach that employs the digestive fluid of deposit feeders to solubilize contaminants in vitro. The proportion of contaminant solubilized by digestive fluid of the polychaete Arenicola brasiliensis was considered a measure of bioavailability and was contrasted with other, more traditional measures (i.e., uptake clearance, bioaccumulation factor, and absorption efficiency). There was generally good agreement among the four methods on the relative bioavailability of benzo[a]pyrene from six sandy sediments. Measures of phenanthrene bioavailability did not show strong correlations due to both a more limited data set and perhaps greater importance ofmore » uptake from the dissolved phase. The bioavailability of spiked polycyclic aromatic hydrocarbons (PAHs) differed from that of equivalent in situ-contaminated PAH but not in a predictable and consistent manner. By direct measurement of PAH content of recently ingested sediments collected from the foregut the authors were able to quantify the importance of particle-selective feeding in increasing PAH content of ingested material relative to the bulk, ambient sediments. In most instances, the effect of selective feeding by A. brasiliensis was minimal, increasing PAH content of ingested material <20% above the ambient sediments. Absorption efficiencies of PAH during gut passage were determined by direct measurement of PAH concentration in sediments at various points along the digestive tract. Overall digestive absorption efficiencies were similar to the extent of in vitro solubilization by digestive fluids from the same sediments. These data suggest that extent of solubilization of sediment-bound contaminants during gut passage is a critical constraint on uptake and that absorption efficiency, with respect to the solubilized fraction, approaches 100%.« less

Characterization of a phosphate solubilizing and antagonistic strain of Pseudomonas putida (B0) isolated from a sub-alpine location in the Indian Central Himalaya.

PubMed

Pandey, Anita; Trivedi, Pankaj; Kumar, Bhavesh; Palni, Lok Man S

2006-08-01

The morphological, biochemical, and physiological characteristics of a phosphate solubilizing and antagonistic bacterial strain, designated as B0, isolated from a sub-alpine Himalayan forest site have been described. The isolate is gram negative, rod shaped, 0.8 x 1.6 microm in size, and psychrotrophic in nature that could grow from 0 to 35 degrees C (optimum temp. 25 degrees C). It exhibited tolerance to a wide pH range (3-12; optimum 8.0) and salt concentration up to 4% (w/v). Although it was sensitive to kanamycin, gentamicin, and streptomycin (<10 microg mL(-1)), it showed resistance to higher concentrations of ampicillin, penicillin, and carbenicillin (>1000 microg mL(-1)). The isolate showed maximum similarity with Pseudomonas putida based on 16S rRNA analysis. It solubilized tricalcium phosphate under in vitro conditions. The phosphate solubilization was estimated along a temperature range (4-28 degrees C), and maximum activity (247 microg mL(-1)) was recorded at 21 degrees C after 15 days of incubation. The phosphate solubilizing activity coincided with a concomitant decrease in pH of the medium. The isolate also exhibited antifungal activity against phytopathogenic fungi in Petri dish assays and produced chitinase, ss-l,3-glucanase, salicylic acid, siderophore, and hydrogen cyanide. The plant growth promotion and antifungal properties were demonstrated through a maize-based bioassay under greenhouse conditions. Although the bacterial inoculation was found to result in significant increment in plant biomass, it stimulated bacterial and suppressed fungal counts in the rhizosphere. The present study is important with respect to enumerating microbial diversity of the colder regions as well as understanding the potential biotechnological applications of native microbes.

Characterization of the coal biosolubilization process using gel permeation chromatography and CPMAS NMR

DOE Office of Scientific and Technical Information (OSTI.GOV)

Linehan, J.C.; Bean, R.M.; Franz, J.A.

1990-05-01

Leonardite, an oxidized lignite, and Illinois {number sign}6 coal were treated with Trametes versicolor and Penicillium sp., respectively, and separately with aqueous base to yield soluble and insoluble products. The products and starting materials were analyzed by gel permeation chromatography (GPC), using both aqueous and organic eluents, and by high-field, high-speed-pinning (>10.0 kHz) {sup 13}C cross polarization/magic angle spinning (CPMAS) nuclear magnetic resonance spectroscopy (NMR). The weight average molecular weights (M{sub w}) of the fungal-and base-solubilized products determined by GPC using acidic tetrahydrofuran (THF) eluent were found to be consistently lower than the M{sub w} determined using basic aqueous eluents.more » The M{sub w} of the leonardite product was measured to be 1800 and 6100 daltons using the THF and aqueous eluents, respectively. The aqueous eluent (phosphate buffered at pH 11.5) was found to be superior to the THF eluent in its solubilizing power, with 10% more material analyzed with the basic eluent. The solubility of the biotreated products in aqueous base was greater than either the starting coal or the chemically solubilized product. The Trametes-solubilized leonardite was found to contain a higher percentage of aliphatic carbon than the raw lignite; the Penicillium- solubilized Illinois {number sign}6 contained more aromatic carbon than before fungal treatment as determined by {sup 13}C CPMAS NMR. Pre-oxidation of Illinois {number sign}6 decreases the relative amount of aliphatic carbon. The high-field, high-speed-spinning CPMAS NMR technique was quantitatively evaluated using Argonne premium coals,International Humic Society Standards, and model compounds at various temperatures. 7 refs., 4 figs., 3 tabs.« less

Influence of biochar application on potassium-solubilizing Bacillus mucilaginosus as potential biofertilizer.

PubMed

Liu, Sainan; Tang, Wenzhu; Yang, Fan; Meng, Jun; Chen, Wenfu; Li, Xianzhen

2017-01-02

Biochar can enhance soil fertility to increase agricultural productivity, whereas its improvement in soil microbial activity is still unclear. In this article, the influence of biochar on the cell growth and the potassium-solubilizing activity of Bacillus mucilaginosus AS1153 was examined. The impact on cell growth is related to the biochar-derived feedstocks and the particle size of biochar. Both intrinsic features and inner component fraction can promote the cell growth of B. mucilaginosus AS1153. The potassium-solubilizing activity was increased by 80% when B. mucilaginosus was incubated in conjunction with the biochar derived from corn stover. The survival time of B. mucilaginosus also was prolonged by adsorption in biochar. The experimental results suggested that the biochar containing B. mucilaginosus could be used as a potential biofertilizer to sustain crop production.

Toxicologic evaluation of the migration of a plasticizer, DI (2-ethylhexyl) phthalate (DEHP) from vinyl plastics. [pathology of vinyl plastics poisoning

NASA Technical Reports Server (NTRS)

Rubin, R. J.

1975-01-01

The intravenous administration of DEHP solubilized by means of a number of different detergents leads to respiratory distress and death in rats. At autopsy the lungs are grossly enlarged, edamatous, and hemorrhagic. Light and electron microscopic evaluation of the lungs indicate engorgement of the interalveolar septa with edema fluid and polymorphonuclear leucocytes, degranulation of the leukocytes, and progessive destruction of the endothelial and epithelial cells. Consistent with the conclusion that solubilized DEHP results in a syndrome of "shock lung" is the associated massive fall in arterial blood pressure and the prevention of the lung pathology by pretreatment with pharmacologic doses of an antiinflammatory steroid, methylprednisolone. Evidence is also presented that suggests that the DEHP inadvertently administered to humans during transfusions is also in a solubilized state in the plasma.

DNA-Assisted Solubilization of Carbon Nanotubes and Construction of DNA-MWCNT Cross-Linked Hybrid Hydrogels

PubMed Central

Zinchenko, Anatoly; Taki, Yosuke; Sergeyev, Vladimir G.; Murata, Shizuaki

2015-01-01

A simple method for preparation of DNA-carbon nanotubes hybrid hydrogel based on a two-step procedure including: (i) solubilization of multi-walled carbon nanotubes (MWCNT) in aqueous solution of DNA, and (ii) chemical cross-linking between solubilized MWCNT via adsorbed DNA and free DNA by ethylene glycol diglycidyl ether is reported. We show that there exists a critical concentration of MWCNT below which a homogeneous dispersion of MWCNT in hybrid hydrogel can be achieved, while at higher concentrations of MWCNT the aggregation of MWCNT inside hydrogel occurs. The strengthening effect of carbon nanotube in the process of hydrogel shrinking in solutions with high salt concentration was demonstrated and significant passivation of MWCNT adsorption properties towards low-molecular-weight aromatic binders due to DNA adsorption on MWCNT surface was revealed. PMID:28347011

DNA-Assisted Solubilization of Carbon Nanotubes and Construction of DNA-MWCNT Cross-Linked Hybrid Hydrogels.

PubMed

Zinchenko, Anatoly; Taki, Yosuke; Sergeyev, Vladimir G; Murata, Shizuaki

2015-03-03

A simple method for preparation of DNA-carbon nanotubes hybrid hydrogel based on a two-step procedure including: (i) solubilization of multi-walled carbon nanotubes (MWCNT) in aqueous solution of DNA, and (ii) chemical cross-linking between solubilized MWCNT via adsorbed DNA and free DNA by ethylene glycol diglycidyl ether is reported. We show that there exists a critical concentration of MWCNT below which a homogeneous dispersion of MWCNT in hybrid hydrogel can be achieved, while at higher concentrations of MWCNT the aggregation of MWCNT inside hydrogel occurs. The strengthening effect of carbon nanotube in the process of hydrogel shrinking in solutions with high salt concentration was demonstrated and significant passivation of MWCNT adsorption properties towards low-molecular-weight aromatic binders due to DNA adsorption on MWCNT surface was revealed.

Nanolipoprotein particles and related methods and systems for protein capture, solubilization, and/or purification

DOEpatents

Chromy, Brett A; Henderson, Paul; Hoeprich, Jr., Paul D

2014-12-09

Provided herein are methods and systems for assembling, solubilizing and/or purifying a membrane associated protein in a nanolipoprotein particle, which comprise a temperature transition cycle performed in presence of a detergent, wherein during the temperature transition cycle the nanolipoprotein components are brought to a temperature above and below the gel to liquid crystalling transition temperature of the membrane forming lipid of the nanolipoprotein particle.

Nanolipoprotein particles and related methods and systems for protein capture, solubilization, and/or purification

DOEpatents

Chromy, Brett A.; Henderson, Paul; Hoeprich, Jr, Paul D.

2016-10-04

Provided herein are methods and systems for assembling, solubilizing and/or purifying a membrane associated protein in a nanolipoprotein particle, which comprise a temperature transition cycle performed in presence of a detergent, wherein during the temperature transition cycle the nanolipoprotein components are brought to a temperature above and below the gel to liquid crystalling transition temperature of the membrane forming lipid of the nanolipoprotein particle.

Purification of the Alpha Glycerophosphate Oxidase from African Trypanosomes

DTIC Science & Technology

1987-02-02

oxidase (GPO). This enzyme has not been purified or characterize in detail. Inhibition of this enzyme coupled with inhibition of the anaerobic...more manageable afterwards and remained in the procedure although it only slightly increased the yield. The stability of the solubilized enzyme was...whether the detergent was added during the assay or in the solubilization procedure. However, the successful assay for the enzyme was ubiquinol oxidase

High temperature flow-through device for rapid solubilization and analysis

DOEpatents

West, Jason A. A. [Castro Valley, CA; Hukari, Kyle W [San Ramon, CA; Patel, Kamlesh D [Dublin, CA; Peterson, Kenneth A [Albuquerque, NM; Renzi, Ronald F [Tracy, CA

2009-09-22

Devices and methods for thermally lysing of biological material, for example vegetative bacterial cells and bacterial spores, are provided. Hot solution methods for solubilizing bacterial spores are described. Systems for direct analysis are disclosed including thermal lysers coupled to sample preparation stations. Integrated systems capable of performing sample lysis, labeling and protein fingerprint analysis of biological material, for example, vegetative bacterial cells, bacterial spores and viruses are provided.

High temperature flow-through device for rapid solubilization and analysis

DOEpatents

West, Jason A. A.; Hukari, Kyle W.; Patel, Kamlesh D.; Peterson, Kenneth A.; Renzi, Ronald F.

2013-04-23

Devices and methods for thermally lysing of biological material, for example vegetative bacterial cells and bacterial spores, are provided. Hot solution methods for solubilizing bacterial spores are described. Systems for direct analysis are disclosed including thermal lysers coupled to sample preparation stations. Integrated systems capable of performing sample lysis, labeling and protein fingerprint analysis of biological material, for example, vegetative bacterial cells, bacterial spores and viruses are provided.

Fluorinated diglucose detergents for membrane-protein extraction.

PubMed

Boussambe, Gildas Nyame Mendendy; Guillet, Pierre; Mahler, Florian; Marconnet, Anaïs; Vargas, Carolyn; Cornut, Damien; Soulié, Marine; Ebel, Christine; Le Roy, Aline; Jawhari, Anass; Bonneté, Françoise; Keller, Sandro; Durand, Grégory

2018-05-29

Fluorinated surfactants have scarcely been explored for the direct extraction of proteins from membranes because fluorination is believed to abrogate detergency. However, we have recently shown that a commercially available fluorinated surfactant readily solubilizes lipid membranes, thereby suggesting that fluorination per se does not interfere with detergent activity. In this work, we developed new fluorinated surfactants that exhibit detergency in terms of both lipid-vesicle solubilization and membrane-protein extraction. The compounds made and tested contain two glucose moieties as polar headgroup, a hydrogenated thioether linker, and a perfluorinated alkyl tail with either 4, 6, or 8 carbon atoms. The physicochemical properties of the micelles formed by the three fluorinated surfactants were evaluated by NMR spectroscopy, surface tensiometry, isothermal titration calorimetry, dynamic light scattering, small-angle X-ray scattering, and analytical ultracentrifugation. At 25°C, micellization was mainly entropy-driven, and the CMC values were found to decrease with chain length of the fluorinated tail, whereas the aggregation number increased with chain length. Remarkably, all three surfactants were found to solubilize lipid vesicles and extract a broad range of proteins from Escherichiacoli membranes. These findings demonstrate, for the first time, that nonionic fluorinated surfactants could be further exploited for the direct extraction and solubilization of membrane proteins. Copyright © 2018. Published by Elsevier Inc.

Development and preventative effect against pine wilt disease of a novel liquid formulation of emamectin benzoate.

PubMed

Takai, Kazuya; Suzuki, Toshio; Kawazu, Kazuyoshi

2003-03-01

Injection of the poorly water-soluble emamectin benzoate (EB) into pine trunks required the development of an efficient liquid formulation. For injection into big trees in forests a good rate of injection and a high active content were required. Tests on the viscosity and EB-solubilizing ability of 14 various solubilizers in diethylene glycol monobutyl ether (DGMBE) led to the selection of Sorpol SM-100PM as the solubilizer of the formulation. Relationships between the solubilizing ability and amounts of Sorpol SM-100PM and DGMBE relative to that of EB, and between the concentration of the latter and the viscosity or the injection rate of the formulation led to a novel 40 g litre(-1) emamectin benzoate formulation (Shot Wan Liquid Formulation), which was composed of EB (40), Sorpol SM-100PM (120), DGMBE (160) and distilled water (50 g litre(-1)) in methanol. Injection of this formulation at a dose of 10 g EB per unit volume of pine tree prevented over 90% of the trees from wilting caused by pine wood nematode, and this preventative effect continued for 3 years. Neither discolouration of the leaves nor injury around the injection hole on the trees was observed after injection of the formulation.

Surfactant mediated enhanced biodegradation of hexachlorocyclohexane (HCH) isomers by Sphingomonas sp. NM05.

PubMed

Manickam, Natesan; Bajaj, Abhay; Saini, Harvinder S; Shanker, Rishi

2012-09-01

Environmental biodegradation of several chlorinated pesticides is limited by their low solubility and sorption to soil surfaces. To mitigate this problem we quantified the effect of three biosurfactant viz., rhamnolipid, sophorolipid and trehalose-containing lipid on the dissolution, bioavailability, and biodegradation of HCH-isomers in liquid culture and in contaminated soil. The effect of biosurfactants was evaluated through the critical micelle concentration (CMC) value as determined for each isomer. The surfactant increased the solubilization of HCH isomers by 3-9 folds with rhamnolipid and sophorolipid being more effective and showing maximum solubilization of HCH isomers at 40 μg/mL, compared to trehalose-containing lipid showing peak solubilization at 60 μg/mL. The degradation of HCH isomers by Sphingomonas sp. NM05 in surfactant-amended liquid mineral salts medium showed 30% enhancement in 2 days as compared to degradation in 10 days in the absence of surfactant. HCH-spiked soil slurry incubated with surfactant also showed around 30-50% enhanced degradation of HCH which was comparable to the corresponding batch culture experiments. Among the three surfactants, sophorolipid offered highest solubilization and enhanced degradation of HCH isomers both in liquid medium and soil culture. The results of this study suggest the effectiveness of surfactants in improving HCH degradation by increased bioaccessibility.

Solvent recyclability in a multistep direct liquefaction process

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hetland, M.D.; Rindt, J.R.

1995-12-31

Direct liquefaction research at the Energy & Environmental Research Center (EERC) has, for a number of years, concentrated on developing a direct liquefaction process specifically for low-rank coals (LRCs) through the use of hydrogen-donating solvents and solvents similar to coal-derived liquids, the water/gas shift reaction, and lower-severity reaction conditions. The underlying assumption of all of the research was that advantage could be taken of the reactivity and specific qualities of LRCs to produce a tetrahydrofuran (THF)-soluble material that might be easier to upgrade than the soluble residuum produced during direct liquefaction of high-rank coals. A multistep approach was taken tomore » produce the THF-soluble material, consisting of (1) preconversion treatment to prepare the coal for solubilization, (2) solubilization of the coal in the solvent, and (3) polishing to complete solubilization of the remaining material. The product of these three steps can then be upgraded during a traditional hydrotreatment step. The results of the EERC`s research indicated that additional studies to develop this process more fully were justified. Two areas were targeted for further research: (1) determination of the recyclability of the solvent used during solubilization and (2) determination of the minimum severity required for hydrotreatment of the liquid product. The current project was funded to investigate these two areas.« less

Employment of colorimetric enzyme assay for monitoring expression and solubility of GST fusion proteins targeted to inclusion bodies.

PubMed

Mačinković, Igor S; Abughren, Mohamed; Mrkic, Ivan; Grozdanović, Milica M; Prodanović, Radivoje; Gavrović-Jankulović, Marija

2013-12-01

High levels of recombinant protein expression can lead to the formation of insoluble inclusion bodies. These complex aggregates are commonly solubilized in strong denaturants, such as 6-8M urea, although, if possible, solubilization under milder conditions could facilitate subsequent refolding and purification of bioactive proteins. Commercially available GST-tag assays are designed for quantitative measurement of GST activity under native conditions. GST fusion proteins accumulated in inclusion bodies are considered to be undetectable by such assays. In this work, solubilization of recombinantly produced proteins was performed in 4M urea. The activity of rGST was assayed in 2M urea and it was shown that rGST preserves 85% of its activity under such denaturing conditions. A colorimetric GST activity assay with 1-chloro-2, 4-dinitrobenzene (CDNB) was examined for use in rapid detection of expression targeted to inclusion bodies and for the identification of inclusion body proteins which can be solubilized in low concentrations of chaotropic agents. Applicability of the assay was evaluated by tracking protein expression of two GST-fused allergens of biopharmaceutical value in E. coli, GST-Der p 2 and GST-Mus a 5, both targeted to inclusion bodies. Copyright © 2013 Elsevier B.V. All rights reserved.

Application of saponin biosurfactant and its recovery in the MEUF process for removal of methyl violet from wastewater.

PubMed

Samal, Kulbhushan; Das, Chandan; Mohanty, Kaustubha

2017-12-01

The potential of saponin, a biosurfactant, in the micellar enhanced ultrafiltration (MEUF) process was tested systematically for removal of methyl violet from wastewater. For this, the aqueous extract of reetha (Sapindus mukorossi) pericarp which contains saponin was used as the biosurfactant. First, the micellar solubilization of methyl violet in saponin micelles was investigated in terms of molar solubilization power (SP) of saponin. It was observed that the adsorption of methyl violet on the agglomerates of saponin micelles was mainly responsible for the enhanced solubilization. The Gibbs free energy of solubilization (calculated as -29.63 kJ mol -1 ) suggested that process was feasible and spontaneous. The MEUF experiments were performed in batch as well as continuous mode using saponin biosurfactant, and the effect of operating parameters on permeate flux and solute retention were evaluated. The removal of methyl violet in MEUF process was >99% achieved with 10 kDa polyethersulfone (PES) membrane for feed dye concentration of 250 mg L -1 at studied conditions. Finally, the saponin in permeate was recovered using n-heptane and n-butanol by solvent extraction process. The solvent n-butanol showed better extraction efficiency as compared to n-heptane for saponin extraction. Copyright © 2017 Elsevier Ltd. All rights reserved.

S. aureus MscL is a pentamer in vivo but of variable stoichiometries in vitro: implications for detergent-solubilized membrane proteins.

PubMed

Dorwart, Michael R; Wray, Robin; Brautigam, Chad A; Jiang, Youxing; Blount, Paul

2010-12-07

While the bacterial mechanosensitive channel of large conductance (MscL) is the best studied biological mechanosensor and serves as a paradigm for how a protein can sense and respond to membrane tension, the simple matter of its oligomeric state has led to debate, with models ranging from tetramers to hexamers. Indeed, two different oligomeric states of the bacterial mechanosensitive channel MscL have been resolved by X-ray crystallography: The M. tuberculosis channel (MtMscL) is a pentamer, while the S. aureus protein (SaMscL) forms a tetramer. Because several studies suggest that, like MtMscL, the E. coli MscL (EcoMscL) is a pentamer, we re-investigated the oligomeric state of SaMscL. To determine the structural organization of MscL in the cell membrane we developed a disulfide-trapping approach. Surprisingly, we found that virtually all SaMscL channels in vivo are pentameric, indicating this as the physiologically relevant and functional oligomeric state. Complementing our in vivo results, we purified SaMscL and assessed its oligomeric state using three independent approaches (sedimentation equilibrium centrifugation, crosslinking, and light scattering) and established that SaMscL is a pentamer when solubilized in Triton X-100 and C(8)E(5) detergents. However, performing similar experiments on SaMscL solubilized in LDAO, the detergent used in the crystallographic study, confirmed the tetrameric oligomerization resolved by X-ray crystallography. We further demonstrate that this stoichiometric shift is reversible by conventional detergent exchange experiments. Our results firmly establish the pentameric organization of SaMscL in vivo. Furthermore they demonstrate that detergents can alter the subunit stoichiometry of membrane protein complexes in vitro; thus, in vivo assays are necessary to firmly establish a membrane protein's true functionally relevant oligomeric state.

Antimicrobial photodynamic therapy with photosensitizer in ethanol improves oxidative status and gingival collagen in a short-term in periodontitis.

