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Sample records for acid hydrolysis revealed

  1. Acid hydrolysis of cellulose

    SciTech Connect

    Salazar, H.

    1980-12-01

    One of the alternatives to increase world production of etha nol is by the hydrolysis of cellulose content of agricultural residues. Studies have been made on the types of hydrolysis: enzimatic and acid. Data obtained from the sulphuric acid hydrolysis of cellulose showed that this process proceed in two steps, with a yield of approximately 95% glucose. Because of increases in cost of alternatives resources, the high demand of the product and the more economic production of ethanol from cellulose materials, it is certain that this technology will be implemented in the future. At the same time further studies on the disposal and reuse of the by-products of this production must be undertaken.

  2. Structure of Human Acid Sphingomyelinase Reveals the Role of the Saposin Domain in Activating Substrate Hydrolysis.

    PubMed

    Xiong, Zi-Jian; Huang, Jingjing; Poda, Gennady; Pomès, Régis; Privé, Gilbert G

    2016-07-31

    Acid sphingomyelinase (ASM) is a lysosomal phosphodiesterase that catalyzes the hydrolysis of sphingomyelin to produce ceramide and phosphocholine. While other lysosomal sphingolipid hydrolases require a saposin activator protein for full activity, the ASM polypeptide incorporates a built-in N-terminal saposin domain and does not require an external activator protein. Here, we report the crystal structure of human ASM and describe the organization of the three main regions of the enzyme: the N-terminal saposin domain, the proline-rich connector, and the catalytic domain. The saposin domain is tightly associated along an edge of the large, bowl-shaped catalytic domain and adopts an open form that exposes a hydrophobic concave surface approximately 30Å from the catalytic center. The calculated electrostatic potential of the enzyme is electropositive at the acidic pH of the lysosome, consistent with the strict requirement for the presence of acidic lipids in target membranes. Docking studies indicate that sphingomyelin binds with the ceramide-phosphate group positioned at the binuclear zinc center and molecular dynamic simulations indicate that the intrinsic flexibility of the saposin domain is important for monomer-dimer exchange and for membrane interactions. Overall, ASM uses a combination of electrostatic and hydrophobic interactions to cause local disruptions of target bilayers in order to bring the lipid headgroup to the catalytic center in a membrane-bound reaction. PMID:27349982

  3. Acid hydrolysis of cellulose to yield glucose

    DOEpatents

    Tsao, George T.; Ladisch, Michael R.; Bose, Arindam

    1979-01-01

    A process to yield glucose from cellulose through acid hydrolysis. Cellulose is recovered from cellulosic materials, preferably by pretreating the cellulosic materials by dissolving the cellulosic materials in Cadoxen or a chelating metal caustic swelling solvent and then precipitating the cellulose therefrom. Hydrolysis is accomplished using an acid, preferably dilute sulfuric acid, and the glucose is yielded substantially without side products. Lignin may be removed either before or after hydrolysis.

  4. Acid Hydrolysis of Trioxalatocobaltate (III) Ion

    ERIC Educational Resources Information Center

    Wiggans, P. W.

    1975-01-01

    Describes an investigation involving acid hydrolysis and using both volumetric and kinetic techniques. Presents examples of the determination of the rate constant and its variation with temperature. (GS)

  5. Optimization of dilute acid hydrolysis of Enteromorpha

    NASA Astrophysics Data System (ADS)

    Feng, Dawei; Liu, Haiyan; Li, Fuchao; Jiang, Peng; Qin, Song

    2011-11-01

    Acid hydrolysis is a simple and direct way to hydrolyze polysaccharides in biomass into fermentable sugars. To produce fermentable sugars effectively and economically for fuel ethanol, we have investigated the hydrolysis of Enteromorpha using acids that are typically used to hydrolyze biomass: H2SO4, HCl, H3PO4 and C4H4O4 (maleic acid). 5%(w/w) Enteromorpha biomass was treated for different times (30, 60, and 90 min) and with different acid concentrations (0.6, 1.0, 1.4, 1.8, and 2.2%, w/w) at 121°C. H2SO4 was the most effective acid in this experiment. We then analyzed the hydrolysis process in H2SO4 in detail using high performance liquid chromatography. At a sulfuric acid concentration of 1.8% and treatment time of 60 min, the yield of ethanol fermentable sugars (glucose and xylose) was high, (230.5 mg/g dry biomass, comprising 175.2 mg/g glucose and 55.3 mg/g xylose), with 48.6% of total reducing sugars being ethanol fermentable. Therefore, Enteromorpha could be a good candidate for production of fuel ethanol. In future work, the effects of temperature and biomass concentration on hydrolysis, and also the fermentation of the hydrolysates to ethanol fuel should be focused on.

  6. Mechanisms of lactone hydrolysis in acidic conditions.

    PubMed

    Gómez-Bombarelli, Rafael; Calle, Emilio; Casado, Julio

    2013-07-19

    The acid-catalyzed hydrolysis of linear esters and lactones was studied using a hybrid supermolecule-polarizable continuum model (PCM) approach including up to six water molecules. The compounds studied included two linear esters, four β-lactones, two γ-lactones, and one δ-lactone: ethyl acetate, methyl formate, β-propiolactone, β-butyrolactone, β-isovalerolactone, diketene (4-methyleneoxetan-2-one), γ-butyrolactone, 2(5H)-furanone, and δ-valerolactone. The theoretical results are in good quantitative agreement with the experimental measurements reported in the literature and also in excellent qualitative agreement with long-held views regarding the nature of the hydrolysis mechanisms at molecular level. The present results help to understand the balance between the unimolecular (A(AC)1) and bimolecular (A(AC)2) reaction pathways. In contrast to the experimental setting, where one of the two branches is often occluded by the requirement of rather extreme experimental conditions, we have been able to estimate both contributions for all the compounds studied and found that a transition from A(AC)2 to A(AC)1 hydrolysis takes place as acidity increases. A parallel work addresses the neutral and base-catalyzed hydrolysis of lactones. PMID:23731203

  7. Acid-functionalized nanoparticles for biomass hydrolysis

    NASA Astrophysics Data System (ADS)

    Pena Duque, Leidy Eugenia

    Cellulosic ethanol is a renewable source of energy. Lignocellulosic biomass is a complex material composed mainly of cellulose, hemicellulose, and lignin. Biomass pretreatment is a required step to make sugar polymers liable to hydrolysis. Mineral acids are commonly used for biomass pretreatment. Using acid catalysts that can be recovered and reused could make the process economically more attractive. The overall goal of this dissertation is the development of a recyclable nanocatalyst for the hydrolysis of biomass sugars. Cobalt iron oxide nanoparticles (CoFe2O4) were synthesized to provide a magnetic core that could be separated from reaction using a magnetic field and modified to carry acid functional groups. X-ray diffraction (XRD) confirmed the crystal structure was that of cobalt spinel ferrite. CoFe2O4 were covered with silica which served as linker for the acid functions. Silica-coated nanoparticles were functionalized with three different acid functions: perfluoropropyl-sulfonic acid, carboxylic acid, and propyl-sulfonic acid. Transmission electron microscope (TEM) images were analyzed to obtain particle size distributions of the nanoparticles. Total carbon, nitrogen, and sulfur were quantified using an elemental analyzer. Fourier transform infra-red spectra confirmed the presence of sulfonic and carboxylic acid functions and ion-exchange titrations accounted for the total amount of catalytic acid sites per nanoparticle mass. These nanoparticles were evaluated for their performance to hydrolyze the beta-1,4 glycosidic bond of the cellobiose molecule. Propyl-sulfonic (PS) and perfluoropropyl-sulfonic (PFS) acid functionalized nanoparticles catalyzed the hydrolysis of cellobiose significantly better than the control. PS and PFS were also evaluated for their capacity to solubilize wheat straw hemicelluloses and performed better than the control. Although PFS nanoparticles were stronger acid catalysts, the acid functions leached out of the nanoparticle during

  8. Characterization of acidic polysaccharides from the mollusks through acid hydrolysis.

    PubMed

    Cao, Jiuling; Wen, Chengrong; Lu, Jiaojiao; Teng, Nan; Song, Shuang; Zhu, Beiwei

    2015-10-01

    Uronic acid-containing polysaccharides (UACPs) including glycosaminoglycans (GAGs) exist widely in nature. Herein we propose an elegant methodology to identify UACPs by analyzing their disaccharides produced from the acid hydrolysis using HPLC-MS(n) upon 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization. Based on the optimization of experimental conditions by the single factor experiment and orthogonal test design, the combination of 1.3M TFA at 105°C for 3h is found to be the optimum. Subsequently, these conditions were applied to investigate the distribution of UACPs in 20 selected species of edible Bivalvia and Gastropoda. PMP-disaccharides derived from UACPs in mollusks were identified by comparing the retention time and mass spectra with those of the reference PMP-disaccharides from hyaluronic acid (HA), chondroitin sulfate (CS), heparin (HP), and AGSP with →4)-GlcA(1→2)-Man(1→ repeating units. The analysis reveals the prevalence of CS in the shellfishes as well as the HP, and existence of three non-GAG UACPs in 7 mollusks. PMID:26076626

  9. Enhanced functional properties of tannic acid after thermal hydrolysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Thermal hydrolysis processing of fresh tannic acid was carried out in a closed reactor at four different temperatures (65, 100, 150 and 200°C). Pressures reached in the system were 1.3 and 4.8 MPa at 150 and 200°C, respectively. Hydrolysis products (gallic acid and pyrogallol) were separated and qua...

  10. ESTIMATION OF CARBOXYLIC ACID ESTER HYDROLYSIS RATE CONSTANTS

    EPA Science Inventory

    SPARC chemical reactivity models were extended to calculate hydrolysis rate constants for carboxylic acid esters from molecular structure. The energy differences between the initial state and the transition state for a molecule of interest are factored into internal and external...

  11. Furfural/ethanol coproduction from biomass feedstocks using acid hydrolysis

    SciTech Connect

    Barrier, J.W.; Bulls, M.M.; Broder, J.D.

    1996-12-31

    The Tennessee Valley Authority (TVA) has been involved in research and development to produce high-value chemicals from biomass for over 15 years. Use of biomass releases less carbon dioxide than use of fossil fuels, and thus represents a more environmentally friendly source of chemicals and fuels. Two biomass conversion processes have been developed as a result of TVA`s work--concentrated acid hydrolysis and dilute acid hydrolysis. Both processes use sulfuric acid as a catalyst. Early hydrolysis research focused on improving ethanol yields through hydrolysis and five-carbon sugar fermentation research. Both processes have been demonstrated at the pilot plant scale. Current work is focused on the development of integrated systems for producing ethanol and a variety of other chemicals and products from biomass. Production of furfural and ethanol from high hemicellulose feedstocks has been identified by TVA as an integrated system with technical and economic potential for commercial success. A system design has been developed to produce ethanol and furfural using dilute acid hydrolysis of sycamore. Furfural yields for the system are estimated at 180--240 pound/ton. Ethanol process yields are 25--38 gallon/ton. Capital and operating costs for a 4,500 ton/day facility are estimated to be $609 million and $183 million, respectively. The dilute acid hydrolysis process proposed by TVA will be described along with additional process economics and potential furfural markets.

  12. ESTIMATION OF PHOSPHATE ESTER HYDROLYSIS RATE CONSTANTS. II. ACID AND GENERAL BASE CATALYZED HYDROLYSIS

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to calculate acid and neutral hydrolysis rate constants of phosphate esters in water. The rate is calculated from the energy difference between the initial and transition states of a ...

  13. Acid hydrolysis of Jerusalem artichoke for ethanol fermentation

    SciTech Connect

    Kim, K.; Hamdy, M.K.

    1986-01-01

    An excellent substrate for ethanol production is the Jerusalem artichoke (JA) tuber (Helianthus tuberosus). This crop contains a high level of inulin that can be hydrolyzed mainly to D-fructose and has several distinct advantages as an energy source compared to others. The potential ethanol yield of ca. 4678 L/ha on good agricultural land is equivalent to that obtained from sugar beets and twice that of corn. When JA is to be used for ethanol fermentation by conventional yeast, it is first converted to fermentable sugars by enzymes or acids although various strains of yeast were used for the direct fermentation of JA extracts. Fleming and GrootWassink compared various acids (hydrochloric, sulfuric, citric, and phosphoric) and strong cation exchange resin for their effectiveness on inulin hydrolysis and reported that no differences were noted among the acids or resin in their influence on inulin hydrolysis. Undesirable side reactions were noted during acid hydrolysis leading to the formation of HMF and 2-(2-hydroxy acetyl) furan. The HMF at a level of 0.1% is known to inhibit growth and ethanol fermentation by yeast. In this study the authors established optimal conditions for complete acid-hydrolysis of JA with minimum side reactions and maximum sugar-ethanol production. A material balance for the ethanol production was also determined.

  14. Acid hydrolysis of cellulose in zinc chloride solution

    SciTech Connect

    Cao, N.J.; Xu, Q.; Chen, L.F.

    1995-12-31

    The efficient conversion of cellulosic materials to ethanol has been hindered by the low yield of sugars, the high energy consumption in pretreatment processes, and the difficulty of recycling the pre-treatment agents. Zinc chloride may provide an alternative for pre-treating biomass prior to the hydrolysis of cellulose. The formation of a zinc-cellulose complex during the pretreatment of cellulose improves the yield of glucose in both the enzymatic and acid hydrolysis of cellulose. Low-temperature acid hydrolysis of cellulose in zinc chloride solution is carried out in two stages, a liquefaction stage and a saccharification stage. Because of the formation of zinc-cellulose complex in the first stage, the required amount of acid in the second stage has been decreased significantly. In 67% zinc chloride solution, a 99.5% yield of soluble sugars has been obtained at 70{degrees}C and 0.5M acid concentration. The ratio of zinc chloride to cellulose has been reduced from 4.5 to 1.5, and the yield of soluble sugars is kept above 80%. The rate of hydrolysis is affected by the ratio of zinc chloride to cellulose, acid concentration, and temperature.

  15. Role of bifidobacteria in the hydrolysis of chlorogenic acid

    PubMed Central

    Raimondi, Stefano; Anighoro, Andrew; Quartieri, Andrea; Amaretti, Alberto; Tomás-Barberán, Francisco A; Rastelli, Giulio; Rossi, Maddalena

    2015-01-01

    This study aimed to explore the capability of potentially probiotic bifidobacteria to hydrolyze chlorogenic acid into caffeic acid (CA), and to recognize the enzymes involved in this reaction. Bifidobacterium strains belonging to eight species occurring in the human gut were screened. The hydrolysis seemed peculiar of Bifidobacterium animalis, whereas the other species failed to release CA. Intracellular feruloyl esterase activity capable of hydrolyzing chlorogenic acid was detected only in B. animalis. In silico research among bifidobacteria esterases identified Balat_0669 as the cytosolic enzyme likely responsible of CA release in B. animalis. Comparative modeling of Balat_0669 and molecular docking studies support its role in chlorogenic acid hydrolysis. Expression, purification, and functional characterization of Balat_0669 in Escherichia coli were obtained as further validation. A possible role of B. animalis in the activation of hydroxycinnamic acids was demonstrated and new perspectives were opened in the development of new probiotics, specifically selected for the enhanced bioconversion of phytochemicals into bioactive compounds. PMID:25515139

  16. Role of bifidobacteria in the hydrolysis of chlorogenic acid.

    PubMed

    Raimondi, Stefano; Anighoro, Andrew; Quartieri, Andrea; Amaretti, Alberto; Tomás-Barberán, Francisco A; Rastelli, Giulio; Rossi, Maddalena

    2015-02-01

    This study aimed to explore the capability of potentially probiotic bifidobacteria to hydrolyze chlorogenic acid into caffeic acid (CA), and to recognize the enzymes involved in this reaction. Bifidobacterium strains belonging to eight species occurring in the human gut were screened. The hydrolysis seemed peculiar of Bifidobacterium animalis, whereas the other species failed to release CA. Intracellular feruloyl esterase activity capable of hydrolyzing chlorogenic acid was detected only in B. animalis. In silico research among bifidobacteria esterases identified Balat_0669 as the cytosolic enzyme likely responsible of CA release in B. animalis. Comparative modeling of Balat_0669 and molecular docking studies support its role in chlorogenic acid hydrolysis. Expression, purification, and functional characterization of Balat_0669 in Escherichia coli were obtained as further validation. A possible role of B. animalis in the activation of hydroxycinnamic acids was demonstrated and new perspectives were opened in the development of new probiotics, specifically selected for the enhanced bioconversion of phytochemicals into bioactive compounds. PMID:25515139

  17. A Kinetic Study of DDGS Hemicellulose Acid Hydrolysis and NMR Characterization of DDGS Hydrolysate.

    PubMed

    Chen, Hanchi; Liu, Shijie

    2015-09-01

    Liquid hot water (LHW) extraction was used as a pretreatment method to separate the hemicellulose fraction from dried distiller's grain with solubles (DDGS) into liquid phase. Acid hydrolysis using 3.264 % w/w sulfuric acid at 130 °C was performed to convert polysaccharides in LHW extract to monosaccharides. The structure characterization of DDGS in anomeric carbon region based on proton NMR and heteronuclear single quantum coherence (HSQC) during acid hydrolysis was studied in this work. It reveals that the sugar units in DDGS hemicelluloses are constructed with (1-4)-β-D-xylopyranose and α-L-arabinofuranosyl residues. A kinetic model is included to explain the changing concentration of monomer, oligomer, and sugar units. The model was further tested based on the changing concentration of five carbon sugar units during hydrolysis. PMID:26198022

  18. Ethanol production with dilute acid hydrolysis using partially dried lignocellulosics

    DOEpatents

    Nguyen, Quang A.; Keller, Fred A.; Tucker, Melvin P.

    2003-12-09

    A process of converting lignocellulosic biomass to ethanol, comprising hydrolyzing lignocellulosic materials by subjecting dried lignocellulosic material in a reactor to a catalyst comprised of a dilute solution of a strong acid and a metal salt to lower the activation energy (i.e., the temperature) of cellulose hydrolysis and ultimately obtain higher sugar yields.

  19. Acetic acid oxidation and hydrolysis in supercritical water

    SciTech Connect

    Meyer, J.C.; Marrone, P.A.; Tester, J.W.

    1995-09-01

    Acetic acid (CH{sub 3}COOH) hydrolysis and oxidation in supercritical water were examined from 425--600 C and 246 bar at reactor residence times of 4.4 to 9.8 s. Over the range of conditions studied, acetic acid oxidation was globally 0.72 {+-} 0.15 order in acetic acid and 0.27 {+-} 0.15 order in oxygen to a 95% confidence level, with an activation energy of 168 {+-} 21 kJ/mol, a preexponential factor of 10{sup 9.9{+-}1.7}, and an induction time of about 1.5 s at 525 C. Isothermal kinetic measurements at 550 C over the range 160 to 263 bar indicated that pressure or density did not affect the rate of acetic acid oxidation as much as was previously observed in the oxidation of hydrogen or carbon monoxide in supercritical water. Major products of acetic acid oxidation in supercritical water are carbon dioxide, carbon monoxide, methane, and hydrogen. Trace amounts of propenoic acid were occasionally detected. Hydrolysis or hydrothermolysis in the absence of oxygen resulted in approximately 35% conversion of acetic acid at 600 C, 246 bar, and 8-s reactor residence time. Regression of the limited hydrolysis runs assuming a reaction rate first-order in organic gave a global rate expression with a preexponential factor of 10{sup 4.4{+-}1.1} and an activation energy of 94 {+-} 17 kJ/mol.

  20. Enhancement of hydrolysis of Chlorella vulgaris by hydrochloric acid.

    PubMed

    Park, Charnho; Lee, Ja Hyun; Yang, Xiaoguang; Yoo, Hah Young; Lee, Ju Hun; Lee, Soo Kweon; Kim, Seung Wook

    2016-06-01

    Chlorella vulgaris is considered as one of the potential sources of biomass for bio-based products because it consists of large amounts of carbohydrates. In this study, hydrothermal acid hydrolysis with five different acids (hydrochloric acid, nitric acid, peracetic acid, phosphoric acid, and sulfuric acid) was carried out to produce fermentable sugars (glucose, galactose). The hydrothermal acid hydrolysis by hydrochloric acid showed the highest sugar production. C. vulgaris was hydrolyzed with various concentrations of hydrochloric acid [0.5-10 % (w/w)] and microalgal biomass [20-140 g/L (w/v)] at 121 °C for 20 min. Among the concentrations examined, 2 % hydrochloric acid with 100 g/L biomass yielded the highest conversion of carbohydrates (92.5 %) into reducing sugars. The hydrolysate thus produced from C. vulgaris was fermented using the yeast Brettanomyces custersii H1-603 and obtained bioethanol yield of 0.37 g/g of algal sugars. PMID:26899601

  1. Carbon-based strong solid acid for cornstarch hydrolysis

    SciTech Connect

    Nata, Iryanti Fatyasari; Irawan, Chairul; Mardina, Primata; Lee, Cheng-Kang

    2015-10-15

    Highly sulfonated carbonaceous spheres with diameter of 100–500 nm can be generated by hydrothermal carbonization of glucose in the presence of hydroxyethylsulfonic acid and acrylic acid at 180 °C for 4 h. The acidity of the prepared carbonaceous sphere C4-SO{sub 3}H can reach 2.10 mmol/g. It was used as a solid acid catalyst for the hydrolysis of cornstarch. Total reducing sugar (TRS) concentration of 19.91 mg/mL could be obtained by hydrolyzing 20 mg/mL cornstarch at 150 °C for 6 h using C4-SO{sub 3}H as solid acid catalyst. The solid acid catalyst demonstrated good stability that only 9% decrease in TRS concentration was observed after five repeat uses. The as-prepared carbon-based solid acid catalyst can be an environmentally benign replacement for homogeneous catalyst. - Highlights: • Carbon solid acid was successfully prepared by one-step hydrothermal carbonization. • The acrylic acid as monomer was effectively reduce the diameter size of particle. • The solid acid catalyst show good catalytic performance of starch hydrolysis. • The solid acid catalyst is not significantly deteriorated after repeated use.

  2. Influence of acid precursors on physicochemical properties of nanosized titania synthesized by thermal-hydrolysis method

    SciTech Connect

    Rajesh, B.; Sasirekha, N.R.; Chen, Y.-W.

    2008-03-04

    The influence of nature and concentration of acid species on surface morphology and physicochemical properties of titania particles synthesized by direct thermal hydrolysis of titanium tetrachloride was investigated. The acids used were hydrochloric acid, nitric acid, sulfuric acid, and perchloric acid with a concentration of 3 M. Thermal hydrolysis of titanium tetrachloride in hydrochloric acid and perchloric acid with molar ratios of [H{sup +}]/[Ti{sup 4+}] = 0.5, 1.0, 1.5, and 2.0, respectively, was used to study the effect of acid concentration. The synthesized materials were characterized by X-ray diffraction, scanning electron microscopy, transmission electron microscopy, dynamic light scattering, and thermogravimetric analysis. Characterization of the samples by X-ray diffraction studies revealed the influence of acid species on the phase transformation of titania. Samples prepared by hydrochloric acid, nitric acid, and perchloric acid formed rutile phase with rhombus primary particles, while sulfuric acid resulted in anatase phase with flake-shaped primary particles. Transmission electron microscopy and dynamic light scattering results confirmed the nanosized titania particles and the agglomeration of primary particles to form secondary particles in spherical shape. The particle size of titania prepared using perchloric acid was smaller than those prepared with other acid sources. A direct correlation between [H{sup +}]/[Ti{sup 4+}] ratio and particle size of titania was observed.

  3. Acid-catalyzed hydrolysis of BMS-582664: degradation product identification and mechanism elucidation.

    PubMed

    Zhao, Fang; Derbin, George; Miller, Scott; Badawy, Sherif; Hussain, Munir

    2012-09-01

    BMS-582664 is an investigational drug intended for cancer treatment through oral administration. The preformulation studies revealed two unexpected degradation products under acidic conditions by reversed-phase high-performance liquid chromatography with ultraviolet detection. Additional liquid chromatography-mass spectrometry results suggested that these were cleavage (hydrolysis) products of a diaryl ether. To further understand the degradation mechanism, the reaction was carried out in (18) O-labeled water. The (18) O was found to be incorporated in only one of the two hydrolysis products. The results suggest that the corresponding α carbon in the heterocycle was unusually eletrophilic in acidic conditions probably because of the protonation of the neighboring nitrogen. This led to the selective attack by water and the consequent hydrolysis products. The study provides a new example of hydrolytic degradation of pharmaceutical compounds, and the reaction center is an aromatic heterocyclic carbon with an aryloxy substitution. PMID:22189636

  4. Hydrolysis of cellulose catalyzed by novel acidic ionic liquids.

    PubMed

    Zhuo, Kelei; Du, Quanzhou; Bai, Guangyue; Wang, Congyue; Chen, Yujuan; Wang, Jianji

    2015-01-22

    The conversion of cellulosic biomass directly into valuable chemicals becomes a hot subject. Six novel acidic ionic liquids (ILs) based on 2-phenyl-2-imidazoline were synthesized and characterized by UV-VIS, TGA, and NMR. The novel acidic ionic liquids were investigated as catalysts for the hydrolysis of cellulose in 1-butyl-3-methylimidazolium chloride ([Bmim]Cl). The acidic ionic liquids with anions HSO4(-) and Cl(-) showed better catalytic performance for the hydrolysis of cellulose than those with H2PO4(-). The temperature and dosage of water affect significantly the yield of total reducing sugar (TRS). When the hydrolysis of cellulose was catalyzed by 1-propyl sulfonic acid-2-phenyl imidazoline hydrogensulfate (IL-1) and the dosage of water was 0.2g, the TRS yield was up to 85.1% within 60 min at 100°C. These new acidic ionic liquids catalysts are expected to have a wide application in the conversion of cellulose into valuable chemicals. PMID:25439867

  5. Thermal synthesis and hydrolysis of polyglyceric acid. [in orgin of life studying

    NASA Technical Reports Server (NTRS)

    Weber, Arthur L.

    1989-01-01

    Polyglyceric acid was synthesized by thermal condensation of glyceric acid at 80 C in the presence and absence of two mole percent of sulfuric acid catalyst. The acid catalyst accelerated the polymerization over 100-fold and made possible the synthesis of insoluble polymers of both L- and DL-glyceric acid by heating for less than 1 day. Racemization of L-glyceric acid yielded less than 1 percent D-glyceric acid in condensations carried out at 80 C with catalyst for 1 day and without catalyst for 12 days. The condensation of L-glyceric acid yielded an insoluble polymer much more readily than condensation of DL-glyceric acid. Studies of the hydrolysis of poly-DL-glyceric acid revealed that it was considerably more stable under mild acidic conditions compared to neutral pH. The relationship of this study to the origin of life is discussed.

  6. Acid hydrolysis of sweet potato for ethanol production

    SciTech Connect

    Kim, K.; Hamdy, M.K.

    1985-01-01

    Studies were conducted to establish optimal conditions for the acid hydrolysis of sweet potato for maximal ethanol yield. The starch contents of two sweet potato cultivars (Georgia Red and TG-4), based on fresh weight, were 21.1 +/- 0.6% and 27.5 +/- 1.6%, respectively. The results of acid hydrolysis experiments showed the following: (1) both hydrolysis rate and hydroxymethylfurfural (HMF) concentration were a function of HCL concentration, temperature, and time; (2) the reducing sugars were rapidly formed with elevated concentrations of HCl and temperature, but also destroyed quickly; and (3) HMF concentration increased significantly with the concentration of HCl, temperature, and hydrolysis time. Maximum reducing sugar value of 84.2 DE and 0.056% HMF (based on wet weight) was achieved after heating 8% SPS for 15 min in 1N HCl at 110/sup 0/C. Degraded 8% SPS (1N HCl, 97/sup 0/C for 20 min or 110/sup 0/C for 10 min) was utilized as substrate for ethanol fermentation and 3.8% ethanol (v/v) was produced from 1400 mL fermented wort. This is equal to 41.6 g ethanol (200 proof) from 400 g of fresh sweet potato tuber (Georgia Red) or an ethanol yield potential of 431 gal of 200-proof ethanol/acre (from 500 bushel tubers/acre).

  7. Carbon-based strong solid acid for cornstarch hydrolysis

    NASA Astrophysics Data System (ADS)

    Nata, Iryanti Fatyasari; Irawan, Chairul; Mardina, Primata; Lee, Cheng-Kang

    2015-10-01

    Highly sulfonated carbonaceous spheres with diameter of 100-500 nm can be generated by hydrothermal carbonization of glucose in the presence of hydroxyethylsulfonic acid and acrylic acid at 180 °C for 4 h. The acidity of the prepared carbonaceous sphere C4-SO3H can reach 2.10 mmol/g. It was used as a solid acid catalyst for the hydrolysis of cornstarch. Total reducing sugar (TRS) concentration of 19.91 mg/mL could be obtained by hydrolyzing 20 mg/mL cornstarch at 150 °C for 6 h using C4-SO3H as solid acid catalyst. The solid acid catalyst demonstrated good stability that only 9% decrease in TRS concentration was observed after five repeat uses. The as-prepared carbon-based solid acid catalyst can be an environmentally benign replacement for homogeneous catalyst.

  8. Nitric acid catalyzed hydrolysis of SO3 in the formation of sulfuric acid: A theoretical study

    NASA Astrophysics Data System (ADS)

    Long, Bo; Chang, Chun-Ran; Long, Zheng-Wen; Wang, Yi-Bo; Tan, Xing-Feng; Zhang, Wei-Jun

    2013-08-01

    The gas-phase hydrolysis of SO3 in the presence of one water molecule, two water molecules, and nitric acid is investigated utilizing high level quantum chemical methods and transition state theory. The calculated results demonstrate that nitric acid exerts a strong catalytic role in the hydrolysis of SO3 because the activated barrier of hydrolysis of SO3 with the assistance of nitric acid is reduced to about 3.7 kcal/mol, which is about 20 kcal/mol lower than that of the SO3 reaction with water relative to the respective pre-reactive complex.

  9. Extraterrestrial material analysis: loss of amino acids during liquid-phase acid hydrolysis

    NASA Astrophysics Data System (ADS)

    Buch, Arnaud; Brault, Amaury; Szopa, Cyril; Freissinet, Caroline

    2015-04-01

    Searching for building blocks of life in extraterrestrial material is a way to learn more about how life could have appeared on Earth. With this aim, liquid-phase acid hydrolysis has been used, since at least 1970 , in order to extract amino acids and other organic molecules from extraterrestrial materials (e.g. meteorites, lunar fines) or Earth analogues (e.g. Atacama desert soil). This procedure involves drastic conditions such as heating samples in 6N HCl for 24 h, either under inert atmosphere/vacuum, or air. Analysis of the hydrolyzed part of the sample should give its total (free plus bound) amino acid content. The present work deals with the influence of the 6N HCl hydrolysis on amino acid degradation. Our experiments have been performed on a standard solution of 17 amino acids. After liquid-phase acid hydrolysis (6N HCl) under argon atmosphere (24 h at 100°C), the liquid phase was evaporated and the dry residue was derivatized with N-Methyl-N-(t-butyldimethylsilyl)trifluoroacetamide (MTBSTFA) and dimethylformamide (DMF), followed by gas chromatography-mass spectrometry analysis. After comparison with derivatized amino acids from the standard solution, a significant reduction of the chromatographic peak areas was observed for most of the amino acids after liquid-phase acid hydrolysis. Furthermore, the same loss pattern was observed when the amino acids were exposed to cold 6N HCl for a short amount of time. The least affected amino acid, i.e. glycine, was found to be 73,93% percent less abundant compared to the non-hydrolyzed standard, while the most affected, i.e. histidine, was not found in the chromatograms after hydrolysis. Our experiments thereby indicate that liquid-phase acid hydrolysis, even under inert atmosphere, leads to a partial or total loss of all of the 17 amino acids present in the standard solution, and that a quick cold contact with 6N HCl is sufficient to lead to a loss of amino acids. Therefore, in the literature, the reported increase

  10. Hydrolysis of aceto-hydroxamic acid under UREX+ conditions

    SciTech Connect

    Alyapyshev, M.; Paulenova, A.; Tkac, P.; Cleveland, M.A.; Bruso, J.E.

    2007-07-01

    Aceto-hydroxamic acid (AHA) is used as a stripping agent In the UREX process. While extraction yields of uranium remain high upon addition of AHA, hexavalent plutonium and neptunium are rapidly reduced to the pentavalent state while the tetravalent species and removed from the product stream. However, under acidic conditions, aceto-hydroxamic acid undergoes hydrolytic degradation. In this study, the kinetics of the hydrolysis of aceto-hydroxamic acid in nitric and perchloric acid media was investigated at several temperatures. The decrease of the concentration of AHA was determined via its ferric complex using UV-Vis spectroscopy. The data obtained were analyzed using the method of initial rates. The data follow the pseudo-first order reaction model. Gamma irradiation of AHA/HNO{sub 3} solutions with 33 kGy/s caused two-fold faster degradation of AHA. The rate equation and thermodynamic data will be presented for the hydrolysis reaction with respect to the concentrations of aceto-hydroxamic acid, nitrate and hydronium ions, and radiation dose. (authors)

  11. Catalytic conversion of carbohydrates to 5-hydroxymethylfurfural from the waste liquid of acid hydrolysis NCC.

    PubMed

    Sun, Yonghui; Liu, Pengtao; Liu, Zhong

    2016-05-20

    The principal goal of this work was to reuse the carbohydrates and recycle sulfuric acid in the waste liquid of acid hydrolysis nanocrystalline cellulose (NCC). Therefore, in this work, the optimizations of further hydrolysis of waste liquid of acid hydrolysis NCC and catalytic conversion of L4 to 5-hydroxymethylfurfural (5-HMF) were studied. Sulfuric acid was separated by spiral wound diffusion dialysis (SWDD). The results revealed that cellulose can be hydrolyze to glucose absolutely under the condition of temperature 35 °C, 3 h, and sulfuric acid's concentration 62 wt%. And 78.3% sulfuric acid was recovered by SWDD. The yield of 5-HMF was highest in aqueous solution under the optimal condition was as follows, temperature 160 °C, 3 h, and sulfuric acid's concentration 12 wt%. Then the effect of biphasic solvent systems catalytic conversion and inorganic salt as additives were still examined. The results showed that both of them contributed to prepare 5-HMF. The yield and selectivity of 5-HMF was up to 21.0% and 31.4%, respectively. PMID:26917388

  12. Quantitation of Indoleacetic Acid Conjugates in Bean Seeds by Direct Tissue Hydrolysis 1

    PubMed Central

    Bialek, Krystyna; Cohen, Jerry D.

    1989-01-01

    Gas chromatography-selected ion monitoring-mass spectral analysis using [13C6]indole-3-acetic acid (IAA) as an internal standard provides an effective means for quantitation of IAA liberated during direct strong basic hydrolysis of bean (Phaseolus vulgaris L.) seed powder, provided that extra precautions are undertaken to exclude oxygen from the reaction vial. Direct seed powder hydrolysis revealed that the major portion of amide IAA conjugates in bean seeds are not extractable by aqueous acetone, the solvent used commonly for IAA conjugate extraction from seeds and other plant tissues. Strong basic hydrolysis of plant tissue can be used to provide new information on IAA content. Images Figure 1 PMID:16666783

  13. Fatty acid hydrolysis of acyl marinobactin siderophores by Marinobacter acylases.

    PubMed

    Kem, Michelle P; Naka, Hiroaki; Iinishi, Akira; Haygood, Margo G; Butler, Alison

    2015-01-27

    The marine bacteria Marinobacter sp. DS40M6 and Marinobacter nanhaiticus D15-8W produce a suite of acyl peptidic marinobactin siderophores to acquire iron under iron-limiting conditions. During late-log phase growth, the marinobactins are hydrolyzed to form the marinobactin headgroup with release of the corresponding fatty acid tail. The bntA gene, a homologue of the Pseudomonas aeruginosa pyoverdine acylase gene, pvdQ, was identified from Marinobacter sp. DS40M6. A bntA knockout mutant of Marinobacter sp. DS40M6 produced the suite of acyl marinobactins A-E, without the usual formation of the marinobactin headgroup. Another marinobactin-producing species, M. nanhaiticus D15-8W, is predicted to have two pvdQ homologues, mhtA and mhtB. MhtA and MhtB have 67% identical amino acid sequences. MhtA catalyzes hydrolysis of the apo-marinobactin siderophores as well as the quorum sensing signaling molecule, dodecanoyl-homoserine lactone. In contrast to hydrolysis of the suite of apo-marinobactins by MhtA, hydrolysis of the iron(III)-bound marinobactins was not observed. PMID:25588131

  14. Hydrolysis kinetics of lead silicate glass in acid solution

    NASA Astrophysics Data System (ADS)

    Rahimi, Rafi Ali; Sadrnezhaad, Sayed Khatibuleslam; Raisali, Gholamreza; Hamidi, Amir

    2009-06-01

    Hydrolysis kinetics of the lead silicate glass (LSG) with 40 mol% PbO in 0.5 N HNO 3 aqueous acid solution was investigated. The surface morphology and the gel layer thickness were studied by scanning electron microscopy (SEM) micrographs. Energy dispersive X-ray spectroscopy (EDS) and inductively coupled plasma spectroscopy (ICP) were used to determine the composition of the gel layer and the aqueous solution, respectively. The silicon content of the dissolution products was determined by using weight-loss data and compositions of the gel layer and the solution. The kinetic parameters were determined using the shrinking-core-model (SCM) for rate controlling step. The activation energy obtained for hydrolysis reaction was Qche = 56.07 kJ/mole. The diffusion coefficient of the Pb ions from the gel layer was determined by using its concentration in solution and in LSG. The shrinkage of the sample and the gel layer thickness during dissolution process were determined.

  15. Dehydration and oxidation of cellulose hydrolysis products in acidic solution

    SciTech Connect

    Garves, K.

    1981-01-01

    The dehydration of cotton cellulose in aqueous solutions in the presence of Ac/sub 2/O, AcOH, HCl, H/sub 2/SO/sub 4/ or HBr proceeded by hydrolysis to carbohydrates with acetate groups, followed by conversion to 5-(hydroxymethyl)furfural (I) and then, to levulinic acid (II) accompanied by humic acids. For the formation of I, HCl was a more efficient and selective catalyst than H/sub 2/SO/sub 4/, and the formation of II was promoted by high acid and H/sub 2/O concentrations in the medium. The addition of FeCl/sub 3/ to the dehydration mixture with HCl and continuous distillation led to the isolation of furfural.

  16. Amino Acid-β-Naphthylamide Hydrolysis by Pseudomonas aeruginosa Arylamidase

    PubMed Central

    Riley, P. S.; Behal, Francis J.

    1971-01-01

    The intracellular and constitutive arylamidase from Pseudomonas aeruginosa was purified 528-fold by salt fractionation, ion-exchange chromatography, gel filtration, and adsorption chromatography. This enzyme hydrolyzed basic and neutral N-terminal amino acid residues from amino-β-naphthylamides, dipeptide-β-naphthylamides, and a variety of polypeptides. Only those substrates having an l-amino acid with an unsubstituted α-amino group as the N-terminal residue were susceptible to enzymatic hydrolysis. The molecular weight was estimated to be 71,000 daltons. The lowest Km values were associated with substrates having neutral or basic amino acid residues with large side chains with no substitution or branching on the β carbon atom. Images PMID:5001871

  17. Chemical evolution. XXI - The amino acids released on hydrolysis of HCN oligomers

    NASA Technical Reports Server (NTRS)

    Ferris, J. P.; Wos, J. D.; Nooner, D. W.; Oro, J.

    1974-01-01

    Major amino acids released by hydrolysis of acidic and basic HCN oligomers are identified by chromatography as Gly, Asp, and diaminosuccinic acid. Smaller amounts of Ala, Ile and alpha-aminoisobutyric acid are also detected. The amino acids released did not change appreciably when the hydrolysis medium was changed from neutral to acidic or basic. The presence of both meso and d, l-diaminosuccinic acids was established by paper chromatography and on an amino acid analyzer.