PubMed

Pillusky, Fernanda Maia; Barcelos, Raquel Cristine Silva; Vey, Luciana Taschetto; Barin, Luisa Machado; de Mello Palma, Victor; Maciel, Roberto Marinho; Kantorski, Karla Zanini; Bürger, Marilise Escobar; Danesi, Cristiane Cademartori

2017-09-01

This study evaluated the antimicrobial photodynamic therapy (aPDT) effects using the methylene blue (MB) in ethanol 20% on systemic oxidative status and collagen content from gingiva of rats with periodontitis. Rats were divided into five experimental groups: NC (negative control; no periodontitis); PC (positive control; periodontitis without any treatment); SRP (periodontitis and scaling and root planing), aPDT I (periodontitis and SRP+aPDT+MB solubilized in water), and aPDT II (periodontitis and SRP+aPDT+MB solubilized in ethanol 20%). After 7days of removal of the ligature, the periodontal treatments were performed. At 7/15/30days, gingival tissue was removed for morphometric analysis. The erythrocytes were used to evaluate systemic oxidative status. PC group showed higher lipoperoxidation levels at 7/15/30days. aPDT indicated a protective influence in erythrocytes at 15days observed by the elevation in levels of systemic antioxidant defense. aPDT II group was the only one that restored the total collagen area in 15days, and recovered the type I collagen area at the same time point. aPDT as an adjunct to the SRP can induce the systemic protective response against oxidative stress periodontitis-induced and recover the gingival collagen, thus promoting the healing periodontal, particularly when the MB is dissolved in ethanol 20%. Copyright © 2017 Elsevier B.V. All rights reserved.

Dissolution and solubility behavior of fenofibrate in sodium lauryl sulfate solutions.

PubMed

Granero, Gladys E; Ramachandran, Chandrasekharan; Amidon, Gordon L

2005-10-01

The solubility of fenofibrate in pH 6.8 McIlvaine buffers containing varying concentrations of sodium lauryl sulfate was determined. The dissolution behavior of fenofibrate was also examined in the same solutions with rotating disk experiments. It was observed that the enhancement in intrinsic dissolution rate was approximately 500-fold and the enhancement in solubility was approximately 2000-fold in a pH 6.8 buffer containing 2% (w/v) sodium lauryl sulfate compared to that in buffer alone. The micellar solubilization equilibrium coefficient (k*) was estimated from the solubility data and found to be 30884+/-213 L/mol. The diffusivity for the free solute, 7.15x10(-6) cm2/s, was calculated using Schroeder's additive molal volume estimates and Hayduk-Laurie correlation. The diffusivity of the drug-loaded micelle, estimated from the experimental solubility and dissolution data and the calculated value for free solute diffusivity, was 0.86x10(-6) cm2/s. Thus, the much lower enhancement in dissolution of fenofibrate compared to its enhancement in solubility in surfactant solutions appears to be consistent with the contribution to the total transport due to enhanced micellar solubilization as well as a large decrease (approximately 8-fold) in the diffusivity of the drug-loaded micelle.

Large transient nonproton ion movements in purple membrane suspensions are abolished by solubilization in Triton X-100.

PubMed Central

Marinetti, T; Mauzerall, D

1986-01-01

Light-induced release/uptake of both protons and other ions cause transient changes in conductivity in suspensions of purple membrane (PM) fragments (Marinetti, Tim, and David Mauzerall, 1983, Proc. Natl. Acad. Sci. USA, 80:178-180). We find that the release/uptake of nonproton ions with quantum yield greater than 1 is observed at most pHs and ionic strengths. Only at both low pH and low ionic strength is the conductivity transient mostly due to protons. Our hypothesis is that during the photocycle, changes occur in the PM's dense surface charge distribution that result in changes in the number of counterions bound or condensed at the membrane surface. To test this, the PM structure was perturbed with the nonionic detergent Triton X-100. Immediately after addition, Triton does not abolish the nonproton ion movements; in fact at low detergent concentrations (0.02% vol/vol) the signal amplitudes increased considerably. However, when PM is completely solubilized into monomers in Triton, the conductivity transients are due to protons alone, though at lower quantum yield compared with native PM. These results suggest that changes in the surface charge distribution in native PM's photocycle could contribute to proton transfer between the aqueous phase and bR itself. PMID:3019444

Process analytical technology to understand the disintegration behavior of alendronate sodium tablets.

PubMed

Xu, Xiaoming; Gupta, Abhay; Sayeed, Vilayat A; Khan, Mansoor A

2013-05-01

Various adverse events including esophagus irritations have been reported with the use of alendronate tablets, likely attributed to the rapid tablet disintegration in the mouth or esophagus. Accordingly, the disintegration of six alendronate tablet drug products was studied using a newly developed testing device equipped with in-line sensors, in addition to the official compendial procedure for measuring the disintegration time. The in-line sensors were used to monitor the particle count and solution pH change to assess the onset and duration of disintegration. A relatively large variation was observed in the disintegration time of the tested drug products using the compendial method. The data collected using the in-line sensors suggested that all tested drug products exhibited almost instantaneous onset of disintegration, under 2 s, and a sharp drop in solution pH. The drop in pH was slower for tablets with slower disintegration. The in-house prepared alendronate test tablets also showed similar trends suggesting rapid solubilization of the drug contributed to the fast tablet disintegration. This research highlights the usefulness of the newly developed in-line analytical method in combination with the compendial method in providing a better understanding of the disintegration and the accompanying drug solubilization processes for fast disintegrating tablet drug products. Copyright © 2013 Wiley Periodicals, Inc.

[Renaturation with simultaneous purification of the recombinant human Flt3 ligand from inclusion bodies by high performance hydrophobic interaction chromatography].

PubMed

Jia, Jia; Wang, Lili; Gao, Dong; Geng, Xindu

2010-06-01

Flt3 ligand (FL) is a class of cytokines with the functions of promoting early hematopoiesis. It has important clinical value in promoting growth and development of hematopoietic cells and hematopoietic mobilization. In order to obtain large quantities of recombinant human FL (rhFL) by genetic engineering methods for clinic and research, in this work, rhFL was expressed in E. coli as inclusion bodies. The inclusion bodies were recovered, cleaned and solubilized in 8 mol/L urea, the solubilized rhFL was renatured by high performance hydrophobic interaction chromatography (HPHIC) with simultaneous purification, the retention feature and renaturation regularity were studied. The results showed that when the denatured protein concentration was 8.51 g/L, and the end group of stationary phase was PEG800, under the conditions of mobile phase of pH 7.0 and with the addition of 4 mol/L urea, 1.8 mmol/L glutathione (GSH) and 0.3 mmol/L oxidative glutathione (GSSG), a mass recovery of 36.9% and a purity of 94.5% were obtained after refolding with simultaneous purification. The obtained rhFL was successfully renatured with simultaneous purification in only one step of HPHIC, and it provided a foundation for the manufacturing of high quality rhFL.

Enhanced Cr bioleaching efficiency from tannery sludge with coinoculation of Acidithiobacillus thiooxidans TS6 and Brettanomyces B65 in an air-lift reactor.

PubMed

Fang, Di; Zhou, Li-Xiang

2007-09-01

Bioleaching process has been demonstrated to be an effective technology in removing Cr from tannery sludge, but a large quantity of dissolved organic matter (DOM) present in tannery sludge often exhibits a marked toxicity to chemolithoautotrophic bioleaching bacteria such as Acidithiobacillus thiooxidans. The purpose of the present study was therefore to enhance Cr bioleaching efficiencies through introducing sludge DOM-degrading heterotrophic microorganism into the sulfur-based sludge bioleaching system. An acid-tolerant DOM-degrading yeast strain Brettanomyces B65 was successfully isolated from a local Haining tannery sludge and it could metabolize sludge DOM as a source of energy and carbon for growth. A combined bioleaching experiment (coupling Brettanomyces B65 and A. thiooxidans TS6) performed in an air-lift reactor indicated that the rates of sludge pH reduction and ORP increase were greatly improved, resulting in enhanced Cr solubilization. Compared with the 5 days required for maximum solubilization of Cr for the control (single bioleaching process without inoculation of Brettanomyces B65), the bioleaching period was significantly shorten to 3 days for the combined bioleaching system. Moreover, little nitrogen and phosphorous were lost and the content of Cr was below the permitted levels for land application after 3 days of bioleaching treatment.

Effect of a new regeneration process by adsorption-coagulation and flocculation on the physicochemical properties and the detergent efficiency of regenerated cleaning solutions.

PubMed

Blel, Walid; Dif, Mehdi; Sire, Olivier

2015-05-15

Reprocessing soiled cleaning-in-place (CIP) solutions has large economic and environmental costs, and it would be cheaper and greener to recycle them. In food industries, recycling of CIP solutions requires a suitable green process engineered to take into account the extreme physicochemical conditions of cleaning while not altering the process efficiency. To this end, an innovative treatment process combining adsorption-coagulation with flocculation was tested on multiple recycling of acid and basic cleaning solutions. In-depth analysis of time-course evolutions was carried out in the physicochemical properties (concentration, surface tension, viscosity, COD, total nitrogen) of these solutions over the course of successive regenerations. Cleaning and disinfection efficiencies were assessed based on both microbiological analyses and organic matter detachment and solubilization from fouled stainless steel surfaces. Microbiological analyses using a resistant bacterial strain (Bacillus subtilis spores) highlighted that solutions regenerated up to 20 times maintained the same bactericidal efficiency as de novo NaOH solutions. The cleanability of stainless steel surfaces showed that regenerated solutions allow better surface wettability, which goes to explain the improved detachment and solubilization found on different types of organic and inorganic fouling. Copyright © 2015 Elsevier Ltd. All rights reserved.

Preparation, characterization and in vitro evaluation of a new nucleotide analogue prodrug cyclodextrin inclusion complexes.

PubMed

Diab, Roudayna; Jordheim, Lars P; Degobert, Ghania; Peyrottes, Suzanne; Périgaud, Christian; Dumontet, Charles; Fessi, Hatem

2009-01-01

Bis(tbutyl-S-acyl-2-thioethyl)-cytidine monophosophate is a new cytotoxic mononucleotide prodrug which have been developed to reverse the cellular resistance to nucleoside analogues. Unfortunately, its in vivo utilisation was hampered by its poor water solubility, raising the need of a molecular vector capable to mask its physicochemical characteristics although without affecting its cytotoxic activity. Hydroxypropyl-beta-cyclodextrin was used to prepare the prodrug inclusion complexes, allowing it to be solubilized in water and hence to be used for in vitro and in vivo experiments. A molar ratio of the cyclodextrin: prodrug of 3 was sufficient to obtain complete solubilization of the prodrug. The inclusion complex was characterized by differential scanning calorimetry, which revealed the disappearance of the melting peak of the prodrug suggesting the formation of inclusion complex. Proton Nuclear Magnetic Resonance spectroscopy provided a definitive proof of the inclusion complex formation, which was evidenced by the large chemical shift displacements observed for protons located in the interior of the hydrophobic cyclodextrin cavity. The complex retained its cytotoxic activity as shown by in vitro cell survival assays on murine leukemia cells. These results provided a basis for potential therapeutic applications of co-formulation of this new nucleotide analogue with hydroxypropyl-beta-CD in cancer therapy.

Characteristics and adaptability of iron- and sulfur-oxidizing microorganisms used for the recovery of metals from minerals and their concentrates

PubMed Central

Rawlings, Douglas E

2005-01-01

Microorganisms are used in large-scale heap or tank aeration processes for the commercial extraction of a variety of metals from their ores or concentrates. These include copper, cobalt, gold and, in the past, uranium. The metal solubilization processes are considered to be largely chemical with the microorganisms providing the chemicals and the space (exopolysaccharide layer) where the mineral dissolution reactions occur. Temperatures at which these processes are carried out can vary from ambient to 80°C and the types of organisms present depends to a large extent on the process temperature used. Irrespective of the operation temperature, biomining microbes have several characteristics in common. One shared characteristic is their ability to produce the ferric iron and sulfuric acid required to degrade the mineral and facilitate metal recovery. Other characteristics are their ability to grow autotrophically, their acid-tolerance and their inherent metal resistance or ability to acquire metal resistance. Although the microorganisms that drive the process have the above properties in common, biomining microbes usually occur in consortia in which cross-feeding may occur such that a combination of microbes including some with heterotrophic tendencies may contribute to the efficiency of the process. The remarkable adaptability of these organisms is assisted by several of the processes being continuous-flow systems that enable the continual selection of microorganisms that are more efficient at mineral degradation. Adaptability is also assisted by the processes being open and non-sterile thereby permitting new organisms to enter. This openness allows for the possibility of new genes that improve cell fitness to be selected from the horizontal gene pool. Characteristics that biomining microorganisms have in common and examples of their remarkable adaptability are described. PMID:15877814

Characteristics and adaptability of iron- and sulfur-oxidizing microorganisms used for the recovery of metals from minerals and their concentrates.

PubMed

Rawlings, Douglas E

2005-05-06

Microorganisms are used in large-scale heap or tank aeration processes for the commercial extraction of a variety of metals from their ores or concentrates. These include copper, cobalt, gold and, in the past, uranium. The metal solubilization processes are considered to be largely chemical with the microorganisms providing the chemicals and the space (exopolysaccharide layer) where the mineral dissolution reactions occur. Temperatures at which these processes are carried out can vary from ambient to 80 degrees C and the types of organisms present depends to a large extent on the process temperature used. Irrespective of the operation temperature, biomining microbes have several characteristics in common. One shared characteristic is their ability to produce the ferric iron and sulfuric acid required to degrade the mineral and facilitate metal recovery. Other characteristics are their ability to grow autotrophically, their acid-tolerance and their inherent metal resistance or ability to acquire metal resistance. Although the microorganisms that drive the process have the above properties in common, biomining microbes usually occur in consortia in which cross-feeding may occur such that a combination of microbes including some with heterotrophic tendencies may contribute to the efficiency of the process. The remarkable adaptability of these organisms is assisted by several of the processes being continuous-flow systems that enable the continual selection of microorganisms that are more efficient at mineral degradation. Adaptability is also assisted by the processes being open and non-sterile thereby permitting new organisms to enter. This openness allows for the possibility of new genes that improve cell fitness to be selected from the horizontal gene pool. Characteristics that biomining microorganisms have in common and examples of their remarkable adaptability are described.

Theory of microemulsions in a gravitational field

NASA Technical Reports Server (NTRS)

Jeng, J. F.; Miller, Clarence A.

1989-01-01

A theory of microemulsions developed previously is extended to include the effect of a gravitational field. It predicts variation with position of drop size, drop volume fraction, and area per molecule in the surfactant films within a microemulsion phase. Variation in volume fraction is greatest and occurs in such a way that oil content increases with increasing elevation, as has been found experimentally. Large composition variations are predicted within a middle phase microemulsion near optimal conditions because inversion from the water-continuous to the oil-continuous arrangement occurs with increasing elevation. Generally speaking, gravity reduces solubilization within microemulsions and promotes separation of excess phases.

Complete genome sequence of Pseudomonas rhizosphaerae IH5T (=DSM 16299T), a phosphate-solubilizing rhizobacterium for bacterial biofertilizer.

PubMed

Kwak, Yunyoung; Jung, Byung Kwon; Shin, Jae-Ho

2015-01-10

Pseudomonas rhizosphaerae IH5(T) (=DSM 16299(T)), isolated from the rhizospheric soil of grass growing in Spain, has been reported as a novel species of the genus Pseudomonas harboring insoluble phosphorus solubilizing activity. To understanding the multifunctional biofertilizer better, we report the complete genome sequence of P. rhizosphaerae IH5(T). Copyright © 2014 Elsevier B.V. All rights reserved.

Effect of rhamnolipid biosurfactant on solubilization of polycyclic aromatic hydrocarbons.

PubMed

Li, Shudong; Pi, Yongrui; Bao, Mutai; Zhang, Cong; Zhao, Dongwei; Li, Yiming; Sun, Peiyan; Lu, Jinren

2015-12-15

Rhamnolipid biosurfactant-producing bacteria, Bacillus Lz-2, was isolated from oil polluted water collected from Dongying Shengli oilfield, China. The factors that influence PAH solubilization such as biosurfactant concentration, pH, ionic strength and temperature were discussed. The results showed that the solubilities of naphthalene, phenanthrene and pyrene increased linearly with the rise of rhamnolipid biosurfactant dose above the biosurfactant critical micelle concentration (CMC). Furthermore, the molar solubilization ratio (MSR) values decreased in the following order: naphthalene>phenanthrene>pyrene. However, the solubility percentage increased and followed the opposite order: pyrene>phenanthrene>naphthalene. The solubilities of PAHs in rhamnolipid biosurfactant solution increased with the rise of pH and ionic strength, and reached the maximum values under the conditions of pH11 and NaCl concentration 8 g · L(-1). The solubility of phenanthrene and pyrene increased with the rise of temperature. Copyright © 2015 Elsevier Ltd. All rights reserved.

Effect of low temperature thermal pre-treatment on the solubilization of organic matter, pathogen inactivation and mesophilic anaerobic digestion of poultry sludge.

PubMed

Ruiz-Espinoza, Juan E; Méndez-Contreras, Juan M; Alvarado-Lassman, Alejandro; Martínez-Delgadillo, Sergio A

2012-01-01

Treatment of poultry industry effluents produces wastewater sludge with high levels of organic compounds and pathogenic microorganisms. In this research, the thermal pre-treatment of poultry slaughterhouse sludge (PSS) was evaluated for low temperatures in combination with different exposure times as a pre-hydrolysis strategy to improve the anaerobic digestion process. Organic compounds solubilization and inactivation of pathogenic microorganisms were evaluated after treatment at 70, 80 or 90°C for 30, 60 or 90 min. The results showed that 90°C and 90 min were the most efficient conditions for solubilization of the organic compounds (10%). In addition, the bacteria populations and the more resistant structures, such as helminth eggs (HE), were completely inactivated. Finally, the thermal pre-treatment applied to the sludge increased methane yield by 52% and reduced hydraulic retention time (HRT) by 52%.

Comparison of the fluorescence kinetics of detergent-solubilized and membrane-reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides.

PubMed

Pflock, Tobias; Dezi, Manuela; Venturoli, Giovanni; Cogdell, Richard J; Köhler, Jürgen; Oellerich, Silke

2008-01-01

Picosecond time-resolved fluorescence spectroscopy has been used in order to compare the fluorescence kinetics of detergent-solubilized and membrane-reconstituted light-harvesting 2 (LH2) complexes from the purple bacteria Rhodopseudomonas (Rps.) acidophila and Rhodobacter (Rb.) sphaeroides. LH2 complexes were reconstituted in phospholipid model membranes at different lipid:protein-ratios and all samples were studied exciting with a wide range of excitation densities. While the detergent-solubilized LH2 complexes from Rps. acidophila showed monoexponential decay kinetics (tau(f )= 980 ps) for excitation densities of up to 3.10(13) photons/(pulse.cm(2)), the membrane-reconstituted LH2 complexes showed multiexponential kinetics even at low excitation densities and high lipid:protein-ratios. The latter finding indicates an efficient clustering of LH2 complexes in the phospholipid membranes. Similar results were obtained for the LH2 complexes from Rb. sphaeroides.

Thermodynamic study on competitive solubilization of cholesterol and beta-sitosterol in bile salt micelles.

PubMed

Matsuoka, Keisuke; Hirosawa, Takashi; Honda, Chikako; Endo, Kazutoyo; Moroi, Yoshikiyo; Shibata, Osamu

2007-07-01

Differences in the preferential solubilization of cholesterol and competitive solubilizates (beta-sitosterol and aromatic compounds) in bile salt micelles was systematically studied by changing the molar ratio of cholesterol to competitive solubilizates. The cholesterol solubility in a mixed binary system (cholesterol and beta-sitosterol) was almost half that of the cholesterol alone system, regardless of the excess beta-sitosterol quantity added. On the other hand, the mutual solubilities of cholesterol and pyrene were not inhibited by their presence in binary mixed crystals. Finally, the cholesterol solubility was measured by changing the alkyl chain length of n-alkylbenzenes. When tetradecylbenzene was added to the bile solution, the cholesterol solubility decreased slightly and was below the original cholesterol solubility. Based on Gibbs energy change (DeltaG degrees ) for solubilization, chemicals that inhibit cholesterol solubility in their combined crystal systems showed a larger negative DeltaG degrees value than cholesterol alone.

Effective solubilization of chalcones in micellar phase: Conductivity and voltammetric study

NASA Astrophysics Data System (ADS)

Ahmed, Safeer; Khan, Gul Tiaz; Shah, Syed Sakhawat

2013-12-01

The solubilization of four chalcones, between aqueous and micellar phases of ionic surfactants (SDS and CTAB), was investigated by conductivity and cyclic voltammetry (CV) techniques. From conductivity data, a decrease in the critical micellar concentration (CMC) of the surfactants, in presence of the chalcones was ascribed to the decreased charge density over the surfactants. The results were seconded by thermodynamic parameters including degree of ionization (α), counter ion binding (β), and standard Gibbs free energy of micellization (Δ G {m/○}). The added surfactant decreased the peak current of the oxidized chalcone and shifted the peak potential either positively (in presence of SDS) or negatively (in presence of CTAB). The effect is rationalized as chalcone-surfactant interaction and quantitated as binding constant ( K b) assorting values from 8.78 to 552.97 M-1. The preferred solubilization of the chalcones in the micellar phase has been inferred.

Obtaining Soluble Folded Proteins from Inclusion Bodies Using Sarkosyl, Triton X-100, and CHAPS: Application to LB and M9 Minimal Media.

PubMed

Massiah, Michael A; Wright, Katharine M; Du, Haijuan

2016-04-01

This unit describes a straightforward and efficient method of using sarkosyl to solubilize and recover difficult recombinant proteins, such as GST- and His6 -tagged fusion proteins, that are overexpressed in E. coli. This protocol is especially useful for rescuing recombinant proteins overexpressed in M9 minimal medium. Sarkosyl added to lysis buffers helps with both protein solubility and cell lysis. Higher percentage sarkosyl (up to 10%) can extract >95% of soluble protein from inclusion bodies. In the case of sarkosyl-solubilized GST-fusion proteins, batch-mode affinity purification requires addition of a specific ratio of Triton X-100 and CHAPS, while sarkosyl-solubilized His6 -tagged fusion proteins can be directly purified on Ni(2+) resin columns. Proteins purified by this method could be widely used in biological assays, structure analysis and mass spectrum assay. Copyright © 2016 John Wiley & Sons, Inc.

Microwave heating of tea residue yields polysaccharides, polyphenols, and plant biopolyester.

PubMed

Tsubaki, Shuntaro; Iida, Hiroyuki; Sakamoto, Masahiro; Azuma, Jun-ichi

2008-12-10

Microwave heating was used to produce aqueous-soluble components from green, oolong, and black tea residues. Heating at 200-230 degrees C for 2 min extracted 40-50% of polysaccharides and 60-70% of the polyphenols. Solubilization of arabinose and galactose by autohydrolysis occurred with heating above 170 degrees C, whereas heating above 200 degrees C was necessary to solubilize xylose. Catechins were soluble in water by heating at low temperature (110 degrees C); however, new polyphenols having strong antioxidant activity were produced above 200 degrees C. The amount of solubilized materials and antioxidant activity increased with increased fermentation of harvested tea leaves (green tea < oolong tea < black tea). Cutin, a plant biopolyester, remained in the residue after heating as did cellulose and lignin/tannin. The predominant cutin monomer that was recovered was 9,10-epoxy-18-hydroxyoctadecanoic acid, followed by dihydroxyhexadecanoic acid and 9,10,18-trihydroxyoctadecanoic acid.

Therapeutic surfactant-stripped frozen micelles

NASA Astrophysics Data System (ADS)

Zhang, Yumiao; Song, Wentao; Geng, Jumin; Chitgupi, Upendra; Unsal, Hande; Federizon, Jasmin; Rzayev, Javid; Sukumaran, Dinesh K.; Alexandridis, Paschalis; Lovell, Jonathan F.

2016-05-01

Injectable hydrophobic drugs are typically dissolved in surfactants and non-aqueous solvents which can induce negative side-effects. Alternatives like `top-down' fine milling of excipient-free injectable drug suspensions are not yet clinically viable and `bottom-up' self-assembled delivery systems usually substitute one solubilizing excipient for another, bringing new issues to consider. Here, we show that Pluronic (Poloxamer) block copolymers are amenable to low-temperature processing to strip away all free and loosely bound surfactant, leaving behind concentrated, kinetically frozen drug micelles containing minimal solubilizing excipient. This approach was validated for phylloquinone, cyclosporine, testosterone undecanoate, cabazitaxel and seven other bioactive molecules, achieving sizes between 45 and 160 nm and drug to solubilizer molar ratios 2-3 orders of magnitude higher than current formulations. Hypertonic saline or co-loaded cargo was found to prevent aggregation in some cases. Use of surfactant-stripped micelles avoided potential risks associated with other injectable formulations. Mechanistic insights are elucidated and therapeutic dose responses are demonstrated.

Decreased solubilization of Pu(IV) polymers by humic acids under anoxic conditions

NASA Astrophysics Data System (ADS)

Xie, Jinchuan; Lin, Jianfeng; Liang, Wei; Li, Mei; Zhou, Xiaohua

2016-11-01

Pu(IV) polymer has a very low solubility (log[Pu(IV)aq]total = -10.4 at pH 7.2 and I = 0). However, some aspects of their environmental fate remain unclear. Humic acids are able to complex with Pu4+ ions and their dissolved species (<10 kD) in the groundwater (neutral to alkaline pH) may cause solubilization of the polymers. Also, humic acids have the native reducing capacity and potentially reduce the polymeric Pu(IV) to Pu(III)aq (log[Pu(III)aq]total = -5.3 at pH 7.2 and I = 0). Solubilization and reduction of the polymers can enhance their mobility in subsurface environments. Nevertheless, humic acids readily coat the surfaces of metal oxides via electrostatic interaction and ligand exchange mechanisms. The humic coatings are expected to prevent both solubilization and reduction of the polymers. Experiments were conducted under anoxic and slightly alkaline (pH 7.2) conditions in order to study whether humic acids have effects on stability of the polymers. The results show that the polymeric Pu(IV) was almost completely transformed into aqueous Pu(IV) in the presence of EDTA ligands. In contrast, the dissolved humic acids did not solubilize the polymers but in fact decreased their solubility by one order of magnitude. The humic coatings were responsible for the decreased solubilization. Such coatings limited the contact between the polymers and EDTA ligands, especially at the relatively high concentrations of humic acids (>0.57 mg/L). Solubilization of the humic-coated polymers was thus inhibited to a significant extent although EDTA, having the great complexation ability, was present in the humic solutions. Reduction of Pu(IV) polymers by the humic acids was also not observed in the absence of EDTA. In the presence of EDTA, the polymers were partially reduced to Pu(III)aq by the humic acids of 0.57 mg/L and the percentage of Pu(III)aq accounted for 51.7% of the total aqueous Pu. This demonstrates that the humic acids were able to reduce the aqueous Pu(IV), instead of the polymeric Pu(IV). Such a demonstration is supported by the very positive redox potential of aqueous Pu(IV)-EDTA complex: Eho ‧ (PuL24-/PuL25-) = 154.3 mV >>Eh (PuO2 (am) /Pu3+) = -182.7 mV calculated at 10-10 mol/L Pu3+ and pH 7.2. At the higher humic concentrations (>0.57 mg/L), the polymers were reduced to a lesser extent because the much denser humic coatings resulted in lower concentrations of the aqueous Pu(IV). Consequently, humic acids make Pu(IV) polymers pretty stable unless the artificial ligands such as EDTA are present in the groundwater.

Competitive and Cooperative Effects during Nickel Adsorption to Iron Oxides in the Presence of Oxalate

DOE Office of Scientific and Technical Information (OSTI.GOV)

Flynn, Elaine D.; Catalano, Jeffrey G.

Iron oxides are ubiquitous in soils and sediments and play a critical role in the geochemical distribution of trace elements and heavy metals via adsorption and coprecipitation. The presence of organic acids may potentially alter how metals associate with iron oxide minerals through a series of cooperative or competitive processes: solution complexation, ternary surface complexation, and surface site competition. The macroscopic and molecular-scale effects of these processes were investigated for Ni adsorption to hematite and goethite at pH 7 in the presence of oxalate. The addition of this organic acid suppresses Ni uptake on both minerals. Aqueous speciation suggests thatmore » this is dominantly the result of oxalate complexing and solubilizing Ni. Comparison of the Ni surface coverage to the concentration of free (uncomplexed) Ni 2+ in solution suggests that the oxalate also alters Ni adsorption affinity. EXAFS and ATR-FTIR spectroscopies indicate that these changes in binding affinity are due to the formation of Ni–oxalate ternary surface complexes. These observations demonstrate that competition between dissolved oxalate and the mineral surface for Ni overwhelms the enhancement in adsorption associated with ternary complexation. Oxalate thus largely enhances Ni mobility, thereby increasing micronutrient bioavailability and inhibiting contaminant sequestration.« less

Fluorescence-based optimization of human bitter taste receptor expression in Saccharomyces cerevisiae

DOE Office of Scientific and Technical Information (OSTI.GOV)

Sugawara, Taishi; Ito, Keisuke; Shiroishi, Mitsunori

2009-05-15

Human TAS2 receptors (hTAS2Rs) perceive bitter tastants, but few studies have explored the structure-function relationships of these receptors. In this paper, we report our trials on the large-scale preparations of hTAS2Rs for structural analysis. Twenty-five hTAS2Rs were expressed using a GFP-fusion yeast system in which the constructs and the culture conditions (e.g., the signal sequence, incubation time and temperature after induction) were optimized by measuring GFP fluorescence. After optimization, five hTAS2Rs (hTAS2R7, hTAS2R8, hTAS2R16, hTAS2R41, and hTAS2R48) were expressed at levels greater than 1 mg protein/L of culture, which is a preferable level for purification and crystallization. Among these fivemore » bitter taste receptors, hTAS2R41 exhibited the highest detergent solubilization efficiency of 87.1% in n-dodecyl-{beta}-D-maltopyranoside (DDM)/cholesteryl hemisuccinate (CHS). Fluorescence size-exclusion chromatography showed that hTAS2R41 exhibited monodispersity in DDM/CHS without aggregates, suggesting that hTAS2R41 is a good target for future crystallization trials.« less

Thermal precipitation fluorescence assay for protein stability screening.

PubMed

Fan, Junping; Huang, Bo; Wang, Xianping; Zhang, Xuejun C

2011-09-01

A simple and reliable method of protein stability assessment is desirable for high throughput expression screening of recombinant proteins. Here we described an assay termed thermal precipitation fluorescence (TPF) which can be used to compare thermal stabilities of recombinant protein samples directly from cell lysate supernatants. In this assay, target membrane proteins are expressed as recombinant fusions with a green fluorescence protein tag and solubilized with detergent, and the fluorescence signals are used to report the quantity of the fusion proteins in the soluble fraction of the cell lysate. After applying a heat shock, insoluble protein aggregates are removed by centrifugation. Subsequently, the amount of remaining protein in the supernatant is quantified by in-gel fluorescence analysis and compared to samples without a heat shock treatment. Over 60 recombinant membrane proteins from Escherichia coli were subject to this screening in the presence and absence of a few commonly used detergents, and the results were analyzed. Because no sophisticated protein purification is required, this TPF technique is suitable to high throughput expression screening of recombinant membrane proteins as well as soluble ones and can be used to prioritize target proteins based on their thermal stabilities for subsequent large scale expression and structural studies. Copyright © 2011 Elsevier Inc. All rights reserved.

Impact of Gut Microbiota-Mediated Bile Acid Metabolism on the Solubilization Capacity of Bile Salt Micelles and Drug Solubility.

PubMed

Enright, Elaine F; Joyce, Susan A; Gahan, Cormac G M; Griffin, Brendan T

2017-04-03

In recent years, the gut microbiome has gained increasing appreciation as a determinant of the health status of the human host. Bile salts that are secreted into the intestine may be biotransformed by enzymes produced by the gut bacteria. To date, bile acid research at the host-microbe interface has primarily been directed toward effects on host metabolism. The aim of this work was to investigate the effect of changes in gut microbial bile acid metabolism on the solubilization capacity of bile salt micelles and consequently intraluminal drug solubility. First, the impact of bile acid metabolism, mediated in vivo by the microbial enzymes bile salt hydrolase (BSH) and 7α-dehydroxylase, on drug solubility was assessed by comparing the solubilization capacity of (a) conjugated vs deconjugated and (b) primary vs secondary bile salts. A series of poorly water-soluble drugs (PWSDs) were selected as model solutes on the basis of an increased tendency to associate with bile micelles. Subsequently, PWSD solubility and dissolution was evaluated in conventional biorelevant simulated intestinal fluid containing host-derived bile acids, as well as in media modified to contain microbial bile acid metabolites. The findings suggest that deconjugation of the bile acid steroidal core, as dictated by BSH activity, influences micellar solubilization capacity for some PWSDs; however, these differences appear to be relatively minor. In contrast, the extent of bile acid hydroxylation, regulated by microbial 7α-dehydroxylase, was found to significantly affect the solubilization capacity of bile salt micelles for all nine drugs studied (p < 0.05). Subsequent investigations in biorelevant media containing either the trihydroxy bile salt sodium taurocholate (TCA) or the dihydroxy bile salt sodium taurodeoxycholate (TDCA) revealed altered drug solubility and dissolution. Observed differences in biorelevant media appeared to be both drug- and amphiphile (bile salt/lecithin) concentration-dependent. Our studies herein indicate that bile acid modifications occurring at the host-microbe interface could lead to alterations in the capacity of intestinal bile salt micelles to solubilize drugs, providing impetus to consider the gut microbiota in the drug absorption process. In the clinical setting, disruption of the gut microbial ecosystem, through disease or antibiotic treatment, could transform the bile acid pool with potential implications for drug absorption and bioavailability.

Winter rye as a bioenergy feedstock: impact of crop maturity on composition, biological solubilization and potential revenue

DOE PAGES

Shao, Xiongjun; DiMarco, Kay; Richard, Tom L.; ...

2015-02-27

We report that winter annual crops such as winter rye (Secale cereale L) can produce biomass feedstock on seasonally fallow land that continues to provide high-value food and feed from summer annuals such as corn and soybeans. As energy double crops, winter grasses are likely to be harvested while still immature and thus structurally different from the fully senesced plant material typically used for biofuels. This study investigates the dynamic trends in biomass yield, composition, and biological solubilization over the course of a spring harvest season. The water soluble fraction decreased with increasing maturity while total carbohydrate content stayed roughlymore » constant at about 65%. The protein mass fraction decreased with increasing maturity, but was counterbalanced by increasing harvest yield resulting in similar total protein across harvest dates. Winter rye was ground and autoclaved then fermented at 15 g/L total solids by either (1) Clostridium thermocellum or (2) simultaneous saccharification and cofermentation (SSCF) using commercial cellulases (CTec2 and HTec2) and a xylose-fermenting Saccharomyces cerevisiae strain. Solubilization of total carbohydrate dropped significantly as winter rye matured for both C. thermocellum (from approximately 80% to approximately 50%) and SSCF (from approximately 60% to approximately 30%). C. thermocellum achieved total solubilization 33% higher than that of SSCF for the earliest harvest date and 50% higher for the latest harvest date. Potential revenue from protein and bioethanol was stable over a range of different harvest dates, with most of the revenue due to ethanol. In a crop rotation with soybean, recovery of the soluble protein from winter rye could increase per hectare protein production by 20 to 35%. Double-cropping winter rye can produce significant biomass for biofuel production and feed protein as coproduct without competing with the main summer crop. During a 24-day harvest window, the total carbohydrate content remained relatively constant while the early-harvest yielded much higher carbohydrate solubilization for both C. thermocellum fermentation and SSCF. C. thermocellum fermentation achieved higher carbohydrate solubilization than SSCF across all growth stages tested. In conclusion, although winter rye’s yield, composition, and biological reactivity change rapidly in the spring, it offers a substantial and stable income across the harvest season and thus flexibility for the farmer.« less

[Filamentous and phosphate solubilizing fungi relationships with some edaphic parameters and coffee plantations management].

PubMed

Posada, Raúl Hernando; Sánchez de Prager, Marina; Sieverding, Ewald; Aguilar Dorantes, Karla; Heredia-Abarca, Gabriela Patricia

2012-09-01

Soil properties and the environment have multiple outcomes on fungal communities. Although, the interaction effects between management intensity, pH, available phosphorus, organic carbon, soil texture and different fractions of water stable macro-aggregates on the communities of microscopic filamentous fungi (MFF), iron phosphate solubilizing fungi (PSF-Fe), and iron and calcium phosphate solubilizing fungi (PSF-(Fe+Ca)), have been previously evaluated in field conditions, this has never been performed in terms of their combined effects, neither with phosphate solubilizing fungi. To assess this, we collected 40 composite soil samples from eight Mexican and Colombian coffee plantations, with different management intensities and physico-chemical edaphic parameters, during 2008-2009. We isolated different communities of MFF, PSF-Fe and PSF-(Fe+Ca), by wet sieving and soil particles culture in Potato-Dextrose-Agar from soil samples, and we classified isolates in terms of their phosphate solubilizing ability. Following the principal component analysis results, we decided to analyze fungal communities and abiotic factors interactions for each country separately. Structural Equation Models revealed that organic carbon was positively associated to MFF richness and number of isolates (lambda>0.58), but its relationship with PSF-Fe and PSF-(Fe+Ca) were variable; while the available phosphorus, pH and water stable macro-aggregate fractions did not show a clear pattern. Management intensity was negatively related to PSF-Fe (lambda < or = -0.21) morphotype richness and the number of isolates in Colombian coffee plantations. We found that the relationships of clay and organic carbon content, and available phosphorus and soil pH, with the species richness and number of isolates of MFF, PSF-Fe and PSF-(Fe+Ca) were highly variable; this made impossible to generalize the responses between saprotrophic fungal groups and geographic zones. The management intensity was not related to species richness and number of isolates of MFF in any coffee areas, while for PSF the relationship could not be defined. The different water stable macro-aggregates fractions did not show a defined pattern in relation to the species richness and the number of isolates of saprophytic and phosphate solubilizing fungi (PSF). This study highlights the need to take into account edaphic and geographic context in order to reach a better understanding of the intensity management effects on MFF and PSF function in agroecosystems.

Winter rye as a bioenergy feedstock: impact of crop maturity on composition, biological solubilization and potential revenue

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shao, Xiongjun; DiMarco, Kay; Richard, Tom L.

We report that winter annual crops such as winter rye (Secale cereale L) can produce biomass feedstock on seasonally fallow land that continues to provide high-value food and feed from summer annuals such as corn and soybeans. As energy double crops, winter grasses are likely to be harvested while still immature and thus structurally different from the fully senesced plant material typically used for biofuels. This study investigates the dynamic trends in biomass yield, composition, and biological solubilization over the course of a spring harvest season. The water soluble fraction decreased with increasing maturity while total carbohydrate content stayed roughlymore » constant at about 65%. The protein mass fraction decreased with increasing maturity, but was counterbalanced by increasing harvest yield resulting in similar total protein across harvest dates. Winter rye was ground and autoclaved then fermented at 15 g/L total solids by either (1) Clostridium thermocellum or (2) simultaneous saccharification and cofermentation (SSCF) using commercial cellulases (CTec2 and HTec2) and a xylose-fermenting Saccharomyces cerevisiae strain. Solubilization of total carbohydrate dropped significantly as winter rye matured for both C. thermocellum (from approximately 80% to approximately 50%) and SSCF (from approximately 60% to approximately 30%). C. thermocellum achieved total solubilization 33% higher than that of SSCF for the earliest harvest date and 50% higher for the latest harvest date. Potential revenue from protein and bioethanol was stable over a range of different harvest dates, with most of the revenue due to ethanol. In a crop rotation with soybean, recovery of the soluble protein from winter rye could increase per hectare protein production by 20 to 35%. Double-cropping winter rye can produce significant biomass for biofuel production and feed protein as coproduct without competing with the main summer crop. During a 24-day harvest window, the total carbohydrate content remained relatively constant while the early-harvest yielded much higher carbohydrate solubilization for both C. thermocellum fermentation and SSCF. C. thermocellum fermentation achieved higher carbohydrate solubilization than SSCF across all growth stages tested. In conclusion, although winter rye’s yield, composition, and biological reactivity change rapidly in the spring, it offers a substantial and stable income across the harvest season and thus flexibility for the farmer.« less

Formulation of a Product Containing the Multifunctional Corrosion Inhibitor System DNBM

DTIC Science & Technology

1989-12-22

corrosion areas are shown in Figs. 14 through 16. The order of anticorrosion effectiveness via the method of time-to-failure was: Best 1. DNBM/Epoxy (No Cr...toluene. In the program described herein, a number of methods were investigated for solubilizing the salts in organic solvents. These included (1...NADC-90049-60 Section 2 DEVELOPMENT OF DNBM CORROSION INHIBITORS The two methods investigated for the solubilization of salts in organic systems were

Plant-associated fluorescent Pseudomonas from red lateritic soil: Beneficial characteristics and their impact on lettuce growth.

PubMed

Maroniche, Guillermo A; Rubio, Esteban J; Consiglio, Adrián; Perticari, Alejandro

2016-11-25

Fluorescent Pseudomonas are ubiquitous soil bacteria that usually establish mutualistic associations with plants, promoting their growth and health by several mechanisms. This makes them interesting candidates for the development of crop bio-inoculants. In this work, we isolated phosphate-solubilizing fluorescent Pseudomonas from the rhizosphere and inner tissues of different plant species growing in red soil from Misiones, Argentina. Seven isolates displaying strong phosphate solubilization were selected for further studies. Molecular identification by rpoD genotyping indicated that they belong to different species within the P. fluorescens and P. putida phylogenetic groups. Screening for in vitro traits such as phosphate solubilization, growth regulators synthesis or degradation, motility and antagonism against phytopathogens or other bacteria, revealed a unique profile of characteristics for each strain. Their plant growth-promoting potential was assayed using lettuce as a model for inoculation under controlled and greenhouse conditions. Five of the strains increased the growth of lettuce plants. Overall, the strongest lettuce growth promoter under both conditions was strain ZME4, isolated from inner tissues of maize. No clear association between lettuce growth promotion and in vitro beneficial traits was detected. In conclusion, several phosphate solubilizing pseudomonads from red soil were isolated that display a rich array of plant growth promotion traits, thus showing a potential for the development of new inoculants.

Metabolic channeling of glucose towards gluconate in phosphate-solubilizing Pseudomonas aeruginosa P4 under phosphorus deficiency.

PubMed

Buch, Aditi; Archana, G; Naresh Kumar, G

2008-01-01

Most phosphate-solubilizing bacteria (PSB), including the Pseudomonas species, release P from sparingly soluble mineral phosphates by producing high levels of gluconic acid from extracellular glucose, in a reaction catalyzed by periplasmic glucose dehydrogenase, which is an integral component of glucose catabolism of pseudomonads. To investigate the differences in the glucose metabolism of gluconic acid-producing PSB pseudomonads and low gluconic acid-producing/non-PSB strains, several parameters pertaining to growth and glucose utilization under P-sufficient and P-deficient conditions were monitored for the PSB isolate Pseudomonas aeruginosa P4 (producing approximately 46 mM gluconic acid releasing 437 microM P) and non-PSB P. fluorescens 13525. Our results show interesting differences in the channeling of glucose towards gluconate and other catabolic end-products like pyruvate and acetate with respect to P status for both strains. However, PSB strain P. aeruginosa P4, apart from exhibiting better growth under both low and high Pi conditions, differed from P. fluorescens 13525 in its ability to accumulate gluconate under P-solubilizing conditions. These alterations in growth, glucose utilization and acid secretion are correlated with glucose dehydrogenase, glucose-6-phosphate dehydrogenase and pyruvate carboxylase activities. The ability to shift glucose towards a direct oxidative pathway under P deficiency is speculated to underlie the differential gluconic acid-mediated P-solubilizing ability observed amongst pseudomonads.

A comparative spectroscopic and kinetic study of photoexcitations in detergent-isolated and membrane-embedded LH2 light-harvesting complexes.

PubMed

Freiberg, Arvi; Rätsep, Margus; Timpmann, Kõu

2012-08-01

Integral membrane proteins constitute more than third of the total number of proteins present in organisms. Solubilization with mild detergents is a common technique to study the structure, dynamics, and catalytic activity of these proteins in purified form. However beneficial the use of detergents may be for protein extraction, the membrane proteins are often denatured by detergent solubilization as a result of native lipid membrane interactions having been modified. Versatile investigations of the properties of membrane-embedded and detergent-isolated proteins are, therefore, required to evaluate the consequences of the solubilization procedure. Herein, the spectroscopic and kinetic fingerprints have been established that distinguish excitons in individual detergent-solubilized LH2 light-harvesting pigment-protein complexes from them in the membrane-embedded complexes of purple photosynthetic bacteria Rhodobacter sphaeroides. A wide arsenal of spectroscopic techniques in visible optical range that include conventional broadband absorption-fluorescence, fluorescence anisotropy excitation, spectrally selective hole burning and fluorescence line-narrowing, and transient absorption-fluorescence have been applied over broad temperature range between physiological and liquid He temperatures. Significant changes in energetics and dynamics of the antenna excitons upon self-assembly of the proteins into intracytoplasmic membranes are observed, analyzed, and discussed. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial. Copyright © 2011. Published by Elsevier B.V.

The effect of switchgrass loadings on feedstock solubilization and biofuel production by Clostridium thermocellum

DOE Office of Scientific and Technical Information (OSTI.GOV)

Verbeke, Tobin J.; Garcia, Gabriela M.; Elkins, James G.

High solids loading fermentations are necessary for the industrialization of lignocellulosic ethanol. To date, only a few studies have investigated the effect of solids loadings on microorganisms of interest for consolidated bioprocessing (CBP). Here, the effect that various switchgrass loadings have on Clostridium thermocellum solubilization and bioconversion are investigated. C. thermocellum was grown for ten days on 10, 25 or 50 g/L switchgrass or Avicel at equivalent glucan loadings. Avicel was completely consumed at all loadings, but total cellulose solubilization decreased from 63% to 37% as switchgrass loadings increased from 10 g/L to 50 g/L. Washed, spent switchgrass could bemore » additionally hydrolyzed and fermented in second-round fermentations suggesting access to fermentable substrates was not the limiting factor at higher feedstock loadings. Fermentations of Avicel or cellobiose using culture medium supplemented with 50% spent fermentation broth identified that compounds present in the samples collected from the 25 or 50 g/L switchgrass loadings were the most inhibitory to continued fermentation. Finally, recalcitrance alone cannot fully account for differences in solubilization and end-production formation between switchgrass and Avicel at increased substrate loadings. Effort to decouple metabolic inhibition from inhibition of hydrolysis suggest that C. thermocellum’s hydrolytic machinery is more vulnerable to inhibition from switchgrass-derived inhibitors than is the bacterium’s metabolism.« less

Colonization and Maize Growth Promotion Induced by Phosphate Solubilizing Bacterial Isolates.