  18. Morphological characteristics, oxidative stability and enzymic hydrolysis of amylose-fatty acid complexes.

    PubMed

    Marinopoulou, Anna; Papastergiadis, Efthimios; Raphaelides, Stylianos N; Kontominas, Michael G

    2016-05-01

    Complexes of amylose with fatty acids varying in carbon chain length and degree of unsaturation were prepared at 30, 50 or 70°C by dissolving amylose in 0.1N KOH and mixing with fatty acid potassium soap solution. The complexes were obtained in solid form as precipitates after neutralization. SEM microscopy revealed that the morphology of the complexes was that of ordered lamellae separated from amorphous regions whereas confocal laser scanning microscopy showed images of the topography of the guest molecules in the complex matrix. FTIR spectroscopy revealed that the absorption peak attributed to carbonyl group of free fatty acid was shifted when the fatty acid was in the form of amylose complex. Thermo-gravimetry showed that the unsaturated fatty acids were effectively protected from oxidation when they were complexed with amylose whereas enzymic hydrolysis experiments showed that the guest molecules were quantitatively released from the amylose complexes. PMID:26877002

  19. Conformational Footprint in Hydrolysis-Induced Nanofibrillation and Crystallization of Poly(lactic acid).

    PubMed

    Xu, Huan; Yang, Xi; Xie, Lan; Hakkarainen, Minna

    2016-03-14

    The origin of hydrolysis-induced nanofibrillation and crystallization, at the molecular level, was revealed by mapping the conformational ordering during long-term hydrolytic degradation of initially amorphous poly(lactic acid) (PLA), a representative model for degradable aliphatic polyesters generally displaying strong interplay between crystallization and hydrolytic erosion. The conformational regularization of chain segments was essentially the main driving force for the morphological evolution of PLA during hydrolytic degradation. For hydrolysis at 37 °C, no significant structural variations were observed due to the immobilization of "frozen" PLA chains. In contrast, conformational ordering in PLA was immediately triggered during hydrolysis at 60 °C and was responsible for the transition from random coils to disordered trans and, further, to quasi-crystalline nanospheres. On the surfaces, the head-by-head absorption and joining of neighboring nanospheres led to nanofibrillar assemblies following a "gluttonous snake"-like manner. The length and density of nanofibers formed were in close relation to the hydrolytic evolution, both of which showed a direct rise in the initial 60 days and then a gradual decline. In the interior, presumably the high surface energy of the nanospheres allowed for the preferential anchoring and packing of conformationally ordered chains into lamellae. In accordance with the well-established hypothesis, the amorphous regions were attacked prior to the erosion of crystalline entities, causing a rapid increase of crystallinity during the initial 30 days, followed by a gradual fall until 90 days. In addition to adequate illustration of hydrolysis-induced variations of crystallinity, our proposed model elucidates the formation of spherulitic nuclei featuring an extremely wide distribution of diameters ranging from several nanometers to over 5 μm, as well as the inferior resistance to hydrolysis observed for the primary nuclei. Our work

  20. Simulation of acid hydrolysis of lignocellulosic residues to fermentable sugars for bioethanol production

    NASA Astrophysics Data System (ADS)

    Sidiras, Dimitris

    2012-12-01

    The dilute acid hydrolysis of fir sawdust with sulfuric acid was undertaken in a batch reactor system (autoclave). The experimental data and reaction kinetic analysis indicate that this is a potential process for cellulose and hemicelluloses hydrolysis, due to a rapid hydrolysis reaction for acid concentration 0.045 N at 160-180°C. It was found that significant sugar degradation occurred at these conditions. The optimum conditions gave a yield of 38% total fermentable sugars. The kinetics of dilute acid hydrolysis of cellulose and hemicelluloses (polysaccharides) were simulated using four pseudo-kinetic models. The reaction rate constants were calculated in each case.

  1. Robustness of two-step acid hydrolysis procedure for composition analysis of poplar.

    PubMed

    Bhagia, Samarthya; Nunez, Angelica; Wyman, Charles E; Kumar, Rajeev

    2016-09-01

    The NREL standard procedure for lignocellulosic biomass composition has two steps: primary hydrolysis in 72% wt sulfuric acid at 30°C for 1h followed by secondary hydrolysis of the slurry in 4wt% acid at 121°C for 1h. Although pointed out in the NREL procedure, the impact of particle size on composition has never been shown. In addition, the effects of primary hydrolysis time and separation of solids prior to secondary hydrolysis on composition have never been shown. Using poplar, it was found that particle sizes less than 0.250mm significantly lowered the glucan content and increased the Klason lignin but did not affect xylan, acetate, or acid soluble lignin contents. Composition was unaffected for primary hydrolysis time between 30 and 90min. Moreover, separating solids prior to secondary hydrolysis had negligible effect on composition suggesting that lignin and polysaccharides are completely separated in the primary hydrolysis stage. PMID:27282557

  2. Effect of the Presence of Carbohydrates during Acid Hydrolysis upon the Subsequent Recovery Free Amino Acids

    PubMed Central

    Schulze, John; Presley, Jack

    2012-01-01

    Amino acid analysis is a classic biochemical technique which has been well characterized with pure protein samples. Much less is known about the effect of the presence other substances in hydrolyzed heterogeneous samples upon the final results. At the Molecular Structure Facility at UCDavis (MSF), we examined amino acid recovery following hydrolysis to determine if the presence of carbohydrates during hydrolysis influenced the recovery of the amino acids. The MSF works with a diverse sample set for different customers and have a keen interest if any of the many substances present in the sample can influence the final results. A typical sample is subjected to 6N HCl for 24hr at 110°C then brought back up in an acidic diluent to run on our dedicated Hitachi 8800 which utilizes ion-exchange chromatography with a post-column reaction with ninhydrin for quantification of each amino acid. We used this typical/routine hydrolysis in our experiments. One technique which is often employed to promote complete hydrolysis is to presoak the sample in dilute acid was also examined. Using various model proteins as well as varying percentages of carbohydrates we demonstrated that the presence of carbohydrates has little effect on the recovery of protein across a wide range of concentrations.

  3. Kinetic Study of Acid Hydrolysis of Rice Straw

    PubMed Central

    Sarkar, Nibedita; Aikat, Kaustav

    2013-01-01

    Rice straw is a renewable, cheap, and abundant waste in tropical countries. The pentose content of rice straw can be used as a substrate for many types of value-added products such as xylitol and biofuel. Dilute acid hydrolysis mainly releases pentose from rice straw. The objective of the study was to determine the effect of H2SO4 concentration and reaction time on the xylose production. The variation of the main product xylose with the reaction time was described by a kinetic model and kinetic parameters were calculated to describe the variation of the xylose production with time. The optimum yield (19.35 g/L) was obtained at 0.24 mol/L H2SO4 and 30 minutes. PMID:25969789

  4. Hydrolysis of organosolv wheat pulp in formic acid at high temperature for glucose production.

    PubMed

    Kupiainen, Laura; Ahola, Juha; Tanskanen, Juha

    2012-07-01

    Organosolv methods can be used to delignify lignocellulosic crop residues for pulp production or to pretreat them prior to enzymatic hydrolysis for bioethanol production. In this study, organic solvent was used as an acidic hydrolysis catalyst to produce glucose. Hydrolysis experiments were carried out in 5-20% formic acid at 180-220 °C. Wheat straw pulp delignified with a formicodeli™ method was used as a raw material. It was found that glucose yields from pulp are significantly higher than yields from microcrystalline cellulose, a model component for cellulose hydrolysis. The results indicate that cellulose hydrolysis of real fibers takes place more selectively to glucose than hydrolysis of microcrystalline cellulose particles does. The effect of the particle size on pulp hydrolysis was investigated, the crystallinity of hydrolyzed pulp was measured by XRD analysis, and the product distribution and its influence on the process was discussed. PMID:22609651

  5. Low-molecular-weight solutes released during mild acid hydrolysis of the lipopolysaccharide of Pseudomonas aeruginosa. Identification of ethanolamine triphosphate

    PubMed Central

    Drewry, David T.; Gray, George W.; Wilkinson, Stephen G.

    1972-01-01

    A careful examination of the low-molecular-weight solutes released during mild acid hydrolysis of the lipopolysaccharide of Pseudomonas aeruginosa (N.C.T.C. 1999) revealed the presence of ethanolamine triphosphate. During storage, the compound decomposed to give ethanolamine pyrophosphate, identified in a previous study (Drewry et al., 1971); PPi may be a further decomposition product. Evidence for the attachment of ethanolamine triphosphate to a polysaccharide fraction was obtained, but the possibility that some was attached to the lipid A moiety was not excluded. Basic compounds released during the hydrolysis of lipopolysaccharide included amino acids, polyamines and oligopeptides. PMID:4632171

  6. Effect of hydrolysis conditions on hydrous TiO2 polymorphs precipitated from a titanyl sulfate and sulfuric acid solution

    NASA Astrophysics Data System (ADS)

    Song, Hao; Liang, Bin; Lü, Li; Wu, Pan; Li, Chun

    2012-07-01

    The relationship between hydrolysis conditions and hydrous titania polymorphs obtained in a titanyl sulfate and sulfuric acid solution was investigated by X-ray diffraction (XRD), scanning electron microscopy (SEM), and high-resolution transmission electron microscopy (HRTEM). The results revealed that the feeding rate of the titanyl sulfate stock solution, the concentration of sulfuric acid, and the seed dosage of rutile crystal could significantly affect the hydrolysis rate, thus influencing the titania crystal phase. Hydrous TiO2 in the form of rutile, anatase, or the mixture of both could be obtained in solutions of low titanium concentrations and 2.5wt% to 15wt% sulfuric acid at 100°C. When the hydrolysis rate of titanium expressed by TiO2 was more than or equal to 0.04 g/(L·min), the hydrolysate was almost phase-pure anatase, while the main phase state was rutile when the hydrolysis rate was less than or equal to 0.01 g/(L·min). With the hydrolysis rate between 0.02 and 0.03 g/(L·min), the hydrolysate contained almost equal magnitude of rutile and anatase. It seems that although rutile phase is thermodynamically stable in very acidic solutions, anatase is a kinetically stable phase.

  7. Value of furfural/ethanol coproduction from acid hydrolysis processes

    SciTech Connect

    Parker, S.; Calnon, M.; Feinberg, D.; Power, A.; Weiss, L.

    1983-08-01

    The benefits of two modifications in the acid hydrolysis of cellulosic feedstocks for the production of ethanol fuels were investigated: marketing of the by-product furfural and xylose fermentation. Preliminary analysis indicate that the furfural by-product furfural and xylose fermentation. Perliminary analyses indicate that the furfural by-product credit is more beneficial at a minimum net profit of $0.08/lb of furfural. For this credit to have a major impact on ethanol production costs, new markets for large quantities of furfural must be identified. Furfural can be an alternative feedstock for hydrocarbon-based commodity chemicals. The costs and profitabilities of producing five chemicals from furfurals as opposed to conventional hydrocarbon-based feedstocks were studied. The furfural processes for production of styrene and butadiene were found to be marginally competitive or not competitive. The furfural processes for adipic acid, maleic anhydride, and butanol could be competitive. Results of analysis by a computer model of the petrochemical industry indicate that with furfural markets additional to these three furfural processes, over 2.5 x 10/sup 9/ gal of ethanol could be marketed at about $1.00/gal. Converting the xylose stream to ethanol has about the same effect on the selling price of ethanol as the furfural credit. The greatest ethanol production will result from xylose fermentation, but the furfural credit offers large near-term profits and has a more diversified impact on reducing petroleum product demand. 6 figures, 17 tables.

  8. Comparison of Enzymatic Hydrolysis and Acid Hydrolysis of Sterol Glycosides from Foods Rich in Δ(7)-Sterols.

    PubMed

    Münger, Linda H; Jutzi, Sabrina; Lampi, Anna-Maija; Nyström, Laura

    2015-08-01

    In this study, we present the difference in sterol composition of extracted steryl glycosides (SG) hydrolyzed by either enzymatic or acid hydrolysis. SG were analyzed from foods belonging to the plant families Cucurbitaceae (melon and pumpkin seeds) and Amaranthaceae (amaranth and beetroot), both of which are dominated by Δ(7)-sterols. Released sterols were quantified by gas chromatography with a flame ionization detector (GC-FID) and identified using gas chromatography/mass spectrometry (GC-MS). All Δ(7)-sterols identified (Δ(7)-stigmastenyl, spinasteryl, Δ(7)-campesteryl, Δ(7)-avenasteryl, poriferasta-7,25-dienyl and poriferasta-7,22,25-trienyl glucoside) underwent isomerization under acidic conditions and high temperature. Sterols with an ethylidene or methylidene side chain were found to form multiple artifacts. The artifact sterols coeluted with residues of incompletely isomerized Δ(7)-sterols, or Δ(5)-sterols if present, and could be identified as Δ(8(14))-sterols on the basis of relative retention time, and their MS spectra as trimethylsilyl (TMS) and acetate derivatives. For instance, SG from melon were composed of 66% Δ(7)-stigmastenol when enzymatic hydrolysis was performed, whereas with acid hydrolysis only 8% of Δ(7)-stigmastenol was determined. The artifact of Δ(7)-stigmastenol coeluted with residual non-isomerized spinasterol, demonstrating the high risk of misinterpretation of compositional data obtained after acid hydrolysis. Therefore, the accurate composition of SG from foods containing sterols with a double bond at C-7 can only be obtained by enzymatic hydrolysis or by direct analysis of the intact SG. PMID:25757602

  9. Effect of citric acid concentration and hydrolysis time on physicochemical properties of sweet potato starches.

    PubMed

    Surendra Babu, Ayenampudi; Parimalavalli, Ramanathan; Rudra, Shalini Gaur

    2015-09-01

    Physicochemical properties of citric acid treated sweet potato starches were investigated in the present study. Sweet potato starch was hydrolyzed using citric acid with different concentrations (1 and 5%) and time periods (1 and 11 h) at 45 °C and was denoted as citric acid treated starch (CTS1 to CTS4) based on their experimental conditions. The recovery yield of acid treated starches was above 85%. The CTS4 sample displayed the highest amylose (around 31%) and water holding capacity its melting temperature was 47.66 °C. The digestibility rate was slightly increased for 78.58% for the CTS3 and CTS4. The gel strength of acid modified starches ranged from 0.27 kg to 1.11 kg. RVA results of acid thinned starches confirmed a low viscosity profile. CTS3 starch illustrated lower enthalpy compared to all other modified starches. All starch samples exhibited a shear-thinning behavior. SEM analysis revealed that the extent of visible degradation was increased at higher hydrolysis time and acid concentration. The CTS3 satisfied the criteria required for starch to act as a fat mimetic. Overall results conveyed that the citric acid treatment of sweet potato starch with 5% acid concentration and 11h period was an ideal condition for the preparation of a fat replacer. PMID:26188303

  10. Pyrolytic characteristics of biomass acid hydrolysis residue rich in lignin.

    PubMed

    Huang, Yanqin; Wei, Zhiguo; Yin, Xiuli; Wu, Chuangzhi

    2012-01-01

    Pyrolytic characteristics of acid hydrolysis residue (AHR) of corncob and pinewood (CAHR, WAHR) were investigated using a thermo-gravimetric analyzer (TGA) and a self-designed pyrolysis apparatus. Gasification reactivity of CAHR char was then examined using TGA and X-ray diffractometer. Result of TGA showed that thermal degradation curves of AHR descended smoothly along with temperature increasing from 150 °C to 850 °C, while a "sharp mass loss stage" for original biomass feedstock (OBF) was observed. Char yield from AHR (42.64-30.35 wt.%) was found to be much greater than that from OBF (26.4-19.15 wt.%). In addition, gasification reactivity of CAHR char was lower than that of corncob char, and there was big difference in micro-crystallite structure. It was also found that CAHR char reactivity decreased with pyrolysis temperature, but increased with pyrolysis heating rate and gasification temperature at 850-950 °C. Furthermore, CAHR char reactivity performed better under steam atmosphere than under CO2 atmosphere. PMID:22055106

  11. Comparison of enzymatic and acid hydrolysis of bound flavor compounds in model system and grapes.

    PubMed

    Dziadas, Mariusz; Jeleń, Henryk H

    2016-01-01

    Four synthesized terpenyl-ß-D-glycopyranosides (geranyl, neryl, citronellyl, myrtenyl) were subjected to enzymatic (AR 2000, pH 5.5) and acid (citric buffer, pH 2.5) hydrolysis. Decrease of glycosides was measured by HPLC and the volatiles released--by comprehensive gas chromatography-mass spectrometry (GC × GC-ToF-MS). Enzymatic hydrolysis performed for 21 h yielded 100% degree of hydrolysis for all glycosides but citronellyl (97%). Degree of acid hydrolysis was highly dependent on type of aglycone and the conditions. The highest degree was achieved for geraniol, followed by citronellol and nerol. Myrtenylo-ß-D-glycopyranoside was the most resistant glycoside to hydrolysis. Acid hydrolysis degree was also related to temperature/time combination, the highest being for 100 °C and 2 h. In a result of enzymatic hydrolysis 85-91% of total peak areas was terpene aglycone, whereas for acid hydrolysis the area of released terpene aglycone did not exceed 1.3% of total peak area indicating almost complete decomposition/transformation of terpenyl aglycone. PMID:26212990

  12. Investigating Mass Transport Limitations on Xylan Hydrolysis During Dilute Acid Pretreatment of Poplar

    SciTech Connect

    Mittal, Ashutosh; Pilath, Heid M.; Parent, Yves; Chatterjee, Siddharth G.; Donohoe, Bryon S.; Yarbrough, John M.; Himmel, Michael E.; Nimlos, Mark R.; Johnson, David K.

    2014-04-28

    Mass transport limitations could be an impediment to achieving high sugar yields during biomass pretreatment and thus be a critical factor in the economics of biofuels production. The objective of this work was to study the mass transfer restrictions imposed by the structure of biomass on the hydrolysis of xylan during dilute acid pretreatment of biomass. Mass transfer effects were studied by pretreating poplar wood at particle sizes ranging from 10 micrometers to 10 mm. This work showed a significant reduction in the rate of xylan hydrolysis in poplar when compared to the intrinsic rate of hydrolysis for isolated xylan that is possible in the absence of mass transfer. In poplar samples we observed no significant difference in the rates of xylan hydrolysis over more than two orders of magnitude in particle size. It appears that no additional mass transport restrictions are introduced by increasing particle size from 10 micrometers to 10 mm. This work suggests that the rates of xylan hydrolysis in biomass particles are limited primarily by the diffusion of hydrolysis products out of plant cell walls. A mathematical description is presented to describe the kinetics of xylan hydrolysis that includes transport of the hydrolysis products through biomass into the bulk solution. The modeling results show that the effective diffusion coefficient of the hydrolysis products in the cell wall is several orders of magnitude smaller than typical values in other applications signifying the role of plant cell walls in offering resistance to diffusion of the hydrolysis products.

  13. Effect of an acid filler on hydrolysis and biodegradation of poly-lactic acid (PLA)

    NASA Astrophysics Data System (ADS)

    Iozzino, Valentina; Speranza, Vito; Pantani, Roberto

    2015-12-01

    The use of biodegradable polymers is certainly an excellent strategy to solve many of the problems related to the disposal of the traditional polymers, whose accumulation in the environment is harmful and damaging. In order to optimize the use of biodegradable polymers, it is very important to understand and control the transformation processes, the structures and the morphologies resulting from the process conditions used to produce the articles and, not least, the biodegradation. The latter is strictly dependent on the just mentioned variables. The poly-lactic acid, PLA, is a biodegradable polymer. Many studies have been carried out on the degradation process of this polymer. In the course of this work we performed degradation tests on the PLA, with a specific D-isomer content, having amorphous structure, and in particular of biodegradation and hydrolysis. An acid chemical, fumaric acid, was added to PLA with the objective of controlling the rate of hydrolysis and of biodegradation. The hydrolysis process was followed, as function of time, by means of different techniques: pH variation, variation of weight of samples and variation of crystallinity degree and glass transition temperature using DSC analysis. The samples were also analyzed in terms of biodegradability by means of a homemade respirometer apparatus, in controlled composting conditions.

  14. Origin of Glycine from Acid Hydrolysis of the β-Lactam Antibiotic A16886B

    PubMed Central

    Brannon, D. R.; Mabe, J. A.; Ellis, R.; Whitney, J. G.; Nagarajan, R.

    1972-01-01

    Structural analysis of two new β-lactam antibiotics, A16884A and A16886B, indicated that they, like cephalosporin C, were composed of modified valine and cysteine residues, and α-aminoadipic acid. However, acid hydrolysis of A16886B and A16884A produced three times as much glycine as did hydrolysis of cephalosporin C under the same conditions. Samples of A16886B-14C-6 and A16886B-14C-8 were prepared by the addition of cysteine-14C-3 and cystine-14C-1 to fermentations of Streptomyces clavuligerus. The specific activity of glycine obtained from hydrolysis of A16886B-14C-6 was considerably higher than that from hydrolysis of A16886B-14C-8. An explanation for the difference in amounts of glycine obtained from hydrolysis of these antibiotics is discussed. PMID:5045470

  15. Optimization of dilute acid pretreatment of water hyacinth biomass for enzymatic hydrolysis and ethanol production

    PubMed Central

    Idrees, Muhammad; Adnan, Ahmad; Sheikh, Shahzad; Qureshic, Fahim Ashraf

    2013-01-01

    The present study was conducted for the optimization of pretreatment process that was used for enzymatic hydrolysis of lignocellulosic biomass (Water Hyacinth, WH), which is a renewable resource for the production of bioethanol with decentralized availability. Response surface methodology has been employed for the optimization of temperature (oC), time (hr) and different concentrations of maleic acid (MA), sulfuric acid (SA) and phosphoric acid (PA) that seemed to be significant variables with P < 0.05. High F and R2 values and low P-value for hydrolysis yield indicated the model predictability. The pretreated biomass producing 39.96 g/l, 39.86 g/l and 37.9 g/l of reducing sugars during enzymatic hydrolysis with yield 79.93, 78.71 and 75.9 % from PA, MA and SA treated respectively. The order of catalytic effectiveness for hydrolysis yield was found to be phosphoric acid > maleic acid > sulfuric acid. Mixture of sugars was obtained during dilute acid pretreatment with glucose being the most prominent sugar while pure glucose was obtained during enzymatic hydrolysis. The resulting sugars, obtained during enzymatic hydrolysis were finally fermented to ethanol, with yield 0.484 g/g of reducing sugars which is 95 % of theoretical yield (0.51 g/g glucose) by using commercial baker's yeast (Sacchromyces cerveasiae). PMID:26417215

  16. HIGH TEMPERATURE DILUTE ACID HYDROLYSIS OF WASTE CELLULOSE: BATCH AND CONTINUOUS PROCESSES

    EPA Science Inventory

    The 5-year investigation on the dilute acid hydrolysis of waste cellulose to glucose has emphasized the crucial step of continuously converting cellulose to glucose. Initial batch studies emphasized pretreatments to improve accessibility of the cellulose, and established suitable...

  17. Hydrolysis of tRNA(sup Phe) on Suspensions of Amino Acids

    NASA Technical Reports Server (NTRS)

    Gao, Kui; Orgel, Leslie E.

    2001-01-01

    RNA is adsorbed strongly on suspensions of many moderately soluble organic solids. In some cases, the hydrolysis of tRNA(sup Phe) is greatly accelerated by adsorption, and the major sites of hydrolysis are changed from those that are important in homogeneous solution. Here we show that the hydrolysis is greatly accelerated by suspensions of aspartic acid and beta-glutamic acid but not by suspensions of alpha-glutamic acid, asparagine, or glutamine. The non-enzymatic hydrolysis of RNA has been studied extensively, especially because of its relevance to the mechanisms of action of ribozymes and to biotechnology and therapy. Many ribonucleases, ribozymes, and non-biological catalysts function via acid-base catalysis of an intramolecular transesterification mechanism in which the 2'-OH group attacks the adjacent phosphate group. The pentacoordinated phosphorane intermediate may collapse back to starting material, or yield isomerized or cleaved products.

  18. Acid hydrolysis of cellulosic fibres: Comparison of bleached kraft pulp, dissolving pulps and cotton textile cellulose.

    PubMed

    Palme, Anna; Theliander, Hans; Brelid, Harald

    2016-01-20

    The behaviour of different cellulosic fibres during acid hydrolysis has been investigated and the levelling-off degree of polymerisation (LODP) has been determined. The study included a bleached kraft pulp (both never-dried and once-dried) and two dissolving pulps (once-dried). Additionally, cotton cellulose from new cotton sheets and sheets discarded after long-time use was studied. Experimental results from the investigation, together with results found in literature, imply that ultrastructural differences between different fibres affect their susceptibility towards acid hydrolysis. Drying of a bleached kraft pulp was found to enhance the rate of acid hydrolysis and also result in a decrease in LODP. This implies that the susceptibility of cellulosic fibres towards acid hydrolysis is affected by drying-induced stresses in the cellulose chains. In cotton cellulose, it was found that use and laundering gave a substantial loss in the degree of polymerisation (DP), but that the LODP was only marginally affected. PMID:26572472

  19. The influence of solid/liquid separation techniques on the sugar yield in two-step dilute acid hydrolysis of softwood followed by enzymatic hydrolysis

    PubMed Central

    Monavari, Sanam; Galbe, Mats; Zacchi, Guido

    2009-01-01

    Background Two-step dilute acid hydrolysis of softwood, either as a stand-alone process or as pretreatment before enzymatic hydrolysis, is considered to result in higher sugar yields than one-step acid hydrolysis. However, this requires removal of the liquid between the two steps. In an industrial process, filtration and washing of the material between the two steps is difficult, as it should be performed at high pressure to reduce energy demand. Moreover, the application of pressure leads to more compact solids, which may affect subsequent processing steps. This study was carried out to investigate the influence of pressing the biomass, in combination with the effects of not washing the material, on the sugar yield obtained from two-step dilute acid hydrolysis, with and without subsequent enzymatic digestion of the solids. Results Washing the material between the two acid hydrolysis steps, followed by enzymatic digestion, resulted in recovery of 96% of the mannose and 81% of the glucose (% of the theoretical) in the liquid fraction, regardless of the choice of dewatering method (pressing or vacuum filtration). Not washing the solids between the two acid hydrolysis steps led to elevated acidity of the remaining solids during the second hydrolysis step, which resulted in lower yields of mannose, 85% and 74% of the theoretical, for the pressed and vacuum-filtered slurry, respectively, due to sugar degradation. However, this increase in acidity resulted in a higher glucose yield (94.2%) from pressed slurry than from filtered slurry (77.6%). Conclusion Pressing the washed material between the two acid hydrolysis steps had no significant negative effect on the sugar yields of the second acid hydrolysis step or on enzymatic hydrolysis. Not washing the material resulted in a harsher second acid hydrolysis step, which caused greater degradation of the sugars during subsequent acid hydrolysis of the solids, particularly in case of the vacuum-filtered solids. However

  20. Kinetic studies on the acid hydrolysis of the methyl ketoside of unsubstituted and O-acetylated N-acetylneuraminic acid

    PubMed Central

    Neuberger, A.; Ratcliffe, Wendy A.

    1973-01-01

    The hydrolysis of the model compound 2-O-methyl-4,7,8,9-tetra-O-acetyl-N-acetyl-α-d-neuraminic acid and neuraminidase (Vibrio cholerae) closely resembled that of the O-acetylated sialic acid residues of rabbit Tamm–Horsfall glycoprotein. This confirmed that O-acetylation was responsible for the unusually slow rate of acid hydrolysis of O-acetylated sialic acid residues observed in rabbit Tamm–Horsfall glycoprotein and their resistance to hydrolysis by neuraminidase. The first-order rate constant of hydrolysis of 2-methyl-N-acetyl-α-d-neuraminic acid by 0.05m-H2SO4 was 56-fold greater than that of 2-O-methyl-4,7,8,9-tetra-O-acetyl-N-acetyl -α-d-neuraminic acid. Kinetic studies have shown that in the pH range 1.00–3.30, the observed rate of hydrolysis of 2-methyl-N-acetyl-α-d-neuraminic acid can be attributed to acid-catalysed hydrolysis of the negatively charged CO2− form of the methyl ketoside. PMID:4748825

  1. Developmental Transition from Enzymatic to Acid Hydrolysis of Sucrose in Acid Limes (Citrus aurantifolia) 1

    PubMed Central

    Echeverria, Ed

    1990-01-01

    The sucrose breakdown mechanisms in juice sacs of acid lime (Citrus aurantifolia [Christm.] Swing.) were investigated throughout fruit development. All three enzymes of sucrose catabolism (sucrose synthase, acid, and alkaline invertase) are present during the initial stages. The activities of these enzymes declined rapidly and disappeared by stage 5 (80% development) but not before vacuolar pH had decreased to approximately 2.5. At this stage, sucrose breakdown occurs by acid hydrolysis. By attaining a vacuolar pH of 2.5 prior to enzyme disappearance, the cell maintains a continuous ability to break down sucrose throughout ontogeny. Thus, acid limes possess a unique and coordinated system for sucrose breakdown that involves both enzymatic and nonenzymatic pathways. PMID:16667241

  2. Enhanced xylose recovery from oil palm empty fruit bunch by efficient acid hydrolysis.

    PubMed

    Tan, Hooi Teng; Dykes, Gary A; Wu, Ta Yeong; Siow, Lee Fong

    2013-08-01

    Oil palm empty fruit bunch (EFB) is abundantly available in Malaysia and it is a potential source of xylose for the production of high-value added products. This study aimed to optimize the hydrolysis of EFB using dilute sulfuric acid (H2SO4) and phosphoric acid (H3PO4) via response surface methodology for maximum xylose recovery. Hydrolysis was carried out in an autoclave. An optimum xylose yield of 91.2 % was obtained at 116 °C using 2.0 % (v/v) H2SO4, a solid/liquid ratio of 1:5 and a hydrolysis time of 20 min. A lower optimum xylose yield of 24.0 % was observed for dilute H3PO4 hydrolysis at 116 °C using 2.4 % (v/v) H3PO4, a solid/liquid ratio of 1:5 and a hydrolysis time of 20 min. The optimized hydrolysis conditions suggested that EFB hydrolysis by H2SO4 resulted in a higher xylose yield at a lower acid concentration as compared to H3PO4. PMID:23709290

  3. Chemical structures of corn stover and its residue after dilute acid prehydrolysis and enzymatic hydrolysis: Insight into factors limiting enzymatic hydrolysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Advanced solid-state NMR techniques and wet chemical analyses were applied to investigate untreated corn stover (UCS) and its residues after dilute acid prehydrolysis (DAP) and enzymatic hydrolysis (RES) to provide evidence for the limitations to the effectiveness of enzyme hydrolysis. Advanced soli...

  4. Ultrasound-assisted dilute acid hydrolysis of tea processing waste for production of fermentable sugar.

    PubMed

    Germec, Mustafa; Tarhan, Kübra; Yatmaz, Ercan; Tetik, Nedim; Karhan, Mustafa; Demirci, Ali; Turhan, Irfan

    2016-03-01

    Lignocellulosic materials that are the most abundant plant biomass in the world have the potential to become sustainable sources of the produced value added products. Tea processing waste (TPW) is a good lignocellulosic source to produce the value added products from fermentable sugars (FSs). Therefore, the present study is undertaken to produce FSs by using ultrasound-assisted dilute acid (UADA) and dilute acid (DA) hydrolysis of TPW followed by enzymatic hydrolysis. UADA hydrolysis of TPW was optimized by response surface methodology (RSM) at maximum power (900 W) for 2 h. The optimum conditions were determined as 50°C, 1:6 (w/v) solid:liquid ratio, and 1% (w/v) DA concentration, which yielded 20.34 g/L FS concentration. Furthermore, its DA hydrolysis was also optimized by using RSM for comparison and the optimized conditions were found as 120°C, 1:8 solid:liquid ratio, and 1% acid concentration, which produced 25.3 g/L FS yield. Even though the produced sugars with UADA hydrolysis are slightly less, but it can provide significant cost saving due to the lower temperature requirement and less liquid consumption. Besides, enzymatic hydrolysis applied after pretreatments of TPW were very more economic than the conventional enzymatic hydrolysis in the literature due to shorter time requiring. In conclusion, ultrasound-assisted is a promising technology that can be successfully applied for hydrolysis of biomass and can be an alternative to the other hydrolysis procedures and also TPW can be considered as suitable carbon source for the production of value-added products like biofuels, organic acids, and polysaccharides. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:393-403, 2016. PMID:26749037

  5. Studying Cellulose Fiber Structure by SEM, XRD, NMR and Acid Hydrolysis

    SciTech Connect

    Zhao, Haibo; Kwak, Ja Hun; Zhang, Z. Conrad; Brown, Heather M.; Arey, Bruce W.; Holladay, John E.

    2007-03-21

    Cotton linters were partially hydrolyzed in dilute acid and the morphology of remaining macrofibrils studied with Scanning Electron Microscopy (SEM) under various magnifications. The crystal region (microfibril bundles) in the macrofibrils was not altered by hydrolysis, and only amorphous cellulose was hydrolyzed and leached out from the macrofibrils. The diameter of microfibril bundles was 20-30 nm after the amorphous cellulose was removed by hydrolysis. XRD experiments confirm the unaltered diameter of the microfibrils after hydrolysis. The strong stability of these microfibril bundles in hydrolysis limits both the total sugar monomer yield and the size of nano particles or rods produced in hydrolysis. The large surface potential on the remaining microfibril bundles drives the agglomeration of macrofibrils.

  6. Reversion and dehydration reactions of glucose during the dilute sulfuric acid hydrolysis of cellulose

    SciTech Connect

    Helm, R.F.

    1987-01-01

    The inaccessibility of all glycosidic bonds necessitates industrial conversion schemes which employ a dilute acid catalyst at high temperatures. Process conditions also promote further reactions of glucose via the reversion and dehydration pathways. Quantitative determination of the yields of the major reversion and dehydration products is important for understanding and predicting the amounts of these materials expected under envisioned industrial operating conditions. Microcrystalline cellulose (Avicel) was hydrolyzed with sulfuric acid (0.0-1.25 wt.%), at high temperatures (160-250/sup 0/C), and at a 3:1 liquid-to-solid ratio. The hydrolysis was monitored by evaluating the amount of cellulose remaining and the yields of glucose, solid humin, levulinic acid, formic acid, hydroxymethylfurfural (HMF), and reversion products as a function of the aforementioned reaction conditions. Analysis of the reversion products required the development of a technique for the quantitation of trace carbohydrates in complex mixtures and led to the development of a reduction/permethylation gas chromatographic procedure. Cellulose hydrolysis followed pseudo-homogeneous first-order kinetics. Glucose yield was adequately described as consecutive first-order reactions. Anhydrosugars formed via reversion followed equilibrium reaction kinetics whereas the disaccharides did not. Total reversion product yields approached 10% at 250/sup 0/C. Quantitative determination of the major dehydration products provided important information concerning the destruction of glucose. HMF was produced in up to 12% yields based on the theoretical amount of glucose available, and furfural was detected in up to 5% yields. A carbon mass balance based on the determined product yields revealed that approximately 90% of all carbon was accounted for at maximum glucose yields.

  7. Optimization of acid hydrolysis from the hemicellulosic fraction of Eucalyptus grandis residue using response surface methodology.

    PubMed

    Canettieri, Eliana Vieira; de Moraes Rocha, George Jackson; de Carvalho, João Andrade; de Almeida e Silva, João Batista

    2007-01-01

    Biotechnological conversion of biomass into fuels and chemicals requires hydrolysis of the polysaccharide fraction into monomeric sugars. Hydrolysis can be performed enzymatically and with dilute or concentrate mineral acids. The present study used dilute sulfuric acid as a catalyst for hydrolysis of Eucalyptus grandis residue. The purpose of this paper was to optimize the hydrolysis process in a 1.4 l pilot-scale reactor and investigate the effects of the acid concentration, temperature and residue/acid solution ratio on the hemicellulose removal and consequently on the production of sugars (xylose, glucose and arabinose) as well as on the formation of by-products (furfural, 5-hydroxymethylfurfural and acetic acid). This study was based on a model composition corresponding to a 2(3) orthogonal factorial design and employed the response surface methodology (RSM) to optimize the hydrolysis conditions, aiming to attain maximum xylose extraction from hemicellulose of residue. The considered optimum conditions were: H(2)SO(4) concentration of 0.65%, temperature of 157 degrees C and residue/acid solution ratio of 1/8.6 with a reaction time of 20 min. Under these conditions, 79.6% of the total xylose was removed and the hydrolysate contained 1.65 g/l glucose, 13.65 g/l xylose, 1.55 g/l arabinose, 3.10 g/l acetic acid, 1.23 g/l furfural and 0.20 g/l 5-hydroxymethylfurfural. PMID:16473004

  8. Efficient hydrogen generation from sodium borohydride hydrolysis using silica sulfuric acid catalyst

    NASA Astrophysics Data System (ADS)

    Manna, Joydev; Roy, Binayak; Sharma, Pratibha

    2015-02-01

    A heterogeneous acid catalyst, silica sulfuric acid, was prepared from silica gel (SiO2) and sulfuric acid (H2SO4). Addition of SO3H functional group to SiO2 has been confirmed through various characterization techniques. The effect of this heterogeneous acid catalyst on hydrogen generation from sodium borohydride hydrolysis reaction was studied for different ratios of catalyst to NaBH4 and at different temperatures. The catalyst exhibited high catalytic activity towards sodium borohydride hydrolysis reaction. The activation energy of the NaBH4 hydrolysis reaction in the presence of silica sulfuric acid was calculated to be the lowest (17 kJ mol-1) among reported heterogeneous catalysts till date.

  9. Obtaining fermentable sugars by dilute acid hydrolysis of hemicellulose and fast pyrolysis of cellulose.

    PubMed

    Jiang, Liqun; Zheng, Anqing; Zhao, Zengli; He, Fang; Li, Haibin; Liu, Weiguo

    2015-04-01

    The objective of this study was to get fermentable sugars by dilute acid hydrolysis of hemicellulose and fast pyrolysis of cellulose from sugarcane bagasse. Hemicellulose could be easily hydrolyzed by dilute acid as sugars. The remained solid residue of acid hydrolysis was utilized to get levoglucosan by fast pyrolysis economically. Levoglucosan yield from crystalline cellulose could be as high as 61.47%. Dilute acid hydrolysis was also a promising pretreatment for levoglucosan production from lignocellulose. The dilute acid pretreated sugarcane bagasse resulted in higher levoglucosan yield (40.50%) in fast pyrolysis by micropyrolyzer, which was more effective than water washed (29.10%) and un-pretreated (12.84%). It was mainly ascribed to the effective removal of alkali and alkaline earth metals and the accumulation of crystalline cellulose. This strategy seems a promising route to achieve inexpensive fermentable sugars from lignocellulose for biorefinery. PMID:25690683

  10. Extraction of protein and amino acids from deoiled rice bran by subcritical water hydrolysis.

    PubMed

    Sereewatthanawut, Issara; Prapintip, Surawit; Watchiraruji, Katemanee; Goto, Motonobu; Sasaki, Mitsuru; Shotipruk, Artiwan

    2008-02-01

    This study investigated the production of value-added protein and amino acids from deoiled rice bran by hydrolysis in subcritical water (SW) in the temperature range between 100 and 220 degrees C for 0-30 min. The results suggested that SW could effectively be used to hydrolyze deoiled rice bran to produce useful protein and amino acids. The amount of protein and amino acids produced are higher than those obtained by conventional alkali hydrolysis. The yields generally increased with increased temperature and hydrolysis time. However, thermal degradation of the product was observed when hydrolysis was carried out at higher temperature for extended period of time. The highest yield of protein and amino acids were 219 +/- 26 and 8.0 +/- 1.6 mg/g of dry bran, and were obtained at 200 degrees C at hydrolysis time of 30 min. Moreover, the product obtained at 200 degrees C after 30 min of hydrolysis exhibited high antioxidant activity and was shown to be suitable for use as culture medium for yeast growth. PMID:17320384

  11. Hydrolysis of acid and alkali presoaked lignocellulosic biomass exposed to electron beam irradiation.

    PubMed

    Karthika, K; Arun, A B; Melo, J S; Mittal, K C; Kumar, Mukesh; Rekha, P D

    2013-02-01

    In this study, synergetic effect of mild acid and alkali with electron beam irradiation (EBI) on the enzymatic hydrolysis of a selected grass biomass was assessed. Biomass samples prepared by soaking with 1% H2SO4, or 1% NaOH, were exposed to 75 and 150 kGy of EBI. Water presoaked biomass was used as control. Hydrolysis of pretreated samples was carried out using cellulase (15 FPU/g biomass) for 120 h. Structural changes were studied by FTIR and XRD analyses. Reducing sugar and glucose yields from enzymatic hydrolysis were significantly higher in acid and alkali presoaked EBI exposed samples. Theoretical glucose yield showed 40% increase from control in alkali presoaked EBI exposed (150 kGy) samples. Removal of hemicellulose, decreased crystallinity and structural changes were major factors for the combined treatment effect favoring the hydrolysis. PMID:23298772

  12. Acid and enzymatic hydrolysis of pretreated cellulosic materials as an analytical tool

    SciTech Connect

    Ladisch, C.M.; Chiasson, C.M.; Tsao, G.T.