PubMed

Li, Yongbin; Liu, Xiaomeng; Hao, Tianyi; Chen, Sanfeng

2017-06-29

Phosphorus (P) limits the production of maize, one of the major food crops in China. Phosphate-solubilizing bacteria (PSB) have the capacity to solubilize phosphate complexes into plant absorbable and utilizable forms by the process of acidification, chelation, and exchange reactions. In this study, six bacteria, including one Paenibacillus sp. B1 strain, four Pseudomonas sp. strains (B10, B14, SX1, and SX2) and one Sphingobium sp. SX14 strain, were those isolated from the maize rhizosphere and identified based on their 16S rRNA sequences. All strains could solubilize inorganic P (Ca₃(PO₄)₂, FePO₄ and AlPO₄), and only B1 and B10 organic P (lecithin). All strains, except of SX1, produced IAA, and SX14 and B1 showed the highest level. B1 incited the highest increase in root length and the second increase in shoot and total dry weight, shoot length, and total P and nitrogen (N), along with increased root length. In addition, by confocal laser scanning microscopy (CLSM), we found that green fluorescent protein (GFP)-labeled B1 mainly colonized root surfaces and in epidermal and cortical tissue. Importantly, B1 can survive through forming spores under adverse conditions and prolong quality guarantee period of bio-fertilizer. Therefore, it can act as a good substitute for bio-fertilizer to promote agricultural sustainability.

Deep Eutectic Solvent Aqueous Solutions as Efficient Media for the Solubilization of Hardwood Xylans.

PubMed

Morais, Eduarda S; Mendonça, Patrícia V; Coelho, Jorge F J; Freire, Mara G; Freire, Carmen S R; Coutinho, João A P; Silvestre, Armando J D

2018-02-22

This work contributes to the development of integrated lignocellulosic-based biorefineries by the pioneering exploitation of hardwood xylans by solubilization and extraction in deep eutectic solvents (DES). DES formed by choline chloride and urea or acetic acid were initially evaluated as solvents for commercial xylan as a model compound. The effects of temperature, molar ratio, and concentration of the DES aqueous solutions were evaluated and optimized by using a response surface methodology. The results obtained demonstrated the potential of these solvents, with 328.23 g L -1 of xylan solubilization using 66.7 wt % DES in water at 80 °C. Furthermore, xylans could be recovered by precipitation from the DES aqueous media in yields above 90 %. The detailed characterization of the xylans recovered after solubilization in aqueous DES demonstrated that 4-O-methyl groups were eliminated from the 4-O-methylglucuronic acids moieties and uronic acids (15 %) were cleaved from the xylan backbone during this process. The similar M w values of both pristine and recovered xylans confirmed the success of the reported procedure. DES recovery in four additional extraction cycles was also demonstrated. Finally, the successful extraction of xylans from Eucalyptus globulus wood by using aqueous solutions of DES was demonstrated. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

The role of glycerol and phosphatidylcholine in solubilizing and enhancing insulin stability in reverse hexagonal mesophases.

PubMed

Amar-Yuli, Idit; Azulay, Doron; Mishraki, Tehila; Aserin, Abraham; Garti, Nissim

2011-12-15

The potential of reverse hexagonal mesophases based on monoolein (GMO) and glycerol (as cosolvent) to facilitate the solubilization of proteins, such as insulin was explored. H(II) mesophases composed of GMO/decane/water were compared to GMO/decane/glycerol/water and GMO/phosphatidylcholine (PC)/decane/glycerol/water systems. The stability of insulin was tested, applying external physical modifications such as low pH and heat treatment (up to 70°C), in which insulin is known to form ordered amyloid-like aggregates (that are associated with several neurodegenerative diseases) with a characteristic cross β-pleated sheet structure. The impact of insulin confinement within these carriers on its stability, unfolding, and aggregation pathways was studied by combining SAXS, FTIR, and AFM techniques. These techniques provided a better insight into the molecular level of the "component interplay" in solubilizing and stabilizing insulin and its conformational modifications that dictate its final aggregate morphology. PC enlarged the water channels while glycerol shrank them, yet both facilitated insulin solubilization within the channels. The presence of glycerol within the mesophase water channels led to the formation of stronger hydrogen bonds with the hosting medium that enhanced the thermal stability of the protein and remarkably affected the unfolding process even after heat treatment (at 70°C for 60 min). Copyright © 2011 Elsevier Inc. All rights reserved.

Impact of Detergents on Membrane Protein Complex Isolation.

PubMed

Lee, Yu-Chen; Bååth, Jenny Arnling; Bastle, Ryan M; Bhattacharjee, Sonali; Cantoria, Mary Jo; Dornan, Mark; Gamero-Estevez, Enrique; Ford, Lenzie; Halova, Lenka; Kernan, Jennifer; Kürten, Charlotte; Li, Siran; Martinez, Jerahme; Sachan, Nalani; Sarr, Medoune; Shan, Xiwei; Subramanian, Nandhitha; Rivera, Keith; Pappin, Darryl; Lin, Sue-Hwa

2018-01-05

Detergents play an essential role during the isolation of membrane protein complexes. Inappropriate use of detergents may affect the native fold of the membrane proteins, their binding to antibodies, or their interaction with partner proteins. Here we used cadherin-11 (Cad11) as an example to examine the impact of detergents on membrane protein complex isolation. We found that mAb 1A5 could immunoprecipitate Cad11 when membranes were solubilized by dodecyl maltoside (DDM) but not by octylglucoside, suggesting that octylglucoside interferes with Cad11-mAb 1A5 interaction. Furthermore, we compared the effects of Brij-35, Triton X-100, cholate, CHAPSO, Zwittergent 3-12, Deoxy BIG CHAP, and digitonin on Cad11 solubilization and immunoprecipitation. We found that all detergents except Brij-35 could solubilize Cad11 from the membrane. Upon immunoprecipitation, we found that β-catenin, a known cadherin-interacting protein, was present in Cad11 immune complex among the detergents tested except Brij-35. However, the association of p120 catenin with Cad11 varied depending on the detergents used. Using isobaric tag for relative and absolute quantitation (iTRAQ) to determine the relative levels of proteins in Cad11 immune complexes, we found that DDM and Triton X-100 were more efficient than cholate in solubilization and immunoprecipitation of Cad11 and resulted in the identification of both canonical and new candidate Cad11-interacting proteins.

Comparative analysis of the ability of Clostridium clariflavum strains and Clostridium thermocellumto utilize hemicellulose and unpretreated plant material

DOE Office of Scientific and Technical Information (OSTI.GOV)

Izquierdo, Javier A.; Pattathil, Sivakumar; Guseva, Anna

2014-11-18

Among themophilic consolidated bioprocessing (CBP) candidate organisms, environmental isolates of Clostridium clariflavum have demonstrated the ability to grow on xylan, and the genome of C. clariflavum DSM 19732 has revealed a number of mechanisms that foster solubilization of hemicellulose that are distinctive relative to the model cellulolytic thermophile Clostridium thermocellum. Growth experiments on xylan, xylooligosaccharides, and xylose reveal that C. clariflavum strains are able to completely break down xylan to xylose and that the environmental strain C. clariflavum sp. 4-2a is able to grow on monomeric xylose. C. clariflavum strains were able to utilize a larger proportion of unpretreated switchgrass,more » and solubilize a higher proportion of glucan, xylan, and arabinan, with strain 4-2a reaching the highest extent of solubilization of these components (64.7 to 69.4%) compared to C. thermocellum (29.5 to 42.5%). In addition, glycome immunoanalyses of residual plant biomass reveal differences in the extent of degradation of easily accessible xylans, with C. clariflavum strains having increased solubilization of this fraction of xylans relative to C. thermocellum. In conclusion, C. clariflavum strains exhibit higher activity than C. thermocellum in the breakdown of hemicellulose and are capable of degrading xylan to xylooligomers and xylose. This capability seems to also play a role in the higher levels of utilization of unpretreated plant material.« less

The effect of switchgrass loadings on feedstock solubilization and biofuel production by Clostridium thermocellum

DOE PAGES

Verbeke, Tobin J.; Garcia, Gabriela M.; Elkins, James G.

2017-11-30

High solids loading fermentations are necessary for the industrialization of lignocellulosic ethanol. To date, only a few studies have investigated the effect of solids loadings on microorganisms of interest for consolidated bioprocessing (CBP). Here, the effect that various switchgrass loadings have on Clostridium thermocellum solubilization and bioconversion are investigated. C. thermocellum was grown for ten days on 10, 25 or 50 g/L switchgrass or Avicel at equivalent glucan loadings. Avicel was completely consumed at all loadings, but total cellulose solubilization decreased from 63% to 37% as switchgrass loadings increased from 10 g/L to 50 g/L. Washed, spent switchgrass could bemore » additionally hydrolyzed and fermented in second-round fermentations suggesting access to fermentable substrates was not the limiting factor at higher feedstock loadings. Fermentations of Avicel or cellobiose using culture medium supplemented with 50% spent fermentation broth identified that compounds present in the samples collected from the 25 or 50 g/L switchgrass loadings were the most inhibitory to continued fermentation. Finally, recalcitrance alone cannot fully account for differences in solubilization and end-production formation between switchgrass and Avicel at increased substrate loadings. Effort to decouple metabolic inhibition from inhibition of hydrolysis suggest that C. thermocellum’s hydrolytic machinery is more vulnerable to inhibition from switchgrass-derived inhibitors than is the bacterium’s metabolism.« less

Hydrotropic Solubilization of Lipophilic Drugs for Oral Delivery: The Effects of Urea and Nicotinamide on Carbamazepine Solubility–Permeability Interplay

PubMed Central

Beig, Avital; Lindley, David; Miller, Jonathan M.; Agbaria, Riad; Dahan, Arik

2016-01-01

Hydrotropy refers to increasing the water solubility of otherwise poorly soluble compound by the presence of small organic molecules. While it can certainly increase the apparent solubility of a lipophilic drug, the effect of hydrotropy on the drugs’ permeation through the intestinal membrane has not been studied. The purpose of this work was to investigate the solubility–permeability interplay when using hydrotropic drug solubilization. The concentration-dependent effects of the commonly used hydrotropes urea and nicotinamide, on the solubility and the permeability of the lipophilic antiepileptic drug carbamazepine were studied. Then, the solubility–permeability interplay was mathematically modeled, and was compared to the experimental data. Both hydrotropes allowed significant concentration-dependent carbamazepine solubility increase (up to ∼30-fold). A concomitant permeability decrease was evident both in vitro and in vivo (∼17-fold for nicotinamide and ∼9-fold for urea), revealing a solubility–permeability tradeoff when using hydrotropic drug solubilization. A relatively simplified simulation approach based on proportional opposite correlation between the solubility increase and the permeability decrease at a given hydrotrope concentration allowed excellent prediction of the overall solubility–permeability tradeoff. In conclusion, when using hydrotropic drug solubilization it is prudent to not focus solely on solubility, but to account for the permeability as well; achieving optimal solubility–permeability balance may promote the overall goal of the formulation to maximize oral drug exposure. PMID:27826241

Solubilization of docetaxel in poly(ethylene oxide)-block-poly(butylene/styrene oxide) micelles.

PubMed

Elsabahy, Mahmoud; Perron, Marie-Eve; Bertrand, Nicolas; Yu, Ga-Er; Leroux, Jean-Christophe

2007-07-01

Poly(ethylene oxide)-block-poly(styrene oxide) (PEO-b-PSO) and PEO-b-poly(butylene oxide) (PEO-b-PBO) of different chain lengths were synthesized and characterized for their self-assembling properties in water by dynamic/static light scattering, spectrofluorimetry, and transmission electron microscopy. The resulting polymeric micelles were evaluated for their ability to solubilize and protect the anticancer drug docetaxel (DCTX) from degradation. The drug release kinetics as well as the cytotoxicity of the loaded micelles were assessed in vitro. All polymers formed micelles with a highly viscous core at low critical association concentrations (<10 mg/L). Micelle morphology depended on the nature of the hydrophobic block, with PBO- and PSO-based micelles yielding monodisperse spherical and cylindrical nanosized aggregates, respectively. The maximum solubilization capacity for DCTX ranged from 0.7 to 4.2% and was the highest for PSO micelles exhibiting the longest hydrophobic segment. Despite their high affinity for DCTX, PEO-b-PSO micelles were not able to efficiently protect DCTX against hydrolysis under accelerated stability testing conditions. Only PEO-b-PBO bearing 24 BO units afforded significant protection against degradation. In vitro, DCTX was released slower from the latter micelles, but all formulations possessed a similar cytotoxic effect against PC-3 prostate cancer cells. These data suggest that PEO-b-P(SO/BO) micelles could be used as alternatives to conventional surfactants for the solubilization of taxanes.

A comparative study of vitamin E TPGS/HPMC supersaturated system and other solubilizer/polymer combinations to enhance the permeability of a poorly soluble drug through the skin.

PubMed

Ghosh, Indrajit; Michniak-Kohn, Bozena

2012-11-01

In transdermal drug delivery systems (TDDS), it is a challenge to achieve stable and prolonged high permeation rates across skin, because the concentration of the drug dissolved in the matrix has to be high in order to maintain zero order release kinetics of the drug. In case of poorly soluble drugs, due to thermodynamic challenges, there is a high tendency for the drug to nucleate immediately after formulating or even during storage. The present study focuses on the efficiency of vitamin E TPGS/HPMC supersaturated solution and other solubilizer/polymer systems to improve the solubility of the drug and inhibit crystal growth in the transdermal formulation. Effect of several solubilizers, for example, Pluronic F-127, vitamin E TPGS and co-solvent, for example, propylene glycol (PG) were studied on the supersaturated systems of ibuprofen as model drug. Various stabilizers such as hydroxylpropyl methylcellulose (HPMC 3 cps) and polyvinylpyrrolidone (PVP K-30) were examined to evaluate their crystal inhibitory effects. Different analytical tools were used in this study to detect the growth of crystals in the systems. Vitamin E TPGS and HPMC 3 cps formulation produced the highest permeation rate of the drug as compared to other systems. In addition, the onset of crystallization time was shown to be longer with this formulation as compared to other solubilizer/polymer combinations.

Design and characterization of submicron formulation for a poorly soluble drug: the effect of Vitamin E TPGS and other solubilizers on skin permeability enhancement.

PubMed

Ghosh, Indrajit; Michniak-Kohn, Bozena

2012-09-15

In transdermal drug delivery systems (TDDS), it is a challenge to achieve stable and prolonged high permeation rates across the skin since the concentrations of the drug dissolved in the matrix have to be high in order to maintain zero order release kinetics. Several attempts have been reported to improve the permeability of poorly soluble drug compounds using supersaturated systems, however, due to thermodynamic challenges, there was a high tendency for the drug to nucleate immediately after formulating or even during storage. The present study focuses on the efficiency of drug crystals at the submicron/nano range in presence of different solubilizers to improve the permeation rate. Effect of several solubilizers, e.g. Pluronic F-127, Vitamin E TPGS, propylene glycol were studied on the submicron suspension systems of ibuprofen as a model drug. Various stabilizers such as hydroxylpropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP) were examined to evaluate their crystal inhibitory effects on particle growth of the drug compound at submicron range. The overall permeation enhancement process through the skin seems to be influenced by the presence of solubilizers and also the presence of submicron drug crystal. The most promising stable formulation was developed with Vitamin E TPGS+HPMC submicron suspension, which produced higher permeation rate compared to other vehicles. Copyright © 2012 Elsevier B.V. All rights reserved.

The effect of cyclodextrin-solubilized curcuminoids on amyloid plaques in Alzheimer transgenic mice: brain uptake and metabolism after intravenous and subcutaneous injection

PubMed Central

2013-01-01

Introduction Curcuminoids may improve pathological conditions associated with Alzheimer's disease. However, their therapeutic potential is limited by their exceedingly low bioavailability after oral administration. A method to deliver solubilized curcuminoids by injection was evaluated in Alzheimer transgenic mice. Methods Amyloid protein precursor (APP)SWE, PS1dE9 mice were intravenously or subcutaneously injected at weekly intervals between the ages of 4 and 12 months with serum- or cyclodextrin-solubilized curcuminoids to assess their potential for plaque prevention. Alternatively, mice between the ages of 11 and 12 months were intravenously injected with cyclodextrin-solubilized curcuminoids at biweekly intervals to evaluate their ability to eliminate existing plaques. Plasma and brain levels of curcuminoids and their metabolites were also determined after subcutaneous and intravenous injection. Results Weekly long-term injections did not result in a significant plaque load reduction. However, intravenous injection of cyclodextrin-solubilized curcuminoids at higher curcuminoid concentrations and at a biweekly frequency between the ages of 11 and 12 months reduced the plaque load to approximately 70% of the control value. After intravenous injection, plasma levels of 100 μM curcuminoids and brain levels of 47 nmol/g could initially be achieved that declined to essentially undetectable levels within 20 minutes. The primary curcuminoid metabolites in plasma were the conjugates of glucuronide or sulfate and hexahydrocurcuminoids as reduction products. In the brain, both hexahydrocurcuminoids and octahydrocurcuminoids were detected as major metabolites. After subcutaneous injection, maximal curcuminoid plasma levels of 23 μM and brain levels of 8 nmol/g were observed at 30 minutes after injection and curcuminoids remained detectable for 2 to 3 h. Conclusion Curcuminoids are rapidly metabolized after injection and their effect on reducing plaque load associated with Alzheimer's disease may be dependent on the frequency of administration. PMID:23537472

Differential solubility of curcuminoids in serum and albumin solutions: implications for analytical and therapeutic applications

PubMed Central

Quitschke, Wolfgang W

2008-01-01

Background Commercially available curcumin preparations contain a mixture of related polyphenols, collectively referred to as curcuminoids. These encompass the primary component curcumin along with its co-purified derivatives demethoxycurcumin and bisdemethoxycurcumin. Curcuminoids have numerous biological activities, including inhibition of cancer related cell proliferation and reduction of amyloid plaque formation associated with Alzheimer disease. Unfortunately, the solubility of curcuminoids in aqueous solutions is exceedingly low. This restricts their systemic availability in orally administered formulations and limits their therapeutic potential. Results Methods are described that achieve high concentrations of soluble curcuminoids in serum. Solid curcuminoids were either mixed directly with serum, or they were predissolved in dimethyl sulfoxide and added as aliquots to serum. Both methods resulted in high levels of curcuminoid-solubility in mammalian sera from different species. However, adding aliquots of dimethyl sulfoxide-dissolved curcuminoids to serum proved to be more efficient, producing soluble curcuminoid concentrations of at least 3 mM in human serum. The methods also resulted in the differential solubility of individual curcuminoids in serum. The addition of dimethyl sulfoxide-dissolved curcuminoids to serum preferentially solubilized curcumin, whereas adding solid curcuminoids predominantly solubilized bisdemethoxycurcumin. Either method of solubilization was equally effective in inhibiting dose-dependent HeLa cell proliferation in culture. The maximum concentration of curcuminoids achieved in serum was at least 100-fold higher than that required for inhibiting cell proliferation in culture and 1000-fold higher than the concentration that has been reported to prevent amyloid plaque formation associated with Alzheimer disease. Curcuminoids were also highly soluble in solutions of purified albumin, a major component of serum. Conclusion These results suggest the possibility of alternative therapeutic approaches by injection or infusion of relatively small amounts of curcuminoid-enriched serum. They also provide tools to reproducibly solubilize curcuminoids for analysis in cell culture applications. The differential solubility of curcuminoids achieved by different methods of solubilization offers convenient alternatives to assess the diverse biological effects contributed by curcumin and its derivatives. PMID:18990234

Temperature-dependent phase behaviour of tetrahydrofuran–water alters solubilization of xylan to improve co-production of furfurals from lignocellulosic biomass

DOE PAGES

Smith, Micholas Dean; Cai, Charles M.; Cheng, Xiaolin; ...

2018-03-06

Xylose, Xylan, Hemicellulose, CELF, THF, Co-solvent, Pretreatment, Biomass ABSTRACT: Xylan is an important polysaccharide found in the hemicellulose fraction of lignocellulosic biomass that can be hydrolysed to xylose and further dehydrated to the furfural, an important renewable platform fuel precursor. Here, pairing molecular simulation and experimental evidences, we reveal how the unique temperature-dependent phase behaviour of water-tetrahydrofuran (THF) co-solvent can delay xylan solubilization to synergistically improve catalytic co-processing of biomass to furfural and 5-HMF. Our results indicate, based on polymer correlations between polymer conformational behaviour and solvent quality, that both co-solvent and aqueous environments serve as ‘good’ solvents for xylan.more » Interestingly, the simulations also revealed that unlike other cell-wall components (i.e., lignin and cellulose), the make-up of the solvation shell of xylan in THF-water is dependent on the temperature-phase behaviour. At temperatures between 333K and 418K, THF and water become immiscible, and THF is evacuated from the solvation shell of xylan, while above and below this temperature range, THF and water are both present in the polysaccharide’s solvation shell. This suggested that the solubilization of xylan in THF-water may be similar to aqueous-only solutions at temperatures between 333K and 418K and different outside this range. Experimental reactions on beachwood xylan corroborate this hypothesis by demonstrating 2-fold reduction of xylan solubilization in THF-water within a miscible temperature regime (445K) and unchanged solubilization within an immiscible regime (400K). Translating this phase-dependent behaviour to processing of maple wood chips, we demonstrate how the weaker xylan solvation in THF-water under miscible conditions can delay furfural production from xylose, allowing 5-HMF production from cellulose to “catch-up” such that their high yield production from biomass can be synergized in a single pot reaction.« less

Creating Drug Solubilization Compartments via Phase Separation in Multicomponent Buccal Patches Prepared by Direct Hot Melt Extrusion-Injection Molding.

PubMed

Alhijjaj, Muqdad; Bouman, Jacob; Wellner, Nikolaus; Belton, Peter; Qi, Sheng

2015-12-07

Creating in situ phase separation in solid dispersion based formulations to allow enhanced functionality of the dosage form, such as improving dissolution of poorly soluble model drug as well as being mucoadhesive, can significantly maximize the in vitro and in vivo performance of the dosage form. This formulation strategy can benefit a wide range of solid dosage forms for oral and alternative routes of delivery. This study using buccal patches as an example created separated phases in situ of the buccal patches by selecting the excipients with different miscibility with each other and the model drug. The quaternary dispersion based buccal patches containing PEG, PEO, Tween 80, and felodipine were prepared by direct hot melt extrusion-injection molding (HME-IM). The partial miscibility between Tween 80 and semicrystalline PEG-PEO led to the phase separation after extrusion. The Tween phases acted as drug solubilization compartments, and the PEG-PEO phase had the primary function of providing mucoadhesion and carrier controlled dissolution. As felodipine was preferably solubilized in the amorphous regions of PEG-PEO, the high crystallinity of PEG-PEO resulted in an overall low drug solubilizing capacity. Tween 80 was added to improve the solubilization capacity of the system as the model drug showed good solubility in Tween. Increasing the drug loading led to the supersaturation of drug in Tween compartments and crystalline drug dispersed in PEG-PEO phases. The spatial distribution of these phase-separated compartments was mapped using X-ray micro-CT, which revealed that the domain size and heterogeneity of the phase separation increased with increasing the drug loading. The outcome of this study provides new insights into the applicability of in situ formed phase separation as a formulation strategy for the delivery of poorly soluble drugs and demonstrated the basic principle of excipient selection for such technology.

Differential bioavailability of polychlorinated biphenyls associated with environmental particles: Microplastic in comparison to wood, coal and biochar.

PubMed

Beckingham, B; Ghosh, U

2017-01-01

Microplastic particles are increasingly being discovered in diverse habitats and a host of species are found to ingest them. Since plastics are known to sorb hydrophobic organic contaminants (HOCs) there is a question of what risk of chemical exposure is posed to aquatic biota from microplastic-associated contaminants. We investigate bioavailability of polychlorinated biphenyls (PCBs) from polypropylene microplastic by measuring solid-water distribution coefficients, gut fluid solubilization, and bioaccumulation using sediment invertebrate worms as a test system. Microplastic-associated PCBs are placed in a differential bioavailability framework by comparing the results to several other natural and anthrogenic particles, including wood, coal, and biochar. PCB distribution coefficients for polypropylene were higher than natural organic materials like wood, but in the range of lipids and sediment organic carbon, and smaller than black carbons like coal and biochars. Gut fluid solubilization potential increased in the order: coal < polypropylene < biochar < wood. Interestingly, lower gut fluid solubilization for polypropylene than biochar infers that gut fluid micelles may have solubilized part of the biochar matrix while bioaccessibility from plastic can be limited by the solubilizing potential of gut fluids dependent on the solid to liquid ratio or renewal of fluids in the gut. Biouptake in worms was lower by 76% when PCBs were associated with polypropylene compared to sediment. The presence of microplastics in sediments had an overall impact of reducing bioavailability and transfer of HOCs to sediment-ingesting organisms. Since the vast majority of sediment and suspended particles in the environment are natural organic and inorganic materials, pollutant transfer through particle ingestion will be dominated by these particles and not microplastics. Therefore, these results support the conclusion that in most cases the transfer of organic pollutants to aquatic organisms from microplastic in the diet is likely a small contribution compared to other natural pathways of exposure. Copyright © 2016 Elsevier Ltd. All rights reserved.

Temperature-dependent phase behaviour of tetrahydrofuran–water alters solubilization of xylan to improve co-production of furfurals from lignocellulosic biomass

DOE Office of Scientific and Technical Information (OSTI.GOV)

Smith, Micholas Dean; Cai, Charles M.; Cheng, Xiaolin

Xylose, Xylan, Hemicellulose, CELF, THF, Co-solvent, Pretreatment, Biomass ABSTRACT: Xylan is an important polysaccharide found in the hemicellulose fraction of lignocellulosic biomass that can be hydrolysed to xylose and further dehydrated to the furfural, an important renewable platform fuel precursor. Here, pairing molecular simulation and experimental evidences, we reveal how the unique temperature-dependent phase behaviour of water-tetrahydrofuran (THF) co-solvent can delay xylan solubilization to synergistically improve catalytic co-processing of biomass to furfural and 5-HMF. Our results indicate, based on polymer correlations between polymer conformational behaviour and solvent quality, that both co-solvent and aqueous environments serve as ‘good’ solvents for xylan.more » Interestingly, the simulations also revealed that unlike other cell-wall components (i.e., lignin and cellulose), the make-up of the solvation shell of xylan in THF-water is dependent on the temperature-phase behaviour. At temperatures between 333K and 418K, THF and water become immiscible, and THF is evacuated from the solvation shell of xylan, while above and below this temperature range, THF and water are both present in the polysaccharide’s solvation shell. This suggested that the solubilization of xylan in THF-water may be similar to aqueous-only solutions at temperatures between 333K and 418K and different outside this range. Experimental reactions on beachwood xylan corroborate this hypothesis by demonstrating 2-fold reduction of xylan solubilization in THF-water within a miscible temperature regime (445K) and unchanged solubilization within an immiscible regime (400K). Translating this phase-dependent behaviour to processing of maple wood chips, we demonstrate how the weaker xylan solvation in THF-water under miscible conditions can delay furfural production from xylose, allowing 5-HMF production from cellulose to “catch-up” such that their high yield production from biomass can be synergized in a single pot reaction.« less

Role of phosphate solubilizing Burkholderia spp. for successful colonization and growth promotion of Lycopodium cernuum L. (Lycopodiaceae) in lateritic belt of Birbhum district of West Bengal, India.

PubMed

Ghosh, Ranjan; Barman, Soma; Mukherjee, Rajib; Mandal, Narayan C

2016-02-01

Profuse growth of Lycpodium cernuum L. was found in phosphate deficient red lateritic soil of West Bengal, India. Interaction of vesicular-arbuscular mycorrhiza (VAM) with Lycopodium rhizoids were described earlier but association of PGPR with their rhizoids were not studied. Three potent phosphate solubilizing bacterial strains (P4, P9 and P10) associated with L. cernuum rhizoids were isolated and identified by 16S rDNA homologies on Ez-Taxon database as Burkholderia tropica, Burkholderia unamae and Burkholderia cepacia respectively. Day wise kinetics of phosphate solubilization against Ca3(PO4)2 suggested P4 (580.56±13.38 μg ml(-1)) as maximum mineral phosphate solubilizer followed by P9 (517.12±17.15 μg ml(-1)) and P10 (485.18±14.23 μg ml(-1)) at 28 °C. Release of bound phosphates by isolated strains from ferric phosphate (FePO4), aluminum phosphate (AlPO4) and four different complex rock phosphates indicated their very good phosphate solubilizng efficacy. Nitrogen independent solubilizition also supports their nitrogen fixing capabilities. Inhibition of P solubilization by calcium salts and induction by EDTA suggested pH dependent chelation of metal cations by all of the isolates. Rhizoidal colonization potentials of Burkholderia spp. were confirmed by in planta experiment and also using scanning electron microscope (SEM). Increases of total phosphate content in Lycopodium plants upon soil treatment with these isolates were also recorded. In addition siderophore production on CAS agar medium, tryptophan dependent IAA production and antifungal activities against pathogenic fungi by rhizospheric isolates deep-rooted that they have definite role in nutrient mobilization for successful colonization of L. cernuum in nutrient deficient lateritic soil. Copyright © 2015 Elsevier GmbH. All rights reserved.

In Vitro Reactivity of 3-M KC1-Solubilized Murine Histocompatibility (H-2) Antigens

DTIC Science & Technology

1974-01-01

experiment, varying concentra- mice were immunized with DBA/2 spleen tions of cells from day 5 BALB/c sensitized cells and subsequently skin grafted . Syn... skin grafts , spleen in MLC with mitomycin-C treated DBA/2 cells were removed and cultured with spleen cells. As seen in Table 4, increasing DBA/2...was solubilized antigens were assayed for levels produced by spleen cells from animals 5 of lymphotoxin (LT), it was seen that nor- days post skin graft sensitization

Biological lignocellulose solubilization: Comparative evaluation of biocatalysts and enhancement via cotreatment

DOE PAGES

Paye, Julie M. D.; Guseva, Anna; Hammer, Sarah K.; ...

2016-01-12

Feedstock recalcitrance is the most important barrier impeding cost-effective production of cellulosic biofuels. Pioneer commercial cellulosic ethanol facilities employ thermochemical pretreatment and addition of fungal cellulase, reflecting the main research emphasis in the field. However, it has been suggested that it may be possible to process cellulosic biomass without thermochemical pretreatment using thermophilic, cellulolytic bacteria. Thus, to further explore this idea, we examine the ability of various biocatalysts to solubilize autoclaved but otherwise unpretreated cellulosic biomass under controlled but not industrial conditions.

Investigation of the micropolarity of reverse micelles using quinolinium betaine compounds as probes

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ueda, Mitsuo; Kimura, Akimune; Wakida, Tomoji

1994-03-15

There is considerable interest in the utilization of reverse micelle and microemulsion systems in a variety of applications such as reactivity control, tertiary oil recovery, solar energy conversion, enzyme mediated synthesis, etc. Fundamental to understanding improved applications of such systems are questions concerning solubilization; thus substantial efforts have been focused on the investigation of the solubilizing state of the assemblies. N-octyl-quinolinium betaine is introduced as an absorption probe for the micropolarity of the interior of reverse micelles. its solubilization by reverse micelles and water/oil microemulsions of Aerosol-OT in isooctane is compared with that of N-methyl-quinolinium betaine at various water contentsmore » of the solution. Analysis of the excitation energies in the visible range of the spectrum indicates that the methyl derivative probes the polarity of the aqueous pool of the micelle, whereas the octyl derivative behaves as a cosurfactant probe that reports on the polarity of the water/oil interfacial region.« less

Solubilization of Therapeutic Agents in Micellar Nanomedicines

PubMed Central

Vuković, Lela; Madriaga, Antonett; Kuzmis, Antonina; Banerjee, Amrita; Tang, Alan; Tao, Kevin; Shah, Neil; Král, Petr; Onyuksel, Hayat

2014-01-01

We use atomistic molecular dynamics simulations to reveal the binding mechanisms of therapeutic agents in PEG-ylated micellar nanocarriers (SSM). In our experiments, SSM in buffer solutions can solubilize either ≈ 11 small bexarotene molecules or ≈ 6 (2 in low ionic strength buffer) human vasoactive intestinal peptide (VIP) molecules. Free energy calculations reveal that molecules of the poorly water soluble drug bexarotene can reside at the micellar ionic interface of the PEG corona, with their polar ends pointing out. Alternatively, they can reside in the alkane core center, where several bexarotene molecules can self-stabilize by forming a cluster held together by a network of hydrogen bonds. We also show that highly charged molecules, such as VIP, can be stabilized at the SSM ionic interface by Coulombic coupling between their positively charged residues and the negatively charged phosphate head-groups of the lipids. The obtained results illustrate that atomistic simulations can reveal drug solubilization character in nanocarriers and be used in efficient optimization of novel nanomedicines. PMID:24283508

Effect of the herbicides oxadiazon and oxyfluorfen on phosphates solubilizing microorganisms and their persistence in rice fields.

PubMed

Das, Amal Chandra; Debnath, Anjan; Mukherjee, Debatosh

2003-10-01

A field experiment has been conducted with two herbicides viz. oxadiazon [5-terbutyl-3-(2,4-dichloro-5-isopropoxyphenyl)-1,3,4-oxadiazol-2-one] and oxyfluorfen [2-chloro-1-(3-ethoxy-4-nitrophenyl)-4-(trifluoromethyl) benzene] at rates of 0.4 and 0.12 kg a.i. ha(-1), respectively, to investigate their effect on the growth and activities of phosphate solubilizing microorganisms in relation to availability of phosphorus as well as persistence of the herbicides in the rhizosphere soil of wetland rice (Oryza sativa L. variety IR-36). Application of herbicides stimulated the population and activities of phosphate solubilizing microorganisms and also the availability of phosphorus in the rhizosphere soil. Oxyfluorfen provided greater microbial stimulation than oxadiazon. Dissipation of oxyfluorfen and oxadiazon followed first order reaction kinetics with half-life (T(1/2)) of 8.8 and 12 days, respectively. Sixty days after application 0.5% and 3% of the applied oxadiazon and oxyfluorfen residues persisted, respectively, in the rhizosphere soil of rice.

Serum albumin forms a lactoferrin-like soluble iron-binding complex in presence of hydrogen carbonate ions.

PubMed

Ueno, Hiroshi M; Urazono, Hiroshi; Kobayashi, Toshiya

2014-02-15

The iron-lactoferrin complex is a common food ingredient because of its iron-solubilizing capability in the presence of hydrogen carbonate ions. However, it is unclear whether the formation of a stable iron-binding complex is limited to lactoferrin. In this study, we investigated the effects of bovine serum albumin (BSA) on iron solubility and iron-catalyzed lipid oxidation in the presence of hydrogen carbonate ions. BSA could solubilize >100-fold molar equivalents of iron at neutral pH, exceeding the specific metal-binding property of BSA. This iron-solubilizing capability of BSA was impaired by thermally denaturing BSA at ≥ 70 °C for 10 min at pH 8.5. The resulting iron-BSA complex inhibited iron-catalyzed oxidation of soybean oil in a water-in-oil emulsion measured using the Rancimat test. Our study is the first to show that BSA, like lactoferrin, forms a soluble iron-binding complex in the presence of hydrogen carbonate ions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Mechanistic Analysis of Cocrystal Dissolution as a Function of pH and Micellar Solubilization

PubMed Central

2016-01-01

The purpose of this work is to provide a mechanistic understanding of the dissolution behavior of cocrystals under the influence of ionization and micellar solubilization. Mass transport models were developed by applying Fick’s law of diffusion to dissolution with simultaneous chemical reactions in the hydrodynamic boundary layer adjacent to the dissolving cocrystal surface to predict the pH at the dissolving solid–liquid interface (i.e., interfacial pH) and the flux of cocrystals. To evaluate the predictive power of these models, dissolution studies of carbamazepine–saccharin (CBZ-SAC) and carbamazepine–salicylic acid (CBZ-SLC) cocrystals were performed at varied pH and surfactant concentrations above the critical stabilization concentration (CSC), where the cocrystals were thermodynamically stable. The findings in this work demonstrate that the pH dependent dissolution behavior of cocrystals with ionizable components is dependent on interfacial pH. This mass transport analysis demonstrates the importance of pH, cocrystal solubility, diffusivity, and micellar solubilization on the dissolution rates of cocrystals. PMID:26877267

Mechanistic Analysis of Cocrystal Dissolution as a Function of pH and Micellar Solubilization.

PubMed

Cao, Fengjuan; Amidon, Gordon L; Rodriguez-Hornedo, Nair; Amidon, Gregory E

2016-03-07

The purpose of this work is to provide a mechanistic understanding of the dissolution behavior of cocrystals under the influence of ionization and micellar solubilization. Mass transport models were developed by applying Fick's law of diffusion to dissolution with simultaneous chemical reactions in the hydrodynamic boundary layer adjacent to the dissolving cocrystal surface to predict the pH at the dissolving solid-liquid interface (i.e., interfacial pH) and the flux of cocrystals. To evaluate the predictive power of these models, dissolution studies of carbamazepine-saccharin (CBZ-SAC) and carbamazepine-salicylic acid (CBZ-SLC) cocrystals were performed at varied pH and surfactant concentrations above the critical stabilization concentration (CSC), where the cocrystals were thermodynamically stable. The findings in this work demonstrate that the pH dependent dissolution behavior of cocrystals with ionizable components is dependent on interfacial pH. This mass transport analysis demonstrates the importance of pH, cocrystal solubility, diffusivity, and micellar solubilization on the dissolution rates of cocrystals.

Crystallographic, hyperfine and magnetic characterization of a maraging-400 alloy

NASA Astrophysics Data System (ADS)

Alves, T. J. B.; Nunes, G. C. S.; Sarvezuk, P. W. C.; Ivashita, F. F.; de Andrade, A. M. H.; Viegas, A.; Paesano, A.

2017-11-01

Maraging400-like alloys were made by arc-melting iron with the alloy elements (i.e., Ni, Co, Ti and Mo), followed by a high temperature heat-treatment for solubilization. The solubilized alloys were further heat-treated (480 °C and 580 °C, by 3 h), for aging. The samples were finely characterized by X-ray diffraction (Rietveld refinement), Mössbauer spectroscopy and magnetization techniques. The results revealed that the as-solubilized sample is martensitic and ferromagnetic. Its residual induction and coercive field increase monotonically with the maximum applied field of a magnetization minor loop and both curves presented very similar shapes. The area of the minor loops varies parabolically with this maximum applied field. The aging induced an atomic rearrangement in the martensite phase, involving change in the composition and lattice parameters, reversion of austenite and the formation of the Fe 3 Mo 2 intermetallic compound. Comparisons are presented between the results obtained by us for these alloys and those obtained for Maraging-350 steel samples.

Crystallization from microemulsions ? a novel method for the preparation of new crystal forms of aspartame

NASA Astrophysics Data System (ADS)

Füredi-Milhofer, Helga; Garti, N.; Kamyshny, A.

1999-03-01

Solubilization and crystallization of the artificial sweetener aspartame (APM), in water/isooctane microemulsions stabilized with sodium diisooctyl sulfosuccinate (AOT) has been investigated. The amount of aspartame that could be solubilized depended primarily on the amount of surfactant and on the temperature. The maximum AOT/aspartame molar ratio at the w/o interface is shown to be 6.2 at 25°C. It was concluded that the dipeptide is located at the w/o interface interspersed between surfactant molecules and that it acts as a cosurfactant. A new crystal form, APM III, was obtained by cooling of hot w/isooctane/AOT microemulsions containing solubilized aspartame. The new crystal form exhibits a distinct X-ray diffraction powder pattern, as well as changes in the FTIR spectra, thermogravimetric and DSC patterns. H-NMR spectra of APM III dissolved in D 2O were identical to the spectrum of commercial aspartame recorded under the same conditions. The new crystal form has greatly improved dissolution kinetics.

Stable and solubilized active Au atom clusters for selective epoxidation of cis-cyclooctene with molecular oxygen

DOE PAGES

Qian, Linping; Wang, Zhen; Beletskiy, Evgeny V.; ...

2017-03-28

Here, the ability of Au catalysts to effect the challenging task of utilizing molecular oxygen for the selective epoxidation of cyclooctene is fascinating. Although supported nanometre-size Au particles are poorly active, here we show that solubilized atomic Au clusters, present in ng ml –1 concentrations and stabilized by ligands derived from the oxidized hydrocarbon products, are active. They can be formed from various Au sources. They generate initiators and propagators to trigger the onset of the auto-oxidation reaction with an apparent turnover frequency of 440 s –1, and continue to generate additional initiators throughout the auto-oxidation cycle without direct participationmore » in the cycle. Spectroscopic characterization suggests that 7–8 atom clusters are effective catalytically. Extension of work based on these understandings leads to the demonstration that these Au clusters are also effective in selective oxidation of cyclohexene, and that solubilized Pt clusters are also capable of generating initiators for cyclooctene epoxidation.« less

Solubilization of flurbiprofen within non-ionic Tween 20 surfactant micelles: a 19F and 1H NMR study.

PubMed

Saveyn, Pieter; Cocquyt, Ellen; Zhu, Wuxin; Sinnaeve, Davy; Haustraete, Katrien; Martins, José C; Van der Meeren, Paul

2009-07-14

The solubilization of the poorly water soluble anti-inflammatory drug flurbiprofen in non-ionic Tween 20 surfactant micellar solutions was studied by both (19)F and (1)H NMR spectroscopy in an acidic environment. These non-destructive techniques allowed us to investigate the effect of temperature cycling in situ. Using (19)F NMR, an increased solubilisation capacity was observed as the temperature increased. This effect became more pronounced above the cloud point, which was reduced by more than 30 degrees C in the presence of an excess of flurbiprofen. Upon clouding, peak splitting was observed in the (19)F spectrum, which indicates that two pools of solubilised flurbiprofen exist that are in slow exchange on the NMR frequency timescale. The clouding and solubilization processes were found to be reversible, albeit with slow kinetics. Based on chemical shift differences of both Tween 20 and flurbiprofen, as well as NOESY experiments, the flurbiprofen was found to be accumulated within the palisade layer of the Tween 20 micelles.

Analytical evaluation of current starch methods used in the international sugar industry: Part I.

PubMed

Cole, Marsha; Eggleston, Gillian; Triplett, Alexa

2017-08-01

Several analytical starch methods exist in the international sugar industry to mitigate starch-related processing challenges and assess the quality of traded end-products. These methods use iodometric chemistry, mostly potato starch standards, and utilize similar solubilization strategies, but had not been comprehensively compared. In this study, industrial starch methods were compared to the USDA Starch Research method using simulated raw sugars. Type of starch standard, solubilization approach, iodometric reagents, and wavelength detection affected total starch determination in simulated raw sugars. Simulated sugars containing potato starch were more accurately detected by the industrial methods, whereas those containing corn starch, a better model for sugarcane starch, were only accurately measured by the USDA Starch Research method. Use of a potato starch standard curve over-estimated starch concentrations. Among the variables studied, starch standard, solubilization approach, and wavelength detection affected the sensitivity, accuracy/precision, and limited the detection/quantification of the current industry starch methods the most. Published by Elsevier Ltd.

Designer lipid-like peptides: a class of detergents for studying functional olfactory receptors using commercial cell-free systems.

PubMed

Corin, Karolina; Baaske, Philipp; Ravel, Deepali B; Song, Junyao; Brown, Emily; Wang, Xiaoqiang; Wienken, Christoph J; Jerabek-Willemsen, Moran; Duhr, Stefan; Luo, Yuan; Braun, Dieter; Zhang, Shuguang

2011-01-01

A crucial bottleneck in membrane protein studies, particularly G-protein coupled receptors, is the notorious difficulty of finding an optimal detergent that can solubilize them and maintain their stability and function. Here we report rapid production of 12 unique mammalian olfactory receptors using short designer lipid-like peptides as detergents. The peptides were able to solubilize and stabilize each receptor. Circular dichroism showed that the purified olfactory receptors had alpha-helical secondary structures. Microscale thermophoresis suggested that the receptors were functional and bound their odorants. Blot intensity measurements indicated that milligram quantities of each olfactory receptor could be produced with at least one peptide detergent. The peptide detergents' capability was comparable to that of the detergent Brij-35. The ability of 10 peptide detergents to functionally solubilize 12 olfactory receptors demonstrates their usefulness as a new class of detergents for olfactory receptors, and possibly other G-protein coupled receptors and membrane proteins.

Designer Lipid-Like Peptides: A Class of Detergents for Studying Functional Olfactory Receptors Using Commercial Cell-Free Systems

PubMed Central

Corin, Karolina; Baaske, Philipp; Ravel, Deepali B.; Song, Junyao; Brown, Emily; Wang, Xiaoqiang; Wienken, Christoph J.; Jerabek-Willemsen, Moran; Duhr, Stefan; Luo, Yuan; Braun, Dieter; Zhang, Shuguang

2011-01-01

A crucial bottleneck in membrane protein studies, particularly G-protein coupled receptors, is the notorious difficulty of finding an optimal detergent that can solubilize them and maintain their stability and function. Here we report rapid production of 12 unique mammalian olfactory receptors using short designer lipid-like peptides as detergents. The peptides were able to solubilize and stabilize each receptor. Circular dichroism showed that the purified olfactory receptors had alpha-helical secondary structures. Microscale thermophoresis suggested that the receptors were functional and bound their odorants. Blot intensity measurements indicated that milligram quantities of each olfactory receptor could be produced with at least one peptide detergent. The peptide detergents' capability was comparable to that of the detergent Brij-35. The ability of 10 peptide detergents to functionally solubilize 12 olfactory receptors demonstrates their usefulness as a new class of detergents for olfactory receptors, and possibly other G-protein coupled receptors and membrane proteins. PMID:22132066

A novel lipoprotein nanoparticle system for membrane proteins

PubMed Central

Frauenfeld, Jens; Löving, Robin; Armache, Jean-Paul; Sonnen, Andreas; Guettou, Fatma; Moberg, Per; Zhu, Lin; Jegerschöld, Caroline; Flayhan, Ali; Briggs, John A.G.; Garoff, Henrik; Löw, Christian; Cheng, Yifan; Nordlund, Pär

2016-01-01

Membrane proteins are of outstanding importance in biology, drug discovery and vaccination. A common limiting factor in research and applications involving membrane proteins is the ability to solubilize and stabilize membrane proteins. Although detergents represent the major means for solubilizing membrane proteins, they are often associated with protein instability and poor applicability in structural and biophysical studies. Here, we present a novel lipoprotein nanoparticle system that allows for the reconstitution of membrane proteins into a lipid environment that is stabilized by a scaffold of Saposin proteins. We showcase the applicability of the method on two purified membrane protein complexes as well as the direct solubilization and nanoparticle-incorporation of a viral membrane protein complex from the virus membrane. We also demonstrate that this lipid nanoparticle methodology facilitates high-resolution structural studies of membrane proteins in a lipid environment by single-particle electron cryo-microscopy (cryo-EM) and allows for the stabilization of the HIV-envelope glycoprotein in a functional state. PMID:26950744

Identification and quantification of the rat hepatocyte asialoglycoprotein receptor.