    1982-07-01

    A rapid and accurate procedure for the quantitative analysis of cellulose in textiles based on acid and enzymatic hydrolysis was investigated. Total hydrolysis was achieved by a two-step procedure: the cellulose in the sample was first dissolved in cadoxen and then reprecipitated. The material, thus pretreated, was then hydrolyzed with acid or enzyme catalytic agents. Hydrolysis products were detected and quantified by colorimetric, enzymic, and liquid chromatographic methods of analysis. Samples examined included cotton, rayon, Avicel, CF-11, and cotton/polyester blends. The specificity of the enzyme hydrolysis method allowed analysis of raw cotton without prior purification. Results of the analyses were compared to those obtained by existing methods of analysis.

  13. Hydrolysis of Sulfur Dioxide in Small Clusters of Sulfuric Acid: Mechanistic and Kinetic Study.

    PubMed

    Liu, Jingjing; Fang, Sheng; Wang, Zhixiu; Yi, Wencai; Tao, Fu-Ming; Liu, Jing-Yao

    2015-11-17

    The deposition and hydrolysis reaction of SO2 + H2O in small clusters of sulfuric acid and water are studied by theoretical calculations of the molecular clusters SO2-(H2SO4)n-(H2O)m (m = 1,2; n = 1,2). Sulfuric acid exhibits a dramatic catalytic effect on the hydrolysis reaction of SO2 as it lowers the energy barrier by over 20 kcal/mol. The reaction with monohydrated sulfuric acid (SO2 + H2O + H2SO4 - H2O) has the lowest energy barrier of 3.83 kcal/mol, in which the cluster H2SO4-(H2O)2 forms initially at the entrance channel. The energy barriers for the three hydrolysis reactions are in the order SO2 + (H2SO4)-H2O > SO2 + (H2SO4)2-H2O > SO2 + H2SO4-H2O. Furthermore, sulfurous acid is more strongly bonded to the hydrated sulfuric acid (or dimer) clusters than the corresponding reactant (monohydrated SO2). Consequently, sulfuric acid promotes the hydrolysis of SO2 both kinetically and thermodynamically. Kinetics simulations have been performed to study the importance of these reactions in the reduction of atmospheric SO2. The results will give a new insight on how the pre-existing aerosols catalyze the hydrolysis of SO2, leading to the formation and growth of new particles. PMID:26450714

  14. Specificity of hydrolysis of phytic acid by alkaline phytase from lily pollen.

    PubMed Central

    Barrientos, L; Scott, J J; Murthy, P P

    1994-01-01

    Phytases are the primary enzymes responsible for the hydrolysis of phytic acid, myo-inositol-1,2,3,4,5,6-hexakisphosphate (I-1,2,3,4,5,6-P6). A number of phytases with varying specificities, properties, and localizations hydrolyze phytic acid present in cells. The specificity of hydrolysis of phytic acid by alkaline phytase from lily (Lilium longiflorum L.) pollen is described. Structures of the intermediate inositol phosphates and the final product were established by a variety of nuclear magnetic resonance techniques (1H-, 31P-, and 31P-1H-detected multiple quantum coherence spectroscopy, and total correlation spectroscopy). On the basis of the structures identified we have proposed a scheme of hydrolysis of phytic acid. Initial hydrolysis of the phosphate ester occurs at the D-5 position of phytic acid to yield the symmetrical I-1,2,3,4,6-P5. The two subsequent dephosphorylations occur adjacent to the D-5 hydroxyl group to yield I-1,2,3-P3 as the final product. Alkaline phytase differs from other phytases in the specificity of hydrolysis of phosphate esters on the inositol ring, its high substrate specificity for phytic acid, and biochemical properties such as susceptibility to activation by calcium and inhibition by fluoride. The physiological significance of alkaline phytase and the biological role of I-1,2,3-P3 remain to be identified. PMID:7846160

  15. High-yield production of biosugars from Gracilaria verrucosa by acid and enzymatic hydrolysis processes.

    PubMed

    Kim, Se Won; Hong, Chae-Hwan; Jeon, Sung-Wan; Shin, Hyun-Jae

    2015-11-01

    Gracilaria verrucosa, the red alga, is a suitable feedstock for biosugar production. This study analyzes biosugar production by the hydrolysis of G. verrucosa conducted under various conditions (i.e., various acid concentrations, substrate concentrations, reaction times, and enzyme dosages). The acid hydrolysates of G. verrucosa yielded a total of 7.47g/L (37.4%) and 10.63g/L (21.26%) of reducing sugars under optimal small (30mL) and large laboratory-scale (1L) hydrolysis processes, respectively. Reducing sugar obtained from acid and enzymatic hydrolysates were 10% higher, with minimum by-products, than those reported in other studies. The mass balance for the small laboratory-scale process showed that the acid and enzymatic hydrolysates had a carbohydrate conversion of 57.2%. The mass balance approach to the entire hydrolysis process of red seaweed for biosugar production can be applied to other saccharification processes. PMID:26299978

  16. Preparation of κ-carra-oligosaccharides with microwave assisted acid hydrolysis method

    NASA Astrophysics Data System (ADS)

    Li, Guangsheng; Zhao, Xia; Lv, Youjing; Li, Miaomiao; Yu, Guangli

    2015-04-01

    A rapid method of microwave assisted acid hydrolysis was established to prepare κ-carra-oligosaccharides. The optimal hydrolysis condition was determined by an orthogonal test. The degree of polymerization (DP) of oligosaccharides was detected by high performance thin layer chromatography (HPTLC) and polyacrylamide gel electrophoresis (PAGE). Considering the results of HPTLC and PAGE, the optimum condition of microwave assisted acid hydrolysis was determined. The concentration of κ-carrageenan was 5 mg mL-1; the reaction solution was adjusted to pH 3 with diluted hydrochloric acid; the solution was hydrolyzed under microwave irradiation at 100 for 15 °C min. Oligosaccharides were separated by a Superdex 30 column (2.6 cm × 90 cm) using AKTA Purifier UPC100 and detected with an online refractive index detector. Each fraction was characterized by electrospray ionization mass spectrometry (ESI-MS). The data showed that odd-numbered κ-carra-oligosaccharides with DP ranging from 3 to 21 could be obtained with this method, and the structures of the oligosaccharides were consistent with those obtained by traditional mild acid hydrolysis. The new method was more convenient, efficient and environment-friendly than traditional mild acid hydrolysis. Our results provided a useful reference for the preparation of oligosaccharides from other polysaccharides.

  17. The Acid Hydrolysis Mechanism of Acetals Catalyzed by a Supramolecular Assembly in Basic Solution

    SciTech Connect

    Pluth, Michael D.; Bergman, Robert G.; Raymond, Kenneth N.

    2008-09-24

    A self-assembled supramolecular host catalyzes the hydrolysis of acetals in basic aqueous solution. The mechanism of hydrolysis is consistent with the Michaelis-Menten kinetic model. Further investigation of the rate limiting step of the reaction revealed a negative entropy of activation ({Delta}S{double_dagger} = -9 cal mol{sup -1}K{sup -1}) and an inverse solvent isotope effect (k(H{sub 2}O)/k(D{sub 2}O) = 0.62). These data suggest that the mechanism of hydrolysis that takes place inside the assembly proceeds through an A-2 mechanism, in contrast to the A-1 mechanism operating in the uncatalyzed reaction. Comparison of the rates of acetal hydrolysis in the assembly with the rate of the reaction of unencapsulated substrates reveals rate accelerations of up to 980 over the background reaction for the substrate diethoxymethane.

  18. Analysis of Phosphorus in Soil Humic Acid Fractions by Enzymatic Hydrolysis and Ultraviolet Irradiation

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Humic acid is an important soil component which influences chemical, biological, and physical soil properties. In this study, we investigated lability of phosphorus (P) in the mobile humic acid (MHA) and calcium humate (CaHA) fractions of four soils by orthophosphate-releasing enzymatic hydrolysis a...

  19. Effect of Varying Acid Hydrolysis Condition in Gracilaria Sp. Fermentation Using Sasad

    NASA Astrophysics Data System (ADS)

    Mansuit, H.; Samsuri, M. D. C.; Sipaut, C. S.; Yee, C. F.; Yasir, S. M.; Mansa, R.

    2015-04-01

    Macroalgae or seaweed is being considered as promising feedstock for bioalcohol production due to high polysaccharides content. Polysaccharides can be converted into fermentable sugar through acid hydrolysis pre-treatment. In this study, the potential of using carbohydrate-rich macroalgae, Gracilaria sp. as feedstock for bioalcohol production via various acid hydrolysis conditions prior to the fermentation process was investigated and evaluated. The seaweed used in this research was from the red algae group, using species of Gracilaria sp. which was collected from Sg. Petani Kedah, Malaysia. Pre-treatment of substrate was done using H2SO4 and HCl with molarity ranging from 0.2M to 0.8M. The pretreatment time were varied in the range of 15 to 30 minutes. Fermentation was conducted using Sasad, a local Sabahan fermentation agent as a starter culture. Alcohol extraction was done using a distillation unit. Reducing sugar analysis was done by Benedict test method. Alcohol content analysis was done using specific gravity test. After hydrolysis, it was found out that acid hydrolysis at 0.2M H2SO4 and pre-treated for 20 minutes at 121°C has shown the highest reducing sugar content which has yield (10.06 mg/g) of reducing sugar. It was followed by other samples hydrolysis using 0.4M HCl with 30 minutes pre-treatment and 0.2M H2SO4, 15 minutes pre-treatment with yield of 8.06 mg/g and 5.75 mg/g reducing sugar content respectively. In conclusion, acid hydrolysis of Gracilaria sp. can produce higher reducing sugar yield and thus it can further enhance the bioalcohol production yield. Hence, acid hydrolysis of Gracilaria sp. should be studied more as it is an important step in the bioalcohol production and upscaling process.

  20. Combined heat treatment and acid hydrolysis of cassava grate waste (CGW) biomass for ethanol production

    SciTech Connect

    Agu, R.C.; Amadife, A.E.; Ude, C.M.; Onyia, A.; Ogu, E.O.; Okafor, M.; Ezejiofor, E.

    1997-12-31

    The effect of combined heat treatment and acid hydrolysis (various concentrations) on cassava grate waste (CGW) biomass for ethanol production was investigated. At high concentrations of H{sub 2}SO{sub 4} (1--5 M), hydrolysis of the CGW biomass was achieved but with excessive charring or dehydration reaction. At lower acid concentrations, hydrolysis of CGW biomass was also achieved with 0.3--0.5 M H{sub 2}SO{sub 4}, while partial hydrolysis was obtained below 0.3 M H{sub 2}SO{sub 4} (the lowest acid concentration that hydrolyzed CGW biomass) at 120 C and 1 atm pressure for 30 min. A 60% process efficiency was achieved with 0.3 M H{sub 2}SO{sub 4} in hydrolyzing the cellulose and lignin materials present in the CGW biomass. High acid concentration is therefore not required for CGW biomass hydrolysis. The low acid concentration required for CGW biomass hydrolysis, as well as the minimal cost required for detoxification of CGW biomass because of low hydrogen cyanide content of CGW biomass would seem to make this process very economical. From three liters of the CGW biomass hydrolysate obtained from hydrolysis with 0.3M H{sub 2}SO{sub 4}, ethanol yield was 3.5 (v/v%) after yeast fermentation. However, although the process resulted in gainful utilization of CGW biomass, additional costs would be required to effectively dispose new by-products generated from CGW biomass processing.

  1. Kinetics of hydrolysis of PET powder in nitric acid by a modified shrinking-core model

    SciTech Connect

    Yoshioka, Toshiaki; Okayama, Nobuchika; Okuwaki, Akitsugu

    1998-02-01

    Poly(ethylene terephthalate) (PET) powder from waste bottles was degraded at atmospheric pressure in 7--13 M nitric acid at 70--100 C for 72 h, to clarify the mechanism of a feed stock recycling process. Terephthalic acid (TPA) and ethylene glycol (EG) were produced by the acid-catalyzed heterogeneous hydrolysis of PET in nitric acid, and the resulting EG was simultaneously oxidized to oxalic acid. The kinetics of the hydrolysis of PET in nitric acid could be explained by a modified shrinking core model of chemical reaction control, in which the effective surface area is proportional to the degree of unreacted PET, affected by the deposition of the product TPA. The apparent rate constant was inversely proportional to particle size and to the concentration of the nitric acid. The activation energy of the reaction was 101.3 kJ/mol.

  2. Evaluation of hyper thermal acid hydrolysis of Kappaphycus alvarezii for enhanced bioethanol production.

    PubMed

    Ra, Chae Hun; Nguyen, Trung Hau; Jeong, Gwi-Taek; Kim, Sung-Koo

    2016-06-01

    Hyper thermal (HT) acid hydrolysis of Kappaphycus alvarezii, a red seaweed, was optimized to 12% (w/v) seaweed slurry content, 180mM H2SO4 at 140°C for 5min. The maximum monosaccharide concentration of 38.3g/L and 66.7% conversion from total fermentable monosaccharides of 57.6g/L with 120gdw/L K. alvarezii slurry were obtained from HT acid hydrolysis and enzymatic saccharification. HT acid hydrolysis at a severity factor of 0.78 efficiently converted the carbohydrates of seaweed to monosaccharides and produced a low concentration of inhibitory compounds. The levels of ethanol production by separate hydrolysis and fermentation with non-adapted and adapted Kluyveromyces marxianus to high concentration of galactose were 6.1g/L with ethanol yield (YEtOH) of 0.19 at 84h and 16.0g/L with YEtOH of 0.42 at 72h, respectively. Development of the HT acid hydrolysis process and adapted yeast could enhance the overall ethanol fermentation yields of K. alvarezii seaweed. PMID:26950757

  3. Factors affecting the rate of hydrolysis of phenylboronic acid in lab-scale precipitate reactor studies

    SciTech Connect

    Bannochie, C.J.; Marek, J.C.; Eibling, R.E.; Baich, M.A.

    1992-10-01

    Removing aromatic carbon from an aqueous slurry of cesium-137 and other alkali tetraphenylborates by acid hydrolysis will be an important step in preparing high-level radioactive waste for vitrification at the Savannah River Site`s Defense Waste Processing Facility (DWPF). Kinetic data obtained in bench-scale precipitate hydrolysis reactors suggest changes in operating parameters to improve product quality in the future plant-scale radioactive operation. The rate-determining step is the removal of the fourth phenyl group, i.e. hydrolysis of phenylboronic acid. Efforts to maximize this rate have established the importance of several factors in the system, including the ratio of copper(II) catalyst to formic acid, the presence of nitrite ion, reactions of diphenylmercury, and the purge gas employed in the system.

  4. Factors affecting the rate of hydrolysis of phenylboronic acid in lab-scale precipitate reactor studies

    SciTech Connect

    Bannochie, C.J.; Marek, J.C.; Eibling, R.E.; Baich, M.A.

    1992-01-01

    Removing aromatic carbon from an aqueous slurry of cesium-137 and other alkali tetraphenylborates by acid hydrolysis will be an important step in preparing high-level radioactive waste for vitrification at the Savannah River Site's Defense Waste Processing Facility (DWPF). Kinetic data obtained in bench-scale precipitate hydrolysis reactors suggest changes in operating parameters to improve product quality in the future plant-scale radioactive operation. The rate-determining step is the removal of the fourth phenyl group, i.e. hydrolysis of phenylboronic acid. Efforts to maximize this rate have established the importance of several factors in the system, including the ratio of copper(II) catalyst to formic acid, the presence of nitrite ion, reactions of diphenylmercury, and the purge gas employed in the system.

  5. Efficient production of glucose by microwave-assisted acid hydrolysis of cellulose hydrogel.

    PubMed

    Sun, Binzhe; Duan, Lian; Peng, Gege; Li, Xiaoxia; Xu, Aihua

    2015-09-01

    To improve the production of glucose from cellulose, a simple and effective route was developed. This process uses a combination of a step of cellulose dissolution in aqueous NaOH/urea solution and then regeneration with water, followed by an acid hydrolysis step under microwave irradiation. The method is effective to obtain glucose from α-cellulose, microcrystalline cellulose, filter paper, ramie fiber and absorbent cotton. Increased with the acid concentration the glucose yield from hydrogel hydrolysis increased from 0.42% to 44.6% at 160 °C for 10 min. Moreover, the ozone treatment of cellulose in NaOH/urea solution before regeneration significantly enhanced the hydrolysis efficiency with a glucose yield of 59.1%. It is believed that the chains in cellulose hydrogel are relatively free approached, making that the acids easily access the β-glycosidic bonds. PMID:26038330

  6. Contribution of acetic acid to the hydrolysis of lignocellulosic biomass under abiotic conditions.

    PubMed

    Trzcinski, Antoine P; Stuckey, David C

    2015-06-01

    Acetic acid was used in abiotic experiments to adjust the solution pH and investigate its influence on the chemical hydrolysis of the Organic Fraction of Municipal Solid Waste (OFMSW). Soluble chemical oxygen demand (SCOD) was used to measure the hydrolysis under oxidative conditions (positive oxidation-reduction potential values), and pH 4 allowed for 20% (±2%) of the COD added to be solubilized, whereas only 12% (±1%) was solubilized at pH7. Under reducing conditions (negative oxidation-reduction potential values) and pH 4, 32.3% (±3%) of the OFMSW was solubilized which shows that acidogenesis at pH 4 during the anaerobic digestion of solid waste can result in chemical hydrolysis. In comparison, bacterial hydrolysis resulted in 54% (±6%) solubilization. PMID:25794810

  7. Fatty acid specificity of hormone-sensitive lipase. Implication in the selective hydrolysis of triacylglycerols.

    PubMed

    Raclot, T; Holm, C; Langin, D

    2001-12-01

    The selective mobilization of fatty acids from white fat cells depends on their molecular structure, in particular the degree of unsaturation. The present study was designed to examine if the release of fatty acids by hormone-sensitive lipase (HSL) in vitro i) is influenced by the amount of unsaturation, ii) depends on the temperature, and iii) could explain the selective pattern of fatty acid mobilization and notably the preferential mobilization of certain highly unsaturated fatty acids. Recombinant rat and human HSL were incubated with a lipid emulsion. The hydrolysis of 35 individual fatty acids, ranging in chain length from 12 to 24 carbon atoms and in unsaturation from 0 to 6 double bonds was measured. Fatty acid composition of in vitro released NEFA was compared with that of fat cell triacylglycerols (TAG), the ratio % NEFA/% TAG being defined as the relative hydrolysis. The relative hydrolysis of individual fatty acids differed widely, ranging from 0.44 (24:1n-9) to 1.49 (18:1n-7) with rat HSL, and from 0.38 (24:1n-9) to 1.67 (18:1n-7) with human HSL. No major difference was observed between rat and human HSL. The relative release was dependent on the number of double bonds according to chain length. The amount of fatty acid released by recombinant rat HSL was decreased but remained robust at 4 degrees C compared with 37 degrees C, and the relative hydrolysis of some individual fatty acids was affected. The relative hydrolysis of fatty acids moderately, weakly, and highly mobilized by adipose tissue in vivo was similar and close to unity in vitro. We conclude that i) the release of fatty acids by HSL is only slightly affected by their degree of unsaturation, ii) the ability of HSL to efficiently and selectively release fatty acids at low temperature could reflect a cold adaptability for poikilotherms or hibernators when endogenous lipids are needed, and iii) the selectivity of fatty acid hydrolysis by HSL does not fully account for the selective pattern of

  8. Determination of DNA adducts by combining acid-catalyzed hydrolysis and chromatographic analysis of the carcinogen-modified nucleobases.

    PubMed

    Leung, Elvis M K; Deng, Kailin; Wong, Tin-Yan; Chan, Wan

    2016-01-01

    The commonly used method of analyzing carcinogen-induced DNA adducts involves the hydrolysis of carcinogen-modified DNA samples by using a mixture of enzymes, followed by (32)P-postlabeling or liquid chromatography (LC)-based analyses of carcinogen-modified mononucleotides/nucleosides. In the present study, we report the development and application of a new approach to DNA adduct analysis by combining the H(+)/heat-catalyzed release of carcinogen-modified nucleobases and the use of LC-based methods to analyze DNA adducts. Results showed that heating the carcinogen-modified DNA samples at 70 °C for an extended period of 4 to 6 h in the presence of 0.05% HCl can efficiently induce DNA depurination, releasing the intact carcinogen-modified nucleobases for LC analyses. After optimizing the hydrolysis conditions, DNA samples with C8- and N (2) -modified 2'-deoxyguanosine, as well as N (6) -modified 2'-deoxyadenosine, were synthesized by reacting DNA with 1-nitropyrene, acetaldehyde, and aristolochic acids, respectively. These samples were then hydrolyzed, and the released nucleobase adducts were analyzed using LC-based analytical methods. Analysis results demonstrated a dose-dependent release of target DNA adducts from carcinogen-modified DNA samples, indicating that the developed H(+)/heat-catalyzed hydrolysis method was quantitative. Comparative studies with enzymatic digestion method on carcinogen-modified DNA samples revealed that the two hydrolysis methods did not yield systematically different results. PMID:26581621

  9. Studies on the carbohydrate moiety of α1-acid glycoprotein (orosomucoid) by using alkaline hydrolysis and deamination by nitrous acid

    PubMed Central

    Isemura, M.; Schmid, K.

    1971-01-01

    Alkaline hydrolysis followed by deamination with nitrous acid was applied for the first time to a glycoprotein, human plasma α1-acid glycoprotein (orosomucoid). This procedure, which specifically cleaves the glycosaminidic bonds, yielded well-defined oligosaccharides. The trisaccharides, which were obtained from the native protein, consisted of a sialic acid derivative, galactose and 2,5-anhydromannose. The linkage between galactose and 2,5-anhydromannose is most probably a (1→4)-glycosidic bond. A hitherto unknown linkage between N-acetylneuraminic acid and galactose was also established, namely a (2→2)-linkage. The three linkages between sialic acid and galactose described in this paper appear to be about equally resistant to mild acid hydrolysis. The disaccharide that was derived from the desialized glycoprotein consisted of galactose and 2,5-anhydromannose. Evidence was obtained for the presence of a new terminal sialyl→N-acetylglucosamine disaccharide accounting for approximately 1mol/mol of protein. The presence of this disaccharide may explain the relatively severe requirements for the complete acid hydrolysis of the sialyl residues. The present study indicates that alkaline hydrolysis followed by nitrous acid deamination in conjunction with gas–liquid chromatography will afford relatively rapid determination of the partial structure of the complex carbohydrate moiety of glycoproteins. PMID:5135244

  10. Fatty acid stimulation of membrane phosphatidylinositol hydrolysis by brain phosphatidylinositol phosphodiesterase.

    PubMed Central

    Irvine, R F; Letcher, A J; Dawson, R M

    1979-01-01

    The hydrolysis of membrane-bound phosphatidylinositol in rat liver microsomal fraction by the soluble phosphatidylinositol phosphodiesterase from rat brain was markedly stimulated by oleic acid or arachidonic acid. The stimulation did not require added calcium, although it was abolished by EDTA. Lysophosphatidylcholine also totally suppressed the stimulation. A possible role for the fatty acid content of a membrane in controlling phosphatidylinositol turnover is suggested. PMID:220968

  11. Acidic 1,3-propanediaminetetraacetato lanthanides with luminescent and catalytic ester hydrolysis properties

    SciTech Connect

    Chen, Mao-Long; Shi, Yan-Ru; Yang, Yu-Chen; Zhou, Zhao-Hui

    2014-11-15

    In acidic solution, a serials of water-soluble coordination polymers (CPs) were isolated as zonal 1D-CPs 1,3-propanediaminetetraacetato lanthanides [Ln(1,3-H{sub 3}pdta)(H{sub 2}O){sub 5}]{sub n}·2Cl{sub n}·3nH{sub 2}O [Ln=La, 1; Ce, 2; Pr, 3; Nd, 4; Sm, 5] (1,3-H{sub 4}pdta=1,3-propanediaminetetraacetic acid, C{sub 11}H{sub 18}N{sub 2}O{sub 8}) in high yields. When 1 eq. mol potassium hydroxide was added to the solutions of 1D-CPs, respectively, two 1D-CPs [Ln(1,3-H{sub 2}pdta)(H{sub 2}O){sub 3}]{sub n}·Cl{sub n}·2nH{sub 2}O [Ln=Sm, 6; Gd, 7] were isolated at room temperature and seven 2D-CPs [Ln(1,3-H{sub 2}pdta)(H{sub 2}O){sub 2}]{sub n}·Cl{sub n}·2nH{sub 2}O [Ln=La, 8; Ce, 9; Pr, 10; Nd, 11; Sm, 12; Eu, 13; Gd, 14] were isolated at 70 °C. When the crystals of 1–4 were hydrothermally heated at 180 °C with 1–2 eq. mol potassium hydroxide, four 3D-CPs [Ln(1,3-Hpdta)]{sub n}·nH{sub 2}O [Ln=La, 15; Ce, 16; Pr, 17; Nd, 18] were obtained. The two 2D-CPs [Ln(1,3-Hpdta)(H{sub 2}O)]{sub n}·4nH{sub 2}O (Sm, 19; Eu, 20) were isolated in similar reaction conditions. With the increments of pH value in the solution and reaction temperature, the structure becomes more complicated. 1–5 are soluble in water and 1 was traced by solution {sup 13}C({sup 1}H) NMR technique, the water-soluble lanthanides 1 and 5 show catalytic activity to ester hydrolysis reaction respectively, which indicate their important roles in the hydrolytic reaction. The europium complexes 13 and 20 show visible fluorescence at an excitation of 394 nm. The structure diversity is mainly caused by the variation of coordinated ligand in different pH values and lanthanide contraction effect. Acidic conditions are favorable for the isolations of lanthanide complexes in different structures and this may helpful to separate different lanthanides. The thermal stability investigations reveal that acidic condition is favorable to obtain the oxides at a lower temperature. - Graphical abstract: A series

  12. Dilute acid/metal salt hydrolysis of lignocellulosics

    DOEpatents

    Nguyen, Quang A.; Tucker, Melvin P.

    2002-01-01

    A modified dilute acid method of hydrolyzing the cellulose and hemicellulose in lignocellulosic material under conditions to obtain higher overall fermentable sugar yields than is obtainable using dilute acid alone, comprising: impregnating a lignocellulosic feedstock with a mixture of an amount of aqueous solution of a dilute acid catalyst and a metal salt catalyst sufficient to provide higher overall fermentable sugar yields than is obtainable when hydrolyzing with dilute acid alone; loading the impregnated lignocellulosic feedstock into a reactor and heating for a sufficient period of time to hydrolyze substantially all of the hemicellulose and greater than 45% of the cellulose to water soluble sugars; and recovering the water soluble sugars.

  13. Selective hydrolysis of hemicellulose from wheat straw by a nanoscale solid acid catalyst.

    PubMed

    Zhong, Chao; Wang, Chunming; Huang, Fan; Wang, Fengxue; Jia, Honghua; Zhou, Hua; Wei, Ping

    2015-10-20

    A nanoscale catalyst, solid acid SO4(2-)/Fe2O3 with both Lewis and Brønsted acidity was found to effectively hydrolyze hemicellulose while keeping cellulose and lignin inactive, and selective hydrolysis of hemicellulose from wheat straw by this catalyst was also confirmed. The factors that significantly affected hydrolysis process were investigated with response surface methodology, and the optimum conditions for time, temperature, and ratio of wheat straw to catalyst (w/w) were calculated to be 4.10h, 141.97°C, and 1.95:1, respectively. A maximum hemicellulose hydrolysis yield of 63.5% from wheat straw could be obtained under these conditions. In addition, the catalyst could be recycled six times with high activity remaining. PMID:26256198

  14. Acid hydrolysis of cellulose as the entry point into biorefinery schemes.

    PubMed

    Rinaldi, Roberto; Schüth, Ferdi

    2009-01-01

    Cellulose is a major source of glucose because it is readily available, renewable, and does not compete with the food supply. Hydrolysis of cellulose is experiencing a new research and development cycle in which this reaction is carried out over solid catalysts and coupled to other reactions for increased efficiency. Cellulose is typically not soluble in conventional solvents and very resistant to chemical and biological transformations. This Review focuses on aspects related to the hydrolysis of cellulose as this process is a significant entry point into the biorefinery scheme based on carbohydrates for the production of biofuels and biochemicals. Structural features of cellulose, conventional acid-catalyzed reactions, and the use of solid acid catalysts for hydrolysis are discussed. The longterm success of the biorefinery concept depends on the development of energetically efficient processes to convert cellulose directly or indirectly into biofuels and chemicals. PMID:19950346

  15. Vacuolar Acid Hydrolysis as a Physiological Mechanism for Sucrose Breakdown 1

    PubMed Central

    Echeverria, Ed; Burns, Jacqueline K.

    1989-01-01

    Sucrose breakdown in mature acidic `Persian' limes (Citrus aurantifolia [Christm.] Swing.) occurred at a rate of 30.6 picomoles per milliliter per day during 9 weeks storage at 15°C. Neither enzyme of sucrose catabolism (sucrose synthase or acid/alkaline invertase) was present in extracts of mature storage tissue. The average vacuolar pH, estimated by direct measurement of sap from isolated vacuoles and by the methylamine method, was about 2.0 to 2.2. In vitro acid hydrolysis of sucrose at physiological concentrations in a buffered solution (pH 2.2) occurred at identical rates as in matured limes. The results indicate that sucrose breakdown in stored mature acidic limes occurs by acid hydrolysis. PMID:16666803

  16. Carboxypeptidase displaying differential velocity in hydrolysis of methotrexate, 5-methyltetrahydrofolic acid, and leucovorin.

    PubMed Central

    Albrecht, A M; Boldizsar, E; Hutchison, D J

    1978-01-01

    An enzyme that catalyzes the hydrolysis of folic acid and the antifolate methotrexate nearly 20 times more rapidly than the hydrolysis of 5-methyltetrahydrofolate was extraced from a gram-negative bacterium tentatively identified as a Flavobacterium sp. The enzyme was purified 500-fold and found to have a molecular weight of about 53,000. Apparently a metallo-enzyme, it is inhibited by citrate and ethylenediaminetetraacetic acid (EDTA). Ca2+, Co2+, Mg2+, and Zn2+ reverse inhibition by EDTA, whereas Ca2+ and Zn2+ are weak activators in the absence of EDTA. The enzymatic reaction releases the carboxy-terminal glutamyl moiety of derivatives of pteroyl-mono-L-glutamic acid. Substituents on N5 of the pteridine ring decrease the velocity of hydrolysis. Some non-specificity for the terminal amino acid is expressed. The strikingly different rates of hydrolysis of methotrexate and 5-methyltetrahydrofolate have stimulated interest in this enzyme for its potential clinical value in improving the therapeutic index of methotrexate. PMID:26657

  17. Chemical Kinetic Analysis of Thermal Decay of Rhodopsin Reveals Unusual Energetics of Thermal Isomerization and Hydrolysis of Schiff Base*

    PubMed Central

    Liu, Jian; Liu, Monica Yun; Fu, Li; Zhu, Gefei Alex; Yan, Elsa C. Y.

    2011-01-01

    The thermal properties of rhodopsin, which set the threshold of our vision, have long been investigated, but the chemical kinetics of the thermal decay of rhodopsin has not been revealed in detail. To understand thermal decay quantitatively, we propose a kinetic model consisting of two pathways: 1) thermal isomerization of 11-cis-retinal followed by hydrolysis of Schiff base (SB) and 2) hydrolysis of SB in dark state rhodopsin followed by opsin-catalyzed isomerization of free 11-cis-retinal. We solve the kinetic model mathematically and use it to analyze kinetic data from four experiments that we designed to assay thermal decay, isomerization, hydrolysis of SB using dark state rhodopsin, and hydrolysis of SB using photoactivated rhodopsin. We apply the model to WT rhodopsin and E181Q and S186A mutants at 55 °C, as well as WT rhodopsin in H2O and D2O at 59 °C. The results show that the hydrogen-bonding network strongly restrains thermal isomerization but is less important in opsin and activated rhodopsin. Furthermore, the ability to obtain individual rate constants allows comparison of thermal processes under various conditions. Our kinetic model and experiments reveal two unusual energetic properties: the steep temperature dependence of the rates of thermal isomerization and SB hydrolysis in the dark state and a strong deuterium isotope effect on dark state SB hydrolysis. These findings can be applied to study pathogenic rhodopsin mutants and other visual pigments. PMID:21921035

  18. Kinetics of acid hydrolysis of hardwood in a continuous plug flow reactor

    SciTech Connect

    Kwarteng, I.K.

    1983-01-01

    This thesis was undertaken to investigate the kinetics of dilute acid catalyzed hydrolysis of hardwood in an isothermally operated continuous plug flow reactor at steady state. Sulphuric acid catalyst concentration ranged from 0.05 to 2.5 wt% and the reaction temperatures were between 160/sup 0/C and 265/sup 0/C. The residence time was limited to between 0.1 and 0.5 minutes while the wood slurry concentration varied from 5 wt% to 10 wt%. Kinetic models, as functions of the reaction conditions, were obtained for the hemicelluloses (xylan) to xylose, xylose to furfural, fufural decomposition, and cellulose (glucan) to glucose reactions. Numerical solutions (Runge-Kutta Methods) to a system of first order differential equations which provided a composite description of the kinetic model for furfural production from xylan were also presented. Furfural yields of up to 87% were verified. Xylose appearance from hardwood xylan could be modelled by a sequence of two irreversible consecutive first-order reactions. The proposed model, however, underestimated xylose yields at the conditions of quantitative yields (1 wt% 2 wt%, 0.25 minutes and 190/sup 0/C to 210/sup 0/C). For the cellulose to glucose reaction, a similar model with different parameters produced yields of up to 56% at 240/sup 0/C to 260/sup 0/C. Two approaches for the carbohydrate recovery and utilization in acid hydrolysis were proposed as follows. 1) Two-stage hydrolysis: in this mode, the first stage is at the conditions for maximum xylose recovery. This is then followed by a second-stage hydrolysis at higher temperatures for glucose recovery. 2) Single-stage hydrolysis: since the conditions for high furfural yields coincide with those for glucose, a single stage hydrolysis for their simultaneous recovery can also be achieved.

  19. Preparative production of colominic acid oligomers via a facile microwave hydrolysis

    PubMed Central

    Patane, Jonathan; Trapani, Vincent; Villavert, Janice; McReynolds, Katherine Dawn

    2009-01-01

    The hydrolysis of colominic acid via microwave irradiation was studied for the production of short chain oligomers with a degree of polymerization (DP) of 1–6. This method was compared to the traditional acid hydrolytic method for the production of preparative quantities of short colominic acid oligomers. The oligomers were purified by size exclusion chromatography and characterized by 1H NMR. Optimal conditions for producing the dimer were found to be 12 minutes at 10% power in a 1000 Watt domestic microwave. This method is advantageous over the traditional technique in that the hydrolysis can be completed in just a few minutes, rather than hours, it is reproducible, and yields large quantities of the desirable short chain oligomers of colominic acid. PMID:19281967

  20. Oxidation and hydrolysis of lactic acid in near-critical water

    SciTech Connect

    Li, L.; Vallejo, D.; Gloyna, E.F.; Portela, J.R.

    1999-07-01

    Hydrothermal reactions (oxidation and hydrolysis) involving lactic acid (LA) were studied at temperatures ranging from 300 to 400 C and a nominal pressure of 27.6 MPa. Kinetic models were developed with respect to concentrations of LA and total organic carbon (TOC), respectively. On the basis of identified liquid and gaseous products, pathways for hydrothermal reactions involving lactic acid were proposed. Acetic acid and acetaldehyde were confirmed as the major liquid intermediates for oxidation and hydrolysis reactions, respectively. Carbon monoxide and methane were identified as the major gaseous byproducts from these reactions. These results demonstrate the potential of completely oxidizing, as well as converting, lactic acid into other organic products, in high-temperature water.

  1. The acid and enzymic hydrolysis of O-acetylated sialic acid residues from rabbit Tamm–Horsfall glycoprotein

    PubMed Central

    Neuberger, A.; Ratcliffe, Wendy A.

    1972-01-01

    Rabbit Tamm–Horsfall glycoprotein and bovine submaxillary glycoprotein were both found to contain sialic acid residues which are released at a slow rate by the standard conditions of acid hydrolysis. These residues are also resistant to neuraminidases from Vibrio cholerae and Clostridium perfringens. This behaviour was attributed to the presence of O-acetylated sialic acid, since the removal of O-acetyl groups by mild alkaline treatment normalized the subsequent release of sialic acid from rabbit Tamm–Horsfall glycoprotein by acid and by enzymic hydrolysis. Determination of the O-acetyl residues in rabbit Tamm–Horsfall glycoprotein indicated that on average two hydroxyl groups of sialic acid are O-acetylated, and these were located on the polyhydroxy side-chain of sialic acid or on C-4 and C-8. These findings confirm the assumption that certain O-acetylated forms of sialic acid are not substrates for bacterial neuraminidases. Several explanations have been suggested to explain the effect of O-acetylation of the side-chain on the rate of acidcatalysed hydrolysis of sialic acid residues. PMID:4349114

  2. Enzymatic hydrolysis and fermentation of dilute acid pretreated cornstalk to biohydrogen

    NASA Astrophysics Data System (ADS)

    Pan, C. M.; Fan, Y. T.; Hou, H. W.

    2010-03-01

    The coupling method of acid pretreatment and enzymatic hydrolysis of cornstalk for hydrogen production was investigated in this study. Experimental results showed that temperature, pH and enzyme loading all had an individual significant influence on soluble sugar yield and Ps. The optimum condition for soluble sugar was close to that for Ps. The maximum hydrogen yield from cornstalk by anaerobic mixed microflora was 209.8 ml/g-TVS on the optimum enzymatic hydrolysis condition which was 52 °C of temperature, pH4.8 and 9.4 IU/g of enzyme loading.

  3. Structure and hydrolysis of p-(2-oxo-1-pyrrolidinyl)- benzenesulfonic acid

    SciTech Connect

    Kukalenko, S.S.; Frolov, S.I.; Lim, I.K.; Putsykina, E.B.; Vasil'ev, A.F.

    1987-11-20

    With the aid of vibrational and PMR spectra of p-(2-oso-1-pyrrolidinyl)benzenesulfonic acid it was shown that in the solid state it exists as an O-protonated dipolar ion in which the protonated amide cation and sulfonate ion are intermolecularly linked by a very strong hydrogen bond. In concentrated hydrochloric acid the dipolar ion is an intermediate link in the chain of processes in the hydrolysis of the amide bond of the lactam ring.

  4. Powerful peracetic acid-ionic liquid pretreatment process for the efficient chemical hydrolysis of lignocellulosic biomass.

    PubMed

    Uju; Goto, Masahiro; Kamiya, Noriho

    2016-08-01

    The aim of this work was to design a new method for the efficient saccharification of lignocellulosic biomass (LB) using a combination of peracetic acid (PAA) pretreatment with ionic liquid (IL)-HCl hydrolysis. The pretreatment of LBs with PAA disrupted the lignin fractions, enhanced the dissolution of LB and led to a significant increase in the initial rate of the IL-HCl hydrolysis. The pretreatment of Bagasse with PAA prior to its 1-buthyl-3-methylimidazolium chloride ([Bmim][Cl])-HCl hydrolysis, led to an improvement in the cellulose conversion from 20% to 70% in 1.5h. Interestingly, the 1-buthyl-3-methylpyridium chloride ([Bmpy][Cl])-HCl hydrolysis of Bagasse gave a cellulose conversion greater than 80%, with or without the PAA pretreatment. For LB derived from seaweed waste, the cellulose conversion reached 98% in 1h. The strong hydrolysis power of [Bmpy][Cl] was attributed to its ability to transform cellulose I to II, and lowering the degree of polymerization of cellulose. PMID:27174616

  5. Fundamental study on kinetics and transport phenomena in low water dilute acid total hydrolysis of cellulosic biomass

    SciTech Connect

    Auburn University

    2004-04-07

    The overall objective of this research is to delineate the process of the dilute-acid hydrolysis of biomass and seek better understanding of the reactions involving dilute-acid treatment of lignocellulosic biomass. Specifically the scope of the work entails the following two primary technical elements: Verification of the heterogeneous nature of the reaction mechanism in dilute-acid hydrolysis of cellulosic component of the biomass. Experimental investigation to identify the overall reaction pattern and the kinetic constants associated with dilute-acid hydrolysis of the cellulosic component of the agricultural residues.