PubMed Central

Schwartz, A L; Marshak-Rothstein, A; Rup, D; Lodish, H F

1981-01-01

The asialoglycoprotein receptor from rat liver was purified by solubilization and affinity chromatography on asialoorosomucoid-Sepharose. The preparation yielded four distinct polypeptides of Mr 40,000-120,000. We prepared a monoclonal antibody that both immunoprecipitates solubilized receptor activity and blocks the binding of galactose-terminal glycoproteins to immobilized receptor. The monoclonal antibody and a rabbit antireceptor antiserum immunoprecipitated all four polypeptide species. Peptide analysis by two-dimensional chromatography of the individual 125I-labeled species showed nearly identical patterns, which also suggested that the four polypeptides have a similar primary structure. To identify and quantitate the asialoglycoprotein receptor on the hepatocyte cell surface, intact cells were iodinated with lactoperoxidase, and the solubilized membranes were treated with antireceptor antibody. The Mr 55,000 and Mr 65,000 species were the major species found. Our results suggest that the Mr of the surface receptor is at least 55,000 and that it comprises between 1-2% of the iodinated hepatocyte surface protein. Images PMID:6267585

Stable and solubilized active Au atom clusters for selective epoxidation of cis-cyclooctene with molecular oxygen

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qian, Linping; Wang, Zhen; Beletskiy, Evgeny V.

Here, the ability of Au catalysts to effect the challenging task of utilizing molecular oxygen for the selective epoxidation of cyclooctene is fascinating. Although supported nanometre-size Au particles are poorly active, here we show that solubilized atomic Au clusters, present in ng ml –1 concentrations and stabilized by ligands derived from the oxidized hydrocarbon products, are active. They can be formed from various Au sources. They generate initiators and propagators to trigger the onset of the auto-oxidation reaction with an apparent turnover frequency of 440 s –1, and continue to generate additional initiators throughout the auto-oxidation cycle without direct participationmore » in the cycle. Spectroscopic characterization suggests that 7–8 atom clusters are effective catalytically. Extension of work based on these understandings leads to the demonstration that these Au clusters are also effective in selective oxidation of cyclohexene, and that solubilized Pt clusters are also capable of generating initiators for cyclooctene epoxidation.« less

Form and function of cnidarian spirocysts. III. Ultrastructure of the thread and the function of spirocysts

DOE Office of Scientific and Technical Information (OSTI.GOV)

Mariscal, R.N.; McLean, R.B.; Hand, C.

1977-01-01

Unlike most nematocysts, undischarged spirocyst threads bear hollow tubules rather than spines. The undischarged tubules are interconnected in hexagonal arrays and appear to be arranged in bundles along the length of the thread. Although the wall of the thread is folded in length and width, the tubules are not. Upon discharge and contact with sea water, the tubules solubilize and adhere to various substrates and prey. Traction between such objects and the everting thread causes the tubules to spin out into a web or meshwork of fine microfibrillae. Lack of contact of the everting thread with objects results in themore » tubules forming small droplets of partially solubilized material, some of which appear to be arranged in a helical pattern around the thread. The web or meshwork formed by the solubilized tubules in contact with various substrates probably serves to increase significantly the surface area and adhesive properties of the everted spirocyst thread.« less

Effects of electron beam irradiation and temperature on the treatment of swine wastewater using an ion exchange biological reactor.

PubMed

Lim, Seung Joo; Kim, Tak-Hyun; Lee, Sang-hun; Kim, Jun-young; Kim, Sun-kyoung

2013-06-01

Swine wastewater was treated using an ion exchange biological reactor (IEBR). Organic matter and nutrient in swine wastewater were pre-treated by electron beam irradiation. The optimal dose for solubilization of organic matter in swine wastewater ranged from 20 kGy to 75 kGy. The carbohydrates, proteins, and lipids were investigated as proteins and lipids mainly contained the solubilized organic matter. The solubilization of organic matter in swine wastewater was affected by the combination effects of temperature and dose. The maximum chemical oxygen demand (COD) and ammonia removal efficiencies were 74.4% and 76.7% at a dose of 0 kGy under room temperatures (23.0°C). The removal of ammonia was significantly affected by low temperature (15.3°C). On the other hand, the removal of phosphorus was not a function of electron beam irradiation or temperature because struvite is one of the main removal mechanisms under anoxic conditions. Published by Elsevier Ltd.

Biosynthetic elongation of isolated teichuronic acid polymers via glucosyl- and N-acetylmannosaminuronosyltransferases from solubilized cytoplasmic membrane fragments of Micrococcus luteus.

PubMed Central

Hildebrandt, K M; Anderson, J S

1990-01-01

Cytoplasmic membrane fragments of Micrococcus luteus catalyze in vitro biosynthesis of teichuronic acid from uridine diphosphate D-glucose (UDP-glucose), uridine diphosphate N-acetyl-D-mannosaminuronic acid (UDP-ManNAcA), and uridine diphosphate N-acetyl-D-glucosamine. Membrane fragments solubilized with Thesit (dodecyl alcohol polyoxyethylene ether) can utilize UDP-glucose and UDP-ManNAcA to effect elongation of teichuronic acid isolated from native cell walls. When UDP-glucose is the only substrate supplied, the detergent-solubilized glucosyltransferase incorporates a single glucosyl residue onto each teichuronic acid acceptor. When both UDP-glucose and UDP-ManNAcA are supplied, the glucosyltransferase and the N-acetylmannosaminuronosyltransferase act cooperatively to elongate the teichuronic acid acceptor by multiple additions of the disaccharide repeat unit. As shown by polyacrylamide gel electrophoresis, low-molecular-weight fractions of teichuronic acid are converted to higher-molecular-weight polymers by the addition of as many as 17 disaccharide repeat units. Images PMID:2118507

Enhancing the anaerobic digestion potential of dairy waste activated sludge by two step sono-alkalization pretreatment.

PubMed

Rani, R Uma; Kumar, S Adish; Kaliappan, S; Yeom, Ick-Tae; Banu, J Rajesh

2014-05-01

High efficiency resource recovery from dairy waste activated sludge (WAS) has been a focus of attention. An investigation into the influence of two step sono-alkalization pretreatment (using different alkaline agents, pH and sonic reaction times) on sludge reduction potential in a semi-continuous anaerobic reactor was performed for the first time in literature. Firstly, effect of sludge pretreatment was evaluated by COD solubilization, suspended solids reduction and biogas production. At optimized condition (4172 kJ/kg TS of supplied energy for NaOH - pH 10), COD solubilization, suspended solids reduction and biogas production was 59%, 46% and 80% higher than control. In order to clearly describe the hydrolysis of waste activated sludge during sono-alkalization pretreatment by a two step process, concentrations of ribonucleic acid (RNA) and bound extracellular polymeric substance (EPS) were also measured. Secondly, semi-continuous process performance was studied in a lab-scale semi-continuous anaerobic reactor (5L), with 4 L working volume. With three operated SRTs, the SRT of 15 d was found to be most appropriate for economic operation of the reactor. Combining pretreatment with anaerobic digestion led to 58% and 62% of suspended solids and volatile solids reduction, respectively, with an improvement of 83% in biogas production. Thus, two step sono-alkalization pretreatment laid the basis in enhancing the anaerobic digestion potential of dairy WAS. Copyright © 2013 Elsevier B.V. All rights reserved.

Binding of (/sup 3/H)forskolin to platelet membranes and solubilized proteins from bovine brain

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nelson, C.A.; Seamon, K.B.

1986-05-01

(/sup 3/H)Forskolin ((/sup 3/H)FSK) bound to platelet membranes with a Kd of 20 nM and a Bmax of 125 fmol/mg protein. The Bmax was increased to 400 fmol/mg protein in the presence of GppNHp (or NaF) and MgCl/sub 2/ with no change in Kd. PGE/sub 1/ decreased the EC50 of GppNHp to increase the Bmax for (/sup 3/H)FSK binding from 600 nM to 35 nM. In contrast, PGE/sub 1/ had no effect on the EC50 of NaF to increase (/sup 3/H)FSK binding. (/sup 3/H)FSK binding increased slowly over 60 min when forskolin and GppNHp were added to membranes simultaneously atmore » 20/sup 0/C. Preincubation of membranes with GppNHp at 20/sup 5/C also caused a linear increase in adenylate cyclase specific activity over 60 minutes. (/sup 3/H)FSK bound to solubilized protein from bovine brain membrane with a Kd of 22 nM. GppNHp increased the number of binding sites in solubilized proteins only if membranes were not preincubated with GppNHp prior to solubilization. In conclusion the number of binding sites for (/sup 3/H)FSK is increased by agents that activate adenylate cyclase through the Ns protein. These sites appear to be associated with an activated complex of the Ns protein and adenylate cyclase.« less

Effects of P limitation and molecules from peanut root exudates on pqqE gene expression and pqq promoter activity in the phosphate-solubilizing strain Serratia sp. S119.

PubMed

Ludueña, Liliana M; Anzuay, Maria S; Magallanes-Noguera, Cynthia; Tonelli, Maria L; Ibañez, Fernando J; Angelini, Jorge G; Fabra, Adriana; McIntosh, Matthew; Taurian, Tania

2017-10-01

The mineral phosphate-solubilizing phenotype in bacteria is attributed predominantly to secretion of gluconic acid produced by oxidation of glucose by the glucose dehydrogenase enzyme and its cofactor, pyrroloquinoline quinone. This study analyzes pqqE gene expression and pqq promoter activity in the native phosphate-solubilizing bacterium Serratia sp S119 growing under P-limitation, and in the presence of root exudates obtained from peanut plants, also growing under P-limitation. Results indicated that Serratia sp. S119 contains a pqq operon composed of six genes (pqqA,B,C,D,E,F) and two promoters, one upstream of pqqA and other between pqqA and pqqB. PqqE gene expression and pqq promoter activity increased under P-limiting growth conditions and not under N-deficient conditions. In the plant-bacteria interaction assay, the activity of the bacterial pqq promoter region varied depending on the concentration and type of root exudates and on the bacterial growth phase. Root exudates from peanut plants growing under P-available and P-limiting conditions showed differences in their composition. It is concluded from this study that the response of Serratia sp. S119 to phosphorus limitation involves an increase in expression of pqq genes, and that molecules exuded by peanut roots modify expression of these phosphate-solubilizing bacterial genes during plant-bacteria interactions. Copyright © 2017 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Unexpected Solubility Enhancement of Drug Bases in the Presence of a Dimethylaminoethyl Methacrylate Copolymer.

PubMed

Saal, Wiebke; Ross, Alfred; Wyttenbach, Nicole; Alsenz, Jochem; Kuentz, Martin

2018-01-02

The methacrylate copolymer Eudragit EPO (EPO) has previously shown to greatly enhance solubilization of acidic drugs via ionic interactions and by multiple hydrophobic contacts with polymeric side chains. The latter type of interaction could also play a role for solubilization of other compounds than acids. The aim of this study was therefore to investigate the solubility of six poorly soluble bases in presence and absence of EPO by quantitative ultrapressure liquid chromatography with concomitant X-ray powder diffraction analysis of the solid state. For a better mechanistic understanding, spectra and diffusion data were obtained by 1 H nuclear magnetic resonance (NMR) spectroscopy. Unexpected high solubility enhancement (up to 360-fold) was evidenced in the presence of EPO despite the fact that bases and polymer were both carrying positive charges. This exceptional and unexpected solubilization was not due to a change in the crystalline solid state. NMR spectra and measured diffusion coefficients indicated both strong drug-polymer interactions in the bulk solution, and diffusion data suggested conformational changes of the polymer in solution. Such conformational changes may have increased the accessibility and extent of hydrophobic contacts thereby leading to increased overall molecular interactions. These initially surprising solubilization results demonstrate that excipient selection should not be based solely on simple considerations of, for example, opposite charges of drug and excipient, but it requires a more refined molecular view. Different solution NMR techniques are especially promising tools to gain such mechanistic insights.

A New Mass Spectrometry-compatible Degradable Surfactant for Tissue Proteomics

PubMed Central

Chang, Ying-Hua; Gregorich, Zachery R.; Chen, Albert J.; Hwang, Leekyoung; Guner, Huseyin; Yu, Deyang; Zhang, Jianyi; Ge, Ying

2015-01-01

Tissue proteomics is increasingly recognized for its role in biomarker discovery and disease mechanism investigation. However, protein solubility remains a significant challenge in mass spectrometry (MS)-based tissue proteomics. Conventional surfactants such as sodium dodecyl sulfate (SDS), the preferred surfactant for protein solubilization, are not compatible with MS. Herein, we have screened a library of surfactant-like compounds and discovered an MS-compatible degradable surfactant (MaSDeS) for tissue proteomics that solubilizes all categories of proteins with performance comparable to SDS. The use of MaSDeS in the tissue extraction significantly improves the total number of protein identifications from commonly used tissues, including tissue from the heart, liver, and lung. Notably, MaSDeS significantly enriches membrane proteins, which are often under-represented in proteomics studies. The acid degradable nature of MaSDeS makes it amenable for high-throughput mass spectrometry-based proteomics. In addition, the thermostability of MaSDeS allows for its use in experiments requiring high temperature to facilitate protein extraction and solubilization. Furthermore, we have shown that MaSDeS outperforms the other MS-compatible surfactants in terms of overall protein solubility and the total number of identified proteins in tissue proteomics. Thus, the use of MaSDeS will greatly advance tissue proteomics and realize its potential in basic biomedical and clinical research. MaSDeS could be utilized in a variety of proteomics studies as well as general biochemical and biological experiments that employ surfactants for protein solubilization. PMID:25589168

Impact of Dendrimers on Solubility of Hydrophobic Drug Molecules

PubMed Central

Choudhary, Sonam; Gupta, Lokesh; Rani, Sarita; Dave, Kaushalkumar; Gupta, Umesh

2017-01-01

Adequate aqueous solubility has been one of the desired properties while selecting drug molecules and other bio-actives for product development. Often solubility of a drug determines its pharmaceutical and therapeutic performance. Majority of newly synthesized drug molecules fail or are rejected during the early phases of drug discovery and development due to their limited solubility. Sufficient permeability, aqueous solubility and physicochemical stability of the drug are important for achieving adequate bioavailability and therapeutic outcome. A number of different approaches including co-solvency, micellar solubilization, micronization, pH adjustment, chemical modification, and solid dispersion have been explored toward improving the solubility of various poorly aqueous-soluble drugs. Dendrimers, a new class of polymers, possess great potential for drug solubility improvement, by virtue of their unique properties. These hyper-branched, mono-dispersed molecules have the distinct ability to bind the drug molecules on periphery as well as to encapsulate these molecules within the dendritic structure. There are numerous reported studies which have successfully used dendrimers to enhance the solubilization of poorly soluble drugs. These promising outcomes have encouraged the researchers to design, synthesize, and evaluate various dendritic polymers for their use in drug delivery and product development. This review will discuss the aspects and role of dendrimers in the solubility enhancement of poorly soluble drugs. The review will also highlight the important and relevant properties of dendrimers which contribute toward drug solubilization. Finally, hydrophobic drugs which have been explored for dendrimer assisted solubilization, and the current marketing status of dendrimers will be discussed. PMID:28559844

In vitro digestive fluid extraction as a measure of the bioavailability of sediment-associated polycyclic aromatic hydrocarbons: Sources of variation and implications for partitioning models

DOE Office of Scientific and Technical Information (OSTI.GOV)

Weston, D.P.; Mayer, L.M.

1998-05-01

In vitro extraction of contaminated sediments using the digestive fluid of a deposit-feeding polychaete has recently been proposed to study contaminant bioaccumulation mechanisms and perhaps to better quantify the bioavailable contaminant fraction. This approach was evaluated using digestive fluid from the polychaete Arenicola brasiliensis and six marine sediments containing both spiked radiolabeled polycyclic aromatic hydrocarbons (PAHs) and in situ-contaminated unlabeled PAHs. The proportion of total contaminant extracted by digestive fluid from each sediment varied from 22 to 71% and 13 to 52%, for phenanthrene and benzo[a]pyrene, respectively. The proportions of contaminant solubilized were inversely correlated with the sediments` organic carbonmore » content. The extent of PAH solubilization among sediments by A. brasiliensis digestive fluid was highly correlated with that of digestive fluid from the echiuran Urechis caupo and appears to be a consequence of surfactant properties of the fluids rather than of their enzymatic activity. The proportion of PAHs solubilized in vitro was similar to in vivo measurements of solubilization for contaminant exposures lasting about 24 h. However, with continued exposure, in vivo PAH concentrations in the digestive fluid increased fivefold, suggesting that digestive fluid is retained in the gut longer than sediment and thus accumulates PAHs through sequential digestion of many gut volumes. This phenomenon may enhance contaminant fugacity in the gut and increase the potential for bioaccumulation or toxicity.« less

Jellyfish mesogloea collagen. Characterization of molecules as alpha 1 alpha 2 alpha 3 heterotrimers.

PubMed

Miura, S; Kimura, S

1985-12-05

The mesogloea collagen of a primitive animal, the jellyfish Stomolophus nomurai, belonging to the class Scyphozoa in the Coelenterata, was studied with respect to its chain structure. Most of the mesogloea collagen was solubilized by limited digestion with pepsin and isolated by selective precipitation at 0.9 m NaCl in 0.5 M acetic acid. Upon denaturation, the pepsin-solubilized collagen produced three distinct alpha chains, alpha 1, alpha 2, and alpha 3, in comparable amounts which were separable by CM-cellulose chromatography. The nonidentity of these alpha chains was confirmed by amino acid and carbohydrate analyses and peptide mapping. Furthermore, the introduction of intramolecular cross-links into native molecules by formaldehyde yielded a large proportion of gamma 123 chain with chain structure alpha 1 alpha 2 alpha 3, as judged by chromatographic behavior and peptide maps. We concluded that mesogloea collagen is comprised of alpha 1 alpha 2 alpha 3 heterotrimers and is chemically like vertebrate Type V collagen. On the other hand, sea anemone mesogloea collagen from the class Anthozoa was previously reported to comprise (alpha)3 homotrimers (Katzman, R. L., and Kang, A. H. (1972) J. Biol. Chem. 247, 5486-5489). On the basis of these findings, we assume that alpha 1 alpha 2 alpha 3 heterotrimers arose in evolution with the divergence of Scyphozoa and Anthozoa.

Determining composition of micron-scale protein deposits in neurodegenerative disease by spatially targeted optical microproteomics.

PubMed

Hadley, Kevin C; Rakhit, Rishi; Guo, Hongbo; Sun, Yulong; Jonkman, James E N; McLaurin, Joanne; Hazrati, Lili-Naz; Emili, Andrew; Chakrabartty, Avijit

2015-09-29

Spatially targeted optical microproteomics (STOMP) is a novel proteomics technique for interrogating micron-scale regions of interest (ROIs) in mammalian tissue, with no requirement for genetic manipulation. Methanol or formalin-fixed specimens are stained with fluorescent dyes or antibodies to visualize ROIs, then soaked in solutions containing the photo-tag: 4-benzoylbenzyl-glycyl-hexahistidine. Confocal imaging along with two photon excitation are used to covalently couple photo-tags to all proteins within each ROI, to a resolution of 0.67 µm in the xy-plane and 1.48 µm axially. After tissue solubilization, photo-tagged proteins are isolated and identified by mass spectrometry. As a test case, we examined amyloid plaques in an Alzheimer's disease (AD) mouse model and a post-mortem AD case, confirming known plaque constituents and discovering new ones. STOMP can be applied to various biological samples including cell lines, primary cell cultures, ex vivo specimens, biopsy samples, and fixed post-mortem tissue.

Superactivity of peroxidase solubilized in reversed micellar systems.

PubMed

Setti, L; Fevereiro, P; Melo, E P; Pifferi, P G; Cabral, J M; Aires-Barros, M R

1995-12-01

Vaccinium mirtyllus peroxidase solubilized in reversed micelles was used for the oxidation of guaiacol. Some relevant parameters for the enzymatic activity, such as pH, w(o) (molar ratio water/surfactant), surfactant type and concentration, and cosurfactant concentration, were investigated. The peroxidase showed higher activities in reversed micelles than in aqueous solution. The stability of the peroxidase in reversed micelles was also studied, namely, the effect of w(o) and temperature on enzyme deactivation. The peroxidase displayed higher stabilities in CTAB/hexanol in isooctane reversed micelles, with half-life times higher than 500 h.

Production and characterization of a novel antifungal chitinase identified by functional screening of a suppressive-soil metagenome.

PubMed

Berini, Francesca; Presti, Ilaria; Beltrametti, Fabrizio; Pedroli, Marco; Vårum, Kjell M; Pollegioni, Loredano; Sjöling, Sara; Marinelli, Flavia

2017-01-31

Through functional screening of a fosmid library, generated from a phytopathogen-suppressive soil metagenome, the novel antifungal chitinase-named Chi18H8 and belonging to family 18 glycosyl hydrolases-was previously discovered. The initial extremely low yield of Chi18H8 recombinant production and purification from Escherichia coli cells (21 μg/g cell) limited its characterization, thus preventing further investigation on its biotechnological potential. We report on how we succeeded in producing hundreds of milligrams of pure and biologically active Chi18H8 by developing and scaling up to a high-yielding, 30 L bioreactor process, based on a novel method of mild solubilization of E. coli inclusion bodies in lactic acid aqueous solution, coupled with a single step purification by hydrophobic interaction chromatography. Chi18H8 was characterized as a Ca 2+ -dependent mesophilic chitobiosidase, active on chitin substrates at acidic pHs and possessing interesting features, such as solvent tolerance, long-term stability in acidic environment and antifungal activity against the phytopathogens Fusarium graminearum and Rhizoctonia solani. Additionally, Chi18H8 was found to operate according to a non-processive endomode of action on a water-soluble chitin-like substrate. Expression screening of a metagenomic library may allow access to the functional diversity of uncultivable microbiota and to the discovery of novel enzymes useful for biotechnological applications. A persisting bottleneck, however, is the lack of methods for large scale production of metagenome-sourced enzymes from genes of unknown origin in the commonly used microbial hosts. To our knowledge, this is the first report on a novel metagenome-sourced enzyme produced in hundreds-of-milligram amount by recovering the protein in the biologically active form from recombinant E. coli inclusion bodies.

Pedogenic formation of montmorillonite from a 2:1-2:2 intergrade clay mineral

USGS Publications Warehouse

Malcolm, R.L.; Nettleton, W.D.; McCracken, R.J.

1968-01-01

Montmorillonite was found to be the dominant clay mineral in surface horizons of certain soils of the North Carolina Coastal Plain whereas a 2:1-2:2 intergrade clay mineral was dominant in subjacent horizons. In all soils where this clay mineral sequence was found, the surface horizon was low in pH (below 4⋅5) and high in organic matter content. In contrast, data from studies of other soils of this region (Weed and Nelson, 1962) show that: (1) montmorillonite occurs infrequently; (2) maximum accumulation of the 2:1-2:2 intergrade normally occurs in the surface horizon and decreases with depth in the profile; (3) organic matter contents are low; and (4) pH values are only moderately acid (pH 5-6).It is theorized that the montmorillonite in the surface horizon of the soils studied originated by pedogenic weathering of the 2:1-2:2 intergrade clay mineral. The combined effects of low pH (below 4⋅5) and high organic matter content in surface horizons are believed to be the agents responsible for this mineral transformation. The protonation and solubilization (reverse of hydrolysis) of Al-polymers in the interlayer of expansible clay minerals will occur at or below pH 4⋅5 depending on the charge and steric effects of the interlayer. A low pH alone may cause this solubilization and thus mineral transformation, but in the soils studied the organic matter is believed to facilitate and accelerage the transformation. The intermediates of organic matter decomposition provide an acid environment, a source of protons, and a source of watersoluble mobile organic substances (principally fulvic acids) which have the ability to complex the solubilized aluminum and move it down the profile. This continuous removal of solubilized aluminum would provide for a favorable gradient for aluminum solubilization.The drainage class or position in a catena is believed to be less important than the chemical factors in formation of montmorillonite from 2:1-2:2 intergrade, because montmorillonite is present in all drainage classes if the surface horizon is low in pH and high in organic matter.

Incineration as a method for resource recovery from inedible biomass in a Controlled Ecological Life Support System.

PubMed

Bubenheim, D L; Wignarajah, K

1995-01-01

Resource recovery from waste streams in a space habitat is essential to minimize the resupply burden and achieve self-sufficiency. In a Controlled Ecological Life Support System (CELSS) human wastes and inedible biomass will represent significant sources of secondary raw materials necessary for support of crop plant production (carbon, water, and inorganic plant nutrients). Incineration, pyrolysis, and water extraction have been investigated as candidate processes for recovery of these important resources from inedible biomass in a CELSS. During incineration CO2 is produced by oxidation of the organic components and this product can be directly utilized by plants. Water is concomitantly produced, requiring only a phase change for recovery. Recovery of inorganics is more difficult, requiring solubilization of the incinerator ash. The process of incineration followed by water solubilization of ash resulted in loss of 35% of the inorganics originally present in the biomass. Losses were attributed to volatilization (8%) and non-water-soluble ash (27%). All of the ash remaining following incineration could be solubilized with acid, with losses resulting from volatilization only. The recovery for individual elements varied. Elemental retention in the ash ranged from 100% of that present in the biomass for Ca, P, Mg, Na, and Si to 10% for Zn. The greatest water solubility was observed for potassium with recovery of approximately 77% of that present in the straw. Potassium represented 80% of the inorganic constituents in the wheat straw, and because of slightly greater solubility made up 86% of the water-soluble ash. Following incineration of inedible biomass from wheat, 65% of the inorganics originally present in the straw were recovered by water solubilization and 92% recovered by acid solubilization. Recovery of resources is more complex for pyrolysis and water extraction. Recovery of carbon, a resource of greater mass than the inorganic component of biomass, is more difficult following pyrolysis and water extraction of biomass. In both cases, additional processors would be required to provide products equivalent to those resulting from incineration alone. The carbon, water, and inorganic resources of inedible biomass are effectively separated and output in usable forms through incineration.

Incineration as a Method for Resource Recovery from Inedible Biomass in a Controlled Ecological Life Support System

NASA Technical Reports Server (NTRS)

Bubenheim, David L.; Wignarajah, Kanapathipillai

1995-01-01

Resource recovery from waste streams in a space habitat is essential to minimize the resupply burden and achieve self-sufficiency. In a Controlled Ecological Life Support System (CELSS) human wastes and inedible biomass will represent significant sources of secondary raw materials necessary for support of crop plant production (carbon, water, and inorganic plant nutrients). Incineration, pyrolysis, and water extraction have been investigated as candidate processes for recovery of these important resources from inedible biomass in a CELSS. During incineration CO2 is produced by oxidation of the organic components and this product can be directly utilized by plants. Water is concomitantly produced, requiring only a phase change for recovery. Recovery of inorganics is more difficult, requiring solubilization of the incinerator ash. The process of incineration followed by water solubilization of ash resulted in loss of 35% of the inorganics originally present in the biomass. Losses were attributed to volatilization (8%) and non-water-soluble ash (27%). All of the ash remaining following incineration could be solubilized with acid, with losses resulting from volatilization only. The recovery for individual elements varied. Elemental retention in the ash ranged from 100% of that present in the biomass for Ca, P, Mg, Na, and Si to 10% for Zn. The greatest water solubility was observed for potassium with recovery of approximately 77% of that present in the straw. Potassium represented 80% of the inorganic constituents in the wheat straw, and because of slightly greater solubility made up 86% of the water-soluble ash. Following incineration of inedible biomass from wheat, 65% of the inorganics originally present in the straw were recovered by water solubilization and 92% recovered by acid solubilization. Recovery of resources is more complex for pyrolysis and water extraction. Recovery of carbon, a resource of greater mass than the inorganic component of biomass, is more difficult following pyrolysis and water extraction of biomass. In both cases, additional processors would be required to provide products equivalent to those resulting from incineration alone. The carbon, water, and organic resources of inedible biomass are effectively separated and output in usable forms through incineration.

Compositional changes in 'Bartlett' pear ( Pyrus communis L.) cell wall polysaccharides as affected by sunlight conditions.

PubMed

Raffo, María D; Ponce, Nora M A; Sozzi, Gabriel O; Vicente, Ariel R; Stortz, Carlos A

2011-11-23

Preharvest conditions can have a great impact on fruit quality attributes and postharvest responses. Firmness is an important quality attribute in pear, and excessive softening increases susceptibility to bruising and decay, thus limiting fruit postharvest life. Textural characteristics of fruits are determined at least in part by cell wall structure and disassembly. Few studies have analyzed the influence of fruit preharvest environment in softening, cell wall composition, and degradation. In the current work 'Bartlett' pears grown either facing the sun (S) or in the shade (H) were harvested and stored for 13 days at 20 °C. An evaluation of fruit soluble solids, acidity, color, starch degradation, firmness, cell wall yield, pectin and matrix glycan solubilization, depolymerization, and monosaccharide composition was carried out. Sun-exposed pears showed more advanced color development and similar levels of starch degradation, sugars, and acids than shaded fruit. Sunlight-grown pears were at harvest firmer than shade-grown pears. Both fruit groups softened during storage at 20 °C, but even after ripening, sun-exposed pears remained firmer. Sunlight exposure did not have a great impact on pectin molecular weight. Instead, at harvest a higher proportion of water-solubilized uronic acids and alkali-solubilized neutral sugars and a larger mean molecular size of tightly bound glycans was found in sun-exposed pears. During ripening cell wall catabolism took place in both sun- and shade-grown pears, but pectin solubilization was clearly delayed in sun-exposed fruit. This was associated with decreased removal of RG I-arabinan side chains rather than with reduced depolymerization.

Glomus fasciculatum Fungi as a Bio-convertor and Bio-activator of Inorganic and Organic P in Dual Symbiosis.

PubMed

Azmat, Rafia; Hamid, Neelofer; Moin, Sumeira; Saleem, Ailyan

2016-01-01

Dual symbiosis played an effective role in drought condition and temperature. Furthermore, performed services in absorption of water and solubilization of chief nutrients specially phosphorus for growth of plants. Phosphorous is essential for plant growth in any climatic condition because of central constituent of ATP providing chemical energy for all metabolic reactions of plants. The goal of this work was to monitor the growth of plant under three climatic conditions in comparison to control plant under Glomus fasciculatum inoculation related with adequate supply of phosphorous. Results demonstrated that Glomus fasciculatum (VAM) activates the solubilization of P into the anionic form H2PO4(-) which is highly consumable form by the plants. Minerals including P in soil most regularly solubilized for fixing in plants and continuously changed to highly soluble forms by reaction with inorganic or organic constituents of the soil which are activated in the presence of fungi for continuous availability. Experimental facts and nonstop growth of plants recommended that VAM fungi act as a bio-convertor and bio-activator of soil nutrients, especially of P and their hyphal interaction absorbs soil nutrients and activates insoluble P to soluble one for plant development. Continuous growth of 18 months old Conocarpus erectus L plant in dual symbiosis supports the proposed idea that phosphorus cycle exists during VAM inoculations, where soil reaction altered in presence of spores that help to solubilize the P which strengthens the plant, activates photo-biological activity and demonstrates the new function of VAM as a recycler for continues growth.

Heterologous expression of pyrroloquinoline quinone (pqq) gene cluster confers mineral phosphate solubilization ability to Herbaspirillum seropedicae Z67.

PubMed

Wagh, Jitendra; Shah, Sonal; Bhandari, Praveena; Archana, G; Kumar, G Naresh

2014-06-01

Gluconic acid secretion mediated by the direct oxidation of glucose by pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenase (GDH) is responsible for mineral phosphate solubilization in Gram-negative bacteria. Herbaspirillum seropedicae Z67 (ATCC 35892) genome encodes GDH apoprotein but lacks genes for the biosynthesis of its cofactor PQQ. In this study, pqqE of Erwinia herbicola (in plasmid pJNK1) and pqq gene clusters of Pseudomonas fluorescens B16 (pOK53) and Acinetobacter calcoaceticus (pSS2) were over-expressed in H. seropedicae Z67. Transformants Hs (pSS2) and Hs (pOK53) secreted micromolar levels of PQQ and attained high GDH activity leading to secretion of 33.46 mM gluconic acid when grown on 50 mM glucose while Hs (pJNK1) was ineffective. Hs (pJNK1) failed to solubilize rock phosphate, while Hs (pSS2) and Hs (pOK53) liberated 125.47 μM and 168.07 μM P, respectively, in minimal medium containing 50 mM glucose under aerobic conditions. Moreover, under N-free minimal medium, Hs (pSS2) and Hs (pOK53) not only released significant P but also showed enhanced growth, biofilm formation, and exopolysaccharide (EPS) secretion. However, indole acetic acid (IAA) production was suppressed. Thus, the addition of the pqq gene cluster, but not pqqE alone, is sufficient for engineering phosphate solubilization in H. seropedicae Z67 without compromising growth under nitrogen-fixing conditions.

Isolation and characterization of phosphofungi, and screening of their plant growth-promoting activities.

PubMed

Wang, Xiaohui; Wang, Changdong; Sui, Junkang; Liu, Zhaoyang; Li, Qian; Ji, Chao; Song, Xin; Hu, Yurong; Wang, Changqian; Sa, Rongbo; Zhang, Jiamiao; Du, Jianfeng; Liu, Xunli

2018-04-20

Rhizospheric microorganisms can increase phosphorus availability in the soil. In this regard, the ability of phosphofungi to dissolve insoluble phosphorus compounds is greater than that of phosphate-solubilizing bacteria. The aim of the current study was to identify efficient phosphofungi that could be developed as commercial microbial agents. Among several phosphate-solubilizing fungal isolates screened, strain CS-1 showed the highest phosphorus-solubilization ability. Based on phylogenetic analysis of the internal transcribed spacer region sequence, it was identified as Aspergillus niger. High-performance liquid chromatography analysis revealed that the mechanism of phosphorus solubilization by CS-1 involved the synthesis and secretion of organic acids, mainly oxalic, tartaric, and citric acids. Furthermore, strain CS-1 exhibited other growth-promoting abilities, including efficient potassium release and degradation of crop straw cellulose. These properties help to returning crop residues to the soil, thereby increasing nutrient availability and sustaining organic matter concentration therein. A pot experiment revealed that CS-1 apparently increased the assessed biometric parameters of wheat seedlings, implying the potential of this strain to be developed as a commercial microbial agent. We used Illumina MiSeq sequencing to investigate the microbial community composition in the rhizosphere of uninoculated wheat plants and wheat plants inoculated with the CS-1 strain to obtain insight into the effect of the CS-1 strain inoculation. The data clearly demonstrated that CS-1 significantly reduced the content of pathogenic fungi, including Gibberella, Fusarium, Monographella, Bipolaris, and Volutella, which cause soil-borne diseases in various crops. Strain CS-1 may hence be developed into a microbial agent for plant growth improvement.

[Combination of phosphorus solubilizing and mobilizing fungi with phosphate rocks and volcanic materials to promote plant growth of lettuce (Lactuca sativa L.)].

PubMed

Velázquez, María S; Cabello, Marta N; Elíades, Lorena A; Russo, María L; Allegrucci, Natalia; Schalamuk, Santiago

Arbuscular mycorrhizal fungi (AMF) increase the uptake of soluble phosphates, while phosphorus solubilizing fungi (S) promote solubilization of insoluble phosphates complexes, favoring plant nutrition. Another alternative to maintaining crop productivity is to combine minerals and rocks that provide nutrients and other desirable properties. The aim of this work was to combine AMF and S with pyroclastic materials (ashes and pumices) from Puyehue volcano and phosphate rocks (PR) from Rio Chico Group (Chubut) - to formulate a substrate for the production of potted Lactuca sativa. A mixture of Terrafertil®:ashes was used as substrate. Penicillium thomii was the solubilizing fungus and Rhizophagus intraradices spores (AMF) was the P mobilizer (AEGIS® Irriga). The treatments were: 1) Substrate; 2) Substrate+AMF; 3) Substrate+S; 4) Substrate+AMF+S; 5) Substrate: PR; 6) Substrate: PR+AMF; 7) Substrate: PR+S and 8) Substrate: PR+AMF+S. Three replicates were performed per treatment. All parameters evaluated (total and assimilable P content in substrate, P in plant tissue and plant dry biomass) were significantly higher in plants grown in substrate containing PR and inoculas with S and AMF. This work confirms that the combination of S/AMF with Puyehue volcanic ashes, PR from the Río Chico Group and a commercial substrate promote the growth of L. sativa, thus increasing the added value of national geomaterials. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Enhancing anaerobic digestibility and phosphorus recovery of dairy manure through microwave-based thermochemical pretreatment.

PubMed

Jin, Ying; Hu, Zhenhu; Wen, Zhiyou

2009-08-01

Anaerobic digestion and struvite precipitation are two effective ways of treating dairy manure for recovering biogas and phosphorus. Anaerobic digestion of dairy manure is commonly limited by slow fiber degradation, while struvite precipitation is limited by the availability of orthophosphate. The aim of this work is to study the possibility of using microwave-based thermochemical pretreatment to simultaneously enhance manure anaerobic digestibility (through fiber degradation) and struvite precipitation (through phosphorus solubilization). Microwave heating combined with different chemicals (NaOH, CaO, H(2)SO(4), or HCl) enhanced solubilization of manure and degradation of glucan/xylan in dairy manure. However, sulfuric acid-based pretreatment resulted in a low anaerobic digestibility, probably due to the sulfur inhibition and Maillard side reaction. The pretreatments released 20-40% soluble phosphorus and 9-14% ammonium. However, CaO-based pretreatment resulted in lower orthophosphate releases and struvite precipitation efficiency as calcium interferes with phosphate to form calcium phosphate. Collectively, microwave heating combined with NaOH or HCl led to a high anaerobic digestibility and phosphorus recovery. Using these two chemicals, the performance of microwave- and conventional-heating in thermochemical pretreatment was further compared. The microwave heating resulted in a better performance in terms of COD solubilization, glucan/xylan reduction, phosphorus solubilization and anaerobic digestibility. Lastly, temperature and heating time used in microwave treatment were optimized. The optimal values of temperature and heating time were 147 degrees C and 25.3 min for methane production, and 135 degrees C and 26 min for orthophosphate release, respectively.

Activity of maize transglutaminase overexpressed in Escherichia coli inclusion bodies: an alternative to protein refolding.

PubMed

Carvajal, Patricia; Gibert, Jordi; Campos, Nefertiti; Lopera, Oriol; Barberà, Eduard; Torné, Jose M; Santos, Mireya

2011-01-01

Transglutaminases (TGases) catalyze protein post-translational modification by ε-(γ-glutamyl) links and covalent polyamine conjugation. In plants, this enzyme is poorly characterized and only the maize plastidial TGase gene (tgz) has been cloned. The tgz gene (Patent WWO03102128) had been subcloned and overexpressed in Escherichia coli cells, and the recombinant protein (TGZp) was present mainly in inclusion bodies (IB) fraction. In this work, after overexpression of TGZ15p and SDS-PAGE IB fraction analysis, bands about 65 and 56 kDa were obtained. Western blot, alkylation and MALDI-TOF/TOF analyses indicated that the 56 kDa band corresponded to a truncated sequence from the native TGZ15p (expected MW 65 kDa), by elimination of a chloroplast signal peptide fragment during expression processing. So that large-scale protein production and protein crystallization can be applied, we characterized the TGZ15p enzyme activity in the IB protein fraction, with and without refolding. Results indicate that it presented the biochemical characteristics of other described TGases, showing a certain plant-substrate preference. Solubilization of the IB fraction with Triton X-100 as nondenaturing detergent yielded active TGZ without the need for refolding, giving activity values comparable to those of the refolded protein, indicating that this is a valuable, faster way to obtain TGZ active protein. Copyright © 2011 American Institute of Chemical Engineers (AIChE).

Phytoremediation of Alberta oil sand tailings using native plants and fungal endophytes

NASA Astrophysics Data System (ADS)

Repas, T.; Germida, J.; Kaminskyj, S.

2012-04-01

Fungal endophytes colonize host plants without causing disease. Some endophytes confer plant tolerance to harsh environments. One such endophyte, Trichoderma harzianum strain TSTh20-1, was isolated from a plant growing on Athabasca oil sand tailings. Tailing sands are a high volume waste product from oil sand extraction that the industry is required to remediate. Tailing sands are low in organic carbon and mineral nutrients, and are hydrophobic due to residual polyaromatic hydrocarbons. Typically, tailing sands are remediated by planting young trees in large quantities of mulch plus mineral fertilizer, which is costly and labour intensive. In greenhouse trials, TSTh20-1 supports growth of tomato seedlings on tailing sands without fertilizer. The potential use of TSTh20-1 in combination with native grasses and forbs to remediate under field conditions is being assessed. Twenty-three commercially available plant species are being screened for seed germination and growth on tailing sands in the presence of TSTh20-1. The best candidates from this group will be used in greenhouse and small scale field trials. Potential mechanisms that contribute to endophyte-induced plant growth promotion, such as plant hormone production, stress tolerance, mineral solubilization, and uptake are also being assessed. As well, TSTh20-1 appears to be remarkably frugal in its nutrient requirements and the possibility that this attribute is characteristic of other plant-fungal endophytes from harsh environments is under study.

Assessment of Cultivation Factors that Affect Biomass and Geraniol Production in Transgenic Tobacco Cell Suspension Cultures

PubMed Central

Vasilev, Nikolay; Schmitz, Christian; Grömping, Ulrike; Fischer, Rainer; Schillberg, Stefan

2014-01-01

A large-scale statistical experimental design was used to determine essential cultivation parameters that affect biomass accumulation and geraniol production in transgenic tobacco (Nicotiana tabacum cv. Samsun NN) cell suspension cultures. The carbohydrate source played a major role in determining the geraniol yield and factors such as filling volume, inoculum size and light were less important. Sucrose, filling volume and inoculum size had a positive effect on geraniol yield by boosting growth of plant cell cultures whereas illumination of the cultures stimulated the geraniol biosynthesis. We also found that the carbohydrates sucrose and mannitol showed polarizing effects on biomass and geraniol accumulation. Factors such as shaking frequency, the presence of conditioned medium and solubilizers had minor influence on both plant cell growth and geraniol content. When cells were cultivated under the screened conditions for all the investigated factors, the cultures produced ∼5.2 mg/l geraniol after 12 days of cultivation in shaking flasks which is comparable to the yield obtained in microbial expression systems. Our data suggest that industrial experimental designs based on orthogonal arrays are suitable for the selection of initial cultivation parameters prior to the essential medium optimization steps. Such designs are particularly beneficial in the early optimization steps when many factors must be screened, increasing the statistical power of the experiments without increasing the demand on time and resources. PMID:25117009

Assessment of cultivation factors that affect biomass and geraniol production in transgenic tobacco cell suspension cultures.

PubMed

Vasilev, Nikolay; Schmitz, Christian; Grömping, Ulrike; Fischer, Rainer; Schillberg, Stefan

2014-01-01

A large-scale statistical experimental design was used to determine essential cultivation parameters that affect biomass accumulation and geraniol production in transgenic tobacco (Nicotiana tabacum cv. Samsun NN) cell suspension cultures. The carbohydrate source played a major role in determining the geraniol yield and factors such as filling volume, inoculum size and light were less important. Sucrose, filling volume and inoculum size had a positive effect on geraniol yield by boosting growth of plant cell cultures whereas illumination of the cultures stimulated the geraniol biosynthesis. We also found that the carbohydrates sucrose and mannitol showed polarizing effects on biomass and geraniol accumulation. Factors such as shaking frequency, the presence of conditioned medium and solubilizers had minor influence on both plant cell growth and geraniol content. When cells were cultivated under the screened conditions for all the investigated factors, the cultures produced ∼ 5.2 mg/l geraniol after 12 days of cultivation in shaking flasks which is comparable to the yield obtained in microbial expression systems. Our data suggest that industrial experimental designs based on orthogonal arrays are suitable for the selection of initial cultivation parameters prior to the essential medium optimization steps. Such designs are particularly beneficial in the early optimization steps when many factors must be screened, increasing the statistical power of the experiments without increasing the demand on time and resources.

Self-Propulsion of Pure Water Droplets by Spontaneous Marangoni-Stress-Driven Motion

NASA Astrophysics Data System (ADS)

Izri, Ziane; van der Linden, Marjolein N.; Michelin, Sébastien; Dauchot, Olivier

2014-12-01

We report spontaneous motion in a fully biocompatible system consisting of pure water droplets in an oil-surfactant medium of squalane and monoolein. Water from the droplet is solubilized by the reverse micellar solution, creating a concentration gradient of swollen reverse micelles around each droplet. The strong advection and weak diffusion conditions allow for the first experimental realization of spontaneous motion in a system of isotropic particles at sufficiently large Péclet number according to a straightforward generalization of a recently proposed mechanism [S. Michelin, E. Lauga, and D. Bartolo, Phys. Fluids 25, 061701 (2013); S. Michelin and E. Lauga, J. Fluid Mech. 747, 572 (2014)]. Experiments with a highly concentrated solution of salt instead of water, and tetradecane instead of squalane, confirm the above mechanism. The present swimming droplets are able to carry external bodies such as large colloids, salt crystals, and even cells.

Self-propulsion of pure water droplets by spontaneous Marangoni-stress-driven motion.

PubMed

Izri, Ziane; van der Linden, Marjolein N; Michelin, Sébastien; Dauchot, Olivier

2014-12-12

We report spontaneous motion in a fully biocompatible system consisting of pure water droplets in an oil-surfactant medium of squalane and monoolein. Water from the droplet is solubilized by the reverse micellar solution, creating a concentration gradient of swollen reverse micelles around each droplet. The strong advection and weak diffusion conditions allow for the first experimental realization of spontaneous motion in a system of isotropic particles at sufficiently large Péclet number according to a straightforward generalization of a recently proposed mechanism [S. Michelin, E. Lauga, and D. Bartolo, Phys. Fluids 25, 061701 (2013); S. Michelin and E. Lauga, J. Fluid Mech. 747, 572 (2014)]. Experiments with a highly concentrated solution of salt instead of water, and tetradecane instead of squalane, confirm the above mechanism. The present swimming droplets are able to carry external bodies such as large colloids, salt crystals, and even cells.