  6. Prediction of acid hydrolysis of lignocellulosic materials in batch and plug flow reactors.

    PubMed

    Jaramillo, Oscar Johnny; Gómez-García, Miguel Ángel; Fontalvo, Javier

    2013-08-01

    This study unifies contradictory conclusions reported in literature on acid hydrolysis of lignocellulosic materials, using batch and plug flow reactors, regarding the influence of the initial liquid ratio of acid aqueous solution to solid lignocellulosic material on sugar yield and concentration. The proposed model takes into account the volume change of the reaction media during the hydrolysis process. An error lower than 8% was found between predictions, using a single set of kinetic parameters for several liquid to solid ratios, and reported experimental data for batch and plug flow reactors. For low liquid-solid ratios, the poor wetting and the acid neutralization, due to the ash presented in the solid, will both reduce the sugar yield. Also, this study shows that both reactors are basically equivalent in terms of the influence of the liquid to solid ratio on xylose and glucose yield. PMID:23770535

  7. Monomeric carbohydrates production from olive tree pruning biomass: modeling of dilute acid hydrolysis.

    PubMed

    Puentes, Juan G; Mateo, Soledad; Fonseca, Bruno G; Roberto, Inês C; Sánchez, Sebastián; Moya, Alberto J

    2013-12-01

    Statistical modeling and optimization of dilute sulfuric acid hydrolysis of olive tree pruning biomass has been performed using response surface methodology. Central composite rotatable design was applied to assess the effect of acid concentration, reaction time and temperature on efficiency and selectivity of hemicellulosic monomeric carbohydrates to d-xylose. Second-order polynomial model was fitted to experimental data to find the optimum reaction conditions by multiple regression analysis. The monomeric d-xylose recovery 85% (as predicted by the model) was achieved under optimized hydrolysis conditions (1.27% acid concentration, 96.5°C and 138 min), confirming the high validity of the developed model. The content of d-glucose (8.3%) and monosaccharide degradation products (0.1% furfural and 0.04% 5-hydroxymethylfurfural) provided a high quality subtract, ready for subsequent biochemical conversion to value-added products. PMID:24096282

  8. The effect of acid hydrolysis pretreatment on crystallinity and solubility of kenaf cellulose membrane

    SciTech Connect

    Saidi, Anis Syuhada Mohd; Zakaria, Sarani; Chia, Chin Hua; Jaafar, Sharifah Nabihah Syed; Padzil, Farah Nadia Mohammad

    2015-09-25

    Cellulose was extracted from kenaf core pulp (KCP) by series of bleaching steps in the sequence (DEED) where D and E are referred as acid and alkali treatment. The bleached kenaf pulp (BKCP) is then pretreated with acid hydrolysis at room temperature for 1 and 3 h respectively. The pretreated cellulose is dissolved in lithium hydroxide/urea (LiOH/urea) and cellulose solution produced was immersed in distilled water bath. BKCP without treatment was also conducted for comparison purpose. The effects of acid hydrolysis pretreatment on solubility and crystallinity are investigated. Higher solubility of cellulose solution is achieved for treated samples. Cellulose II formation and crystallinity index of the cellulose membrane were determined by X-ray diffraction (XRD)

  9. Fast hemicellulose quantification via a simple one-step acid hydrolysis.

    PubMed

    Gao, Xiadi; Kumar, Rajeev; Wyman, Charles E

    2014-06-01

    As the second most common polysaccharides in nature, hemicellulose has received much attention in recent years for its importance in biomass conversion in terms of producing high yields of fermentable sugars and value-added products, as well as its role in reducing biomass recalcitrance. Therefore, a time and labor efficient method that specifically analyzes hemicellulose content would be valuable to facilitate the screening of biomass feedstocks. In this study, a one-step acid hydrolysis method was developed, which applied 4 wt% sulfuric acid at 121°C for 1 h to rapidly quantify XGM (xylan + galactan + mannan) contents in various types of lignocellulosic biomass and model hemicelluloses. This method gave statistically identical results in XGM contents compared to results from conventional two-step acid hydrolysis while significantly shortening analysis time. PMID:24343864

  10. Kinetics of acid hydrolysis and reactivity of some antibacterial hydrophilic iron(II) imino-complexes

    NASA Astrophysics Data System (ADS)

    Shaker, Ali Mohamed; Nassr, Lobna Abdel-Mohsen Ebaid; Adam, Mohamed Shaker Saied; Mohamed, Ibrahim Mohamed Abdelhalim

    2015-05-01

    Kinetic study of acid hydrolysis of some hydrophilic Fe(II) Schiff base amino acid complexes with antibacterial properties was performed using spectrophotometry. The Schiff base ligands were derived from sodium 2-hydroxybenzaldehyde-5-sulfonate and glycine, L-alanine, L-leucine, L-isoleucine, DL-methionine, DL-serine, or L-phenylalanine. The reaction was studied in aqueous media under conditions of pseudo-first order kinetics. Moreover, the acid hydrolysis was studied at different temperatures and the activation parameters were calculated. The general rate equation was suggested as follows: rate = k obs [Complex], where k obs = k 2 [H+]. The evaluated rate constants and activation parameters are consistent with the hydrophilicity of the investigated complexes.

  11. The effect of acid hydrolysis pretreatment on crystallinity and solubility of kenaf cellulose membrane

    NASA Astrophysics Data System (ADS)

    Saidi, Anis Syuhada Mohd; Zakaria, Sarani; Chia, Chin Hua; Jaafar, Sharifah Nabihah Syed; Padzil, Farah Nadia Mohammad

    2015-09-01

    Cellulose was extracted from kenaf core pulp (KCP) by series of bleaching steps in the sequence (DEED) where D and E are referred as acid and alkali treatment. The bleached kenaf pulp (BKCP) is then pretreated with acid hydrolysis at room temperature for 1 and 3 h respectively. The pretreated cellulose is dissolved in lithium hydroxide/urea (LiOH/urea) and cellulose solution produced was immersed in distilled water bath. BKCP without treatment was also conducted for comparison purpose. The effects of acid hydrolysis pretreatment on solubility and crystallinity are investigated. Higher solubility of cellulose solution is achieved for treated samples. Cellulose II formation and crystallinity index of the cellulose membrane were determined by X-ray diffraction (XRD).

  12. Optimization of wastewater microalgae saccharification using dilute acid hydrolysis for acetone, butanol, and ethanol fermentation

    SciTech Connect

    Castro, Yessica; Ellis, Joshua T.; Miller, Charles D.; Sims, Ronald C.

    2015-02-01

    Exploring and developing sustainable and efficient technologies for biofuel production are crucial for averting global consequences associated with fuel shortages and climate change. Optimization of sugar liberation from wastewater algae through acid hydrolysis was determined for subsequent fermentation to acetone, butanol, and ethanol (ABE) by Clostridium saccharoperbutylacetonicum N1-4. Acid concentration, retention time, and temperature were evaluated to determine optimal hydrolysis conditions by assessing the sugar and ABE yield as well as the associated costs. Sulfuric acid concentrations ranging from 0-1.5 M, retention times of 40-120 min, and temperatures from 23°C- 90°C were combined to form a full factorial experiment. Acid hydrolysis pretreatment of 10% dried wastewater microalgae using 1.0 M sulfuric acid for 120 min at 80-90°C was found to be the optimal parameters, with a sugar yield of 166.1 g for kg of dry algae, concentrations of 5.23 g/L of total ABE, and 3.74 g/L of butanol at a rate of USD $12.83 per kg of butanol.

  13. Acid hydrolysis of crude tannins from infructescence of Platycarya strobilacea Sieb. et Zucc to produce ellagic acid.

    PubMed

    Zhang, Liangliang; Wang, Yongmei; Xu, Man

    2014-01-01

    The infructescence of Platycarya strobilacea Sieb. et Zucc is a well-known traditional medicine in China, Japan and Korea. The infructescence of P. strobilacea Sieb. et Zucc is a rich source of ellagitannins that are composed of ellagic acid (EA) and gallic acid, linked to a sugar moiety. The aim of this study was to prepare EA by acid hydrolysis of crude tannins from the infructescence of P. strobilacea Sieb. et Zucc, and establish a new technological processing method for EA. The natural antioxidant EA was prepared by using the water extraction of infructescence of P. strobilacea Sieb. et Zucc, evaporation, condensation, acid hydrolysis and prepared by the process of crystallisation. The yield percentage of EA from crude EA was more than 20% and the purity of the product was more than 98%, as identified by using HPLC. The structure was identified on the basis of spectroscopic analysis and comparison with authentic compound. PMID:24911045

  14. Effects of acid-hydrolysis and hydroxypropylation on functional properties of sago starch.

    PubMed

    Fouladi, Elham; Mohammadi Nafchi, Abdorreza

    2014-07-01

    In this study, sago starch was hydrolyzed by 0.14M HCl for 6, 12, 18, and 24h, and then modified by propylene oxide at a concentration of 0-30% (v/w). The effects of hydrolysis and etherification on molecular weight distribution, physicochemical, rheological, and thermal properties of dually modified starch were estimated. Acid hydrolysis of starch decreased the molecular weight of starch especially amylopectin, but hydroxypropylation had no effect on the molecular weight distribution. The degree of Molar substitution (DS) of hydroxypropylated starch after acid hydrolysis ranged from 0.007 to 0.15. Dually modified starch with a DS higher than 0.1 was completely soluble in cold water at up to 25% concentration of the starch. This study shows that hydroxypropylation and hydrolysis have synergistic effects unlike individual modifications. Dually modified sago starch can be applied to dip-molding for food and pharmaceutical processing because of its high solubility and low tendency for retrogradation. PMID:24832983

  15. Surface Chemical Compositions and Dispersity of Starch Nanocrystals Formed by Sulfuric and Hydrochloric Acid Hydrolysis

    PubMed Central

    Wei, Benxi; Xu, Xueming; Jin, Zhengyu; Tian, Yaoqi

    2014-01-01

    Surface chemical compositions of starch nanocrystals (SNC) prepared using sulfuric acid (H2SO4) and hydrochloric acid (HCl) hydrolysis were analyzed by X-ray photoelectron spectroscopy (XPS) and FT-IR. The results showed that carboxyl groups and sulfate esters were presented in SNC after hydrolysis with H2SO4, while no sulfate esters were detected in SNC during HCl-hydrolysis. TEM results showed that, compared to H2SO4-hydrolyzed sample, a wider size distribution of SNC prepared by HCl-hydrolysis were observed. Zeta-potentials were −23.1 and −5.02 mV for H2SO4- and HCl-hydrolyzed SNC suspensions at pH 6.5, respectively. Nevertheless, the zeta-potential values decreased to −32.3 and −10.2 mV as the dispersion pH was adjusted to 10.6. After placed 48 h at pH 10.6, zeta-potential increased to −24.1 mV for H2SO4-hydrolyzed SNC, while no change was detected for HCl-hydrolyzed one. The higher zeta-potential and relative small particle distribution of SNC caused more stable suspensions compared to HCl-hydrolyzed sample. PMID:24586246

  16. Facile, room-temperature pre-treatment of rice husks with tetrabutylphosphonium hydroxide: Enhanced enzymatic and acid hydrolysis yields.

    PubMed

    Lau, B B Y; Luis, E T; Hossain, M M; Hart, W E S; Cencia-Lay, B; Black, J J; To, T Q; Aldous, L

    2015-12-01

    Aqueous solutions of tetrabutylphosphonium hydroxide have been evaluated as pretreatment media for rice husks, prior to sulphuric acid hydrolysis or cellulase enzymatic hydrolysis. Varying the water:tetrabutylphosphonium hydroxide ratio varied the rate of delignification, as well as silica, lignin and cellulose solubility. Pre-treatment with 60wt% hydroxide dissolved the rice husk and the regenerated material was thus heavily disrupted. Sulphuric acid hydrolysis of 60wt%-treated samples yielded the highest amount of glucose per gram of rice husk. Solutions with good lignin and silica solubility but only moderate to negligible cellulose solubility (10-40wt% hydroxide) were equally effective as pre-treatment media for both acid and enzymatic hydrolysis. However, pre-treatment with 60wt% hydroxide solutions was incompatible with downstream enzymatic hydrolysis. This was due to significant incorporation of phosphonium species in the regenerated biomass, which significantly inhibited the activity of the cellulase enzymes. PMID:26342336

  17. Spore Cortex Hydrolysis Precedes Dipicolinic Acid Release during Clostridium difficile Spore Germination

    PubMed Central

    Francis, Michael B.; Allen, Charlotte A.

    2015-01-01

    ABSTRACT Bacterial spore germination is a process whereby a dormant spore returns to active, vegetative growth, and this process has largely been studied in the model organism Bacillus subtilis. In B. subtilis, the initiation of germinant receptor-mediated spore germination is divided into two genetically separable stages. Stage I is characterized by the release of dipicolinic acid (DPA) from the spore core. Stage II is characterized by cortex degradation, and stage II is activated by the DPA released during stage I. Thus, DPA release precedes cortex hydrolysis during B. subtilis spore germination. Here, we investigated the timing of DPA release and cortex hydrolysis during Clostridium difficile spore germination and found that cortex hydrolysis precedes DPA release. Inactivation of either the bile acid germinant receptor, cspC, or the cortex hydrolase, sleC, prevented both cortex hydrolysis and DPA release. Because both cortex hydrolysis and DPA release during C. difficile spore germination are dependent on the presence of the germinant receptor and the cortex hydrolase, the release of DPA from the core may rely on the osmotic swelling of the core upon cortex hydrolysis. These results have implications for the hypothesized glycine receptor and suggest that the initiation of germinant receptor-mediated C. difficile spore germination proceeds through a novel germination pathway. IMPORTANCE Clostridium difficile infects antibiotic-treated hosts and spreads between hosts as a dormant spore. In a host, spores germinate to the vegetative form that produces the toxins necessary for disease. C. difficile spore germination is stimulated by certain bile acids and glycine. We recently identified the bile acid germinant receptor as the germination-specific, protease-like CspC. CspC is likely cortex localized, where it can transmit the bile acid signal to the cortex hydrolase, SleC. Due to the differences in location of CspC compared to the Bacillus subtilis germinant

  18. Hydrolysis of ketene catalyzed by formic acid: modification of reaction mechanism, energetics, and kinetics with organic acid catalysis.

    PubMed

    Louie, Matthew K; Francisco, Joseph S; Verdicchio, Marco; Klippenstein, Stephen J; Sinha, Amitabha

    2015-05-14

    The hydrolysis of ketene (H2C═C═O) to form acetic acid involving two water molecules and also separately in the presence of one to two water molecules and formic acid (FA) was investigated. Our results show that, while the currently accepted indirect mechanism, involving addition of water across the carbonyl C═O bond of ketene to form an ene-diol followed by tautomerization of the ene-diol to form acetic acid, is the preferred pathway when water alone is present, with formic acid as catalyst, addition of water across the ketene C═C double bond to directly produce acetic acid becomes the kinetically favored pathway for temperatures below 400 K. We find not only that the overall barrier for ketene hydrolysis involving one water molecule and formic acid (H2C2O + H2O + FA) is significantly lower than that involving two water molecules (H2C2O + 2H2O) but also that FA is able to reduce the barrier height for the direct path, involving addition of water across the C═C double bond, so that it is essentially identical with (6.4 kcal/mol) that for the indirect ene-diol formation path involving addition of water across the C═O bond. For the case of ketene hydrolysis involving two water molecules and formic acid (H2C2O + 2H2O + FA), the barrier for the direct addition of water across the C═C double bond is reduced even further and is 2.5 kcal/mol lower relative to the ene-diol path involving addition of water across the C═O bond. In fact, the hydrolysis barrier for the H2C2O + 2H2O + FA reaction through the direct path is sufficiently low (2.5 kcal/mol) for it to be an energetically accessible pathway for acetic acid formation under atmospheric conditions. Given the structural similarity between acetic and formic acid, our results also have potential implications for aqueous-phase chemistry. Thus, in an aqueous environment, even in the absence of formic acid, though the initial mechanism for ketene hydrolysis is expected to involve addition of water across the

  19. Comparison of cell wall polysaccharide hydrolysis by a dilute acid/enzymatic saccharification process and rumen microorganisms

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Evaluation of biomass crops for breeding or pricing purposes requires an assay that predicts performance of biomass in the bioenergy conversion process. Cell wall polysaccharide hydrolysis by dilute sulfuric acid pretreatment at 121 degrees C followed by cellulase hydrolysis for 72 h (CONV) and in v...

  20. Utilization of sugarcane bagasse for bioethanol production: sono-assisted acid hydrolysis approach.

    PubMed

    Velmurugan, Rajendran; Muthukumar, Karuppan

    2011-07-01

    In this study, the production of sugar monomers from sugarcane bagasse (SCB) by sono-assisted acid hydrolysis was performed. The SCB was subjected to sono-assisted alkaline pretreatment. The cellulose and hemicellulose recovery observed in the solid content was 99% and 78.95%, respectively and lignin removal observed during the pretreatment was about 75.44%. The solid content obtained was subjected to sono-assisted acid hydrolysis. Under optimized conditions, the maximum hexose and pentose yield observed was 69.06% and 81.35% of theoretical yield, respectively. The hydrolysate obtained was found to contain very less inhibitors, which improved the bioethanol production and the ethanol yield observed was 0.17 g/g of pretreated SCB. PMID:21570831

  1. Nanocellulose prepared by acid hydrolysis of isolated cellulose from sugarcane bagasse

    NASA Astrophysics Data System (ADS)

    Wulandari, W. T.; Rochliadi, A.; Arcana, I. M.

    2016-02-01

    Cellulose in nanometer range or called by nano-cellulose has attracted much attention from researchers because of its unique properties. Nanocellulose can be obtained by acid hydrolysis of cellulose. The cellulose used in this study was isolated from sugarcane bagasse, and then it was hydrolyzed by 50% sulfuric acid at 40 °C for 10 minutes. Nanocellulose has been characterized by Transmission Electron Microscope (TEM), Particle Size Analyzer (PSA), Fourier Transform Infrared Spectroscopy (FTIR) and X-Ray Diffraction (XRD). Analysis of FTIR showed that there were not a new bond which formed during the hydrolysis process. Based on the TEM analysis, nano-cellulose has a spherical morphology with an average diameter of 111 nm and a maximum distribution of 95.9 nm determined by PSA. The XRD analysis showed that the crystallinity degree of nano-cellulose was higher than cellulose in the amount of 76.01%.

  2. Effects of crystallinity on dilute acid hydrolysis of cellulose by cellulose ball-milling study

    SciTech Connect

    Zhao, Haibo; Kwak, Ja Hun; Wang, Yong; Franz, James A.; White, John M.; Holladay, Johnathan E.

    2005-12-23

    The dilute acid (0.05 M H2SO4) hydrolysis at 175°C of samples comprising varying fractions of crystalline (α-form) and amorphous cellulose was studied. The amorphous content, based on XRD and NMR, and then the product (glucose) yield, based on HPLC, increased by as much as a factor of three upon ball milling. These results are interpreted in terms of a model involving mechanical disruption of crystallinity by breaking hydrogen bonds in α-cellulose, opening up the structure and making more β-1,4 glycosidic bonds readily accessible to the dilute acid. In parallel with hydrolysis to form liquid phase products, there are reactions of amorphous cellulose that form solid degradation products.

  3. Structural Basis for Nucleotide Hydrolysis by the Acid Sphingomyelinase-like Phosphodiesterase SMPDL3A.

    PubMed

    Gorelik, Alexei; Illes, Katalin; Superti-Furga, Giulio; Nagar, Bhushan

    2016-03-18

    Sphingomyelin phosphodiesterase, acid-like 3A (SMPDL3A) is a member of a small family of proteins founded by the well characterized lysosomal enzyme, acid sphingomyelinase (ASMase). ASMase converts sphingomyelin into the signaling lipid, ceramide. It was recently discovered that, in contrast to ASMase, SMPDL3A is inactive against sphingomyelin and, surprisingly, can instead hydrolyze nucleoside diphosphates and triphosphates, which may play a role in purinergic signaling. As none of the ASMase-like proteins has been structurally characterized to date, the molecular basis for their substrate preferences is unknown. Here we report crystal structures of murine SMPDL3A, which represent the first structures of an ASMase-like protein. The catalytic domain consists of a central mixed β-sandwich surrounded by α-helices. Additionally, SMPDL3A possesses a unique C-terminal domain formed from a cluster of four α-helices that appears to distinguish this protein family from other phosphoesterases. We show that SMDPL3A is a di-zinc-dependent enzyme with an active site configuration that suggests a mechanism of phosphodiester hydrolysis by a metal-activated water molecule and protonation of the leaving group by a histidine residue. Co-crystal structures of SMPDL3A with AMP and α,β-methylene ADP (AMPCP) reveal that the substrate binding site accommodates nucleotides by establishing interactions with their base, sugar, and phosphate moieties, with the latter the major contributor to binding affinity. Our study provides the structural basis for SMPDL3A substrate specificity and sheds new light on the function of ASMase-like proteins. PMID:26792860

  4. Hydrolysis of oxaliplatin-evaluation of the acid dissociation constant for the oxalato monodentate complex.

    PubMed

    Jerremalm, Elin; Eksborg, Staffan; Ehrsson, Hans

    2003-02-01

    Alkaline hydrolysis of the platinum anticancer drug oxaliplatin gives the oxalato monodentate complex and the dihydrated oxaliplatin complex in two consecutive steps. The acid dissociation constant for the oxalato monodentate intermediate was determined by a kinetic approach. The pK(a) value was estimated as 7.23. The monodentate intermediate is assumed to rapidly react with endogenous compounds, resulting in a continuous conversion of oxaliplatin via the monodentate form. PMID:12532393

  5. Neutral and acidic hydrolysis reactions of the third generation anticancer drug oxaliplatin.

    PubMed

    Lucas, Maria Fatima A; Pavelka, Mateij; Alberto, Marta E; Russo, Nino

    2009-01-22

    The hydrolysis of oxaliplatin, a third generation anticancer drug, is expected to play an important role in the activation of this compound before it reaches DNA. The first and second hydrolysis corresponding to the addition of the first water molecule concomitant with the ring-opening, followed by addition of a second water and loss of the monodentate oxalato ligand, respectively, were studied combining density functional theory (DFT) with the conductor-like dielectric continuum model (CPCM) approach. The reaction was studied in neutral and acidic conditions, and all stationary points have been identified. The computed potential energy surfaces show that, for the neutral hydrolysis, the ring-opening reaction is the rate-limiting process, with an activation barrier of about 28 kcal/mol. For the acid degradation in water, according to experimental data, the reaction is expected to proceed in a faster biphasic process, and the rate-limiting process is the ligand detachment that occurs with a barriers of about 22 kcal/mol. According to the calculated results, we expect that the reaction is favored in acidic conditions and that the monoaquated complex should be the species reacting with DNA. PMID:19143575

  6. Effects of pressing lignocellulosic biomass on sugar yield in two-stage dilute-acid hydrolysis process.

    PubMed

    Kim, Kyoung Heon; Tucker, Melvin P; Nguyen, Quang A

    2002-01-01

    Dilute sulfuric acid catalyzed hydrolysis of biomass such as wood chips often involves pressing the wood particles in a dewatering step (e.g., after acid impregnation) or in compression screw feeders commonly used in continuous hydrolysis reactors. This study addresses the effects of pressing biomass feedstocks using a hydraulic press on soluble sugar yield obtained from two-stage dilute-acid hydrolysis of softwood. The pressed acid-impregnated feedstock gave significantly lower soluble sugar yields than the never-pressed (i.e., partially air-dried or filtered) feedstock. Pressing acid-impregnated feedstocks before pretreatment resulted in a soluble hemicellulosic sugar yield of 76.9% from first-stage hydrolysis and a soluble glucose yield of 33.7% from second-stage hydrolysis. The dilute-acid hydrolysis of partially air-dried feedstocks having total solids and acid concentrations similar to those of pressed feedstocks gave yields of 87.0% hemicellulosic sugar and 46.9% glucose in the first and second stages, respectively. Microscopic examination of wood structures showed that pressing acid-impregnated wood chips from 34 to 54% total solids (TS) did not cause the wood structure to collapse. However, pressing first-stage pretreated wood chips (i.e., feedstock for second-stage hydrolysis) from approximately 30 to 43% TS caused the porous wood matrix to almost completely collapse. It is hypothesized that pressing alters the wood structure and distribution of acid within the cell cavities, leading to uneven heat and mass transfer during pretreatment using direct steam injection. Consequently, lower hydrolysis yield of soluble sugars results. Dewatering of corn stover by pressing did not impact negatively on the sugar yield from single-stage dilute-acid pretreatment. PMID:12052064

  7. Hydrolysis of Indole-3-Acetic Acid Esters Exposed to Mild Alkaline Conditions 1

    PubMed Central

    Baldi, Bruce G.; Maher, Barbara R.; Cohen, Jerry D.

    1989-01-01

    Ester conjugates of indole-3-acetic acid are hydrolyzed easily in basic solutions; however, quantitative data have not been available on the relationship between pH and rate of hydrolysis of the known ester conjugates. The use of basic conditions during extraction or purification of IAA by several laboratories suggested that a more systematic analysis of this process was needed. In this report we present data indicating: (a) that measurable hydrolysis of IAA-glucose (from standard solutions) and IAA-esters (from maize kernel extracts) occurs with only a few hours of treatment at pH 9 or above; (b) that the lability of some ester conjugates is even greater than that of IAA-glucose; and (c) that ester hydrolysis of standard compounds, IAA-glucose and IAA-p-nitrophenol, occurs in the `three phase extraction system' proposed by Liu and Tillberg ([1983] Physiol Plant 57: 441-447). These data indicate that the potential for problems with inadvertent hydrolysis of ester conjugates of IAA exists even at moderate pH values and in the multiphase system where exposure to basic conditions was thought to be limited. PMID:16667049

  8. Heterogeneous ceria catalyst with water-tolerant Lewis acidic sites for one-pot synthesis of 1,3-diols via Prins condensation and hydrolysis reactions.

    PubMed

    Wang, Yehong; Wang, Feng; Song, Qi; Xin, Qin; Xu, Shutao; Xu, Jie

    2013-01-30

    The use of a heterogeneous Lewis acid catalyst, which is insoluble and easily separable during the reaction, is a promising option for hydrolysis reactions from both environmental and practical viewpoints. In this study, ceria showed excellent catalytic activity in the hydrolysis of 4-methyl-1,3-dioxane to 1,3-butanediol in 95% yield and in the one-pot synthesis of 1,3-butanediol from propylene and formaldehyde via Prins condensation and hydrolysis reactions in an overall yield of 60%. In-depth investigations revealed that ceria is a water-tolerant Lewis acid catalyst, which has seldom been reported previously. The ceria catalysts showed rather unusual high activity in hydrolysis, with a turnover number (TON) of 260, which is rather high for bulk oxide catalysts, whose TONs are usually less than 100. Our conclusion that ceria functions as a Lewis acid catalyst in hydrolysis reactions is firmly supported by thorough characterizations with IR and Raman spectroscopy, acidity measurements with IR and (31)P magic-angle-spinning NMR spectroscopy, Na(+)/H(+) exchange tests, analyses using the in situ active-site capping method, and isotope-labeling studies. A relationship between surface vacancy sites and catalytic activity has been established. CeO(2)(111) has been confirmed to be the catalytically active crystalline facet for hydrolysis. Water has been found to be associatively adsorbed on oxygen vacancy sites with medium strength, which does not lead to water dissociation to form stable hydroxides. This explains why the ceria catalyst is water-tolerant. PMID:23228093

  9. Rapid Online Non-Enzymatic Protein Digestion Combining Microwave Heating Acid Hydrolysis and Electrochemical Oxidation

    PubMed Central

    Basile, Franco; Hauser, Nicolas

    2010-01-01

    We report an online non-enzymatic method for site-specific digestion of proteins to yield peptides that are well suited for collision induced dissociation (CID) tandem mass spectrometry (MS/MS). The method combines online microwave heating acid hydrolysis at aspartic acid and online electrochemical oxidation at tryptophan and tyrosine. The combined microwave/electrochemical (microwave/echem) digestion is reproducible and produces peptides with an average sequence length of 10 amino acids. This peptide length is similar to the average peptide length of 9 amino acids obtained by digestion of proteins with the enzyme trypsin. As a result, the peptides produced by this novel non-enzymatic digestion method, when analyzed by ESI-MS, produce protonated molecules with mostly +1 and +2 charge states. The combination of these two non-enzymatic methods overcomes shortcomings with each individual method in that: i) peptides generated by the microwave-hydrolysis method have an average amino acid length of 16 amino acids, and ii) the inability of the electrochemical-cleavage method to reproducibly digest proteins with molecular masses above 4 kDa. Preliminary results are presented on the application and utility of this rapid online digestion (total of 6 min digestion time) on a series of standard peptides and proteins as well as an E. coli protein extract. PMID:21138252

  10. Acidic 1,3-propanediaminetetraacetato lanthanides with luminescent and catalytic ester hydrolysis properties

    NASA Astrophysics Data System (ADS)

    Chen, Mao-Long; Shi, Yan-Ru; Yang, Yu-Chen; Zhou, Zhao-Hui

    2014-11-01

    In acidic solution, a serials of water-soluble coordination polymers (CPs) were isolated as zonal 1D-CPs 1,3-propanediaminetetraacetato lanthanides [Ln(1,3-H3pdta)(H2O)5]n·2Cln·3nH2O [Ln=La, 1; Ce, 2; Pr, 3; Nd, 4; Sm, 5] (1,3-H4pdta=1,3-propanediaminetetraacetic acid, C11H18N2O8) in high yields. When 1 eq. mol potassium hydroxide was added to the solutions of 1D-CPs, respectively, two 1D-CPs [Ln(1,3-H2pdta)(H2O)3]n·Cln·2nH2O [Ln=Sm, 6; Gd, 7] were isolated at room temperature and seven 2D-CPs [Ln(1,3-H2pdta)(H2O)2]n·Cln·2nH2O [Ln=La, 8; Ce, 9; Pr, 10; Nd, 11; Sm, 12; Eu, 13; Gd, 14] were isolated at 70 °C. When the crystals of 1-4 were hydrothermally heated at 180 °C with 1-2 eq. mol potassium hydroxide, four 3D-CPs [Ln(1,3-Hpdta)]n·nH2O [Ln=La, 15; Ce, 16; Pr, 17; Nd, 18] were obtained. The two 2D-CPs [Ln(1,3-Hpdta)(H2O)]n·4nH2O (Sm, 19; Eu, 20) were isolated in similar reaction conditions. With the increments of pH value in the solution and reaction temperature, the structure becomes more complicated. 1-5 are soluble in water and 1 was traced by solution 13C{1H} NMR technique, the water-soluble lanthanides 1 and 5 show catalytic activity to ester hydrolysis reaction respectively, which indicate their important roles in the hydrolytic reaction. The europium complexes 13 and 20 show visible fluorescence at an excitation of 394 nm. The structure diversity is mainly caused by the variation of coordinated ligand in different pH values and lanthanide contraction effect. Acidic conditions are favorable for the isolations of lanthanide complexes in different structures and this may helpful to separate different lanthanides. The thermal stability investigations reveal that acidic condition is favorable to obtain the oxides at a lower temperature.

  11. A preliminary study of the hydrolysis of hydroxamic acid complexants in the presence of oxidising metal ions

    NASA Astrophysics Data System (ADS)

    Andrieux, Fabrice P. L.; Boxall, Colin; May, Iain; Taylor, Robin J.

    2010-03-01

    Simple hydroxamic acids (XHAs) are salt free, organic compounds with affinities for cations such as Np4+, Pu4+ and Fe3+. As such they have been identified as suitable reagents for the separation of either Pu and/or Np from U in modified or single cycle Purex based solvent extraction processes designed to meet the emerging requirements of Advanced Fuel Cycles. Acid catalyzed hydrolysis of free XHAs is well known and may impact negatively on reprocessing applications. The hydrolysis of metal-bound XHAs within metal ion-XHA complexes is less understood. Using a model derived for the study of hydroxamic acid hydrolysis in the presence of non-oxidising metal ions (Np (IV) and Fe(III)), we review data pertaining to the hydrolysis of hydroxamic acids in the presence of the oxidising Pu4+ ion, under conditions where the influence of the redox processes may potentially be neglected.

  12. Process for enzymatic hydrolysis of fatty acid triglycerides with oat caryopses

    SciTech Connect

    Hammond, E.G.; Lee, I.

    1992-02-18

    This patent describes the process for enzymatic hydrolysis of fatty acid triglycerides to obtain free fatty acids and glycerol. It comprises: increasing the water content of dehulled whole oat caryopses to a total water content of 17 to 44% the thus moistened caryopses having active oat lipase associated with the outer surfaces thereof; contacting the moistened whole caryopses with a liquid medium, continuing the contacting until at least 20% by volume of the triglyceride reactant has been hydrolyzed to free fatty acids and glycerol, most of the free fatty acids dissolving in the oil phase external to the caryopses and most of the glycerol being absorbed into the water within the caryopses; and separating the glycerol-containing caryopses from the fatty acid-containing oil phase.

  13. A combination of two lactic acid bacteria improves the hydrolysis of gliadin during wheat dough fermentation.

    PubMed

    Gerez, Carla Luciana; Dallagnol, Andrea; Rollán, Graciela; Font de Valdez, Graciela

    2012-12-01

    The evaluation of gliadin hydrolysis during dough fermentation by using two lactic acid bacteria, Lactobacillus plantarum CRL 775 and Pediococcus pentosaceus CRL 792, as pooled cell suspension (LAB) or cell free extract (CFE) was undertaken. The CFE pool produced a greater (121%) increase in amino acid concentration than the LAB pool (70-80%). These results were correlated with the decrease (76,100 and 64,300 ppm) in the gliadin concentration of doughs supplemented with CFE and LAB, respectively, compared to control doughs. The use of LAB peptidases seemed to be a viable technologic alternative to reduce the gliadin concentration in wheat dough without using living bacteria as starter. PMID:22986210

  14. Impact of dual temperature profile in dilute acid hydrolysis of spruce for ethanol production

    PubMed Central

    2010-01-01

    Background The two-step dilute acid hydrolysis (DAH) of softwood is costly in energy demands and capital costs. However, it has the advantage that hydrolysis and subsequent removal of hemicellulose-derived sugars can be carried out under conditions of low severity, resulting in a reduction in the level of sugar degradation products during the more severe subsequent steps of cellulose hydrolysis. In this paper, we discuss a single-step DAH method that incorporates a temperature profile at two levels. This profile should simulate the two-step process while removing its major disadvantage, that is, the washing step between the runs, which leads to increased energy demand. Results The experiments were conducted in a reactor with a controlled temperature profile. The total dry matter content of the hydrolysate was up to 21.1% w/w, corresponding to a content of 15.5% w/w of water insoluble solids. The highest measured glucose yield, (18.3 g glucose per 100 g dry raw material), was obtained after DAH cycles of 3 min at 209°C and 6 min at 211°C with 1% H2SO4, which resulted in a total of 26.3 g solubilized C6 sugars per 100 g dry raw material. To estimate the remaining sugar potential, enzymatic hydrolysis (EH) of the solid fraction was also performed. EH of the solid residue increased the total level of solubilized C6 sugars to a maximum of 35.5 g per 100 g dry raw material when DAH was performed as described above (3 min at 210°C and 2 min at 211°C with 1% H2SO4). Conclusion The dual-temperature DAH method did not yield decisively better results than the single-temperature, one-step DAH. When we compared the results with those of earlier studies, the hydrolysis performance was better than with the one-step DAH but not as well as that of the two-step, single-temperature DAH. Additional enzymatic hydrolysis resulted in lower levels of solubilized sugars compared with other studies on one-step DAH and two-step DAH followed by enzymatic hydrolysis. A two-step steam

  15. Mechanism of Orlistat Hydrolysis by the Thioesterase of Human Fatty Acid Synthase

    PubMed Central

    2015-01-01

    Fatty acid synthase (FASN), the sole protein capable of de novo synthesis of free fatty acids, is overexpressed in a wide variety of human cancers and is associated with poor prognosis and aggressiveness of these cancers. Orlistat, an FDA-approved drug for obesity treatment that inhibits pancreatic lipases in the GI tract, also inhibits the thioesterase (TE) of human FASN. The cocrystal structure of TE with orlistat shows a pseudo TE dimer containing two different forms of orlistat in the active site, an intermediate that is covalently bound to a serine residue (Ser2308) and a hydrolyzed and inactivated product. In this study, we attempted to understand the mechanism of TE-catalyzed orlistat hydrolysis by examining the role of the hexyl tail of the covalently bound orlistat in water activation for hydrolysis using molecular dynamics simulations. We found that the hexyl tail of the covalently bound orlistat undergoes a conformational transition, which is accompanied by destabilization of a hydrogen bond between a hydroxyl moiety of orlistat and the catalytic His2481 of TE that in turn leads to an increased hydrogen bonding between water molecules and His2481 and increased chance for water activation to hydrolyze the covalent bond between orlistat and Ser2308. Thus, the conformation of the hexyl tail of orlistat plays an important role in orlistat hydrolysis. Strategies that stabilize the hexyl tail may lead to the design of more potent irreversible inhibitors that target FASN and block TE activity with greater endurance. PMID:25309810

  16. Improved enzymatic hydrolysis of wheat straw by combined use of gamma ray and dilute acid for bioethanol production

    NASA Astrophysics Data System (ADS)

    Hyun Hong, Sung; Taek Lee, Jae; Lee, Sungbeom; Gon Wi, Seung; Ju Cho, Eun; Singh, Sudhir; Sik Lee, Seung; Yeoup Chung, Byung

    2014-01-01

    Pretreating wheat straw with a combination of dilute acid and gamma irradiation was performed in an attempt to enhance the enzymatic hydrolysis for bioethanol production. The glucose yield was significantly affected by combined pretreatment (3% sulfuric acid-gamma irradiation), compared with untreated wheat straw and individual pretreatment. The increasing enzymatic hydrolysis after combined pretreatment is resulting from decrease in crystallinity of cellulose, loss of hemicelluloses, and removal or modification of lignin. Therefore, combined pretreatment is one of the most effective methods for enhancing the enzymatic hydrolysis of wheat straw biomass.

  17. Hydrolysis of sorghum straw using phosphoric acid: evaluation of furfural production.

    PubMed

    Vázquez, Manuel; Oliva, Martha; Téllez-Luis, Simón J; Ramírez, José A

    2007-11-01

    Sorghum straw is a waste that has been studied scarcely. The main application is its use as raw material for xylose production. Xylose is a hemicellulosic sugar mainly used for its bioconversion toward xylitol. An alternative use could be its conversion toward furfural. The objective of this work was to study the furfural production by hydrolysis of sorghum straw with phosphoric acid at 134 degrees C. Several concentrations of H(3)PO(4) in the range 2-6% and reaction time (range 0-300 min) were evaluated. Kinetic parameters of mathematical models for predicting the concentration of xylose, glucose, arabinose, acetic acid and furfural in the hydrolysates were found. Optimal conditions for furfural production by acid hydrolysis were 6% H(3)PO(4) at 134 degrees C for 300 min, which yielded a solution with 13.7 g furfural/L, 4.0 g xylose/L, 2.9 g glucose/L, 1.1g arabinose/L and 1.2g acetic acid/L. The furfural yield of the process was 0.1336 g furfural/g initial dry matter was obtained. The results confirmed that sorghum straw can be used for furfural production when it is hydrolyzed using phosphoric acid. PMID:17145181

  18. Sulfuric Acid Pretreatment and Enzymatic Hydrolysis of Photoperiod Sensitvie Sorghum for Ethanol Production

    SciTech Connect

    F Xu; Y Shi; X Wu

    2011-12-31

    Photoperiod sensitive (PS) sorghum, with high soluble sugar content, high mass yield and high drought tolerance in dryland environments, has great potential for bioethanol production. The effect of diluted sulfuric acid pretreatment on enzymatic hydrolysis was investigated. Hydrolysis efficiency increased from 78.9 to 94.4% as the acid concentration increased from 0.5 to 1.5%. However, the highest total glucose yield (80.3%) occurred at the 1.0% acid condition because of the significant cellulose degradation at the 1.5% concentration. Synchrotron wide-angle X-ray diffraction was used to study changes of the degree of crystallinity. With comparison of cellulosic crystallinity and adjusted cellulosic crystallinity, the crystalline cellulose decreased after low acidic concentration (0.5%) applied, but did not change significantly, as the acid concentration increased. Scanning electron microscopy was also employed to understand how the morphological structure of PS sorghum changed after pretreatment. Under current processing conditions, the total ethanol yield is 74.5% (about 0.2 g ethanol from 1 g PS sorghum). A detail mass balance was also provided.