Influence of biogeochemical interactions on metal bioleaching performance in contaminated marine sediment.

PubMed

Fonti, Viviana; Dell'Anno, Antonio; Beolchini, Francesca

2013-09-15

Bioleaching strategies are still far from finding real applications in sediment clean-up, although metabolic mechanisms governing bioleaching processes have been deeply studied and can be considered well established. In this study, we carried out bioleaching experiments, using autotrophic and heterotrophic acidophilic bacteria strains, and worked with marine sediments characterized by different geochemical properties and metal concentrations and speciations. The solubilization efficiency of the metals was highly variable, with the highest for Zn (40%-76%) and the lowest for Pb (0%-7%). Our data suggest that the role of autotrophic Fe/S oxidizing bacteria is mainly associated with the production and re-cycling of leaching chemical species, mainly as protons and ferric ions. Metal solubilization appears to be more related to establishing environmental conditions that allow each metal or semimetal to remain stable in the solution phase. Thus, the maintenance of acid and oxidative conditions, the chemical behavior in aqueous environment of each metal species and the geochemical characteristics of sediment interact intimately to influence metal solubilization in site-specific and metal-specific way. Copyright © 2013 Elsevier Ltd. All rights reserved.

Statistical thermodynamics unveils the dissolution mechanism of cellobiose.

PubMed

Nicol, Thomas W J; Isobe, Noriyuki; Clark, James H; Shimizu, Seishi

2017-08-30

In the study of the cellulose dissolution mechanism opinion is still divided. Here, the solution interaction components of the most prominent hypotheses for the driving force of cellulose dissolution were evaluated quantitatively. Combining a rigorous statistical thermodynamic theory and cellobiose solubility data in the presence of chloride salts, whose cations progress in the Hofmeister series (KCl, NaCl, LiCl and ZnCl 2 ), we have shown that cellobiose solubilization is driven by the preferential accumulation of salts around the solutes which is stronger than cellobiose hydration. Yet contrary to the classical chaotropy hypothesis, increasing salt concentration leads to cellobiose dehydration in the presence of the strongest solubilizer ZnCl 2 . However, thanks to cellobiose dehydration, cellobiose-salt interaction still remains preferential despite weakening salt accumulation. Based on such insights, the previous hypotheses based on hydrophobicity and polymer charging have also been evaluated quantitatively. Thus, our present study successfully paved a way towards identifying the basic driving forces for cellulose solubilization in a quantitative manner for the first time. When combined with unit additivity methods this quantitative information could lead to a full understanding of cellulose solubility.

Comparison of the solubilizing efficiencies of some pH lowering (sulphur and (NH4)2SO4) amendments on Cd and Zn mobility in soils.

PubMed

Amoakwah, Emmanuel; Van Slycken, Stijn; Essumang, David K

2014-08-01

The use of conventional methods to clean up the soil is very expensive and destructive to the ecosystem. The concept of phytoextraction has been introduced to safely manage soils contaminated with heavy metals. However, using plants to remediate polluted soils is a lengthy process. This has necessitated the use of amendments to potentially enhance solubilization of metals in order to increase their bioavailability in the soil solution. A pot experiment was conducted to study the effect of some selected pH lowering amendments [elemental sulphur and (NH4)2SO4] on the solubility and availability of Cd and Zn. The application of these amendments resulted in a decrease in the pH of the soil. The decrease in pH significantly enhanced the solubilization and the mobility of Cd and Zn into the soil solution. The CaCl2 extraction protocol was employed to study the effects of the various amendments on the mobility of Cd and Zn.

Structure and characteristics of acid and pepsin-solubilized collagens from the skin of cobia (Rachycentron canadum).

PubMed

Zeng, Shaokui; Yin, Juanjuan; Yang, Shuqi; Zhang, Chaohua; Yang, Ping; Wu, Wenlong

2012-12-01

Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin of cobia (Rachycentron canadum). The yields of ASC and PSC were 35.5% and 12.3%, respectively. Based on the protein patterns and carboxymethyl-cellulose chromatography, ASC and PSC were composed of α1α2α3 heterotrimers and were characterised as type I collagen with no disulfide bond. Their amounts of imino acids were 203 and 191 residues per 1000 residues, respectively. LC-MS/MS analysis demonstrated the high sequences similarities of ASC and PSC. Fourier transform infrared spectroscopy spectra showed that the amide I, II and III peaks of PSC were obtained at a lower wave number compared with ASC. The thermal denaturation temperatures of ASC and PSC, as measured by viscometry, were 34.62 and 33.97°C, respectively. The transition temperatures (T(max)) were 38.17 and 36.03°C, respectively, as determined by differential scanning calorimetry (DSC). Both collagens were soluble at acidic pH and below 2% (w/v) NaCl concentration. Copyright © 2012 Elsevier Ltd. All rights reserved.

Enhancement of solubilization and biodegradation of polyaromatic hydrocarbons by the bioemulsifier alasan

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barkay, T.; Navon-Venezia, S.; Ron, E.Z.

Alasan, a high-molecular-weight bioemulsifier complex of an anionic polysaccharide and proteins that is produced by Acinetobacter radioresistent KA53 enhanced the aqueous solubility and biodegradation rates of polyaromatic hydrocarbons (PAHs). In the presence of 500 {micro}g of alasan ml{sup {minus}1}, the apparent aqueous solubilities of phenanthrene, fluoranthene, and pyrene were increased 6.6-, 25.7-, and 19.8-fold, respectively. Physicochemical characterization of the solubilization activity suggested that alasan solubilizes PAHs by a physical interaction, most likely of a hydrophobic nature, and that this interaction is slowly reversible. Moreover, the increase in apparent aqueous solubility of PAHs does not depend on the conformation of alasanmore » and is not affected by the formation of multimolecular aggregates of alasan above its saturation concentration. The presence of alasan more than doubled the rate of [{sup 14}C]fluoranthene mineralization and significantly increased the rate of [{sup 14}C]phenanthrene mineralization by Sphingomonas paucimobilis EPA505. The results suggest that alasan-enhanced solubility of hydrophobic compounds has potential applications in bioremediation.« less

Bioleaching of multiple heavy metals from contaminated sediment by mesophile consortium.

PubMed

Gan, Min; Zhou, Shuang; Li, Mingming; Zhu, Jianyu; Liu, Xinxing; Chai, Liyuan

2015-04-01

A defined mesophile consortium including Acidithiobacillus ferrooxidans, Acidithiobacillus thiooxidans, and Leptospirilum ferriphilum was applied in bioleaching sediments contaminated with multiple heavy metals. Flask experiments showed that sulfur favored the acidification in the early stage while pyrite led to a great acidification potential in the later stage. An equal sulfur/pyrite ratio got the best acidification effect. Substrate utilization started with sulfur in the early stage, and then the pH decline and the community shift give rise to the utilization of pyrite. Solubilization efficiency of Zn, Cu, Mn, and Cd reached 96.1, 93.3, 92.13, and 87.65%, respectively. Bioleaching efficiency of other elements (As, Hg, Pb) was not more than 30%. Heavy metal solubilization was highly negatively correlated with pH variation. Logistic models were well fitted with the solubilization efficiency, which can be used to predict the bioleaching process. The dominant species in the early stage of bioleaching were A. ferrooxidans and A. thiooxidans, and the abundance of L. ferriphilum increased together with pyrite utilization and pH decline.

Statistical key factors optimization of conditions for hydrogen production from S-TE (solubilization by thermophilic enzyme) waste sludge.

PubMed

Guo, Liang; Zhao, Jun; She, Zonglian; Lu, Mingmin; Zong, Yan

2013-06-01

Waste sludge can be solubilized after S-TE (solubilization by thermophilic enzyme) pretreatment as the cryptic growth occurs at the expense of the cell lysate. The hydrogen production from S-TE sludge is greatly influenced by many factors. In this study, factors including pH, C/N, C/P, and Fe(2+) affecting hydrogen production from S-TE sludge were optimized using uniform design. The optimum condition for maximum hydrogen yield of 68.4 ml H2/g VSS (volatile suspended solid) could be predicted from regression model, and the optimum conditions were pH of 6.4, C/N ratio of 38, C/P ratio of 265, and Fe(2+) concentration of 85 mg/L. There was interaction effect of factors on hydrogen production from S-TE sludge. Different pH, C/N, C/P and Fe(2+) conditions could influence the VSS removal rate, carbohydrate and protein utilization. When the highest compositions of acetate and ethanol and lowest propionate were observed in metabolites, effective hydrogen production was also achieved. Copyright © 2013 Elsevier Ltd. All rights reserved.

Stimulating short-chain fatty acids production from waste activated sludge by nano zero-valent iron.

PubMed

Luo, Jingyang; Feng, Leiyu; Chen, Yinguang; Li, Xiang; Chen, Hong; Xiao, Naidong; Wang, Dongbo

2014-10-10

An efficient and green strategy, i.e. adding nano zero-valent iron into anaerobic fermentation systems to remarkably stimulate the accumulation of short-chain fatty acids from waste activated sludge via accelerating the solubilization and hydrolysis processes has been developed. In the presence of nano zero-valent iron, not only the short-chain fatty acids production was significantly improved, but also the fermentation time for maximal short-chain fatty acids was shortened compared with those in the absence of nano zero-valent iron. Mechanism investigations showed that the solubilization of sludge, hydrolysis of solubilized substances and acidification of hydrolyzed products were all enhanced by addition of nano zero-valent iron. Also, the general microbial activity of anaerobes and relative activities of key enzymes with hydrolysis and acidification of organic matters were improved than those in the control. 454 high-throughput pyrosequencing analysis suggested that the abundance of bacteria responsible for waste activated sludge hydrolysis and short-chain fatty acids production was greatly enhanced due to nano zero-valent iron addition. Copyright © 2014 Elsevier B.V. All rights reserved.

Acyclic Cucurbit[n]uril-type Molecular Containers: Influence of Aromatic Walls on their Function as Solubilizing Excipients for Insoluble Drugs

PubMed Central

2015-01-01

We studied the influence of the aromatic sidewalls on the ability of acyclic CB[n]-type molecular containers (1a–1e) to act as solubilizing agents for 19 insoluble drugs including the developmental anticancer agent PBS-1086. All five containers exhibit good water solubility and weak self-association (Ks ≤ 624 M–1). We constructed phase solubility diagrams to extract Krel and Ka values for the container·drug complexes. The acyclic CB[n]-type containers generally display significantly higher Ka values than HP-β-CD toward drugs. Containers 1a–1e bind the steroidal ring system and aromatic moieties of insoluble drugs. Compound 1b displays highest affinity toward most of the drugs studied. Containers 1a and 1b are broadly applicable and can be used to formulate a wider variety of insoluble drugs than was previously possible with cyclodextrin technology. For drugs that are solubilized by both HP-β-CD and 1a–1e, lower concentrations of 1a–1e are required to achieve identical [drug]. PMID:25369565

Acyclic cucurbit[n]uril-type molecular containers: influence of aromatic walls on their function as solubilizing excipients for insoluble drugs.

PubMed

Zhang, Ben; Isaacs, Lyle

2014-11-26

We studied the influence of the aromatic sidewalls on the ability of acyclic CB[n]-type molecular containers (1a-1e) to act as solubilizing agents for 19 insoluble drugs including the developmental anticancer agent PBS-1086. All five containers exhibit good water solubility and weak self-association (Ks ≤ 624 M(-1)). We constructed phase solubility diagrams to extract Krel and Ka values for the container·drug complexes. The acyclic CB[n]-type containers generally display significantly higher Ka values than HP-β-CD toward drugs. Containers 1a-1e bind the steroidal ring system and aromatic moieties of insoluble drugs. Compound 1b displays highest affinity toward most of the drugs studied. Containers 1a and 1b are broadly applicable and can be used to formulate a wider variety of insoluble drugs than was previously possible with cyclodextrin technology. For drugs that are solubilized by both HP-β-CD and 1a-1e, lower concentrations of 1a-1e are required to achieve identical [drug].

Combined treatment of alkaline and disperser for improving solubilization and anaerobic biodegradability of dairy waste activated sludge.

PubMed

Uma Rani, R; Kaliappan, S; Adish Kumar, S; Rajesh Banu, J

2012-12-01

An investigation into the influence of combined alkaline and disperser pretreatment on sludge disintegration was studied. The effects of four variables, alkalines (NaOH, KOH, Ca(OH)(2)), treatment time (15-180 min), pH (8-11) and rpm (4000-24,000) were investigated. The effect of sludge pretreatment was evaluated by COD solubilization, suspended solids reduction and biogas production. The best performances, in terms of COD solubilization, SS reduction and biogas production, were the ones that occurred for specific energy input of 4544 kJ kg(-1) TS for NaOH at pH10, were found to be 24%, 23.3% and 76%, higher than the control, respectively. Not only the increase in biogas production was investigated, excluding protein hydrolysis was also performed successfully by this combined pretreatment even at low specific energy input. Thus, this chemo-mechanical is an effective method for enhancement of biodegradability and it laid the basis to produce higher biogas quantities, to improve clean energy generation from WAS. Copyright © 2012 Elsevier Ltd. All rights reserved.

Effect of Pluronic F-127 on the photosensitizing activity of tetraphenylporphyrins in organic and aqueous phases

NASA Astrophysics Data System (ADS)

Savko, M. A.; Aksenova, N. A.; Akishina, A. K.; Khasanova, O. V.; Glagolev, N. N.; Rumyantseva, V. D.; Zhdanova, K. A.; Spokoinyi, A. L.; Solov'eva, A. B.

2017-11-01

The solubilization of hydrophobic porphyrin photosensitizers (PPSes) to obtain corresponding water-soluble forms is an important line of modern antimicrobial photodynamic therapy. It is shown that a triblock copolymer of ethylene and propylene oxides, Pluronic F-127, one of the most nontoxic and effective polymer surface active substances (SASes), can be used for the conversion of hydrophobic tetraphenylporphyrin (TPP) and monosubstituted and tetrasubstituted hydroxy, amino, and nitro TPPs into water-soluble forms. It is found that Pluronic has a substantially higher solubilizing affinity (defined as the minimum molar concentration of an SAS required for the complete migration of porphyrin with a specific molar concentration to the aqueous phase) toward monosubstituted TPPs than to corresponding tetrasubstituted porphyrins. It is shown that with Pluronic in the organic phase, the activity of tetraphenylporphyrin in a test reaction of the oxidation of anthracene is higher than that of its monosubstituted and tetrasubstituted derivatives. In an aqueous medium, the activity of solubilized mono derivatives of TPP is comparable to that of unsubstituted TPP and is higher than the activity of the corresponding tetra derivatives of TPP.

Transient expression of the influenza A virus PB1-F2 protein using a plum pox virus-based vector in Nicotiana benthamiana.

PubMed

Kamencayová, M; Košík, I; Hunková, J; Subr, Z W

2014-01-01

PB1-F2 protein of influenza A virus (IAV) was cloned in a plum pox virus (PPV) genome-based vector and attempts to express it in biolistically transfected Nicotiana benthamiana plants were performed. The vector-insert construct replicated in infected plants properly and was stable during repeated passage by mechanical inoculation, as demonstrated by disease symptoms and immunoblot detection of PPV capsid protein, while PB1-F2-specific band was more faint. We showed that it was due its low solubility. Modification of sample preparation (denaturation/solubilization preceding the centrifugation of cell debris) led to substantial signal enhancement. Maximal level of PB1-F2 expression in plants was observed 12 days post inoculation (dpi). Only 1% SDS properly solubilized the protein, other detergents were much less efficient. Solubilization with 8M urea released approximately 50% of PB1-F2 from the plant tissues, thus the treatment with this removable chaotropic agent may be a good starting point for the purification of the protein for eventual functional studies in the future.

Solubilization of polycyclic aromatic hydrocarbons in micellar nonionic surfactant solutions

DOE Office of Scientific and Technical Information (OSTI.GOV)

Edwards, D.A.; Luthy, R.G.; Liu, Zhongbao

1991-01-01

Experimental data are presented on the enhanced apparent solubilities of naphthalene, phenanthrene, and pyrene resulting from solubilization in aqueous solutions of four commercial, nonionic surfactants: an alkyl polyoxyethylene (POE) type, two octylphenol POE types, and a nonylphenol POE type. Apparent solubilities of the polycyclic aromatic hydrocarbon (PAH) compounds in surfactant solutions were determined by radiolabeled techniques. Solubilization of each PAH compound commenced at the surfactant critical micelle concentration and was proportional to the concentration of surfactant in micelle form. The partitioning of organic compounds between surfactant micelles and aqueous solution is characterized by a mole fraction micelle-phase/aqueous-phase partition coefficient, K{submore » m}. Values of log K{sub m} for PAH compounds in surfactant solutions of this study range from 4.57 to 6.53. Log K{sub m} appears to be a linear function of log K{sub ow} for a given surfactant solution. A knowledge of partitioning in aqueous surfactant systems is a prerequisite to understanding mechanisms affecting the behavior of hydrophobic organic compounds in soil-water systems in which surfactants play a role in contaminant remediation or facilitated transport.« less

Proteomic Analysis of Human Tendon and Ligament: Solubilization and Analysis of Insoluble Extracellular Matrix in Connective Tissues.

PubMed

Sato, Nori; Taniguchi, Takako; Goda, Yuichiro; Kosaka, Hirofumi; Higashino, Kosaku; Sakai, Toshinori; Katoh, Shinsuke; Yasui, Natsuo; Sairyo, Koichi; Taniguchi, Hisaaki

2016-12-02

Connective tissues such as tendon, ligament and cartilage are mostly composed of extracellular matrix (ECM). These tissues are insoluble, mainly due to the highly cross-linked ECM proteins such as collagens. Difficulties obtaining suitable samples for mass spectrometric analysis render the application of modern proteomic technologies difficult. Complete solubilization of them would not only elucidate protein composition of normal tissues but also reveal pathophysiology of pathological tissues. Here we report complete solubilization of human Achilles tendon and yellow ligament, which is achieved by chemical digestion combined with successive protease treatment including elastase. The digestion mixture was subjected to liquid chromatography-mass spectrometry. The low specificity of elastase was overcome by accurate mass analysis achieved using FT-ICR-MS. In addition to the detailed proteome of both tissues, we also quantitatively determine the major protein composition of samples, by measuring peak area of some characteristic peptides detected in tissue samples and in purified proteins. As a result, differences between human Achilles tendon and yellow ligament were elucidated at molecular level.

Modified denatured lysozyme effectively solubilizes fullerene c60 nanoparticles in water

NASA Astrophysics Data System (ADS)

Siepi, Marialuisa; Politi, Jane; Dardano, Principia; Amoresano, Angela; De Stefano, Luca; Monti, Daria Maria; Notomista, Eugenio

2017-08-01

Fullerenes, allotropic forms of carbon, have very interesting pharmacological effects and engineering applications. However, a very low solubility both in organic solvents and water hinders their use. Fullerene C60, the most studied among fullerenes, can be dissolved in water only in the form of nanoparticles of variable dimensions and limited stability. Here the effect on the production of C60 nanoparticles by a native and denatured hen egg white lysozyme, a highly basic protein, has been systematically studied. In order to obtain a denatured, yet soluble, lysozyme derivative, the four disulfides of the native protein were reduced and exposed cysteines were alkylated by 3-bromopropylamine, thus introducing eight additional positive charges. The C60 solubilizing properties of the modified denatured lysozyme proved to be superior to those of the native protein, allowing the preparation of biocompatible highly homogeneous and stable C60 nanoparticles using lower amounts of protein, as demonstrated by dynamic light scattering, transmission electron microscopy and atomic force microscopy studies. This lysozyme derivative could represent an effective tool for the solubilization of other carbon allotropes.

Sequential ultrasound and low-temperature thermal pretreatment: Process optimization and influence on sewage sludge solubilization, enzyme activity and anaerobic digestion.

PubMed

Neumann, Patricio; González, Zenón; Vidal, Gladys

2017-06-01

The influence of sequential ultrasound and low-temperature (55°C) thermal pretreatment on sewage sludge solubilization, enzyme activity and anaerobic digestion was assessed. The pretreatment led to significant increases of 427-1030% and 230-674% in the soluble concentrations of carbohydrates and proteins, respectively, and 1.6-4.3 times higher enzymatic activities in the soluble phase of the sludge. Optimal conditions for chemical oxygen demand solubilization were determined at 59.3kg/L total solids (TS) concentration, 30,500kJ/kg TS specific energy and 13h thermal treatment time using response surface methodology. The methane yield after pretreatment increased up to 50% compared with the raw sewage sludge, whereas the maximum methane production rate was 1.3-1.8 times higher. An energy assessment showed that the increased methane yield compensated for energy consumption only under conditions where 500kJ/kg TS specific energy was used for ultrasound, with up to 24% higher electricity recovery. Copyright © 2017 Elsevier Ltd. All rights reserved.

Antigenic properties of the envelope of influenza virus rendered soluble by surfactant-solvent systems

PubMed Central

Larin, N. M.; Gallimore, P. H.

1971-01-01

Dissociating chemical treatments employing surfactant-solvent systems were applied to purified influenza A and B viruses to obtain viral preparations possessing a significantly higher or lower haemagglutinating activity than the intact virus. All preparations, whether with high or low haemagglutinating activity, with the exception of envelope protein solubilized by Triton X-100, were significantly lacking in the ability to excite the formation of haemagglutination-inhibiting and virus-neutralizing antibodies in inoculated ferrets. In contrast to other treatments, Triton X-100 treatment of virus significantly enhanced the antigenicity of viral protein as judged by virus neutralization and haemagglutination inhibition tests. Yet the haemagglutinating activity of the envelope protein solubilized with Triton X-100 was about 1% that of the intact virus. Results suggest that the correlation assumed to exist between the haemagglutinating activity of influenza virus and its ability to excite the formation of humoral antibodies is coincidental. Another important point is that the specific antigenicity of viral protein may be lost or enhanced owing to effects, other than solubilization, by surface-active agents. PMID:5291750

Biodegradation and bioconversion of coals by fungi: Technical report No. 6 for the period December 1, 1986-February 28, 1987. [Candida sp. , Cunninghamella sp. , Acremonium sp. , and Trametes versicolor

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ward, B.

We continued our tests for biosolubilization activity on lignite, subbituminous, and bituminous coals, using both surface solubilization and agar diffusion assays for selected lignite-solubilizing strains challenged with untreated and treated coals shown in earlier tests to be resistant to biosolubilization. These latter were subjected to treatments with heat or pressurized oxygen atmosphere, or to combinations of the same. We have observed biosolubilization of some weathered high-rank coals by some of the fungal isolates tested. 1 tab.

Solubilization of water in water-in-oil microemulsions of kerosene

DOE Office of Scientific and Technical Information (OSTI.GOV)

Andheria, A.P.; Bhagwat, S.S.

1995-04-01

The incorporation of water into fuels formulated as microemulsions can offer several advantages such as fire resistance, increased flash point, and improved air-fuel contact. To this end, phase equilibria of kerosene microemulsions employing ionic and nonionic surfactants such as sodium di-(2-ethylhexyl) sulfosuccinate (AOT), lauryl diethanolamide (LDEA), nonylphenol EO-4.5 (NPEO-4.5), sorbitan monolaurate (Span-20), and cetyltrimethylammonium bromide (CTAB), as well as cosurfactants such as n-pentanol, n-hexanol, and n-heptanol, were studied. The effect of the aromaticity of the oil phase on the solubilization of water was also investigated.