  19. Alcohol fermentation of sweet potato. 1. Acid hydrolysis and factors involved

    SciTech Connect

    Azhar, A.; Hamdy, M.K.

    1981-04-01

    Factors affecting acid hydrolysis of sweet potato powder (SPP) to fermentable sugars were examined. These include HCl concentration, temperature, time, and levels of SPP. Maximum reducing sugar, reported as dextrose equivalent (DE), was detected after 24 min hydrolysis (1% SPP) in 0.034N HCl heated at 154 degrees celcius. These samples also had 3.43% hydroxymethylfurfural (HMF) based on dry weight. A high level of HMF (9.2%) was detected in 1% SPP heated at 154 degrees C in 0.10N HCl for 18 min. The lowest concentration of HMF formed (1.8%), at maximal DE of 61%, was established in samples containing 5% SPP and heated at 154 degrees C in 0.034N HCl for 48 min. Aqueous extracts of uncured SPP, examined by HPLC, contained glucose, fructose and sucrose, butdegreaded SPP had only glucose and fructose. Products of degraded SPP, under appropriate conditions, could be used for alcohol fermentation. (Refs. 18).

  20. Characterisation of the products from pyrolysis of residues after acid hydrolysis of Miscanthus.

    PubMed

    Melligan, F; Dussan, K; Auccaise, R; Novotny, E H; Leahy, J J; Hayes, M H B; Kwapinski, W

    2012-03-01

    Platform chemicals such as furfural and hydroxymethylfurfural are major products formed during the acid hydrolysis of lignocellulosic biomass in second generation biorefining processes. Solid hydrolysis residues (HR) can amount to 50 wt.% of the starting biomass materials. Pyrolysis of the HRs gives rise to biochar, bio-liquids, and gases. Time and temperature were variables during the pyrolysis of HRs in a fixed bed tubular reactor, and both parameters have major influences on the amounts and properties of the products. Biochar, with potential for carbon sequestration and soil conditioning, composed about half of the HR pyrolysis product. The amounts (11-20 wt.%) and compositions (up to 77% of phenols in organic fraction) of the bio-liquids formed suggest that these have little value as fuels, but could be sources of phenols, and the gas can have application as a fuel. PMID:22281143

  1. Value of Coproduction of Ethanol and Furfural from Acid Hydrolysis Processes

    SciTech Connect

    Parker, S.; Calnon, M.; Feinberg, D.; Power, A.; Weiss, L.

    1984-05-01

    In the acid hydrolysis of a cellulosic feedstock (wood, wood wastes, or crop residues), up to 3.65 lb of furfural may be coproduced with each gallon of ethanol for only the cost of recovering and purifying it. Each plant producing 50 x 106 gal/yr of ethanol would produce an amount of by-product furfural equal to the total current domestic production. Thus, the need arises for investigation into potentially suitable processes for deriving profitable end products from furfural and thus expanding the market. The objectives of this study were to determine the economic potential of five selected, large volume derivatives of furfural that could displace hydrocarbon-based chemicals, and the consequent value of furfural as a by-product to the cellulose hydrolysis process of ethanol production.

  2. Phosphoric acid pretreatment of Achyranthes aspera and Sida acuta weed biomass to improve enzymatic hydrolysis.

    PubMed

    Siripong, Premjet; Duangporn, Premjet; Takata, Eri; Tsutsumi, Yuji

    2016-03-01

    Achyranthes aspera and Sida acuta, two types of weed biomass are abundant and waste in Thailand. We focus on them as novel feedstock for bio-ethanol production because they contain high-cellulose content (45.9% and 46.9%, respectively) and unutilized material. Phosphoric acid (70%, 75%, and 80%) was employed for the pretreatment to improve by enzymatic hydrolysis. The pretreatment process removed most of the xylan and a part of the lignin from the weeds, while most of the glucan remained. The cellulose conversion to glucose was greater for pretreated A. aspera (86.2 ± 0.3%) than that of the pretreated S. acuta (82.2 ± 1.1%). Thus, the removal of hemicellulose significantly affected the efficiency of the enzymatic hydrolysis. The scanning electron microscopy images showed the exposed fibrous cellulose on the cell wall surface, and this substantial change of the surface structure contributed to improving the enzyme accessibility. PMID:26744804

  3. Kinetic study of enzymatic hydrolysis of acid-pretreated coconut coir

    NASA Astrophysics Data System (ADS)

    Fatmawati, Akbarningrum; Agustriyanto, Rudy

    2015-12-01

    Biomass waste utilization for biofuel production such as bioethanol, has become more prominent currently. Coconut coir is one of lignocellulosic food wastes, which is abundant in Indonesia. Bioethanol production from such materials consists of more than one step. Pretreatment and enzymatic hydrolysis is crucial steps to produce sugar which can then be fermented into bioethanol. In this research, ground coconut coir was pretreated using dilute sulfuric acid at 121°C. This pretreatment had increased the cellulose content and decreased the lignin content of coconut coir. The pretreated coconut coir was hydrolyzed using a mix of two commercial cellulase enzymes at pH of 4.8 and temperature of 50°C. The enzymatic hydrolysis was conducted at several initial coconut coir slurry concentrations (0.1-2 g/100 mL) and reaction times (2-72 hours). The reducing sugar concentration profiles had been produced and can be used to obtain reaction rates. The highest reducing sugar concentration obtained was 1,152.567 mg/L, which was produced at initial slurry concentration of 2 g/100 mL and 72 hours reaction time. In this paper, the reducing sugar concentrations were empirically modeled as a function of reaction time using power equations. Michaelis-Menten kinetic model for enzymatic hydrolysis reaction is adopted. The kinetic parameters of that model for sulfuric acid-pretreated coconut coir enzymatic hydrolysis had been obtained which are Vm of 3.587×104 mg/L.h, and KM of 130.6 mg/L.

  4. Modeling Sucrose Hydrolysis in Dilute Sulfuric Acid Solutions at Pretreatment Conditions for Lignocellulosic Biomass

    SciTech Connect

    Bower, S.; Wickramasinghe, R.; Nagle, N. J.; Schell, D. J.

    2008-01-01

    Agricultural and herbaceous feedstocks may contain appreciable levels of sucrose. The goal of this study was to evaluate the survivability of sucrose and its hydrolysis products, fructose and glucose, during dilute sulfuric acid processing at conditions typically used to pretreat lignocellulose biomass. Solutions containing 25 g/l sucrose with 0.1-2.0% (w/w) sulfuric acid concentrations were treated at temperatures of 160-200 C for 3-12 min. Sucrose was observed to completely hydrolyze at all treatment conditions. However, appreciable concentrations of fructose and glucose were detected and glucose was found to be significantly more stable than fructose. Different mathematical approaches were used to fit the kinetic parameters for acid-catalyzed thermal degradation of these sugars. Since both sugars may survive dilute acid pretreatment, they could provide an additional carbon source for production of ethanol and other bio-based products.

  5. The mechanism of hydrothermal hydrolysis for glycyrrhizic acid into glycyrrhetinic acid and glycyrrhetinic acid 3-O-mono-β-D-glucuronide in subcritical water.

    PubMed

    Fan, Rui; Li, Nan; Xu, Honggao; Xiang, Jun; Wang, Lei; Gao, Yanxiang

    2016-01-01

    To improve the bioactivity and sweetness properties of glycyrrhizic acid (GL), the hydrothermal hydrolysis of GL into glycyrrhetinic acid (GA) and glycyrrhetinic acid 3-O-mono-β-D-glucuronide (GAMG) in subcritical water was investigated. The effects of temperature, time and their interaction on the conversion ratios were analyzed and the reactions were elaborated with kinetics and thermodynamics. The results showed that GL hydrothermal hydrolysis was significantly (P < 0.05) affected by reaction time and temperature, as well as their interaction, and could be fitted into first-order kinetics. The thermodynamic analysis indicated that the hydrolysis of GL was endergonic and non-spontaneous. The hydrolytic pathways were composed of complex consecutive and parallel reactions. It was concluded that subcritical water may be a potential medium for producing GAMG and GA. PMID:26213056

  6. A novel diffusion-biphasic hydrolysis coupled kinetic model for dilute sulfuric acid pretreatment of corn stover.

    PubMed

    Chen, Longjian; Zhang, Haiyan; Li, Junbao; Lu, Minsheng; Guo, Xiaomiao; Han, Lujia

    2015-02-01

    Kinetic experiments on the dilute sulfuric acid pretreatment of corn stover were performed. A high xylan removal and a low inhibitor concentration were achieved by acid pretreatment. A novel diffusion-hydrolysis coupled kinetic model was proposed. The contribution to the xylose yield was analyzed by the kinetic model. Compared with the inhibitor furfural negatively affecting xylose yield, the fast and slow-hydrolyzing xylan significantly contributed to the xylose yield, however, their dominant roles were dependent on reaction temperature and time. The impact of particle size and acid concentration on the xylose yield were also investigated. The diffusion process may significantly influence the hydrolysis of large particles. Increasing the acid concentration from 0.15 M to 0.30 M significantly improved the xylose yield, whereas the extent of improvement decreased to near-quantitative when further increasing acid loading. These findings shed some light on the mechanism for dilute sulfuric acid hydrolysis of corn stover. PMID:25479388

  7. Phosphine and thiophene cyclopalladated complexes: hydrolysis reactions in strong acidic media.

    PubMed

    García, Begoña; Hoyuelos, Francisco J; Ibeas, Saturnino; Muñoz, María S; Navarro, Ana M; Peñacoba, Indalecio A; Leal, José M

    2010-12-01

    The mechanisms for the hydrolysis of organopalladium complexes [Pd(CNN)R]BF(4) (R=P(OPh)(3), PPh(3), and SC(4)H(8)) were investigated at 25 °C by using UV/Vis absorbance measurements in 10 % v/v ethanol/water mixtures containing different sulphuric acid concentrations in the 1.3-11.7 M range. In all cases, a biphasic behavior was observed with rate constants k(1obs), which corresponds to the initial step of the hydrolysis reaction, and k(2obs), where k(1obs)>k(2obs). The plots of k(1obs) and k(2obs) versus sulfuric acid concentration suggest a change in the reaction mechanism. The change with respect to the k(1obs) value corresponds to 35 %, 2 %, and 99 % of the protonated complexes for R=PPh(3), P(OPh)(3), and SC(4)H(8), respectively. Regarding k(2obs), the change occurred in all cases at about 6.5 M H(2)SO(4) and matched up with the results reported for the hydrolysis of the 2-acetylpyridinephenylhydrazone (CNN) ligand. By using the excess acidity method, the mechanisms were elucidated by carefully looking at the variation of k(i),(obs) (i=1,2) versus cH+. The rate-determining constants, k(0,A-1), k(0,A-2), and k(0,A-SE2) were evaluated in all cases. The R=P(OPh)(3) complex was most reactive due to its π-acid character, which favors the rupture of the trans nitrogen-palladium bond in the A-2 mechanism and also that of the pyridine nitrogen-palladium bond in the A-1 mechanism. The organometallic bond exerts no effect on the relative basicity of the complexes, which are strongly reliant on the substituent. PMID:21125574

  8. Heteropoly Acid/Nitrogen Functionalized Onion-like Carbon Hybrid Catalyst for Ester Hydrolysis Reactions.

    PubMed

    Liu, Wei; Qi, Wei; Guo, Xiaoling; Su, Dangsheng

    2016-02-18

    A novel heteropoly acid (HPA)/nitrogen functionalized onion-like carbon (NOLC) hybrid catalyst was synthesized through supramolecular (electrostatic and hydrogen bond) interactions between the two components. The chemical structure and acid strength of the HPA/NOLC hybrid have been fully characterized by thermogravimetric analysis, IR spectroscopy, X-ray photoelectron spectroscopy, NH3 temperature-programmed desorption and acid-base titration measurements. The proposed method for the fabrication of the HPA/NOLC hybrid catalyst is a universal strategy for different types of HPAs to meet various requirements of acidic or redox catalysis. The hydrophobic environment of NOLC effectively prevents the deactivation of HPA in an aqueous system, and the combination of uniformly dispersed HPA clusters and the synergistic effect between NOLC and HPA significantly promotes its activity in ester hydrolysis reactions, which is higher than that of bare PWA as homogeneous catalyst. The kinetics of the hydrolysis reactions indicate that the aggregation status of the catalyst particles has great influence on the apparent activity. PMID:26606266

  9. Identification of amino acid thiohydantoins directly by thin-layer chromatography and indirectly by gas-liquid chromatography after hydrolysis.

    PubMed Central

    Rangarajan, M; Darbre, A

    1975-01-01

    A method is described for the identification of amino acid thiohydantoins by two-dimensional t.l.c. An indirect method for the determination of amino acid thiohydantoins is described which, after hydrolysis, the corresponding amino acids are determined by g.l.c. PMID:1167153

  10. Effect of ultrasonic pre-treatment on low temperature acid hydrolysis of oil palm empty fruit bunch.

    PubMed

    Yunus, Robiah; Salleh, Shanti Faridah; Abdullah, Nurhafizah; Biak, Dyg Radiah Awg

    2010-12-01

    Various pre-treatment techniques change the physical and chemical structure of the lignocellulosic biomass and improve hydrolysis rates. The effect of ultrasonic pre-treatment on oil palm empty fruit bunch (OPEFB) fibre prior to acid hydrolysis has been evaluated. The main objective of this study was to determine if ultrasonic pre-treatment could function as a pre-treatment method for the acid hydrolysis of OPEFB fibre at a low temperature and pressure. Hydrolysis at a low temperature was studied using 2% sulphuric acid; 1:25 solid liquid ratio and 100 degrees C operating temperature. A maximum xylose yield of 58% was achieved when the OPEFB fibre was ultrasonicated at 90% amplitude for 45min. In the absence of ultrasonic pre-treatment only 22% of xylose was obtained. However, no substantial increase of xylose formation was observed for acid hydrolysis at higher temperatures of 120 and 140 degrees C on ultrasonicated OPEFB fibre. The samples were then analysed using a scanning electron microscope (SEM) to describe the morphological changes of the OPEFB fibre. The SEM observations show interesting morphological changes within the OPEFB fibre for different acid hydrolysis conditions. PMID:20719502

  11. Optimization of the Hydrolysis of Safflower Oil for the Production of Linoleic Acid, Used as Flavor Precursor

    PubMed Central

    Aziz, Marya; Husson, Florence; Kermasha, Selim

    2015-01-01

    Commercial lipases, from porcine pancreas (PPL), Candida rugosa (CRL), and Thermomyces lanuginosus (Lipozyme TL IM), were investigated in terms of their efficiency for the hydrolysis of safflower oil (SO) for the liberation of free linoleic acid (LA), used as a flavor precursor. Although PPL, under the optimized conditions, showed a high degree of hydrolysis (91.6%), its low tolerance towards higher substrate concentrations could limit its use for SO hydrolysis. In comparison to the other investigated lipases, Lipozyme TL IM required higher amount of enzyme and an additional 3 h of reaction time to achieve its maximum degree of SO hydrolysis (90.2%). On the basis of the experimental findings, CRL was selected as the most appropriate biocatalyst, with 84.1% degree of hydrolysis. The chromatographic analyses showed that the CRL-hydrolyzed SO is composed mainly of free LA. PMID:26904663

  12. Stimulation of phospholipid hydrolysis and arachidonic acid mobilization in human uterine decidua cells by phorbol ester.

    PubMed Central

    Schrey, M P; Read, A M; Steer, P J

    1987-01-01

    Vasopressin and oxytocin both stimulated inositol phosphate accumulation in isolated uterine decidua cells. Pretreatment of cells with the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) prevented this agonist-induced phosphoinositide hydrolysis. TPA (0.1 microM) alone had no effect on basal inositol phosphate accumulation, but stimulated phosphoinositide deacylation, as indicated by a 2-fold increase in lysophosphatidylinositol and glycerophosphoinositol. TPA also stimulated a dose-related release of arachidonic acid from decidua-cell phospholipid [phosphatidylcholine (PC) much greater than phosphatidylinositol (PI) greater than phosphatidylethanolamine]. The phorbol ester 4 beta-phorbol 12,13-diacetate (PDA) at 0.1 microM had no effect on arachidonic acid mobilization. The TPA-stimulated increase in arachidonic acid release was apparent by 2 1/2 min (116% of control), maximal after 20 min (283% of control), and remained around this value (306% of control) after 120 min incubation. TPA also stimulated significant increases in 1,2-diacylglycerol and monoacylglycerol production at 20 and 120 min. Although the temporal increases in arachidonic acid and monoacylglycerol accumulation in the presence of TPA continued up to 120 min, that of 1,2-diacylglycerol declined after 20 min. In decidua cells prelabelled with [3H]choline, TPA also stimulated a significant decrease in radiolabelled PC after 20 min, which was accompanied by an increased release of water-soluble metabolites into the medium. Most of the radioactivity in the extracellular pool was associated with choline, whereas the main cellular water-soluble metabolite was phosphorylcholine. TPA stimulated extracellular choline accumulation to 183% and 351% of basal release after 5 and 20 min respectively and cellular phosphorylcholine production to 136% of basal values after 20 min. These results are consistent with a model in which protein kinase C activation by TPA leads to arachidonic acid mobilization

  13. Effect of acid hydrolysis on regenerated kenaf core membrane produced using aqueous alkaline-urea systems.

    PubMed

    Padzil, Farah Nadia Mohammad; Zakaria, Sarani; Chia, Chin Hua; Jaafar, Sharifah Nabihah Syed; Kaco, Hatika; Gan, Sinyee; Ng, Peivun

    2015-06-25

    Bleached kenaf core pulps (BKC) were hydrolyzed in H2SO4 (0.5M) at different time (0min to 90min) at room temperature. After the hydrolysis process, the viscosity average molecular weight (Mŋ) for BKC sample has reduced from 14.5×10(4) to 2.55×10(4). The hydrolyzed BKC was then dissolved in NaOH:urea:water and in LiOH:urea:water mixed solvent at the ratio of 7:12:81 and 4.6:15:80.4, respectively. The increased in hydrolysis time has decreased Mŋ of cellulose leading to easy dissolution process. Higher porosity and transparency with lower crystallinity index (CrI) of regenerated membrane produced can be achieved as the Mŋ reduced. The properties of membrane were observed through FESEM, UV-vis spectrophotometer and XRD. This study has proven that acid hydrolysis has reduced the Mŋ of cellulose, thus, enhanced the properties of regenerated membrane produced with assisted by alkaline/urea system. PMID:25839807

  14. Glycosyl conformational and inductive effects on the acid catalysed hydrolysis of purine nucleosides.

    PubMed Central

    Jordan, F; Niv, H

    1977-01-01

    The log kobs vs. pH profiles were determined in the intermediate acidity region for the glycosyl hydrolysis of guanosine and its 8-amino, 8-monomethylamino, 8-dimethylamino and 8-bromo derivatives. The decreased rate of the 8-amino and enhanced rate of the 8-bromo compound compared to guanosine support an A type mechanism: base protonation followed by glycosyl bond cleavage. All three 8-amino guanosines exhibited log kobs - pH profiles clearly showing that both mono and di-base protonated nucleosides undergo hydrolysis. The 700 fold rate acceleration of 8-N(CH3)-guanosine compared to 8-NHCH3-guanosine and the 110 fold rate acceleration of 8-N(CH3)2-adenosine compared to 8-NHCH3-adenosine could be unequivocally attributed to the fixed syn glycosyl conformation of both 8-dimethylamino compounds and relief of steric compression upon hydrolysis in these molecules. The lack of anomerization of all substrates during the course of the reaction supports an A rather than a Schiff-base mechanism. PMID:17100

  15. Housefly larvae hydrolysate: orthogonal optimization of hydrolysis, antioxidant activity, amino acid composition and functional properties

    PubMed Central

    2013-01-01

    Background Antioxidant, one of the most important food additives, is widely used in food industry. At present, antioxidant is mostly produced by chemical synthesis, which would accumulate to be pathogenic. Therefore, a great interest has been developed to identify and use natural antioxidants. It was showed that there are a lot of antioxidative peptides in protein hydrolysates, possessing strong capacity of inhibiting peroxidation of macro-biomolecular and scavenging free redicals in vivo. Enzymatic hydrolysis used for preparation of antioxidative peptides is a new hot-spot in the field of natural antioxidants. It reacts under mild conditions, with accurate site-specific degradation, good repeatability and few damages to biological activity of protein. Substrates for enzymatic hydrolysis are usually plants and aqua-animals. Insects are also gaining attention because of their rich protein and resource. Antioxidative peptides are potential to be exploited as new natural antioxidant and functional food. There is a huge potential market in medical and cosmetic field as well. Result Protein hydrolysate with antioxidant activity was prepared from housefly larvae, by a two-step hydrolysis. Through orthogonal optimization of the hydrolysis conditions, the degree of hydrolysis was determined to be approximately 60%. Fractionated hydrolysate at 25 mg/mL, 2.5 mg/mL and 1 mg/mL exhibited approximately 50%, 60% and 50% of scavenging capacity on superoxide radicals, 1, 1-Diphenyl-2-picrylhydrazyl radicals and hydroxyl radicals, respectively. Hydrolysate did not exhibit substantial ion chelation. Using a linoneic peroxidation system, the inhibition activity of hydrolysate at 20 mg/mL was close to that of 20 μg/mL tertiary butylhydroquinone, suggesting a potential application of hydrolysate in the oil industry as an efficient antioxidant. The lyophilized hydrolysate presented almost 100% solubility at pH 3-pH 9, and maintained nearly 100% activity at pH 5-pH 8 at 0

  16. Hydrolysis of Selected Tropical Plant Wastes Catalyzed by a Magnetic Carbonaceous Acid with Microwave.

    PubMed

    Su, Tong-Chao; Fang, Zhen; Zhang, Fan; Luo, Jia; Li, Xing-Kang

    2015-01-01

    In this study, magnetic carbonaceous acids were synthesized by pyrolysis of the homogeneous mixtures of glucose and magnetic Fe3O4 nanoparticles, and subsequent sulfonation. The synthesis conditions were optimized to obtain a catalyst with both high acid density (0.75 mmol g(-1)) and strong magnetism [magnetic saturation, Ms = 19.5 Am(2) kg(-1)]. The screened catalyst (C-SO3H/Fe3O4) was used to hydrolyze ball-milled cellulose in a microwave reactor with total reducing sugar (TRS) yield of 25.3% under the best conditions at 190 °C for 3.5 h. It was cycled for at least seven times with high catalyst recovery rate (92.8%), acid density (0.63 mmol g(-1)) and magnetism (Ms = 12.9 Am(2) kg(-1)), as well as high TRS yield (20.1%) from the hydrolysis of ball-milled cellulose. The catalyst was further successfully tested for the hydrolysis of tropical biomass with high TRS and glucose yields of 79.8% and 58.3% for bagasse, 47.2% and 35.6% for Jatropha hulls, as well as 54.4% and 35.8% for Plukenetia hulls. PMID:26648414

  17. Hydrolysis of Selected Tropical Plant Wastes Catalyzed by a Magnetic Carbonaceous Acid with Microwave

    NASA Astrophysics Data System (ADS)

    Su, Tong-Chao; Fang, Zhen; Zhang, Fan; Luo, Jia; Li, Xing-Kang

    2015-12-01

    In this study, magnetic carbonaceous acids were synthesized by pyrolysis of the homogeneous mixtures of glucose and magnetic Fe3O4 nanoparticles, and subsequent sulfonation. The synthesis conditions were optimized to obtain a catalyst with both high acid density (0.75 mmol g-1) and strong magnetism [magnetic saturation, Ms = 19.5 Am2 kg-1]. The screened catalyst (C-SO3H/Fe3O4) was used to hydrolyze ball-milled cellulose in a microwave reactor with total reducing sugar (TRS) yield of 25.3% under the best conditions at 190 °C for 3.5 h. It was cycled for at least seven times with high catalyst recovery rate (92.8%), acid density (0.63 mmol g-1) and magnetism (Ms = 12.9 Am2 kg-1), as well as high TRS yield (20.1%) from the hydrolysis of ball-milled cellulose. The catalyst was further successfully tested for the hydrolysis of tropical biomass with high TRS and glucose yields of 79.8% and 58.3% for bagasse, 47.2% and 35.6% for Jatropha hulls, as well as 54.4% and 35.8% for Plukenetia hulls.

  18. Hydrolysis of Selected Tropical Plant Wastes Catalyzed by a Magnetic Carbonaceous Acid with Microwave

    PubMed Central

    Su, Tong-Chao; Fang, Zhen; Zhang, Fan; Luo, Jia; Li, Xing-Kang

    2015-01-01

    In this study, magnetic carbonaceous acids were synthesized by pyrolysis of the homogeneous mixtures of glucose and magnetic Fe3O4 nanoparticles, and subsequent sulfonation. The synthesis conditions were optimized to obtain a catalyst with both high acid density (0.75 mmol g−1) and strong magnetism [magnetic saturation, Ms = 19.5 Am2 kg−1]. The screened catalyst (C-SO3H/Fe3O4) was used to hydrolyze ball-milled cellulose in a microwave reactor with total reducing sugar (TRS) yield of 25.3% under the best conditions at 190 °C for 3.5 h. It was cycled for at least seven times with high catalyst recovery rate (92.8%), acid density (0.63 mmol g−1) and magnetism (Ms = 12.9 Am2 kg−1), as well as high TRS yield (20.1%) from the hydrolysis of ball-milled cellulose. The catalyst was further successfully tested for the hydrolysis of tropical biomass with high TRS and glucose yields of 79.8% and 58.3% for bagasse, 47.2% and 35.6% for Jatropha hulls, as well as 54.4% and 35.8% for Plukenetia hulls. PMID:26648414

  19. Effect of acid concentration and pulp properties on hydrolysis reactions of mercerized sisal.

    PubMed

    Lacerda, Talita M; Zambon, Márcia D; Frollini, Elisabete

    2013-03-01

    The influence of sulfuric acid concentration (H2SO4 5-25%, 100°C), crystallinity and fibers size on the hydrolysis reaction of sisal pulps were investigated, with the goal of evaluating both the liquor composition, as an important step in the production of bioethanol, and the residual non-hydrolyzed pulp, to determine its potential application as materials. Aliquots were withdrawn from the reaction media, and the liquor composition was analyzed by HPLC. The residual non-hydrolyzed pulps were characterized by SEM, their average molar mass and crystallinity index, and their size distribution was determined using a fiber analyzer. Sulfuric acid 25% led to the highest glucose content (approximately 10gL(-1)), and this acid concentration was chosen to evaluate the influence of both the fiber size and crystallinity of the starting pulp on hydrolysis. The results showed that fibers with higher length and lower crystallinity favored glucose production in approximately 12%, with respect to the highly crystalline shorter fibers. PMID:23465940

  20. Nitrification of archaeal ammonia oxidizers in acid soils is supported by hydrolysis of urea

    PubMed Central

    Lu, Lu; Han, Wenyan; Zhang, Jinbo; Wu, Yucheng; Wang, Baozhan; Lin, Xiangui; Zhu, Jianguo; Cai, Zucong; Jia, Zhongjun

    2012-01-01

    The hydrolysis of urea as a source of ammonia has been proposed as a mechanism for the nitrification of ammonia-oxidizing bacteria (AOB) in acidic soil. The growth of Nitrososphaera viennensis on urea suggests that the ureolysis of ammonia-oxidizing archaea (AOA) might occur in natural environments. In this study, 15N isotope tracing indicates that ammonia oxidation occurred upon the addition of urea at a concentration similar to the in situ ammonium content of tea orchard soil (pH 3.75) and forest soil (pH 5.4) and was inhibited by acetylene. Nitrification activity was significantly stimulated by urea fertilization and coupled well with abundance changes in archaeal amoA genes in acidic soils. Pyrosequencing of 16S rRNA genes at whole microbial community level demonstrates the active growth of AOA in urea-amended soils. Molecular fingerprinting further shows that changes in denaturing gradient gel electrophoresis fingerprint patterns of archaeal amoA genes are paralleled by nitrification activity changes. However, bacterial amoA and 16S rRNA genes of AOB were not detected. The results strongly suggest that archaeal ammonia oxidation is supported by hydrolysis of urea and that AOA, from the marine Group 1.1a-associated lineage, dominate nitrification in two acidic soils tested. PMID:22592820

  1. Ferulic acid: a key component in grass lignocellulose recalcitrance to hydrolysis.

    PubMed

    de Oliveira, Dyoni Matias; Finger-Teixeira, Aline; Rodrigues Mota, Thatiane; Salvador, Victor Hugo; Moreira-Vilar, Flávia Carolina; Correa Molinari, Hugo Bruno; Craig Mitchell, Rowan Andrew; Marchiosi, Rogério; Ferrarese-Filho, Osvaldo; Dantas Dos Santos, Wanderley

    2015-12-01

    In the near future, grasses must provide most of the biomass for the production of renewable fuels. However, grass cell walls are characterized by a large quantity of hydroxycinnamic acids such as ferulic and p-coumaric acids, which are thought to reduce the biomass saccharification. Ferulic acid (FA) binds to lignin, polysaccharides and structural proteins of grass cell walls cross-linking these components. A controlled reduction of FA level or of FA cross-linkages in plants of industrial interest can improve the production of cellulosic ethanol. Here, we review the biosynthesis and roles of FA in cell wall architecture and in grass biomass recalcitrance to enzyme hydrolysis. PMID:25417596

  2. Preparation and evaluation of lignosulfonates as a dispersant for gypsum paste from acid hydrolysis lignin.

    PubMed

    Matsushita, Yasuyuki; Yasuda, Seiichi

    2005-03-01

    In order to effectively utilize a by-product of the acid saccharification process of woody materials, the chemical conversion of guaiacyl sulfuric acid lignin (SAL), one of the acid hydrolysis lignins, into water-soluble sulfonated products with high dispersibitity was investigated. At first, SAL was phenolated (P-SAL) to enhance the solubility and reactivity. Lignosulfonates were prepared from P-SAL by three methods of hydroxymethylation followed by neutral sulfonation (two-step method), sulfomethylation (one-step method) and arylsulfonation. Surprisingly, all prepared lignosulfonates possessed 30 to 70% higher dispersibility for gypsum paste than the commercial lignosulfonate. Evaluation of the preparations for gypsum paste suggested that the higher molecular weights and sulfur contents of the preparations increased their dispersibility. PMID:15491828

  3. Antioxidative Peptides Derived from Enzyme Hydrolysis of Bone Collagen after Microwave Assisted Acid Pre-Treatment and Nitrogen Protection

    PubMed Central

    Lin, Yun-Jian; Le, Guo-Wei; Wang, Jie-Yun; Li, Ya-Xin; Shi, Yong-Hui; Sun, Jin

    2010-01-01

    This study focused on the preparation method of antioxidant peptides by enzymatic hydrolysis of bone collagen after microwave assisted acid pre-treatment and nitrogen protection. Phosphoric acid showed the highest ability of hydrolysis among the four other acids tested (hydrochloric acid, sulfuric acid and/or citric acid). The highest degree of hydrolysis (DH) was 9.5% using 4 mol/L phosphoric acid with a ratio of 1:6 under a microwave intensity of 510 W for 240 s. Neutral proteinase gave higher DH among the four protease tested (Acid protease, neutral protease, Alcalase and papain), with an optimum condition of: (1) ratio of enzyme and substrate, 4760 U/g; (2) concentration of substrate, 4%; (3) reaction temperature, 55 °C and (4) pH 7.0. At 4 h, DH increased significantly (P < 0.01) under nitrogen protection compared with normal microwave assisted acid pre-treatment hydrolysis conditions. The antioxidant ability of the hydrolysate increased and reached its maximum value at 3 h; however DH decreased dramatically after 3 h. Microwave assisted acid pre-treatment and nitrogen protection could be a quick preparatory method for hydrolyzing bone collagen. PMID:21151439

  4. Imidase catalyzing desymmetric imide hydrolysis forming optically active 3-substituted glutaric acid monoamides for the synthesis of gamma-aminobutyric acid (GABA) analogs.

    PubMed

    Nojiri, Masutoshi; Hibi, Makoto; Shizawa, Hiroaki; Horinouchi, Nobuyuki; Yasohara, Yoshihiko; Takahashi, Satomi; Ogawa, Jun

    2015-12-01

    The recent use of optically active 3-substituted gamma-aminobutyric acid (GABA) analogs in human therapeutics has identified a need for an efficient, stereoselective method of their synthesis. Here, bacterial strains were screened for enzymes capable of stereospecific hydrolysis of 3-substituted glutarimides to generate (R)-3-substituted glutaric acid monoamides. The bacteria Alcaligenes faecalis NBRC13111 and Burkholderia phytofirmans DSM17436 were discovered to hydrolyze 3-(4-chlorophenyl) glutarimide (CGI) to (R)-3-(4-chlorophenyl) glutaric acid monoamide (CGM) with 98.1% enantiomeric excess (e.e.) and 97.5% e.e., respectively. B. phytofirmans DSM17436 could also hydrolyze 3-isobutyl glutarimide (IBI) to produce (R)-3-isobutyl glutaric acid monoamide (IBM) with 94.9% e.e. BpIH, an imidase, was purified from B. phytofirmans DSM17436 and found to generate (R)-CGM from CGI with specific activity of 0.95 U/mg. The amino acid sequence of BpIH had a 75% sequence identity to that of allantoinase from A. faecalis NBRC13111 (AfIH). The purified recombinant BpIH and AfIH catalyzed (R)-selective hydrolysis of CGI and IBI. In addition, a preliminary investigation of the enzymatic properties of BpIH and AfIH revealed that both enzymes were stable in the range of pH 6-10, with an optimal pH of 9.0, stable at temperatures below 40 °C, and were not metalloproteins. These results indicate that the use of this class of hydrolase to generate optically active 3-substituted glutaric acid monoamide could simplify the production of specific chiral GABA analogs for drug therapeutics. PMID:26205522

  5. Acid-catalyzed carboxylic acid esterification and ester hydrolysis mechanism: acylium ion as a sharing active intermediate via a spontaneous trimolecular reaction based on density functional theory calculation and supported by electrospray ionization-mass spectrometry.

    PubMed

    Shi, Hongchang; Wang, Yilei; Hua, Ruimao

    2015-11-11

    By DFT calculation, we found that acid-catalyzed carboxylic acid esterification and ester hydrolysis are brief two-step reactions. First, the carboxylic acid hydroxyl-oxygen or ester alkyl-oxygen is protonated, which generates a highly active acylium ion. The protonation requires an activation energy (Ea) of 4-10 kcal mol(-1), and is the rate-controlling step of the esterification or hydrolysis. Sequentially, the acylium ion spontaneously reacts with two alcohol or two water molecules to form a neutral product molecule; this is a trimolecular reaction. The acylium ion is the highly active intermediate shared by esterification and hydrolysis. ESI-MS data for several typical carboxylic acids confirmed that their acylium ions are easily generated. For 2,4,6-trialkylbenzoic acid and its ester, the two unsubstituted carbons in the benzene ring are very easily protonated, and we have thus revealed the root of the success of Newman's method. Based on these results, the popular esterification and hydrolysis mechanism in organic chemistry textbooks is incorrect. PMID:26445892

  6. Kinetics and mechanism of the acid-catalyzed hydrolysis of a hypermodified nucleoside wyosine and its 5'-monophosphate.

    PubMed Central

    Golankiewicz, B; Zielonacka-Lis, E; Folkman, W

    1985-01-01

    The rates of acid-catalyzed hydrolysis of a hypermodified nucleoside, wyosine and its 5'-monophosphate were determined at various pH, temperature and buffer concentrations. The results show that despite distinct differences in structure and the glycosyl bond stability, the hydrolysis of wyosine proceeds via cleavage of the C-N bond by A-1 mechanism, analogously to simple nucleosides. Unlike majority of other monophosphates studied so far, wyosine 5'-monophosphate is not more stable than respective nucleoside. PMID:4000960

  7. Peptide sequencing by using a combination of partial acid hydrolysis and fast-atom-bombardment mass spectrometry.

    PubMed Central

    De Angelis, F; Botta, M; Ceccarelli, S; Nicoletti, R

    1986-01-01

    To overcome the limit of the intensity of ions carrying sequence information in structural determinations of peptides by fast-atom-bombardment m.s., we have developed a method that consists in taking spectra of the peptide acid hydrolysates at different hydrolysis times. Peaks correspond to the oligomers arising from the peptide partial hydrolysis. The sequence can then be identified from the structurally overlapping fragments. PMID:2428356

  8. Extraction of cellulose nano-crystals from old corrugated container fiber using phosphoric acid and enzymatic hydrolysis followed by sonication.

    PubMed

    Tang, Yanjun; Shen, Xiaochuang; Zhang, Junhua; Guo, Daliang; Kong, Fangong; Zhang, Nan

    2015-07-10

    Due to its amazing physicochemical properties and high environmental compatibility, cellulose nano-crystals (CNC) hold great promise for serving as a strategic platform for sustainable development. Now, there has been growing interest in the development of processes using waste or residual biomass as CNC source for addressing economic and environmental concerns. In the present work, a combined process involving phosphoric acid hydrolysis, enzymatic hydrolysis and sonication was proposed aiming to efficiently exact CNC from low-cost old corrugated container (OCC) pulp fiber. The effect of enzymatic hydrolysis on the yield and microstructure of resulting CNC was highlighted. Results showed that the enzymatic hydrolysis was effective in enhancing CNC yield after phosphoric acid hydrolysis. CNC was obtained with a yield of 23.98 wt% via the combined process with phosphoric acid concentration of 60 wt%, cellulase dosage of 2 mL (84 EGU) per 2g fiber and sonication intensity of 200 W. Moreover, the presence of enzymatic hydrolysis imparted the obtained CNC with improved dispersion, increased crystallinity and thermal stability. PMID:25857993

  9. Hybrid reuteransucrase enzymes reveal regions important for glucosidic linkage specificity and the transglucosylation/hydrolysis ratio.

    PubMed

    Kralj, Slavko; van Leeuwen, Sander S; Valk, Vincent; Eeuwema, Wieger; Kamerling, Johannis P; Dijkhuizen, Lubbert

    2008-12-01

    The reuteransucrase enzymes of Lactobacillus reuteri strain 121 (GTFA) and L. reuteri strain ATCC 55730 (GTFO) convert sucrose into alpha-d-glucans (labelled reuterans) with mainly alpha-(1-->4) glucosidic linkages (50% and 70%, respectively), plus alpha-(1-->6) linkages. In the present study, we report a detailed analysis of various hybrid GTFA/O enzymes, resulting in the identification of specific regions in the N-termini of the catalytic domains of these proteins as the main determinants of glucosidic linkage specificity. These regions were divided into three equal parts (A1-3; O1-3), and used to construct six additional GTFA/O hybrids. All hybrid enzymes were able to synthesize alpha-glucans from sucrose, and oligosaccharides from sucrose plus maltose or isomaltose as acceptor substrates. Interestingly, not only the A2/O2 regions, with the three catalytic residues, affect glucosidic linkage specificity, but also the upstream A1/O1 regions make a strong contribution. Some GTFO derived hybrid/mutant enzymes displayed strongly increased transglucosylation/hydrolysis activity ratios. The reduced sucrose hydrolysis allowed the much improved conversion of sucrose into oligo- and polysaccharide products. Thus, the glucosidic linkage specificity and transglucosylation/hydrolysis ratios of reuteransucrase enzymes can be manipulated in a relatively simple manner. This engineering approach has yielded clear changes in oligosaccharide product profiles, as well as a range of novel reuteran products differing in alpha-(1-->4) and alpha-(1-->6) linkage ratios. PMID:19016850

  10. Ultrasonic enhance acid hydrolysis selectivity of cellulose with HCl-FeCl3 as catalyst.

    PubMed

    Li, Jinbao; Zhang, Xiangrong; Zhang, Meiyun; Xiu, Huijuan; He, Hang

    2015-03-01

    The effect of ultrasonic pretreatment coupled with HCl-FeCl3 catalyst was evaluated to hydrolyze cellulose amorphous regions. The ultrasonic pretreatment leads to cavitation that affects the morphology and microstructure of fibers, enhancing the accessibility of chemical reagent to the loosened amorphous regions of cellulose. In this work, Fourier transform infrared spectroscopy (FTIR) was used to identify characteristic absorption bands of the constituents and the crystallinity was evaluated by the X-ray diffraction (XRD) technique. The results indicated that appropriate ultrasonic pretreatment assisted with FeCl3 can enhance the acid hydrolysis of amorphous regions of cellulose, thus improving the crystallinity of the remaining hydrocellulose. It was observed that sonication samples that were pretreated for 300 W and 20 min followed by acid hydrolysis had maximum of 78.9% crystallinity. The crystallinity was 9.2% higher than samples that were not subjected to ultrasound. In addition, the average fines length decreased from 49 μm to 37 μm. PMID:25498717

  11. Hydrolysis optimization and characterization study of preparing fatty acids from Jatropha curcas seed oil

    PubMed Central

    2011-01-01

    Background Fatty acids (FAs) are important as raw materials for the biotechnology industry. Existing methods of FAs production are based on chemical methods. In this study potassium hydroxide (KOH)-catalyzed reactions were utilized to hydrolysis Jatropha curcas seed oil. Results The parameters effect of ethanolic KOH concentration, reaction temperature, and reaction time to free fatty acid (FFA%) were investigated using D-Optimal Design. Characterization of the product has been studied using Fourier transforms infrared spectroscopy (FTIR), gas chromatography (GC) and high performance liquid chromatography (HPLC). The optimum conditions for maximum FFA% were achieved at 1.75M of ethanolic KOH concentration, 65°C of reaction temperature and 2.0 h of reaction time. Conclusions This study showed that ethanolic KOH concentration was significant variable for J. curcas seed oil hydrolysis. In a 18-point experimental design, FFA% of hydrolyzed J. curcas seed oil can be raised from 1.89% to 102.2%, which proved by FTIR and HPLC. PMID:22044685

  12. Ferrihydrite dissolution by pyridine-2,6-bis(monothiocarboxylic acid) and hydrolysis products

    NASA Astrophysics Data System (ADS)

    Dhungana, Suraj; Anthony, Charles R.; Hersman, Larry E.

    2007-12-01

    Pyridine-2,6-bis(monothiocarboxylate) (pdtc), a metabolic product of microorganisms, including Pseudomonas putida and Pseudomonas stutzeri was investigated for its ability of dissolve Fe(III)(hydr)oxides at pH 7.5. Concentration dependent dissolution of ferrihydrite under anaerobic environment showed saturation of the dissolution rate at the higher concentration of pdtc. The surface controlled ferrihydrite dissolution rate was determined to be 1.2 × 10 -6 mol m -2 h -1. Anaerobic dissolution of ferrihydrite by pyridine-2,6-dicarboxylic acid or dipicolinic acid (dpa), a hydrolysis product of pdtc, was investigated to study the mechanism(s) involved in the pdtc facilitated ferrihydrite dissolution. These studies suggest that pdtc dissolved ferrihydrite using a reduction step, where dpa chelates the Fe reduced by a second hydrolysis product, H 2S. Dpa facilitated dissolution of ferrihydrite showed very small increase in the Fe dissolution when the concentration of external reductant, ascorbate, was doubled, suggesting the surface dynamics being dominated by the interactions between dpa and ferrihydrite. Greater than stoichiometric amounts of Fe were mobilized during dpa dissolution of ferrihydrite assisted by ascorbate and cysteine. This is attributed to the catalytic dissolution of Fe(III)(hydr)oxides by the in situ generated Fe(II) in the presence of a complex former, dpa.

  13. Quantitative solid state NMR analysis of residues from acid hydrolysis of loblolly pine wood.

    PubMed

    Sievers, Carsten; Marzialetti, Teresita; Hoskins, Travis J C; Valenzuela Olarte, Mariefel B; Agrawal, Pradeep K; Jones, Christopher W

    2009-10-01

    The composition of solid residues from hydrolysis reactions of loblolly pine wood with dilute mineral acids is analyzed by (13)C Cross Polarization Magic Angle Spinning (CP MAS) NMR spectroscopy. Using this method, the carbohydrate and lignin fractions are quantified in less than 3h as compared to over a day using wet chemical methods. In addition to the quantitative information, (13)C CP MAS NMR spectroscopy provides information on the formation of additional extractives and pseudo lignin from the carbohydrates. Being a non-destructive technique, NMR spectroscopy provides unambiguous evidence of the presence of side reactions and products, which is a clear advantage over the wet chemical analytical methods. Quantitative results from NMR spectroscopy and proximate analysis are compared for the residues from hydrolysis of loblolly pine wood under 13 different conditions; samples were treated either at 150 degrees C or 200 degrees C in the presence of various acids (HCl, H(2)SO(4), H(3)PO(4), HNO(3) and TFA) or water. The lignin content determined by both methods differed on averaged by 2.9 wt% resulting in a standard deviation of 3.5 wt%. It is shown that solid degradation products are formed from saccharide precursors under harsh reaction conditions. These degradation reactions limit the total possible yield of monosaccharides from any subsequent reaction. PMID:19477123

  14. Torque measurements reveal large process differences between materials during high solid enzymatic hydrolysis of pretreated lignocellulose

    PubMed Central

    2012-01-01

    Background A common trend in the research on 2nd generation bioethanol is the focus on intensifying the process and increasing the concentration of water insoluble solids (WIS) throughout the process. However, increasing the WIS content is not without problems. For example, the viscosity of pretreated lignocellulosic materials is known to increase drastically with increasing WIS content. Further, at elevated viscosities, problems arise related to poor mixing of the material, such as poor distribution of the enzymes and/or difficulties with temperature and pH control, which results in possible yield reduction. Achieving good mixing is unfortunately not without cost, since the power requirements needed to operate the impeller at high viscosities can be substantial. This highly important scale-up problem can easily be overlooked. Results In this work, we monitor the impeller torque (and hence power input) in a stirred tank reactor throughout high solid enzymatic hydrolysis (< 20% WIS) of steam-pretreated Arundo donax and spruce. Two different process modes were evaluated, where either the impeller speed or the impeller power input was kept constant. Results from hydrolysis experiments at a fixed impeller speed of 10 rpm show that a very rapid decrease in impeller torque is experienced during hydrolysis of pretreated arundo (i.e. it loses its fiber network strength), whereas the fiber strength is retained for a longer time within the spruce material. This translates into a relatively low, rather WIS independent, energy input for arundo whereas the stirring power demand for spruce is substantially larger and quite WIS dependent. By operating the impeller at a constant power input (instead of a constant impeller speed) it is shown that power input greatly affects the glucose yield of pretreated spruce whereas the hydrolysis of arundo seems unaffected. Conclusions The results clearly highlight the large differences between the arundo and spruce materials, both in terms of

  15. Effect of acid hydrolysis and fungal biotreatment on agro-industrial wastes for obtainment of free sugars for bioethanol production

    PubMed Central

    El-Tayeb, T.S.; Abdelhafez, A.A.; Ali, S.H.; Ramadan, E.M.

    2012-01-01

    This study was designed to evaluate selected chemical and microbiological treatments for the conversion of certain local agro-industrial wastes (rice straw, corn stalks, sawdust, sugar beet waste and sugarcane bagasse) to ethanol. The chemical composition of these feedstocks was determined. Conversion of wastes to free sugars by acid hydrolysis varied from one treatment to another. In single-stage dilute acid hydrolysis, increasing acid concentration from 1 % (v/v) to 5 % (v/v) decreased the conversion percentage of almost all treated agro-industrial wastes. Lower conversion percentages for some treatments were obtained when increasing the residence time from 90 to 120 min. The two-stage dilute acid hydrolysis by phosphoric acid (1.0 % v/v) followed by sulphuric acid (1.0 % v/v) resulted in the highest conversion percentage (41.3 % w/w) on treated sugar beet waste. This treatment when neutralized, amended with some nutrients and inoculated with baker’s yeast, achieved the highest ethanol concentration (1.0 % v/v). Formation of furfural and hydroxymethylfurfural (HMF) were functions of type of acid hydrolysis, acid concentration, residence time and feedstock type. The highest bioconversion of 5 % wastes (37.8 % w/w) was recorded on sugar beet waste by Trichoderma viride EMCC 107. This treatment when followed by baker’s yeast fermentation, 0.41 % (v/v) ethanol and 8.2 % (v/w) conversion coefficient were obtained. PMID:24031984

  16. Effect of acid hydrolysis and fungal biotreatment on agro-industrial wastes for obtainment of free sugars for bioethanol production.

    PubMed

    El-Tayeb, T S; Abdelhafez, A A; Ali, S H; Ramadan, E M

    2012-10-01

    This study was designed to evaluate selected chemical and microbiological treatments for the conversion of certain local agro-industrial wastes (rice straw, corn stalks, sawdust, sugar beet waste and sugarcane bagasse) to ethanol. The chemical composition of these feedstocks was determined. Conversion of wastes to free sugars by acid hydrolysis varied from one treatment to another. In single-stage dilute acid hydrolysis, increasing acid concentration from 1 % (v/v) to 5 % (v/v) decreased the conversion percentage of almost all treated agro-industrial wastes. Lower conversion percentages for some treatments were obtained when increasing the residence time from 90 to 120 min. The two-stage dilute acid hydrolysis by phosphoric acid (1.0 % v/v) followed by sulphuric acid (1.0 % v/v) resulted in the highest conversion percentage (41.3 % w/w) on treated sugar beet waste. This treatment when neutralized, amended with some nutrients and inoculated with baker's yeast, achieved the highest ethanol concentration (1.0 % v/v). Formation of furfural and hydroxymethylfurfural (HMF) were functions of type of acid hydrolysis, acid concentration, residence time and feedstock type. The highest bioconversion of 5 % wastes (37.8 % w/w) was recorded on sugar beet waste by Trichoderma viride EMCC 107. This treatment when followed by baker's yeast fermentation, 0.41 % (v/v) ethanol and 8.2 % (v/w) conversion coefficient were obtained. PMID:24031984

  17. Theoretical study of the alkaline hydrolysis of an aza-β-lactam derivative of clavulanic acid

    NASA Astrophysics Data System (ADS)

    Garcías, Rafael C.; Coll, Miguel; Donoso, Josefa; Muñoz, Francisco

    2003-04-01

    DFT calculations based on the hybrid functional B3LYP/6-31+G * were used to study the alkaline hydrolysis of an aza-clavulanic acid, which results from the substitution of the carbon atom at position 6 in clavulanic acid by a nitrogen atom. The presence of the nitrogen atom endows the compound with special properties; in fact, once formed, the tetrahedral intermediate can evolve with cleavage of the N 4-C 7 or N 6-C 7 bond, which obviously leads to different reaction products. These differential bond cleavages may play a central role in the inactivation of β-lactamases, so the compound may be a powerful inactivator of these enzymes.

  18. Hydrolysis reaction of 2,4-dichlorophenoxyacetic acid. A kinetic and computational study

    NASA Astrophysics Data System (ADS)

    Romero, Jorge Marcelo; Jorge, Nelly Lidia; Grand, André; Hernández-Laguna, Alfonso

    2015-10-01

    The degradation of the 2,4-dichlorophenoxyacetic acid in aqueous solution is an hydrolysis reaction. Two products are identified: 2,4-dichlorophenol and glycolic acid. Reaction is investigated as a function of pH and temperature, and it is first-order kinetics and pH-dependent. Reaction is modeled in gas phase, where a proton catalyses the reaction. Critical points of PES are calculated at B3LYP/6-311++G(3df,2p), and aug-cc-pvqz//6-311++G(3df,2p) levels plus ZPE at 6-311++G(3df,2p) level. The activation barrier is 21.2 kcal/mol. Theoretical results agree with the experimental results. A second mechanism related with a Cl2Phsbnd Osbnd CH2sbnd COOH⋯H2O complex is found, but with a rate limiting step of 38.4 kcal/mol.

  19. Preparation of highly charged cellulose nanofibrils using high-pressure homogenization coupled with strong acid hydrolysis pretreatments.

    PubMed

    Tian, Cuihua; Yi, Jianan; Wu, Yiqiang; Wu, Qinglin; Qing, Yan; Wang, Lijun

    2016-01-20

    Cellulose nanofibrils (CNFs) are attracting much attention for the advantages of excellent mechanical strength, good optical transparency, and high surface area. An eco-friendly and energy-saving method was created in this work to produce highly negative charged CNFs using high-pressure mechanical defibrillation coupled with strong acid hydrolysis pretreatments. The morphological development, zeta potential, crystal structure, chemical composition and thermal degradation behavior of the resultant materials were evaluated by transmission electron microscopy (TEM), zeta potential analysis, X-ray diffraction (XRD), Fourier transform infrared spectrometry (FTIR), and thermogravimetric analysis (TGA). These CNFs were fully separated, surface-charged, and highly entangled. They showed a large fiber aspect ratio compared to traditional cellulose nanocrystrals that are produced by strong acid hydrolysis. Compared to hydrochloric acid hydrolysis, the CNFs produced by sulfuric acid pretreatments were completely defibrillated and presented stable suspensions (or gels) even at low fiber content. On the other hand, CNFs pretreated by hydrochloric acid hydrolysis trended to aggregate because of the absence of surface charge. The crystallinity index (CI) of CNFs decreased because of mechanical defibrillation, and then increased dramatically with increased sulfuric acid concentration and reaction time. FTIR analysis showed that the C-O-SO3 group was introduced on the surfaces of CNFs during sulfuric acid hydrolysis. These sulfate groups accelerated the thermal degradation of CNFs, which occurred at lower temperature than wood pulp, indicating that the thermal stability of sulfuric acid hydrolyzed CNFs was decreased. The temperature of the maximum decomposition rate (Tmax) and the maximum weight-loss rates (MWLRmax) were much lower than for wood pulp because of the retardant effect of sulfuric acid during the combustion of CNFs. By contrast, the CNFs treated with hydrochloric acid

  20. Accelerated hydrolysis method to estimate the amino acid content of wheat (Triticum durum Desf.) flour using microwave irradiation.

    PubMed

    Kabaha, Khaled; Taralp, Alpay; Cakmak, Ismail; Ozturk, Levent

    2011-04-13

    The technique of microwave-assisted acid hydrolysis was applied to wholegrain wheat (Triticum durum Desf. cv. Balcali 2000) flour in order to speed the preparation of samples for analysis. The resultant hydrolysates were chromatographed and quantified in an automated amino acid analyzer. The effect of different hydrolysis temperatures, times and sample weights was examined using flour dispersed in 6 N HCl. Within the range of values tested, the highest amino acid recoveries were generally obtained by setting the hydrolysis parameters to 150 °C, 3 h and 200 mg sample weight. These conditions struck an optimal balance between liberating amino acid residues from the wheat matrix and limiting their subsequent degradation or transformation. Compared to the traditional 24 h reflux method, the hydrolysates were prepared in dramatically less time, yet afforded comparable ninhydrin color yields. Under optimal hydrolysis conditions, the total amino acid recovery corresponded to at least 85.1% of the total protein content, indicating the efficient extraction of amino acids from the flour matrix. The findings suggest that this microwave-assisted method can be used to rapidly profile the amino acids of numerous wheat grain samples, and can be extended to the grain analysis of other cereal crops. PMID:21375298

  1. Alcohol fermentation of sweet potato - 1. Acid hydrolysis and factors involved

    SciTech Connect

    Azhar, A.; Hamdy, M.K.

    1981-04-01

    Factors affecting acid hydrolysis of sweet potato powder (SPP) to fermentable sugars were examined. These include HCl concentration, temperature, time, and levels of SPP. Maximum reducing sugar, reported as dextrose equivalent (DE), was detected after 24 min hydrolysis (1% SPP) in 0.034N HCl heated at 154/degree/C. These samples also had 3.43% hydroxymethylfurfural (HMF) based on dry weight. A high level of HMF (9.2%) was detected in 1% SPP heated at 154/degree/C in 0.10N HCl for 18 min. The lowest concentration of HMF formed (1.8%), at maximal DE of 61%, was established in samples containing 5% SPP and heated at 154/degree/C in 0.034N HCl for 48 min. Aqueous extracts of uncured SPP, examined by High Performance Liquid Chromatography, contained glucose, fructose and sucrose, but degraded SPP had only glucose and fructose. Products of degraded SPP, Under appropriate conditions, could be used for alcohol fermentation. 18 refs.

  2. Strategies to achieve high-solids enzymatic hydrolysis of dilute-acid pretreated corn stover.

    PubMed

    Geng, Wenhui; Jin, Yongcan; Jameel, Hasan; Park, Sunkyu

    2015-01-01

    Three strategies were presented to achieve high solids loading while maximizing carbohydrate conversion, which are fed-batch, splitting/thickening, and clarifier processes. Enzymatic hydrolysis was performed at water insoluble solids (WIS) of 15% using washed dilute-acid pretreated corn stover. The carbohydrate concentration increased from 31.8 to 99.3g/L when the insoluble solids content increased from 5% to 15% WIS, while the final carbohydrate conversion was decreased from 78.4% to 73.2%. For the fed-batch process, a carbohydrate conversion efficiency of 76.8% was achieved when solid was split into 60:20:20 ratio, with all enzymes added first. For the splitting/thickening process, a carbohydrate conversion of 76.5% was realized when the filtrate was recycled to simulate a steady-state process. Lastly, the clarifier process was evaluated and the highest carbohydrate conversion of 81.4% was achieved. All of these results suggests the possibility of enzymatic hydrolysis at high solids to make the overall conversion cost-competitive. PMID:25836373

  3. Enantioselective Hydrolysis of Amino Acid Esters Promoted by Bis(β-cyclodextrin) Copper Complexes

    NASA Astrophysics Data System (ADS)

    Xue, Shan-Shan; Zhao, Meng; Ke, Zhuo-Feng; Cheng, Bei-Chen; Su, Hua; Cao, Qian; Cao, Zhen-Kun; Wang, Jun; Ji, Liang-Nian; Mao, Zong-Wan

    2016-02-01

    It is challenging to create artificial catalysts that approach enzymes with regard to catalytic efficiency and selectivity. The enantioselective catalysis ranks the privileged characteristic of enzymatic transformations. Here, we report two pyridine-linked bis(β-cyclodextrin) (bisCD) copper(II) complexes that enantioselectively hydrolyse chiral esters. Hydrolytic kinetic resolution of three pairs of amino acid ester enantiomers (S1-S3) at neutral pH indicated that the “back-to-back” bisCD complex CuL1 favoured higher catalytic efficiency and more pronounced enantioselectivity than the “face-to-face” complex CuL2. The best enantioselectivity was observed for N-Boc-phenylalanine 4-nitrophenyl ester (S2) enantiomers promoted by CuL1, which exhibited an enantiomer selectivity of 15.7. We observed preferential hydrolysis of L-S2 by CuL1, even in racemic S2, through chiral high-performance liquid chromatography (HPLC). We demonstrated that the enantioselective hydrolysis was related to the cooperative roles of the intramolecular flanking chiral CD cavities with the coordinated copper ion, according to the results of electrospray ionization mass spectrometry (ESI-MS), inhibition experiments, rotating-frame nuclear Overhauser effect spectroscopy (ROESY), and theoretical calculations. Although the catalytic parameters lag behind the level of enzymatic transformation, this study confirms the cooperative effect of the first and second coordination spheres of artificial catalysts in enantioselectivity and provides hints that may guide future explorations of enzyme mimics.

  4. The shape and size distribution of crystalline nanoparticles prepared by acid hydrolysis of native cellulose.

    PubMed

    Elazzouzi-Hafraoui, Samira; Nishiyama, Yoshiharu; Putaux, Jean-Luc; Heux, Laurent; Dubreuil, Frédéric; Rochas, Cyrille

    2008-01-01

    The shape and size distribution of crystalline nanoparticles resulting from the sulfuric acid hydrolysis of cellulose from cotton, Avicel, and tunicate were investigated using transmission electron microscopy (TEM) and atomic force microscopy (AFM) as well as small- and wide-angle X-ray scattering (SAXS and WAXS). Images of negatively stained and cryo-TEM specimens showed that the majority of cellulose particles were flat objects constituted by elementary crystallites whose lateral adhesion was resistant against hydrolysis and sonication treatments. Moreover, tunicin whiskers were described as twisted ribbons with an estimated pitch of 2.4-3.2 microm. Length and width distributions of all samples were generally well described by log-normal functions, with the exception of tunicin, which had less lateral aggregation. AFM observation confirmed that the thickness of the nanocrystals was almost constant for a given origin and corresponded to the crystallite size measured from peak broadening in WAXS spectra. Experimental SAXS profiles were numerically simulated, combining the dimensions and size distribution functions determined by the various techniques. PMID:18052127

  5. Enantioselective Hydrolysis of Amino Acid Esters Promoted by Bis(β-cyclodextrin) Copper Complexes.

    PubMed

    Xue, Shan-Shan; Zhao, Meng; Ke, Zhuo-Feng; Cheng, Bei-Chen; Su, Hua; Cao, Qian; Cao, Zhen-Kun; Wang, Jun; Ji, Liang-Nian; Mao, Zong-Wan

    2016-01-01

    It is challenging to create artificial catalysts that approach enzymes with regard to catalytic efficiency and selectivity. The enantioselective catalysis ranks the privileged characteristic of enzymatic transformations. Here, we report two pyridine-linked bis(β-cyclodextrin) (bisCD) copper(II) complexes that enantioselectively hydrolyse chiral esters. Hydrolytic kinetic resolution of three pairs of amino acid ester enantiomers (S1-S3) at neutral pH indicated that the "back-to-back" bisCD complex CuL(1) favoured higher catalytic efficiency and more pronounced enantioselectivity than the "face-to-face" complex CuL(2). The best enantioselectivity was observed for N-Boc-phenylalanine 4-nitrophenyl ester (S2) enantiomers promoted by CuL(1), which exhibited an enantiomer selectivity of 15.7. We observed preferential hydrolysis of L-S2 by CuL(1), even in racemic S2, through chiral high-performance liquid chromatography (HPLC). We demonstrated that the enantioselective hydrolysis was related to the cooperative roles of the intramolecular flanking chiral CD cavities with the coordinated copper ion, according to the results of electrospray ionization mass spectrometry (ESI-MS), inhibition experiments, rotating-frame nuclear Overhauser effect spectroscopy (ROESY), and theoretical calculations. Although the catalytic parameters lag behind the level of enzymatic transformation, this study confirms the cooperative effect of the first and second coordination spheres of artificial catalysts in enantioselectivity and provides hints that may guide future explorations of enzyme mimics. PMID:26916830

  6. Methane production from acid hydrolysates of Agave tequilana bagasse: evaluation of hydrolysis conditions and methane yield.

    PubMed

    Arreola-Vargas, Jorge; Ojeda-Castillo, Valeria; Snell-Castro, Raúl; Corona-González, Rosa Isela; Alatriste-Mondragón, Felipe; Méndez-Acosta, Hugo O

    2015-04-01

    Evaluation of diluted acid hydrolysis for sugar extraction from cooked and uncooked Agave tequilana bagasse and feasibility of using the hydrolysates as substrate for methane production, with and without nutrient addition, in anaerobic sequencing batch reactors (AnSBR) were studied. Results showed that the hydrolysis over the cooked bagasse was more effective for sugar extraction at the studied conditions. Total sugars concentration in the cooked and uncooked bagasse hydrolysates were 27.9 g/L and 18.7 g/L, respectively. However, 5-hydroxymethylfurfural was detected in the cooked bagasse hydrolysate, and therefore, the uncooked bagasse hydrolysate was selected as substrate for methane production. Interestingly, results showed that the AnSBR operated without nutrient addition obtained a constant methane production (0.26 L CH4/g COD), whereas the AnSBR operated with nutrient addition presented a gradual methane suppression. Molecular analyses suggested that methane suppression in the experiment with nutrient addition was due to a negative effect over the archaeal/bacterial ratio. PMID:25647030

  7. Dilute acid pretreatment and enzymatic hydrolysis of sorghum biomass for sugar recovery--a statistical approach.

    PubMed

    Akanksha, Karthik; Prasad, Arjun; Sukumaran, Rajeev K; Nampoothiri, Madhavan; Pandey, Ashok; Rao, S S; Parameswaran, Binod

    2014-11-01

    Sorghum is one of the commercially feasible lignocellulosic biomass and has a great potential of being sustainable feedstock for renewable energy. As with any lignocellulosic biomass, sorghum also requires pretreatment which increases its susceptibility to hydrolysis by enzymes for generating sugars which can be further fermented to alcohol. In the present study, sorghum biomass was evaluated for deriving maximum fermentable sugars by optimizing various pretreatment parameters using statistical optimization methods. Pretreatment studies were done with H2SO4, followed by enzymatic saccharification. The efficiency of the process was evaluated on the basis of production of the total reducing sugars released during the process. Compositional analysis was done for native as well as pretreated biomass and compared. The biomass pretreated with the optimized conditions could yield 0.408 g of reducing sugars /g of pretreated biomass upon enzymatic hydrolysis. The cellulose content in the solid portion obtained after pretreatment using optimised conditions was found to be increased by 43.37% with lesser production of inhibitors in acid pretreated liquor. PMID:25434103

  8. EFFECT OF ANATOMICAL FRACTIONATION ON THE ENZYMATIC HYDROLYSIS OF ACID AND ALKALINE PRETREATED CORN STOVER

    SciTech Connect

    K. B. Duguid; M. D. Montross; C. W. Radtke; C. L. Crofcheck; L. M. Wendt; S. A. Shearer

    2009-11-01

    Due to concerns with biomass collection systems and soil sustainability there are opportunities to investigate the optimal plant fractions to collect for conversion. An ideal feedstock would require low severity pretreatment to release a maximum amount of sugar during enzymatic hydrolysis. Corn stover fractions were separated by hand and analyzed for glucan, xylan, acid soluble lignin, acid insoluble lignin, and ash composition. The stover fractions were also pretreated with either 0, 0.4, or 0.8% NaOH for 2 hours at room temperature, washed, autoclaved and saccharified. In addition, acid pretreated samples underwent simultaneous saccharification and fermentation (SSF) to ethanol. In general, the two pretreatments produced similar trends with cobs, husks, and leaves responding best to the pretreatments, the tops of stalks responding slightly less, and the bottom of the stalks responding the least. For example, corn husks pretreated with 0.8% NaOH released over 90% (standard error of 3.8%) of the available glucan, while only 45% (standard error of 1.1%) of the glucan was produced from identically treated stalk bottoms. Estimates of the theoretical ethanol yield using acid pretreatment followed by SSF were 65% (standard error of 15.9%) for husks and 29% (standard error of 1.8%) for stalk bottoms. This suggests that integration of biomass collection systems to remove sustainable feedstocks could be integrated with the processes within a biorefinery to minimize overall ethanol production costs.

  9. Hydrolysis of N{sub 2}O{sub 5} on submicron sulfuric acid aerosols

    SciTech Connect

    Hallquist, M.; Stewart, D.J.; Baker, J.; Cox, R.A.

    2000-05-04

    The kinetics of reactive uptake of gaseous N{sub 2}O{sub 5} on submicron sulfuric acid aerosol particles has been investigated using a laminar flow reactor coupled with a differential mobility analyzer (DMA) to characterize the aerosol. The particles were generated by homogeneous nucleation of SO{sub 3}/H{sub 2}O mixtures. In the H{sub 2}SO{sub 4} concentration range 26.3--64.5 wt {degree} the uptake coefficient was {gamma} = 0.033 {+-} 0.004, independent of acid strength. For an acid strength of 45 wt % {gamma} was found to decrease with increasing temperature over the range 263--298 K. From this, temperature dependence values of {minus}115 {+-} 30 kJ/mol and {minus}25.5 {+-} 8.4 J/K mol were determined for the changes in enthalpy and entropy of the uptake process, respectively. The results are consistent with a previous model of N{sub 2}O{sub 5} hydrolysis involving both a direct and an acid catalyzed mechanism, with uptake under the experimental conditions limited by mass accommodation.

  10. Determination of amino acids in two Polysiphonia species and study of enzymatic hydrolysis method

    NASA Astrophysics Data System (ADS)

    Zhang, Li-Xin; Fan, Xiao; Wei, Yu-Xi

    2002-09-01

    The total content of the rich amino acids in two common red algae, Polysiphonia urceolata and Polysiphonia japonica growing in the Qingdao seashore were determined. The algae powder was hydrolyzed by 6 mol/L HCl at 110°C for 48 h and determined by amino acid analyzer. The content was 25.35% and 24.16%, respectively, much higher than that of some other species. In addition, a nutritive liquid with abundant amino acids was prepared (by the enzymatic hydrolysis method using Polysiphonia urceolata) as raw material for a kind of health beverage. The dried seaweed was decolored by 0.25% KMnO4 and 0.5% active carbon, then enzymalized. In the selection of enzymalizing condition, the orthogonal experimental design was used. Four factors including kinds of enzyme, quantity of enzyme, temperature and time were studied at 3 levels. According to the orthogonal design results, we can choose an optimal condition: hydrolyzing at 45°C by neutral proteinase (0.25%, w/w) for 2h, adjusting pH to 8.5, then adding trypsin (0.25%, w/w) and hydrolyzing for 2 h. Finally the above solution was alkalized by NaOH and neutralized by casein. After the hydrolyzed liquid was filtered and concentrated, suitable additives were added. The final products contain rich amino acids.

  11. Evolution of the phase content of zirconia powders prepared by sol-gel acid hydrolysis

    SciTech Connect

    Rivas, P.C.; Martinez, J.A.; Caracoche, M.C.; Rodriguez, A.M.; Lopez Garcia, A.R.; Pavlik, R.S. Jr.; Klein, L.C.

    1998-01-01

    The evolution of the phase content in zirconia powders that have been prepared by sol-gel acid hydrolysis has been investigated using the perturbed-angular-correlation (PAC) technique and X-ray diffractometry. As a consequence of performing annealing treatments at increasing temperatures between room temperature and 1,000 C, the amorphous starting material transforms to the tetragonal form and then to the monoclinic form. The metastable tetragonal phase exhibits two hyperfine components, one of which describes very defective zirconium surroundings. The evolution of PAC relative fractions is in agreement with the diffraction results. The durability of the samples in sodium hydroxide seems to increase as the relative amount of the most-defective zirconium surroundings of the tetragonal form increases.

  12. Cellulose nanocrystals prepared via formic acid hydrolysis followed by TEMPO-mediated oxidation.

    PubMed

    Li, Bin; Xu, Wenyang; Kronlund, Dennis; Määttänen, Anni; Liu, Jun; Smått, Jan-Henrik; Peltonen, Jouko; Willför, Stefan; Mu, Xindong; Xu, Chunlin

    2015-11-20

    Cellulose nanocrystals (CNCs) as a renewable and biodegradable nanomaterial have wide application value. In this work, CNCs were extracted from bleached chemical pulp using two stages of isolation (i.e. formic acid (FA) hydrolysis and 2,2,6,6-tetramethyl-piperidine-1-oxyl (TEMPO) mediated oxidation) under mild conditions. In the first stage, FA was used to remove hemicellulose, swell cellulose fibers, and release CNCs. The FA could be readily recovered and reused. In the second stage, the CNCs isolated by FA were further modified by TEMPO-mediated oxidation to increase the surface charge of CNCs. It was found that the modified CNCs with more ordered crystal structure and higher surface charge had better redispersibility and higher viscosity in aqueous phase. Therefore, the modified CNCs could be more effective when used as rheology modifier in the fields of water based coating, paint, food etc. PMID:26344319

  13. Steam gasification of acid-hydrolysis biomass CAHR for clean syngas production.

    PubMed

    Chen, Guanyi; Yao, Jingang; Yang, Huijun; Yan, Beibei; Chen, Hong

    2015-03-01

    Main characteristics of gaseous product from steam gasification of acid-hydrolysis biomass CAHR have been investigated experimentally. The comparison in terms of evolution of syngas flow rate, syngas quality and apparent thermal efficiency was made between steam gasification and pyrolysis in the lab-scale apparatus. The aim of this study was to determine the effects of temperature and steam to CAHR ratio on gas quality, syngas yield and energy conversion. The results showed that syngas and energy yield were better with gasification compared to pyrolysis under identical thermal conditions. Both high gasification temperature and introduction of proper steam led to higher gas quality, higher syngas yield and higher energy conversion efficiency. However, excessive steam reduced hydrogen yield and energy conversion efficiency. The optimal value of S/B was found to be 3.3. The maximum value of energy ratio was 0.855 at 800°C with the optimal S/B value. PMID:25553562

  14. RAPESEED PHOSPHATIDYLCHOLINE HYDROLYSIS TO PHOSPHATIDIC ACID USING PLANT EXTRACTS WITH PHOPSPHOLIPASE D.

    PubMed

    Pasker, Beata; Sosada, Marian; Fraś, Paweł; Boryczka, Monika; Górecki, Michał; Zych, Maria

    2015-01-01

    Phosphatidic acid (PA) has a crucial role in cell membrane structure and function. For that reason it has a possible application in the treatment of some health disorders in humans, can be used as a natural and non toxic emulsifier and the component of drug carriers in pharmaceuticals and cosmetics as well as a component for synthesis of some new phospholipids. PA is short-lived in the cell and is difficult to extract directly from the biological material. PA may be easily prepared by hydrolysis of phospholipids, especially phosphatidylcholine (PC), using cabbage phospholipase D (PLD). Hydrolytic activity of purified by us PLD extracts from cabbage towards rapeseed phosphatidylcholine (RPC) was investigated. Hydrolysis was carried out in the biphasic system (water/diethyl ether) at pH 6,5 and temp 30°C. Influence of enzymatic extracts from three cabbage varieties, reaction time, Ca2+ concentration and enzyme extracts/PC ratio, on activity towards RPC resulting in rapeseed phosphatidic acid (RPA) formation were examined. Our study shows that the PLD extracts from savoy cabbage (PLDsc), white cabbage (PLDwc) and brussels sprouts (PLDbs) used in experiments exhibit hydrolytic activity towards RPC resulting in rapeseed RPA with different yield. The highest activity towards RPC shows PLD extract from PLDsc with the RPC conversion degree to RPA (90%) was observed at 120 mM Ca2+ concentration, reaction time 60 min and ratio of PLD extract to RPC 6 : 1 (w/w). Our study shows that purified by us PLDsc extracts exhibit hydrolytic activity towards RPC giving new RPA with satisfying conversion degree for use in pharmacy, cosmetics and as a standard in analytical chemistry. PMID:26642684

  15. Preparation, analysis and antioxidant evaluation of the controlled product of polysaccharide from Mactra veneriformis by mild acid hydrolysis.

    PubMed

    Wang, Ling Chong; Wu, Hao; Ji, Jing; Xue, Feng; Liu, Rui

    2016-02-10

    The polysaccharides from Mactra veneriformis (MVPS) were degraded by controlled mild acid hydrolysis to produce active oligosaccharides. MVPS can easily be hydrolyzed by H2SO4 or HCl. The hydrolyzing process was investigated in acid addition and reaction time by evaluating the producing content of reducing sugar. Hydrolysis with 1M HCl to MVPS can generate a time-depended behavior that is mild and controllable. HPLC analysis monitored the change of oligosaccharides composition in hydrolyzing. Total nine oligosaccharides are recognizable in the HPLC profile, and their content showed a regular transformation in hydrolysis. Those nine ingredients were identified as glucooligosaccharides with DP from 1 to 7 by MS analysis. Antioxidant activities of the typical hydrolyzates as well as MVPs were further tested in assays of DPPH and hydroxyl radicals scavenging, and reducing power. It was found that HCl hydrolyzate exhibited stronger antioxidant effects than MVPS and H2SO4 hydrolyzate due to its higher content of oligosaccharides. PMID:26686183

  16. /sup 18/O isotope effect in /sup 13/C nuclear magnetic resonance spectroscopy. Part 9. Hydrolysis of benzyl phosphate by phosphatase enzymes and in acidic aqueous solutions

    SciTech Connect

    Parente, J.E.; Risley, J.M.; Van Etten, R.L.

    1984-12-26

    The /sup 18/O isotope-induced shifts in /sup 13/C and /sup 31/P nuclear magnetic resonance (NMR) spectroscopy were used to establish the position of bond cleavage in the phosphatase-catalyzed and acid-catalyzed hydrolysis reactions of benzyl phosphate. The application of the /sup 18/O-isotope effect in NMR spectroscopy affords a continuous, nondestructive assay method for following the kinetics and position of bond cleavage in the hydrolytic process. The technique provides advantages over most discontinuous methods in which the reaction components must be isolated and converted to volatile derivatives prior to analysis. In the present study, (..cap alpha..-/sup 13/C,ester-/sup 18/O)benzyl phosphate and (ester-/sup 18/O)benzyl phosphate were synthesized for use in enzymatic and nonenzymatic studies. Hydrolysis reactions catalyzed by the alkaline phosphatase from E. coli and by the acid phosphatases isolated from human prostate and human liver were all accompanied by cleavage of the substrate phosphorus-oxygen bond consistent with previously postulated mechanisms involving covalent phosphoenzyme intermediates. An extensive study of the acid-catalyzed hydrolysis of benzyl phosphate at 75/sup 0/C revealed that the site of bond cleavage is dependent on pH. At pH less than or equal to 1.3, the hydrolysis proceeds with C-O bond cleavage; at 1.3 < pH < 2.0, there is a mixture of C-O and P-O bond scission, the latter progressively predominating as the pH is raised; at pH greater than or equal to 2.0, the hydrolysis proceeds with exclusive P-O bond scission. (S)-(+)-(..cap alpha..-/sup 2/H)Benzyl phosphate was also synthesized. Hydrolysis of this chiral benzyl derivative demonstrated that the acid-catalyzed C-O bond scission of benzyl phosphate proceeds by an A-1 (S/sub N/1) mechanism with 70% racemization and 30% inversion at carbon. 37 references, 4 figures, 2 tables.

  17. Isolation and structural characterization of sugarcane bagasse lignin after dilute phosphoric acid plus steam explosion pretreatment and its effect on cellulose hydrolysis.

    PubMed

    Zeng, Jijiao; Tong, Zhaohui; Wang, Letian; Zhu, J Y; Ingram, Lonnie

    2014-02-01

    The structure of lignin after dilute phosphoric acid plus steam explosion pretreatment process of sugarcane bagasse in a pilot scale and the effect of the lignin extracted by ethanol on subsequent cellulose hydrolysis were investigated. The lignin structural changes caused by pretreatment were identified using advanced nondestructive techniques such as gel permeation chromatography (GPC), quantitative (13)C, and 2-D nuclear magnetic resonance (NMR). The structural analysis revealed that ethanol extractable lignin preserved basic lignin structure, but had relatively lower amount of β-O-4 linkages, syringyl/guaiacyl units ratio (S/G), p-coumarate/ferulate ratio, and other ending structures. The results also indicated that approximately 8% of mass weight was extracted by pure ethanol. The bagasse after ethanol extraction had an approximate 22% higher glucose yield after enzyme hydrolysis compared to pretreated bagasse without extraction. PMID:24412855

  18. Direct lactic acid fermentation of Jerusalem artichoke tuber extract using Lactobacillus paracasei without acidic or enzymatic inulin hydrolysis.

    PubMed

    Choi, Hwa-Young; Ryu, Hee-Kyoung; Park, Kyung-Min; Lee, Eun Gyo; Lee, Hongweon; Kim, Seon-Won; Choi, Eui-Sung

    2012-06-01

    Lactic acid fermentation of Jerusalem artichoke tuber was performed with strains of Lactobacillus paracasei without acidic or enzymatic inulin hydrolysis prior to fermentation. Some strains of L. paracasei, notably KCTC13090 and KCTC13169, could ferment hot-water extract of Jerusalem artichoke tuber more efficiently compared with other Lactobacillus spp. such as L. casei type strain KCTC3109. The L. paracasei strains could utilize almost completely the fructo-oligosaccharides present in Jerusalem artichoke. Inulin-fermenting L. paracasei strains produced c.a. six times more lactic acid compared with L. casei KCTC3109. Direct lactic fermentation of Jerusalem artichoke tuber extract at 111.6g/L of sugar content with a supplement of 5 g/L of yeast extract by L. paracasei KCTC13169 in a 5L jar fermentor produced 92.5 ce:hsp sp="0.25"/>g/L of lactic acid with 16.8 g/L fructose equivalent remained unutilized in 72 h. The conversion efficiency of inulin-type sugars to lactic acid was 98% of the theoretical yield. PMID:22516247

  19. Effect of gamma interferon on phospholipid hydrolysis and fatty acid incorporation in L929 cells infected with Rickettsia prowazekii.

    PubMed Central

    Winkler, H H; Day, L; Daugherty, R; Turco, J

    1993-01-01

    Treatment of Rickettsia prowazekii-infected L929 cells with gamma interferon (IFN-gamma) immediately after infection altered the lipid metabolism of the host cells as determined by measurement of phospholipid hydrolysis and oleic acid incorporation into phospholipids and neutral lipids. At 48 h postinfection, there was increased phospholipid hydrolysis in infected cultures relative to mock-infected cultures and a further increase in radiolabeled phospholipid hydrolysis in IFN-gamma-treated infected cultures. Oleic acid, the radiolabeled product of hydrolysis, was found in both the free fatty acid and neutral lipid fractions. None of the mock-infected cultures demonstrated increased hydrolysis of their radiolabeled phospholipids in response to treatment with IFN-gamma. Most of the radiolabeled oleic acid incorporated into cultures in the interval between 24 and 48 h after infection and IFN-gamma treatment was present in the phospholipid fraction. However, the neutral lipid fraction from the infected cultures that had been IFN-gamma treated was labeled to a greater extent than that from the untreated cultures. Thin-layer chromatographic analysis of the neutral lipid fractions from both the hydrolysis and incorporation experiments demonstrated that most of the radiolabel was in triglycerides. The infected cultures at 30 h were intact as assessed by the exclusion of trypan blue, but at 48 h postinfection in the IFN-gamma-treated infected cultures more than half of the cells were unable to exclude trypan blue. In no case did the mock-infected cells show substantial damage as a result of IFN-gamma treatment. PMID:8335370

  20. Effect of sulfuric and phosphoric acid pretreatments on enzymatic hydrolysis of corn stover.

    PubMed

    Um, Byung-Hwan; Karim, M; Henk, Linda

    2003-01-01

    The pretreatment of corn stover with H2SO4 and H3PO4 was investigated. Pretreatments were carried out from 30 to 120 min in a batch reactor at 121 degrees C, with acid concentrations ranging from 0 to 2% (w/v) at a solid concentration of 5% (w/v). Pretreated corn stover was washed with distilled water until the filtrate was adjusted to pH 7.0, followed by surfactant swelling of the cellulosic fraction in a 0-10% (w/v) solution of Tween-80 at room temperature for 12 h. The dilute acid treatment proved to be a very effective method in terms of hemicellulose recovery and cellulose digestibility. Hemicellulose recovery was 62-90%, and enzymatic digestibility of the cellulose that remained in the solid was >80% with 2% (w/v) acid. In all cases studied, the performance of H2SO4 pretreatment (hemicellulose recovery and cellulose digestibility) was significantly better than obtained with H3PO4. Enzymatic hydrolysis was more effective using surfactant than without it, producing 10-20% more sugar. Furthermore, digestibility was investigated as a function of hemicellulose removal. It was found that digestibility was more directly related to hemicellulose removal than to delignification. PMID:12721479

  1. Effect of degree of hydrolysis of whey protein on in vivo plasma amino acid appearance in humans.

    PubMed

    Farup, Jean; Rahbek, Stine Klejs; Storm, Adam C; Klitgaard, Søren; Jørgensen, Henry; Bibby, Bo M; Serena, Anja; Vissing, Kristian

    2016-01-01

    Whey protein is generally found to be faster digested and to promote faster and higher increases in plasma amino acid concentrations during the immediate ~60 min following protein ingestion compared to casein. The aim of the present study was to compare three different whey protein hydrolysates with varying degrees of hydrolysis (DH, % cleaved peptide bonds) to evaluate if the degree of whey protein hydrolysis influences the rate of amino acid plasma appearance in humans. A casein protein was included as reference. The three differentially hydrolysed whey proteins investigated were: High degree of hydrolysis (DH, DH = 48 %), Medium DH (DH = 27 %), and Low DH (DH = 23 %). The casein protein was intact. Additionally, since manufacturing of protein products may render some amino acids unavailable for utilisation in the body the digestibility and the biological value of all four protein fractions were evaluated in a rat study. A two-compartment model for the description of the postprandial plasma amino acid kinetics was applied to investigate the rate of postprandial total amino acid plasma appearance of the four protein products. The plasma amino acid appearance rates of the three whey protein hydrolysates (WPH) were all significantly higher than for the casein protein, however, the degree of hydrolysis of the WPH products did not influence plasma total amino acid appearance rate (estimates of DH and 95 % confidence intervals [CI] (mol L(-1) min(-1)): High DH 0.0585 [0.0454, 0.0754], Medium DH 0.0594 [0.0495, 0.0768], Low DH 0.0560 [0.0429, 0.0732], Casein 0.0194 [0.0129, 0.0291]). The four protein products were all highly digestible, while the biological value decreased with increasing degree of hydrolysis. In conclusion, the current study does not provide evidence that the degree of whey protein hydrolysis is a strong determinant for plasma amino acid appearance rate within the studied range of hydrolysis and protein dose. PMID:27065230

  2. Activation Energies for an Enzyme-Catalyzed and Acid-Catalyzed Hydrolysis: An Introductory Interdisciplinary Experiment for Chemists and Biochemists.

    ERIC Educational Resources Information Center

    Adams, K. R.; Meyers, M. B.

    1985-01-01

    Background information, procedures used, and typical results obtained are provided for an experiment in which students determine and compare the Arrhenius activation energies (Ea) for the hydrolysis of salicin. This reaction is subject to catalysis both by acid and by the enzyme emulsin (beta-d-glucoside glycohydrolase). (JN)

  3. Hydrolysis mechanisms for the organopalladium complex [Pd(CNN)P(OMe)3]BF4 in sulfuric acid.

    PubMed

    García, Begoña; Hoyuelos, Francisco J; Ibeas, Saturnino; Muñoz, María S; Peñacoba, Indalecio; Leal, José M

    2009-08-13

    The acid-catalyzed hydrolysis of the organopalladium complex [Pd(CNN)P(OMe)3]BF4 species was monitored spectrophotometrically at different sulfuric acid concentrations (3.9 and 11.0 M) in 10% v:v ethanol-water over the 25-45 degrees C temperature range and in 30% and 50% (v/v) ethanol-water at 25 degrees C. Two acidity regions (I and II) could be differentiated. In each of the two regions the kinetic data pairs yielded two different rate constants, k(1obs) and k(2obs), the former being faster. These constants were fitted by an Excess Acidity analysis to different hydrolyses mechanisms: A-1, A-2, and A-SE2. In region I ([H2SO4] < 7.0 M), the k(1obs) values remained constant k(1obs)(av) = 1.6 x 10(-3) s(-1) and the set of k(2obs) values nicely matched an A-SE2 mechanism, yielding a rate-determining constant k(0,ASE2) = 2.4 x 10(-7) M(-1) s(-1). In region II ([H2SO4] > 7.0 M), a switchover was observed from an A-1 mechanism (k(0,A1) = 1.3 x 10(-4) s(-1)) to an A-2 mechanism (k(0,A2) = 3.6 x 10(-3) M(-1) s(-1)). The temperature effect on the rate constants in 10% (v/v) ethanol-water yielded positive DeltaH and negative DeltaS values, except for the A-1 mechanism, where DeltaS adopted positive values throughout. The solvent permittivity effect, epsilonr, revealed that k(1obs)(av) and k(0,A2) dropped with a fall in epsilonr, whereas the k(0,ASE2) value remained unaffected. The set of results deduced is in line with the schemes put forward. PMID:19621916

  4. Cholesterol efflux from THP-1 macrophages is impaired by the fatty acid component from lipoprotein hydrolysis by lipoprotein lipase

    SciTech Connect

    Yang, Yanbo; Thyagarajan, Narmadaa; Coady, Breanne M.; Brown, Robert J.

    2014-09-05

    Highlights: • Lipoprotein hydrolysis products were produced by lipoprotein lipase. • Hydrolysis products lowers expression of macrophage cholesterol transporters. • Hydrolysis products reduces expression of select nuclear receptors. • Fatty acid products lowers cholesterol transporters and select nuclear receptors. • Fatty acid products reduces cholesterol efflux from macrophages. - Abstract: Lipoprotein lipase (LPL) is an extracellular lipase that primarily hydrolyzes triglycerides within circulating lipoproteins. Macrophage LPL contributes to atherogenesis, but the mechanisms behind it are poorly understood. We hypothesized that the products of lipoprotein hydrolysis generated by LPL promote atherogenesis by inhibiting the cholesterol efflux ability by macrophages. To test this hypothesis, we treated human THP-1 macrophages with total lipoproteins that were hydrolyzed by LPL and we found significantly reduced transcript levels for the cholesterol transporters ATP binding cassette transporter A1 (ABCA1), ABCG1, and scavenger receptor BI. These decreases were likely due to significant reductions for the nuclear receptors liver-X-receptor-α, peroxisome proliferator activated receptor (PPAR)-α, and PPAR-γ. We prepared a mixture of free fatty acids (FFA) that represented the ratios of FFA species within lipoprotein hydrolysis products, and we found that the FFA mixture also significantly reduced cholesterol transporters and nuclear receptors. Finally, we tested the efflux of cholesterol from THP-1 macrophages to apolipoprotein A-I, and we found that the treatment of THP-1 macrophages with the FFA mixture significantly attenuated cholesterol efflux. Overall, these data show that the FFA component of lipoprotein hydrolysis products generated by LPL may promote atherogenesis by inhibiting cholesterol efflux, which partially explains the pro-atherogenic role of macrophage LPL.

  5. Potential of phosphoric acid-catalyzed pretreatment and subsequent enzymatic hydrolysis for biosugar production from Gracilaria verrucosa.

    PubMed

    Kwon, Oh-Min; Kim, Sung-Koo; Jeong, Gwi-Taek

    2016-07-01

    This study combined phosphoric acid-catalyzed pretreatment and enzymatic hydrolysis to produce biosugars from Gracilaria verrucosa as a potential renewable resource for bioenergy applications. We optimized phosphoric acid-catalyzed pretreatment conditions to 1:10 solid-to-liquid ratio, 1.5 % phosphoric acid, 140 °C, and 60 min reaction time, producing a 32.52 ± 0.06 % total reducing sugar (TRS) yield. By subsequent enzymatic hydrolysis, a 68.61 ± 0.90 % TRS yield was achieved. These results demonstrate the potential of phosphoric acid to produce biosugars for biofuel and biochemical production applications. PMID:27003825

  6. Investigating the Properties and Hydrolysis Ability of Poly-Lactic Acid/Chitosan Nanocomposites Using Polycaprolactone.

    PubMed

    Trang, Nguyen Thi Thu; Chinhl, Nguyen Thuy; Thanh, Dinh Thi Mai; Hang, To Thi Xuan; Giang, Nguyen Vu; Hoang, Thai; Quan, Pham Minh; Giang, Le Duc; Thai, Nguyen Viet; Lawrence, Geoffrey

    2015-12-01

    Poly-lactic acid (PLA) has been widely applied in the medical field (in biomedicines such as medical capsules, surgical sutures and suture wounds) owing to its high biodegradability, good biocompatibility and ability to be dissolved in common solvents. Chitosan (CS) is an abundant polysaccharide and a cationic polyelectrolyte present in nature. In this study, the combination of PLA and CS has been used to form PLA/CS nanocomposites having the advantages of both the original components. To enhance the dispersibility and compatibility between PLA and CS in the PLA/CS nanocomposites, polycaprolactone (PCL) is added as a compatibilizer. The Fourier Transform Infrared spectroscopies prove the existence of the interactions of PCL with PLA and CS. A more regular dispersion of CS of 200-400 nm particle size, is observed in the PLA matrix of the PLA/CS nanocomposites containing PCL, through the Field Emission Scanning Electron Microscopy images. The appearance of one glass transition temperature (T(g)) value of PLA/CS/PCL nanocomposites occuring between the T(g) values of PLA and CS in DSC diagrams confirms the improvement in the compatibility between PLA and CS, due to the presence of PCL. The TGA result shows that PCL plays an important role in enhancing the thermal stability of PLA/CS/PCL nanocomposites. The hydrolysis of PLA/CS/PCL nanocomposites in alkaline and phosphate buffer solutions was investigated. The obtained results show that the PLA/CS/PCL nanocomposites have slower hydrolysis ability than the PLA/CS composites. PMID:26682382

  7. Enantioselective Hydrolysis of Amino Acid Esters Promoted by Bis(β-cyclodextrin) Copper Complexes

    PubMed Central

    Xue, Shan-Shan; Zhao, Meng; Ke, Zhuo-Feng; Cheng, Bei-Chen; Su, Hua; Cao, Qian; Cao, Zhen-Kun; Wang, Jun; Ji, Liang-Nian; Mao, Zong-Wan

    2016-01-01

    It is challenging to create artificial catalysts that approach enzymes with regard to catalytic efficiency and selectivity. The enantioselective catalysis ranks the privileged characteristic of enzymatic transformations. Here, we report two pyridine-linked bis(β-cyclodextrin) (bisCD) copper(II) complexes that enantioselectively hydrolyse chiral esters. Hydrolytic kinetic resolution of three pairs of amino acid ester enantiomers (S1–S3) at neutral pH indicated that the “back-to-back” bisCD complex CuL1 favoured higher catalytic efficiency and more pronounced enantioselectivity than the “face-to-face” complex CuL2. The best enantioselectivity was observed for N-Boc-phenylalanine 4-nitrophenyl ester (S2) enantiomers promoted by CuL1, which exhibited an enantiomer selectivity of 15.7. We observed preferential hydrolysis of L-S2 by CuL1, even in racemic S2, through chiral high-performance liquid chromatography (HPLC). We demonstrated that the enantioselective hydrolysis was related to the cooperative roles of the intramolecular flanking chiral CD cavities with the coordinated copper ion, according to the results of electrospray ionization mass spectrometry (ESI-MS), inhibition experiments, rotating-frame nuclear Overhauser effect spectroscopy (ROESY), and theoretical calculations. Although the catalytic parameters lag behind the level of enzymatic transformation, this study confirms the cooperative effect of the first and second coordination spheres of artificial catalysts in enantioselectivity and provides hints that may guide future explorations of enzyme mimics. PMID:26916830

  8. MTBE Hydrolysis in Dilute Aqueous Solution Using Heterogeneous Strong Acid Catalysts

    NASA Astrophysics Data System (ADS)

    Rixey, W. G.

    2003-12-01

    The objective of this research has been the development of a potential in situ catalytic process for the hydrolysis of methyl tertiary butyl ether (MTBE) to tertiary butyl alcohol (TBA) and methanol in ground water. Bench-scale batch reactor studies were conducted over a temperature range of 23 deg C to 50 deg C with several heterogeneous strong acid catalysts to obtain rates of hydrolysis of MTBE to TBA and methanol at dilute concentrations in water. Continuous flow experiments were then conducted to obtain kinetic data over a temperature range of 15 deg C to 50 deg C for various flow rates for the most active catalysts. It was found that the batch and continuous flow experiments yielded similar intrinsic kinetic rate constants when sorption of MTBE to the catalyst was accounted for. Additional fixed-bed experiments were conducted with deionized water and 0.005 M CaCl2 feed solutions containing 100 mg/L MTBE, respectively, to assess the deactivation of the catalyst, and deactivation was found to be controlled by ion exchange of H+ in the catalyst with Ca+2 in the feed. Our results indicate that, for low to moderate groundwater velocities and cation concentrations at ambient temperatures, an in situ reactive barrier process using the most active catalysts studied in this research could be a viable process in terms of both suitable conversion of MTBE and catalyst life. Although application to in situ remediation is emphasized, the results of this research are also applicable to ex-situ groundwater treatment.

  9. Optimization studies on acid hydrolysis of oil palm empty fruit bunch fiber for production of xylose.

    PubMed

    Rahman, S H A; Choudhury, J P; Ahmad, A L; Kamaruddin, A H

    2007-02-01

    Oil palm empty fruit bunch fiber is a lignocellulosic waste from palm oil mills. It is a potential source of xylose which can be used as a raw material for production of xylitol, a high value product. The increasing interest on use of lignocellulosic waste for bioconversion to fuels and chemicals is justifiable as these materials are low cost, renewable and widespread sources of sugars. The objective of the present study was to determine the effect of H(2)SO(4) concentration, reaction temperature and reaction time for production of xylose. Batch reactions were carried out under various reaction temperature, reaction time and acid concentrations and Response Surface Methodology (RSM) was followed to optimize the hydrolysis process in order to obtain high xylose yield. The optimum reaction temperature, reaction time and acid concentration found were 119 degrees C, 60 min and 2%, respectively. Under these conditions xylose yield and selectivity were found to be 91.27% and 17.97 g/g, respectively. PMID:16647852

  10. A simple procedure for preparing chitin oligomers through acetone precipitation after hydrolysis in concentrated hydrochloric acid.

    PubMed

    Kazami, Nao; Sakaguchi, Masayoshi; Mizutani, Daisuke; Masuda, Tatsuhiko; Wakita, Satoshi; Oyama, Fumitaka; Kawakita, Masao; Sugahara, Yasusato

    2015-11-01

    Chitin oligomers are of interest because of their numerous biologically relevant properties. To prepare chitin oligomers containing 4-6 GlcNAc units [(GlcNAc)4-6], α- and β-chitin were hydrolyzed with concentrated hydrochloric acid at 40 °C. The reactant was mixed with acetone to recover the acetone-insoluble material, and (GlcNAc)4-6 was efficiently recovered after subsequent water extraction. Composition analysis using gel permeation chromatography and MALDI-TOF mass spectrometry indicated that (GlcNAc)4-6 could be isolated from the acetone-insoluble material with recoveries of approximately 17% and 21% from the starting α-chitin and β-chitin, respectively. The acetone precipitation method is highly useful for recovering chitin oligomers from the acid hydrolysate of chitin. The changes in the molecular size and higher-order structure of chitin during the course of hydrolysis were also analyzed, and a model that explains the process of oligomer accumulation is proposed. PMID:26256353

  11. Hydrolysis of Miscanthus for bioethanol production using dilute acid presoaking combined with wet explosion pre-treatment and enzymatic treatment.

    PubMed

    Sørensen, Annette; Teller, Philip J; Hilstrøm, Troels; Ahring, Birgitte K

    2008-09-01

    Miscanthus is a high yielding bioenergy crop. In this study we used acid presoaking, wet explosion, and enzymatic hydrolysis to evaluate the combination of the different pre-treatment methods for bioethanol production with Miscanthus. Acid presoaking is primarily carried out in order to remove xylose prior to wet explosion. The acid presoaking extracted 63.2% xylose and 5.2% glucose. Direct enzymatic hydrolysis of the presoaked biomass was found to give only low sugar yields of 24-26% glucose. Wet explosion is a pre-treatment method that combines wet-oxidation and steam explosion. The effect of wet explosion on non-presoaked and presoaked Miscanthus was investigated using both atmospheric air and hydrogen peroxide as the oxidizing agent. All wet explosion pre-treatments showed to have a disrupting effect on the lignocellulosic biomass, making the sugars accessible for enzymatic hydrolysis. The combination of presoaking, wet explosion, and enzymatic hydrolysis was found to give the highest sugar yields. The use of atmospheric air gave the highest xylose yield (94.9% xylose, 61.3% glucose), while hydrogen peroxide gave the highest glucose yield (82.4% xylose, 63.7% glucose). PMID:18164954

  12. Conversion of levulinic acid into γ-valerolactone using Fe3(CO)12: mimicking a biorefinery setting by exploiting crude liquors from biomass acid hydrolysis.

    PubMed

    Metzker, Gustavo; Burtoloso, Antonio C B

    2015-09-28

    The conversion of biomass-derived levulinic acid (LA) into gamma-valerolactone (GVL) using formic acid (FA) and Fe3(CO)12 as the catalyst precursor was achieved in 92% yield. To mimic a biorefinery setting, crude liquor (containing 20% LA) from the acid hydrolysis of sugarcane biomass in a pilot plant facility was directly converted into GVL in good yield (50%), without the need for isolating LA. PMID:26258183

  13. Cassava starch maltodextrinization/monomerization through thermopressurized aqueous phosphoric acid hydrolysis.

    PubMed

    Fontana, J D; Passos, M; Baron, M; Mendes, S V; Ramos, L P

    2001-01-01

    Kinetic conditions were established for the depolymerization of cassava starch for the production of maltodextrins and glucose syrups. Thin-layer chromatography and high-performance liquid chromatography analyses corroborated that the proper H3PO4 strength and thermopressurization range (e.g., 142-170 degrees C; 2.8-6.8 atm) can be successfully explored for such hydrolytic purposes of native starch granules. Because phosphoric acid can be advantageously maintained in the hydrolysate and generates, after controlled neutralization with ammonia, the strategic nutrient triplet for industrial fermentations (C, P, N), this pretreatment strategy can be easily recognized as a recommended technology for hydrolysis and upgrading of starch and other plant polysaccharides. Compared to the classic catalysts, the mandatory desalting step (chloride removal by expensive anion-exchange resin or sulfate precipitation as the calcium-insoluble salt) can be avoided. Furthermore, properly diluted phosphoric acid is well known as an allowable additive in several popular soft drinks such as colas since its acidic feeling in the mouth is compatible and synergistic with both natural and artificial sweeteners. Glycosyrups from phosphorolyzed cassava starch have also been upgraded to high-value single-cell protein such as the pigmented yeast biomass of Xanthophyllomyces dendrorhous (Phaffia rhodozyma), whose astaxanthin (diketo-dihydroxy-beta-carotene) content may reach 0.5-1.0 mg/g of dry yeast cell. This can be used as an ideal complement for animal feeding as well as a natural staining for both fish farming (meat) and poultry (eggs). PMID:11963875

  14. [Determination of docosahexaenoic acid in milk powder by gas chromatography using acid hydrolysis].

    PubMed

    Shao, Shiping; Xiang, Dapeng; Li, Shuang; Xi, Xinglin; Chen, Wenrui

    2015-11-01

    A method to determine docosahexenoic acid (DHA) in milk powder by gas chromatography was established. The milk powder samples were hydrolyzed with hydrochloric acid, extracted to get total fatty acids by Soxhlet extractor, then esterified with potassium hydroxide methanol solution to form methyl esters, and treated with sodium hydrogen sulfate. The optimal experiment conditions were obtained from orthogonal experiment L9(3(3)) which performed with three factors and three levels, and it requires the reaction performed with 1 mol/L potassium hydroxide solution at 25 degrees C for 5 min. The derivative treated with sodium hydrogen sulfate was separated on a column of SP-2560 (100 m x 0.25 mm x 0.20 μm), and determined in 55 min by temperature programming-gas chromatography. Good linearity was obtained in the range 5.0-300 mg/L with the correlation coefficient of 0.999 9. The relative standard deviations (RSDs) were 3.4%, 1.2% and 1.1% for the seven repeated experiments of 10, 50 and 100 mg/L of DHA, respectively. The limit of detection was 2 mg/kg, and the recoveries of DHA were in the range of 90.4%-93.5%. The results are satisfactory through the tests of practical samples. PMID:26939370

  15. Selection of suitable mineral acid and its concentration for biphasic dilute acid hydrolysis of the sodium dithionite delignified Prosopis juliflora to hydrolyze maximum holocellulose.

    PubMed

    Naseeruddin, Shaik; Desai, Suseelendra; Venkateswar Rao, L

    2016-02-01

    Two grams of delignified substrate at 10% (w/v) level was subjected to biphasic dilute acid hydrolysis using phosphoric acid, hydrochloric acid and sulfuric acid separately at 110 °C for 10 min in phase-I and 121 °C for 15 min in phase-II. Combinations of acid concentrations in two phases were varied for maximum holocellulose hydrolysis with release of fewer inhibitors, to select the suitable acid and its concentration. Among three acids, sulfuric acid in combination of 1 & 2% (v/v) hydrolyzed maximum holocellulose of 25.44±0.44% releasing 0.51±0.02 g/L of phenolics and 0.12±0.002 g/L of furans, respectively. Further, hydrolysis of delignified substrate using selected acid by varying reaction time and temperature hydrolyzed 55.58±1.78% of holocellulose releasing 2.11±0.07 g/L and 1.37±0.03 g/L of phenolics and furans, respectively at conditions of 110 °C for 45 min in phase-I & 121 °C for 60 min in phase-II. PMID:26716889

  16. A potentiometric study of the hydrolysis of ethylenediaminetetraacetic acid to 150{degrees}C

    SciTech Connect

    Palmer, D.A.; Nguyen-Trung, Chinh

    1995-02-01

    Ethylenediaminetetraacetate anions, EDTA{sup 4-}, were titrated in a hydrogen-electrode concentration cell with an acidic titrant from 0 to 150{degrees}C at 25{degrees}C intervals. These titrations were carried out in the presence of 0. 1, 0.2, and 1.0 mol{center_dot}kg{sup -1} with the supporting electrolytes, sodium chloride, NaCl, and 1.0 mol{center_dot}kg{sup -1} tetramethylammonium trifluoromethylsulfonate, (CH{sub 3}){sub 4}N(F{sub 3}CSO{sub 3}) {l_brace}TMATFMS{r_brace} in order to assess the effect of both cation complexation by EDTA{sup 4-} and anion activity coefficient variations. The resulting hydrolysis quotients are discussed with reference to applications in boiler and heat exchanger chemical cleaning, as well as chemical and nuclear waste containment. Some recent diverse uses of this emf technique that also pertain to these applications will be mentioned briefly, e.g., surface absorption - zero-point-of-charge - measurements to high temperatures and in situ pH measurements in solubility and kinetic experiments.

  17. Valorisation of food waste via fungal hydrolysis and lactic acid fermentation with Lactobacillus casei Shirota.

    PubMed

    Kwan, Tsz Him; Hu, Yunzi; Lin, Carol Sze Ki

    2016-10-01

    Food waste recycling via fungal hydrolysis and lactic acid (LA) fermentation has been investigated. Hydrolysates derived from mixed food waste and bakery waste were rich in glucose (80.0-100.2gL(-1)), fructose (7.6gL(-1)) and free amino nitrogen (947-1081mgL(-1)). In the fermentation with Lactobacillus casei Shirota, 94.0gL(-1) and 82.6gL(-1) of LA were produced with productivity of 2.61gL(-1)h(-1) and 2.50gL(-1)h(-1) for mixed food waste and bakery waste hydrolysate, respectively. The yield was 0.94gg(-1) for both hydrolysates. Similar results were obtained using food waste powder hydrolysate, in which 90.1gL(-1) of LA was produced with a yield and productivity of 0.92gg(-1) and 2.50gL(-1)h(-1). The results demonstrate the feasibility of an efficient bioconversion of food waste to LA and a decentralized approach of food waste recycling in urban area. PMID:26873283

  18. Solid acid catalysts pretreatment and enzymatic hydrolysis of macroalgae cellulosic residue for the production of bioethanol.

    PubMed

    Tan, Inn Shi; Lee, Keat Teong

    2015-06-25

    The aim of this study is to investigate the technical feasibility of converting macroalgae cellulosic residue (MCR) into bioethanol. An attempt was made to present a novel, environmental friendly and economical pretreatment process that enhances enzymatic conversion of MCR to sugars using Dowex (TM) Dr-G8 as catalyst. The optimum yield of glucose reached 99.8% under the optimal condition for solid acid pretreatment (10%, w/v biomass loading, 4%, w/v catalyst loading, 30min, 120°C) followed by enzymatic hydrolysis (45FPU/g of cellulase, 52CBU/g of β-glucosidase, 50°C, pH 4.8, 30h). The yield of sugar obtained was found more superior than conventional pretreatment process using H2SO4 and NaOH. Biomass loading for the subsequent simultaneous saccharification and fermentation (SSF) of the pretreated MCR was then optimized, giving an optimum bioethanol yield of 81.5%. The catalyst was separated and reused for six times, with only a slight drop in glucose yield. PMID:25839825

  19. Comparative theoretical studies of the phosphomonoester hydrolysis mechanism by purple acid phosphatases.

    PubMed

    Retegan, M; Milet, A; Jamet, H

    2010-07-01

    We present here the first ONIOM (our own n-layered integrated molecular orbital + molecular mechanics method) studies of a purple acid phosphatase enzyme. Our study focused on the structures of the red kidney bean PAP (kbPAP) complexed with phosphate and with phenyl phosphate and on the mechanism of the phenyl phosphate hydrolysis by the enzyme. Density functional theory (DFT) calculations were also performed using models of different sizes for comparison purpose. Results show that the inclusion of three histidine residues, His202, His295, and His296, with their protein surrounding, is crucial to properly describe the coordination of the substrates. They induce a conformation with the substrate closer to the nucleophilic mu-hydroxyde bridge. In the mechanistic study, a transition state is stabilized by a strong hydrogen bond between His202 and the leaving group of the substrate. Consequently, a smaller value for the activation energy barrier is obtained from DFT calculations including this histidine to the same calculations without this histidine. Using the ONIOM method, this activation energy barrier is even more reduced. So the mechanism, which considers the hydroxo group bridging the two metal ions as nucleophile, becomes really convincing, contrary to the results obtained with a small model at the DFT level. PMID:20550096

  20. Hydrolysis of phosphodiesters by diiron complexes: design of nonequivalent iron sites in purple acid phosphatase models.

    PubMed

    Verge, François; Lebrun, Colette; Fontecave, Marc; Ménage, Stéphane

    2003-01-27

    New mu-oxo-diferric complexes have been designed for hydrolysis of phosphodiesters. To mimic the diiron active site of purple acid phosphatase, a combinatorial method has been used to select complexes containing two distinct iron coordination spheres. The introduction of a bidentate ligand, a substituted phenanthroline (L) into complex 1, [Fe2O(bipy)4(OH2)2](NO3)4, generates in solution the complex [Fe2O(bipy)3(L)(OH2)2](NO3)4 as shown by ESI/MS and 1H NMR studies. The latter complex was found to be 20-fold more active than complex 1. On the basis of kinetic studies, we demonstrated that the complex [Fe2O(bipy)3(L)(OH)(OH2)](NO3)3 was the active species and the reaction proceeded through the formation of a ternary complex in which one iron binds a hydroxide and the second, the substrate. At nonsaturating concentrations of the substrate, the increased activity with increased methyl substituents in L was due to an increased affinity of the complex for the substrate. The activity of [Fe2O(bipy)3(33'44'Me2-Phen)(OH2)2](NO3)4 [33'44'Me2Phen = 3,3',4,4'-dimethyl-1,10-phenanthroline] was found to be comparable to that reported for Co(III) or Ce(IV) complexes. PMID:12693232

  1. Investigation of the complex reaction coordinate of acid catalyzed amide hydrolysis from molecular dynamics simulations

    NASA Astrophysics Data System (ADS)

    Zahn, Dirk

    2004-05-01

    The rate-determining step of acid catalyzed peptide hydrolysis is the nucleophilic attack of a water molecule to the carbon atom of the amide group. Therein the addition of the hydroxyl group to the amide carbon atom involves the association of a water molecule transferring one of its protons to an adjacent water molecule. The protonation of the amide nitrogen atom follows as a separate reaction step. Since the nucleophilic attack involves the breaking and formation of several bonds, the underlying reaction coordinate is rather complex. We investigate this reaction step from path sampling Car-Parrinello molecular dynamics simulations. This approach does not require the predefinition of reaction coordinates and is thus particularly suited for investigating reaction mechanisms. From our simulations the most relevant components of the reaction coordinate are elaborated. Though the C⋯O distance of the oxygen atom of the water molecule performing the nucleophilic attack and the corresponding amide carbon atom is a descriptor of the reaction progress, a complete picture of the reaction coordinate must include all three molecules taking part in the reaction. Moreover, the proton transfer is found to depend on favorable solvent configurations. Thus, also the arrangement of non-reacting, i.e. solvent water molecules needs to be considered in the reaction coordinate.

  2. beta-Lactamase-catalyzed hydrolysis of acyclic depsipeptides and acyl transfer to specific amino acid acceptors.

    PubMed Central

    Pratt, R F; Govardhan, C P

    1984-01-01

    beta-Lactamases from all three classes, A, B, and C, catalyze the hydrolysis of specific acyclic depsipeptide (PhCH2CONHCR1R2CO2CHR3CO2H) analogs of acyl-D-alanyl-D-alanine peptides. The depsipeptides investigated, which are chemically as reactive toward nucleophiles as penicillins, are in general poor substrates, although differences between the classes of beta-lactamases have been observed: the order of effectiveness seems to be C greater than B greater than A. Certain class A and C beta-lactamases also catalyze phenylacetylglycyl transfer between phenylacetylglycyl depsipeptides and specific amino acid acceptors, a type of reaction hitherto identified more closely with D-alanyl-D-alanine transpeptidases than with beta-lactamases. Preliminary indications of an acyl-enzyme intermediate in these reactions have been obtained. These results support the suggestion [Tipper, D.J. and Strominger, J.L. (1965) Proc. Natl. Acad. Sci. USA 54, 1133-1141] that beta-lactamases are evolutionary descendants of bacterial cell wall D-alanyl-D-alanine transpeptidases. PMID:6424114

  3. Large scale purification of puerarin from Puerariae Lobatae Radix through resins adsorption and acid hydrolysis.

    PubMed

    Guo, Hai-Dong; Zhang, Qing-Feng; Chen, Ji-Guang; Shangguang, Xin-Cheng; Guo, Yu-Xian

    2015-02-01

    Puerarin is the major isoflavone of Puerariae Lobatae Radix. A method for large scale purification of puerarin was developed through resins adsorption and acid hydrolysis. The adsorption properties of six macroporous resins (D101, S-8, H103, X-5, HPD600, AB-8) were compared through the adsorption kinetics and equilibrium adsorption isotherms. Results showed that H103 resin had the best adsorption rate and capacity. The mass transfer zone motion model was further used for analyzing the fixed bed adsorption of H103 resin. Its length of mass transfer zone with 2mg/ml of puerarin in water and 10% ethanol at flow rate of 10ml/min were 41.6 and 47.5cm, while the equilibrium adsorption capacity was 165.03 and 102.88mg/g, respectively. By using 75% ethanol, puerarin could be well desorbed from the resin with recovery of 97.4%. Subsequently, H103 resin was successfully used for puerarin purification from Puerariae Lobatae Radix. The content of total isoflavones and puerarin in the resin adsorption product were 69.25% and 41.78%, respectively, which were about three times increased compared to the crude extract. Then, the product was hydrolyzed by 2.5M HCl at 90°C for 1h. Puerarin with purity of 90% and a byproduct daidzein with purity of 78% were obtained. PMID:25553536

  4. Lactic acid based PEU/HA and PEU/BCP composites: Dynamic mechanical characterization of hydrolysis.

    PubMed

    Rich, Jaana; Tuominen, Jukka; Kylmä, Janne; Seppälä, Jukka; Nazhat, Showan N; Tanner, K Elizabeth

    2002-01-01

    Lactic acid based poly(ester-urethane) (PEU-BDI) and its composites with 20 and 40 vol.% bioceramic filler were characterized prior to their use as biocompatible and bioabsorbable artificial bone materials. Morphological, dynamic mechanical properties, and degradation of these either hydroxyapatite or biphasic calcium phosphate containing composites were determined. Addition of particulate bioactive filler increased the composite stiffness and the glass transition temperature, indicating strong interactions between the filler and matrix. Materials were sterilized by gamma-irradiation, which reduced the average molecular weights by 30-40%. However, dynamic mechanical properties were not significantly affected by irradiation. Specimens were immersed in 0.85 w/v saline at 37 degrees C for 5 weeks, and changes in molecular weights, mass, water absorption, and dynamic mechanical properties were recorded. All the composite materials showed promising dynamic mechanical performance over the 5 weeks of hydrolysis. Average molecular weights of PEU-BDI and its composites did not change substantially during the test period. PEU-BDI retained its modulus values relatively well, and although the moduli of the composite materials were much higher, especially at high filler content, they exhibited faster loss of mechanical integrity. PMID:12115768

  5. Volatile organic acid adsorption and cation dissociation by porphyritic andesite for enhancing hydrolysis and acidogenesis of solid food wastes.

    PubMed

    Cheng, Fan; Li, Ming; Li, Dawei; Chen, Ling; Jiang, Weizhong; Kitamura, Yutaka; Li, Baoming

    2010-07-01

    Volatile organic acid adsorption, cation dissociation by porphyritic andesite, and their effects on the hydrolysis and acidogenesis of solid food wastes were evaluated through batch experiments. The acetic acid adsorption experiments show that pH was mainly regulated by H(+) adsorption. The mono-layer and multi-layer adsorption were found under the low (8.3-83.2 mmol/L) and high (133.22-532.89 mmol/L) initial acetic acid concentration, respectively. The dissociated cations concentration in acidic solution showed the predominance of Ca(2+). Porphyritic andesite addition elevated the pH levels and accelerated hydrolysis and acidogenesis in the batch fermentation experiment. Leachate of porphyritic andesite addition achieved the highest hydrolysis constant of 22.1 x 10(-3)kgm(-2)d(-1) and VS degradation rates of 3.9 g L(-1)d(-1). The highest activity of microorganisms represented by specific growth rate of ATP, 0.16d(-1), and specific consumption rate of Ca(2+), 0.18d(-1), was obtained by adding leachate of porphyritic andesite. PMID:20156676

  6. Theoretical studies of fundamental pathways for alkaline hydrolysis of carboxylic acid esters in gas phase

    SciTech Connect

    Zhan, C.G.; Landry, D.W.; Ornstein, R.L.

    2000-02-23

    Fundamental reaction pathways for the alkaline hydrolysis of carboxylic acid esters, RCOOR{prime}, were examined through a series of first-principle calculations. The reactions of six representative esters with hydroxide ion were studied in the gas phase. A total of three competing reaction pathways were found and theoretically confirmed for each of the esters examined: bimolecular base-catalyzed acyl-oxygen cleavage (B{sub AC}2), bimolecular base-catalyzed alkyl-oxygen cleavage (B{sub AL}2), and carbonyl oxygen exchange with hydroxide. For the two-step B{sub AC}2 process, this is the first theoretical study to consider the individual sub-steps of the reaction process and to consider substituent effects. For the carbonyl oxygen exchange with hydroxide and for the one-step B{sub AL}2 process, the authors report here the first quantitative theoretical results for the reaction pathways and for the energy barriers. The energy barrier calculated for the second step of the B{sub AC}2 process, that is, the decomposition of the tetrahedral intermediate, is larger in the gas phase than that of the first step, that is, the formation of the tetrahedral intermediate, for all but one of the esters examined. The exception, CH{sub 3}COOC(CH{sub 3}){sub 3}, does not have an {alpha} hydrogen in the leaving group. The highest energy barrier calculated for the B{sub AC}2 process is always lower than the barriers for the oxygen exchange and for the B{sub AL}2 process. The difference between the barrier for the B{sub AL}2 process and the highest barrier for the B{sub AC}2 process is only {approximately}1--3 kcal/mol for the methyl esters, but becomes much larger for the others. Substitution of an {alpha} hydrogen in R{prime} with a methyl group considerably increases the energy barrier for the B{sub AL}2 process, and significantly decreases the energy barrier for the second step of the B{sub AC}2 process. The calculated substituent shifts of the energy barrier for the first step of the

  7. Zirconium(IV) tetramer/octamer hydrolysis equilibrium in aqueous hydrochloric acid solution

    SciTech Connect

    Singhal, A.; Toth, L.M.; Lin, J.S.; Affholter, K.

    1996-11-20

    Small-angle X-ray scattering measurements were performed on zirconium acidic aqueous solutions to investigate the structure and the size of polynuclear species larger than the previously identified tetrameric species. Solutions with [H{sup +}]{sub added} and [OH{sup -}]{sub added} ranging from 0.0 to 1.0 M and 0.0 to 0.02 M, respectively, were analyzed. This study demonstrates that an octameric species, Zr{sub 8}(OH){sub 20}(H{sub 2}O){sub 24}Cl{sub 12}, exists in equilibrium with the tetrameric species, Zr{sub 4}(OH){sub 8}(H{sub 2} O){sub 16}Cl{sub 6}{sup 2+}, such that, for 0.05 M Zr(IV) in highly acidic solutions ([H{sup +}]{sub added} = 0.6 M), the tetramer in the dominant species; and at conditions with [H{sup +}] added =0.05 M, the octameric species becomes predominant. The estimated value of the equilibrium quotient obtained for the tetramer/octamer equilibrium is 0.20 {+-} 0.05 M{sup 3}. The tetramer and octamer have radii of gyration of 3.8 and 5.1 {+-} 0.2 A and a hydrolysis ratio of 2.0 and 2.5, respectively. The octamer forms by stacking two tetramers on top of each other. At conditions where small amounts of NaOH are added, unidentified polymeric Zr(IV) species larger in size than the octameric species appear. 26 refs., 7 figs.

  8. Competition between the hydrolysis and deamination of cytidine and its 5-substituted derivatives in aqueous acid.

    PubMed Central

    Lönnberg, H; Käppi, R

    1985-01-01

    The monocations of a few 5-substituted cytidines have been shown to undergo competitive deamination to the corresponding uridines and hydrolysis to 5-substituted cytosines and D-ribose. The first-order rate constants measured at different temperatures indicate that the proportion of the hydrolysis is considerably increased with the increasing temperature. Electron-withdrawal by a polar substituent at C5 appears to facilitate the hydrolysis to a larger extent that the deamination. The ionic strength has practically no influence on the rate of either reaction. PMID:4000961

  9. Effects of waste activated sludge and surfactant addition on primary sludge hydrolysis and short-chain fatty acids accumulation.

    PubMed

    Ji, Zhouying; Chen, Guanlan; Chen, Yinguang

    2010-05-01

    This paper focused on the effects of waste activated sludge (WAS) and surfactant sodium dodecylbenzene sulfonate (SDBS) addition on primary sludge (PS) hydrolysis and short-chain fatty acids (SCFA) accumulation in fermentation. The results showed that sludge hydrolysis, SCFA accumulation, NH(4)(+)-N and PO(4)(3-)-P release, and volatile suspended solids (VSS) reduction were increased by WAS addition to PS, which were further increased by the addition of SDBS to the mixture of PS and WAS. Acetic, propionic and valeric acids were the top three SCFA in all experiments. Also, the fermentation liquids of PS, PS+WAS, and PS+WAS+SDBS were added, respectively, to municipal wastewater to examine their effects on biological municipal wastewater treatment, and the enhancement of both wastewater nitrogen and phosphorus removals was observed compared with no fermentation liquid addition. PMID:20096564

  10. Preliminary results and economics of the New York University process: continuous acid hydrolysis of cellulose, producing glucose for fermentation

    SciTech Connect

    Rugg, B.; Armstrong, P.; Stanton, R.

    1981-01-01

    The title process for the continuous acid hydrolysis of cellulose to glucose was evaluated in both batch- and pilot plant-scales. The optimal temperature and reaction time for batch-scale dilute acid hydrolysis were 232 degrees and 10-20 s, respectively. Comparison of glucose yield from newspaper pulp (10% solids) with sawdust (95% solids) as feedstock indicated that 50-60% conversions of alpha-cellulose to glucose were possible on a pilot-plant scale. Acceptable recovery of glucose (greater than 90%) was best accomplished by centrifugation at glucose concentrations of less than 4% from a 30% solids cake. In general, favorable results with respect to sugar yield and energy consumption were obtained.

  11. Effects of ultrasonic treatment during acid hydrolysis on the yield, particle size and structure of cellulose nanocrystals.

    PubMed

    Guo, Juan; Guo, Xuxia; Wang, Siqun; Yin, Yafang

    2016-01-01

    Ultrasonic treatment is useful for the isolation of cellulose nanocrystals (CNCs). However, the effects of ultrasonic treatment on the structure and properties of CNCs have not been fully understood. We statistically analyzed the variations in yields and dimensions of CNCs prepared using acid hydrolysis, with or without ultrasonic treatment, and illustrated these variations by investigating the CNC structures. The results demonstrated that ultrasonic treatment promoted an increase in yields only for short hydrolysis times, while resulting in CNCs with shorter lengths and narrower dimension distributions. Furthermore, a prolonged acid attack under ultrasonic treatment caused a partial dissociation of cellulose hydrogen bond networks in the CNCs, ultimately resulting in the delamination and disorder of the cellulose crystalline structure, thus leading to a decrease in the width and thickness of the CNCs. This work provides more insights into the mechanisms of ultrasonic treatment on the structure and properties of CNCs. PMID:26453875

  12. Structural features of dilute acid, steam exploded, and alkali pretreated mustard stalk and their impact on enzymatic hydrolysis.

    PubMed

    Kapoor, Manali; Raj, Tirath; Vijayaraj, M; Chopra, Anju; Gupta, Ravi P; Tuli, Deepak K; Kumar, Ravindra

    2015-06-25

    To overcome the recalcitrant nature of biomass several pretreatment methodologies have been explored to make it amenable to enzymatic hydrolysis. These methodologies alter cell wall structure primarily by removing/altering hemicelluloses and lignin. In this work, alkali, dilute acid, steam explosion pretreatment are systematically studied for mustard stalk. To assess the structural variability after pretreatment, chemical analysis, surface area, crystallinity index, accessibility of cellulose, FT-IR and thermal analysis are conducted. Although the extent of enzymatic hydrolysis varies upon the methodologies used, nevertheless, cellulose conversion increases from <10% to 81% after pretreatment. Glucose yield at 2 and 72h are well correlated with surface area and maximum adsorption capacity. However, no such relationship is observed for xylose yield. Mass balance of the process is also studied. Dilute acid pretreatment is the best methodology in terms of maximum sugar yield at lower enzyme loading. PMID:25839820

  13. In Candida parapsilosis the ATC1 Gene Encodes for an Acid Trehalase Involved in Trehalose Hydrolysis, Stress Resistance and Virulence

    PubMed Central

    Sánchez-Fresneda, Ruth; Martínez-Esparza, María; Maicas, Sergi; Argüelles, Juan-Carlos; Valentín, Eulogio

    2014-01-01

    An ORF named CPAR2-208980 on contig 005809 was identified by screening a Candida parapsilosis genome data base. Its 67% identity with the acid trehalase sequence from C. albicans (ATC1) led us to designate it CpATC1. Homozygous mutants that lack acid trehalase activity were constructed by gene disruption at the two CpATC1 chromosomal alleles. Phenotypic characterization showed that atc1Δ null cells were unable to grow on exogenous trehalose as carbon source, and also displayed higher resistance to environmental challenges, such as saline exposure (1.2 M NaCl), heat shock (42°C) and both mild and severe oxidative stress (5 and 50 mM H2O2). Significant amounts of intracellular trehalose were specifically stored in response to the thermal upshift in both wild type and mutant strains. Analysis of their antioxidant activities revealed that catalase was only triggered in response to heat shock in atc1Δ cells, whereas glutathione reductase was activated upon mild oxidative stress in wild type and reintegrant strains, and in response to the whole set of stress treatments in the homozygous mutant. Furthermore, yeast cells with double CpATC1 deletion were significantly attenuated in non-mammalian infection models, suggesting that CpATC1 is required for the pathobiology of the fungus. Our results demonstrate the involvement of CpAtc1 protein in the physiological hydrolysis of external trehalose in C. parapsilosis, where it also plays a major role in stress resistance and virulence. PMID:24922533

  14. Sulfuric acid hydrolysis and detoxification of red alga Pterocladiella capillacea for bioethanol fermentation with thermotolerant yeast Kluyveromyces marxianus.

    PubMed

    Wu, Chien-Hui; Chien, Wei-Chen; Chou, Han-Kai; Yang, Jungwoo; Lin, Hong-Ting Victor

    2014-09-01

    One-step sulfuric acid saccharification of the red alga Pterocladiella capillacea was optimized, and various detoxification methods (neutralization, overliming, and electrodialysis) of the acid hydrolysate were evaluated for fermentation with the thermotolerant yeast Kluyveromyces marxianus. A proximate composition analysis indicated that P. capillacea was rich in carbohydrates. A significant galactose recovery of 81.1 ± 5% was also achieved under the conditions of a 12% (w/v) biomass load, 5% (v/v) sulfuric acid, 121°C, and hydrolysis for 30 min. Among the various detoxification methods, electrodialysis was identified as the most suitable for fermentable sugar recovery and organic acid removal (100% reduction of formic and levulinic acids), even though it failed to reduce the amount of the inhibitor 5-HMF. As a result, K. marxianus fermentation with the electrodialyzed acid hydrolysate of P. capillacea resulted in the best ethanol levels and fermentation efficiency. PMID:24851812

  15. Production of 2,3-butanediol from cellulose and Jatropha hulls after ionic liquid pretreatment and dilute-acid hydrolysis

    PubMed Central

    2013-01-01

    Abundant Jatropha waste is a promising renewable feedstock for the production of sugars and 2,3-butanediol fermentation. To obtain high yield of water-soluble products and high concentration of reducing-sugars, ionic liquid (IL) pretreatment and dilute acid hydrolysis at 150°C were combined in this work. The destruction of crystalline structure and increase surface area of biomasses after IL-pretreatment, made their hydrolysis more efficient. Compared with original cellulose, after IL-pretreatment, both the yield and concentration of reducing-sugars increased by 139%, and the water-soluble products yield increased by 128% after hydrolysis. Compared with water-washed Jatropha hulls, after IL-pretreatment, the yield and concentration of reducing-sugars increased by 80% and 76%, respectively, and the water-soluble products yield increased by 70% after hydrolysis. IL-pretreatment benefited the fermentation of Jatropha hull hydrolysate with 66.58% diol yield and its productivity increased from 0.35 to 0.40 g/(L · h). PMID:23958155

  16. ESTIMATION OF HYDROLYSIS RATE CONSTANTS OF CARBOXYLIC ACID ESTER AND PHOSPHATE ESTER COMPOUNDS IN AQUEOUS SYSTEMS FROM MOLECULAR STRUCTURE BY SPARC

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to calculate hydrolysis rate constants for carboxylic acid ester and phosphate ester compounds in aqueous non- aqueous and systems strictly from molecular structure. The energy diffe...

  17. Polyphosphate Hydrolysis within Acidic Vacuoles in Response to Amine-Induced Alkaline Stress in the Halotolerant Alga Dunaliella salina

    PubMed Central

    Pick, Uri; Weiss, Meira

    1991-01-01

    The location and mobilization of polyphosphates in response to an amine-induced alkaline stress were studied in the halotolerant alga Dunaliella salina. The following observations suggest that polyphosphates accumulate in acidic vacuoles: (a) Accumulation of large amounts of polyphosphates is manifested as intravacuolar dense osmiophilic bodies in electron micrographs. (b) Uptake of amines into the vacuoles induces massive hydrolysis of polyphosphates, demonstrated by in vivo 31P-nuclear magnetic resonance, and by analysis of hydrolytic products on thin layer chromatograms. The analysis indicates that: (a) Polyphosphate hydrolysis is kinetically correlated with amine accumulation and with the recovery of cytoplasmic pH. (b) The major hydrolytic product is tripolyphosphate. (c) The peak position of the tripolyphosphate terminal phosphate in nuclear magnetic resonance spectra is progressively shifted as the cells recover, indicating that the pH inside the vacuoles increases while the pH in the cytoplasm decreases. (d) In lysed cell preparations, in which vacuoles become exposed to the external pH, mild alkalinization in the absence of amines induces polyphosphate hydrolysis to tripolyphosphates. It is suggested that amine accumulation within vacuoles activates a specific phosphatase, which hydrolyzes long-chain polyphosphates to tripolyphosphates. The hydrolysis increases the capacity of the vacuoles to sequester amines from the cytoplasm probably by releasing protons required to buffer the amine, and leads to recovery of cytoplasmic pH. Thus, polyphosphate hydrolysis provides a high-capacity buffering system that sustains amine compartmentation into vacuoles and protects cytoplasmic pH. ImagesFigure 1Figure 7 PMID:16668514

  18. Acid Catalysis in Basic Solution: A Supramolecular Host PromotesOrthoformate Hydrolysis

    SciTech Connect

    Pluth, Michael D.; Bergman, Robert G.; Raymond, Kenneth N.

    2007-12-12

    Though many enzymes can promote chemical reactions by tuning substrate properties purely through the electrostatic environment of a docking cavity, this strategy has proven challenging to mimic in synthetic host-guest systems. Here we report a highly-charged, water soluble, metal-ligand assembly with a hydrophobic interior cavity that thermodynamically stabilizes protonated substrates and consequently catalyzes the normally acidic hydrolysis of orthoformates in basic solution, with rate accelerations of up to 890-fold. The catalysis reaction obeys Michaelis-Menten kinetics, exhibits competitive inhibition, and the substrate scope displays size selectivity consistent with the constrained binding environment of the molecular host. Synthetic chemists have long endeavored to design host molecules capable of selectively binding slow-reacting substrates and catalyzing their chemical reactions. While synthetic catalysts are often site-specific and require certain properties of the substrate to insure catalysis, enzymes are often able to modify basic properties of the bound substrate such as pK{sub a} in order to enhance reactivity. Two common motifs used by nature to activate otherwise unreactive compounds are the precise arrangement of hydrogen-bonding networks and electrostatic interactions between the substrate and adjacent residues of the protein. Precise arrangement of hydrogen bonding networks near the active sites of proteins can lead to well-tuned pK{sub a}-matching, and can result in pK{sub a} shifts of up to eight units, as shown in bacteriorhodopsin. Similarly, purely electrostatic interactions can greatly favor charged states and have been responsible for pK{sub a} shifts of up to five units for acetoacetate decarboxylase. Attempts have been made to isolate the contributions of electrostatic versus covalent interactions to such pK{sub a} shifts; however this remains a difficult challenge experimentally. This challenge emphasizes the importance of synthesizing

  19. Inhibitory activity of thermal copolymers of amino acids for the metal-catalyzed hydrolysis of an RNA dinucleotide

    NASA Astrophysics Data System (ADS)

    Kawamura, Kunio; Nagahama, Minoru; Yao, Toshio

    2006-01-01

    It is well known that the hydrolysis of RNA is substantially catalyzed by several metal ions. Although this fact poses a problem for the RNA world hypothesis, there may have been unknown pathways for the protection of RNA molecules against the hydrolytic degradation under the primitive earth conditions. Thus, we have investigated whether or not thermal copolymers of amino acids (TCAA) inhibit the catalytic activity of metal ions for the RNA hydrolyses; TCAA is a suitable model material for prebiotic protein-like molecules since TCAA involving peptide bonding is readily prepared by heating amino acid mixtures under prebiotic conditions. The activities of metal ions that Fe(III) and Co(II) enhance somewhat the 3',5'-cytidylylguanosine (CpG) hydrolysis and Ce(III) and Eu(III) accelerate greatly the CpG hydrolysis were notably reduced by TCAA. This fact indicates that protein-like molecules would have played important roles for the accumulation of RNA under the primitive earth conditions.

  20. The use of sub-critical water hydrolysis for the recovery of peptides and free amino acids from food processing wastes. Review of sources and main parameters.

    PubMed

    Marcet, Ismael; Álvarez, Carlos; Paredes, Benjamín; Díaz, Mario

    2016-03-01

    Food industry processing wastes are produced in enormous amounts every year, such wastes are usually disposed with the corresponding economical cost it implies, in the best scenario they can be used for pet food or composting. However new promising technologies and tools have been developed in the last years aimed at recovering valuable compounds from this type of materials. In particular, sub-critical water hydrolysis (SWH) has been revealed as an interesting way for recovering high added-value molecules, and its applications have been broadly referred in the bibliography. Special interest has been focused on recovering protein hydrolysates in form of peptides or amino acids, from both animal and vegetable wastes, by means of SWH. These recovered biomolecules have a capital importance in fields such as biotechnology research, nutraceuticals, and above all in food industry, where such products can be applied with very different objectives. Present work reviews the current state of art of using sub-critical water hydrolysis for protein recovering from food industry wastes. Key parameters as reaction time, temperature, amino acid degradation and kinetic constants have been discussed. Besides, the characteristics of the raw material and the type of products that can be obtained depending on the substrate have been reviewed. Finally, the application of these hydrolysates based on their functional properties and antioxidant activity is described. PMID:26831563

  1. Production of nanocrystalline cellulose from an empty fruit bunches using sulfuric acid hydrolysis: Effect of reaction time on the molecular characteristics

    NASA Astrophysics Data System (ADS)

    Al-Dulaimi, Ahmed A.; R, Rohaizu; D, Wanrosli W.

    2015-06-01

    Nanocrystalline cellulose (NCC) was isolated from OPEFB pulp via sulfuric acid hydrolysis. The influence of reaction time to the molecular weight and surface charge of the NCC was investigated. Characterization of the product was carried out using zeta potential measurement and gel permeation chromatography test. Zeta potential measurement showed that the surface negative charge significantly increases with increasing reaction time. Gel permeation chromatography test indicates that molecular weight of NCC change variably with increasing of hydrolysis time. (Keywords: Nanocrystalline cellulose; acid hydrolysis; sulfate content; molecular weight)

  2. Isolation of bacterial cellulose nanocrystalline from pineapple peel waste: Optimization of acid concentration in the hydrolysis method

    NASA Astrophysics Data System (ADS)

    Anwar, Budiman; Rosyid, Nurul Huda; Effendi, Devi Bentia; Nandiyanto, Asep Bayu Dani; Mudzakir, Ahmad; Hidayat, Topik

    2016-02-01

    Isolation of needle-shaped bacterial cellulose nanocrystalline with a diameter of 16-64 nm, a fiber length of 258-806 nm, and a degree of crystallinity of 64% from pineapple peel waste using an acid hydrolysis process was investigated. Experimental showed that selective concentration of acid played important roles in isolating the bacterial cellulose nanocrystalline from the cellulose source. To achieve the successful isolation of bacterial cellulose nanocrystalline, various acid concentrations were tested. To confirm the effect of acid concentration on the successful isolation process, the reaction conditions were fixed at a temperature of 50°C, a hydrolysis time of 30 minutes, and a bacterial cellulose-to-acid ratio of 1:50. Pineapple peel waste was used as a model for a cellulose source because to the best of our knowledge, there is no report on the use of this raw material for producing bacterial cellulose nanocrystalline. In fact, this material can be used as an alternative for ecofriendly and cost-free cellulose sources. Therefore, understanding in how to isolate bacterial cellulose nanocrystalline from pineapple peel waste has the potential for large-scale production of inexpensive cellulose nanocrystalline.

  3. Co-hydrolysis of hydrothermal and dilute acid pretreated populus slurries to support development of a high-throughput pretreatment system

    PubMed Central

    2011-01-01

    Background The BioEnergy Science Center (BESC) developed a high-throughput screening method to rapidly identify low-recalcitrance biomass variants. Because the customary separation and analysis of liquid and solids between pretreatment and enzymatic hydrolysis used in conventional analyses is slow, labor-intensive and very difficult to automate, a streamlined approach we term 'co-hydrolysis' was developed. In this method, the solids and liquid in the pretreated biomass slurry are not separated, but instead hydrolysis is performed by adding enzymes to the whole pretreated slurry. The effects of pretreatment method, severity and solids loading on co-hydrolysis performance were investigated. Results For hydrothermal pretreatment at solids concentrations of 0.5 to 2%, high enzyme protein loadings of about 100 mg/g of substrate (glucan plus xylan) in the original poplar wood achieved glucose and xylose yields for co-hydrolysis that were comparable with those for washed solids. In addition, although poplar wood sugar yields from co-hydrolysis at 2% solids concentrations fell short of those from hydrolysis of washed solids after dilute sulfuric acid pretreatment even at high enzyme loadings, pretreatment at 0.5% solids concentrations resulted in similar yields for all but the lowest enzyme loading. Conclusions Overall, the influence of severity on susceptibility of pretreated substrates to enzymatic hydrolysis was clearly discernable, showing co-hydrolysis to be a viable approach for identifying plant-pretreatment-enzyme combinations with substantial advantages for sugar production. PMID:21749707

  4. [The use of enzymatic hydrolysis for isolation of barbituric acid derivatives from blood (as exemplified by phenobarbital and barbamyl)].

    PubMed

    Chuvina, N A; Kolupaeva, A S; Strelova, O Iu; Zabolotskaia, I V; Gorbacheva, T V

    2010-01-01

    Modern isolation techniques by direct extraction with organic solvents or after protein precipitation by various sedimenting or salting-out agents are characterized by low efficiency and do not permit to liberate derivatives of barbituric acid from their complexes with blood proteins. The use of enzymatic hydrolysis makes it possible to break bonds between barbiturates and protein and thereby improve the efficiency of isolation. We performed enzymatic hydrolysis of the model phenobarbital-blood and barbamyl-blood complexes with the use of trypsin, pepsin, chymotrypsin, and papain. The degree of phenobarbital extraction with trypsin and barbamyl was estimated at 62.1 +/- 1.2% and 75.1 +/- 1.6% respectively; in other words, it was 32.7 +/- 1.0% and 51.1 +/- 1.0% higher than that achieved by traditional methods. Certain validation characteristics of the new method are presented. PMID:21265178

  5. Assay of phenolic compounds from four species of ber (Ziziphus mauritiana L.) fruits: comparison of three base hydrolysis procedure for quantification of total phenolic acids.

    PubMed

    Memon, Ayaz Ali; Memon, Najma; Bhanger, Muhammad Iqbal; Luthria, Devanand L

    2013-08-15

    The present study was undertaken to investigate the flavonoid profile in four species of ber (Ziziphus mauritiana Lamk.) fruit. The 12 flavonoids identified were quercetin 3-O-robinobioside, quercetin 3-O-rutinoside, quercetin 3'-O-galactoside, quercetin 3'-O-glucoside, quercetin 3'-O-rhamnoside, quercetin 3'-O-pentosylhexoside, quercetin 3-O-6'malonylglucoside, quercetin 3'-O-malonylglucoside, luteolin 7-O-6'malonylglucoside, luteolin 7-O-malonylglucoside, myricetin 3-O-galactoside, and naringenin tri glycoside. This is the first report on extraction of nine additional flavonoids from the ber fruits. In addition, we also compared the impact of three different base hydrolysis techniques namely ultrasonic assisted base hydrolysis (UABH), microwave assisted base hydrolysis (MWABH), and pressurised liquid assisted base hydrolysis (PLABH) for the quantification of total phenolic acids. Nine phenolic acids, protocatechuic acid, p-hydroxybenzoic acid, ferulic acid, chlorogenic acid, vanillic acid, caffeic acid, vanillin, ortho- and para-coumaric acids, were identified and quantified. The three major phenolic acids identified in all four ber species were p-coumaric acid, vanillin and ferulic acids. Higher amounts (p<0.05) of total phenolic acids in all cultivars were obtained with the PLABH technique as compared to other two procedures (UABH and MWABH). PMID:23561136

  6. Antioxidant activities of fucoidan degraded by gamma irradiation and acidic hydrolysis

    NASA Astrophysics Data System (ADS)

    Lim, Sangyong; Choi, Jong-il; Park, Hyun

    2015-04-01

    Low molecular weight fucoidan, prepared by radical degradation using gamma ray was investigated for its antioxidant activities with different assay methods. As the molecular weight of fucoidan decreased with a higher absorbed dose, ferric-reducing antioxidant power values increased, but β-carotene bleaching inhibition did not change significantly. The antioxidant activity of acid-degraded fucoidan was also examined to investigate the effect of different degradation methods. At the same molecular weight, fucoidan degraded by gamma irradiation showed higher 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity than that observed with the acidic method. This result reveals that in addition to molecular weight, the degradation method affects the antioxidant activity of fucoidan.

  7. Acceleration of Acid-Catalyzed Hydrolysis in a Biphasic System by Sodium Tetracyanocyclopentadienides.

    PubMed

    Sakai, Takeo; Bito, Mariko; Itakura, Makoto; Sato, Honami; Mori, Yuji

    2016-01-01

    The hydrolysis of tert-butyldimethylsilyl L-menthyl ether (3) in a CH2Cl2-1 M HCl biphasic solvent system was accelerated by the addition of sodium tetracyanocyclopentadienides 1. Particularly, the reaction rate was enhanced using sodium salt 1a-c with a lipophilic substituent on the cyclopentadienide ring. From the results obtained by a triphasic experiment, hydrolysis proceeds via the formation of hydronium ion 2 in the aqueous phase by ion exchange, followed by the transfer of 2 to the CH2Cl2 phase. PMID:27373648

  8. Hydrolysis of Cellulose by a Mesoporous Carbon-Fe₂(SO₄)₃/γ-Fe₂O₃ Nanoparticle-Based Solid Acid Catalyst.

    PubMed

    Yamaguchi, Daizo; Watanabe, Koki; Fukumi, Shinya

    2016-01-01

    Carbon-based solid acid catalysts have shown significant potential in a wide range of applications, and they have been successfully synthesized using simple processes. Magnetically separable mesoporous carbon composites also have enormous potential, especially in separation and adsorption technology. However, existing techniques have been unable to produce a magnetically separable mesoporous solid acid catalyst because no suitable precursors have been identified. Herein we describe a magnetically separable, mesoporous solid acid catalyst synthesized from a newly developed mesoporous carbon-γ-Fe2O3 nanoparticle composite. This material exhibits an equivalent acid density and catalytic activity in the hydrolysis of microcrystalline cellulose, to that of the cellulose-derived conventional catalyst. Since it is magnetically separable, this material can be readily recovered and reused, potentially reducing the environmental impact of industrial processes to which it is applied. PMID:26856604

  9. Complete amino acid analysis of peptides and proteins after hydrolysis by a mixture of Sepharose-bound peptidases

    PubMed Central

    Bennett, H. P. J.; Elliott, D. F.; Evans, B. E.; Lowry, P. J.; McMartin, C.

    1972-01-01

    Incubation with a mixture of Sepharose-bound peptidases was shown to result in the quantitative release of amino acids from certain peptides and S-aminoethylated proteins. Subtraction of the low background values of amino acids generated by the enzymes enables amino acid ratios of corticotrophin-(1–24)-tetracosapeptide to be determined with a standard deviation on repeat digestions of 3–5%. Good values were obtained for amino acids that are completely or partially destroyed on acid hydrolysis, i.e. tryptophan, tyrosine, serine, asparagine and glutamine. Experiments with peptides containing d-amino acids showed that the enzyme mixture is stereospecific and could therefore be used to detect the presence of d-residues in peptides. The enzyme mixture completely hydrolyses peptide fragments obtained after Edman degradation and should therefore be useful for determining sequences of peptides containing acid-labile amino acid residues. The activities of the bound enzymes were unaltered over a period of 7 months and they provide a simple, reproducible procedure for the quantitative determination of amino acids in peptides and proteins containing l-amino acids. PMID:4349115

  10. Structural studies on colanic acid, the common exopolysaccharide found in the Enterobacteriaceae, by partial acid hydrolysis. Oligosaccharides from colanic acid

    PubMed Central

    Sutherland, I. W.

    1969-01-01

    The exopolysaccharide slime colanic acid has been isolated from representative strains of Escherichia coli, Salmonella typhimurium and Aerobacter cloacae. Analysis showed that each polymer contained glucose, galactose, fucose and glucuronic acid, together with acetate and pyruvate. The molar proportions of these components were 1:1·8:1·9:1:1:1 approximately. On the basis of periodate oxidation of the natural and deacetylated polysaccharide, glucose is proposed as the site of the acetyl groups. The pyruvate is attached to galactose. Three neutral oligosaccharides and ten electrophoretically mobile oligosaccharides were isolated and partially characterized. Four of the fragments were esters of pyruvic acid. Most oligosaccharides were isolated from all three polysaccharide preparations. Three further oligosaccharides were isolated from carboxyl-reduced colanic acid and sodium borotritide was used to label the glucose derived from glucuronic acid in these fragments. One trisaccharide was obtained from periodate-oxidized polysaccharide. On the basis of these oligosaccharides a repeating hexasaccharide unit of the following structure is proposed: [Formula: see text] The significance of this structure in colanic acid biosynthesis is discussed. PMID:4311825

  11. Molecular mechanism of acid-catalyzed hydrolysis of peptide bonds using a model compound.

    PubMed

    Pan, Bin; Ricci, Margaret S; Trout, Bernhardt L

    2010-04-01

    The stability of peptide bonds is a critical aspect of biological chemistry and therapeutic protein applications. Recent studies found elevated nonenzymatic hydrolysis in the hinge region of antibody molecules, but no mechanism was identified. As a first step in providing a mechanistic interpretation, this computational study examines the rate-determining step of the hydrolytic reaction of a peptide bond under acidic pH by a path sampling technique using a model compound N-MAA. Most previous computational studies did not include explicit water molecules, whose effects are significant in solution chemistry, nor did they provide a dynamic picture for the reaction process in aqueous conditions. Because no single trajectory can be used to describe the reaction dynamics due to fluctuations at finite temperatures, a variant version of the transition path sampling technique, the aimless shooting algorithm, was used to sample dynamic trajectories and to generate an ensemble of transition trajectories according to their statistical weights in the trajectory space. Each trajectory was computed as the time evolution of the molecular system using the Car-Parrinello molecular dynamics technique. The likelihood maximization procedure and its modification were used in extracting dynamically relevant degrees of freedom in the system, and approximations of the reaction coordinate were compared. Its low log-likelihood score and poor p(B) histogram indicate that the C-O distance previously assumed as the reaction coordinate for the rate-determining step is inadequate in describing the dynamics of the reaction. More than one order parameter in a candidate set including millions of geometric quantities was required to produce a convergent reaction coordinate model; its involvement of many degrees of freedom suggests that this hydrolytic reaction step is very complex. In addition to affecting atoms directly involved in bond-making and -breaking processes, the water network also has

  12. Accelerated hydrolysis of substituted cellulose for potential biofuel production: kinetic study and modeling.

    PubMed

    Mu, Bingnan; Xu, Helan; Yang, Yiqi

    2015-11-01

    In this work, kinetics of substitution accelerated cellulose hydrolysis with multiple reaction stages was investigated to lay foundation for mechanism study and molecular design of substituting compounds. High-efficiency hydrolysis of cellulose is critical for cellulose-based bioethanol production. It is known that, substitution could substantially decrease activation energy and increase reaction rate of acidic hydrolysis of glycosidic bonds in cellulose. However, reaction kinetics and mechanism of the accelerated hydrolysis were not fully revealed. In this research, it was proved that substitution therefore accelerated hydrolysis only occurred in amorphous regions of cellulose fibers, and was a process with multiple reaction stages. With molar ratio of substitution less than 1%, the overall hydrolysis rate could be increased for around 10 times. We also quantified the relationship between the hydrolysis rate of individual reaction stage and its major influences, including molar ratio of substitution, activation energy of acidic hydrolysis, pH and temperature. PMID:26253917

  13. Kinetic and Modeling Investigation to Provide Design Guidelines for the NREL Dilute-Acid Process Aimed at Total Hydrolysis/Fractionation of Lignocellulosic Biomass: July 1998

    SciTech Connect

    Lee, Y. Y.; Iyer, P.; Xiang, Q.; Hayes, J.

    2004-08-01

    Following up on previous work, subcontractor investigated three aspects of using NREL ''pretreatment'' technology for total hydrolysis (cellulose as well as hemicellulose) of biomass. Whereas historic hydrolysis of biomass used either dilute acid or concentrated acid technology for hydrolysis of both hemicellulose and cellulose, NREL has been pursuing very dilute acid hydrolysis of hemicellulose followed by enzymatic hydrolysis of cellulose. NREL's countercurrent shrinking-bed reactor design for hemicellulose hydrolysis (pretreatment) has, however, shown promise for total hydrolysis. For the first task, subcontractor developed a mathematical model of the countercurrent shrinking bed reactor operation and, using yellow poplar sawdust as a feedstock, analyzed the effect of: initial solid feeding rate, temperature, acid concentration, acid flow rate, Peclet number (a measure of backmixing in liquid flow), and bed shrinking. For the second task, subcontractor used laboratory trials, with yellow poplar sawdust and 0.07 wt% sulfuric acid at various temperatures, to verify the hydrolysis of cellulose to glucose (desired) and decomposition of glucose (undesired) and determine appropriate parameters for use in kinetic models. Unlike cellulose and hemicellulose, lignins, the third major component of biomass, are not carbohydrates that can be broken down into component sugars. They are, however, aromatic complex amorphous phenolic polymers that can likely be converted into low-molecular weight compounds suitable for production of fuels and chemicals. Oxidative degradation is one pathway for such conversion and hydrogen peroxide would be an attractive reagent for this, as it would leave no residuals. For the third task, subcontractor reacted lignin with hydrogen peroxide under various conditions and analyzed the resulting product mix.

  14. Cholesterol efflux from THP-1 macrophages is impaired by the fatty acid component from lipoprotein hydrolysis by lipoprotein lipase.

    PubMed

    Yang, Yanbo; Thyagarajan, Narmadaa; Coady, Breanne M; Brown, Robert J

    2014-09-01

    Lipoprotein lipase (LPL) is an extracellular lipase that primarily hydrolyzes triglycerides within circulating lipoproteins. Macrophage LPL contributes to atherogenesis, but the mechanisms behind it are poorly understood. We hypothesized that the products of lipoprotein hydrolysis generated by LPL promote atherogenesis by inhibiting the cholesterol efflux ability by macrophages. To test this hypothesis, we treated human THP-1 macrophages with total lipoproteins that were hydrolyzed by LPL and we found significantly reduced transcript levels for the cholesterol transporters ATP binding cassette transporter A1 (ABCA1), ABCG1, and scavenger receptor BI. These decreases were likely due to significant reductions for the nuclear receptors liver-X-receptor-α, peroxisome proliferator activated receptor (PPAR)-α, and PPAR-γ. We prepared a mixture of free fatty acids (FFA) that represented the ratios of FFA species within lipoprotein hydrolysis products, and we found that the FFA mixture also significantly reduced cholesterol transporters and nuclear receptors. Finally, we tested the efflux of cholesterol from THP-1 macrophages to apolipoprotein A-I, and we found that the treatment of THP-1 macrophages with the FFA mixture significantly attenuated cholesterol efflux. Overall, these data show that the FFA component of lipoprotein hydrolysis products generated by LPL may promote atherogenesis by inhibiting cholesterol efflux, which partially explains the pro-atherogenic role of macrophage LPL. PMID:25130461

  15. Integration of mild acid hydrolysis in γ-valerolactone/water system for enhancement of enzymatic saccharification from cotton stalk.

    PubMed

    Wu, Miao; Yan, Zhong Ya; Zhang, Xue Ming; Xu, Feng; Sun, Run Cang

    2016-01-01

    In this study, mild acid hydrolysis using γ-valerolactone (GVL)/water system integrated with enzymatic hydrolysis was carried out for the enhancement of enzymatic saccharification efficiency. The quantitative analysis of soluble carbohydrates and structural characterizations of solid residues were conducted. Results showed that the soluble carbohydrates in the water-phase were mainly composed of monomers and oligomers from xylose and glucose, while the contents of which were depended on the ratio of GVL to water. Moreover, the inhibitors were hardly detected due to the moderate pretreatment severity. Compared with the untreated feedstock, the yields of enzymatic hydrolysis from pretreated samples increased by two-fold with the mixture of 80/20 GVL/H2O. Combined with the amount of glucose (14.6%) dissolved in the water-phase, over 92.6% of glucose in cotton stalk was released and recovered. Based on the comprehensive analysis, treatment with GVL/H2O system provided us a more effective approach for sugar production from biomass. PMID:26476160

  16. Pitfalls in protein quantitation using acid-catalyzed O18 labeling: hydrolysis-driven deamidation

    PubMed Central

    Wang, Shunhai; Bobst, Cedric E.; Kaltashov, Igor A.

    2011-01-01

    Proteolysis combined with O18 labeling emerged recently as a powerful tool for quantitation of proteins for which suitable internal standards cannot be produced using molecular biology methods. Several recent reports suggested that acid-catalyzed O18 labeling may be superior to the commonly accepted enzymatic protocol, as it may allow more significant spacing between the isotopic clusters of labeled and unlabeled peptides, thereby eliminating signal interference and enhancing the quality of quantitation. However, careful examination of this procedure reveals that the results of protein quantitation assisted by acid-catalyzed O18 labeling are highly peptide-dependent. The inconsistency was found to be caused by deamidation of Asn, Gln and carbamidomethylated Cys residues during prolonged exposure of the proteolytic fragments to the acidic environment of the labeling reaction, which translates into a loss in signal for theses peptides. Taking deamidation into account leads to a significant improvement in the consistency of quantitation across a range of different proteolytic fragments. PMID:21819098

  17. Impact of hydrolysis conditions on the detection of mannuronic to guluronic acid ratio in alginate and its derivatives.

    PubMed

    Lu, Jiaojiao; Yang, Hai; Hao, Jie; Wu, Chengling; Liu, Li; Xu, Naiyu; Linhardt, Robert J; Zhang, Zhenqing

    2015-05-20

    Alginate is a linear and acidic polysaccharide, composed of (1 → 4) linked β-D-mannuronic acid (ManA) and α-L-guluronic acid (GulA). The ratio of ManA to GulA (M/G) is one of the most important factors for the application of alginate and its derivatives in various areas. In this work, a robust and accurate method was developed to analyze M/G using high-performance anion-exchange chromatography with pulsed amperometric detection (HPAEC-PAD). The impact of hydrolysis conditions on the release patterns of ManA and GulA from alginate and its derivatives was investigated. The release patterns of ManA and GulA need to be considered separately to obtain an accurate M/G. Several hydrolysis conditions were established that released ManA and GulA completely and maintained these saccharide residues intact. The proper M/G of alginates from different sources and its derivatives could then be calculated by integration of the corresponding ManA and GulA peaks. PMID:25817657

  18. Highly Amino Acid Selective Hydrolysis of Myoglobin at Aspartate Residues as Promoted by Zirconium(IV)-Substituted Polyoxometalates.

    PubMed

    Ly, Hong Giang T; Absillis, Gregory; Janssens, Rik; Proost, Paul; Parac-Vogt, Tatjana N

    2015-06-15

    SDS-PAGE/Edman degradation and HPLC MS/MS showed that zirconium(IV)-substituted Lindqvist-, Keggin-, and Wells-Dawson-type polyoxometalates (POMs) selectively hydrolyze the protein myoglobin at Asp-X peptide bonds under mildly acidic and neutral conditions. This transformation is the first example of highly sequence selective protein hydrolysis by POMs, a novel class of protein-hydrolyzing agents. The selectivity is directed by Asp residues located on the surface of the protein and is further assisted by electrostatic interactions between the negatively charged POMs and positively charged surface patches in the vicinity of the cleavage site. PMID:25950869

  19. Enhanced enzymatic hydrolysis of poplar bark by combined use of gamma ray and dilute acid for bioethanol production

    NASA Astrophysics Data System (ADS)

    Chung, Byung Yeoup; Lee, Jae Taek; Bai, Hyoung-Woo; Kim, Ung-Jin; Bae, Hyeun-Jong; Gon Wi, Seung; Cho, Jae-Young

    2012-08-01

    Pretreatment of poplar bark with a combination of sulfuric acid (3%, w/w, H2SO4) and gamma irradiation (0-1000 kGy) was performed in an attempt to enhance enzymatic hydrolysis for bioethanol production. The yields of reducing sugar were slightly increased with an increasing irradiation dose, ranging from 35.4% to 51.5%, with a 56.1% reducing sugar yield observed after dilute acid pretreatment. These results clearly showed that soluble sugars were released faster and to a greater extent in dilute acid-pretreated poplar bark than in gamma irradiation-pretreated bark. When combined pretreatment was carried out, a drastic increase in reducing sugar yield (83.1%) was found compared with individual pretreatment, indicating the possibility of increasing the convertibility of poplar bark following combined pretreatment. These findings are likely associated with cellulose crystallinity, lignin modification, and removal of hemicelluloses.

  20. Substituent Effects on the Hydrolysis of p-Substituted Benzonitriles in Sulfuric Acid Solutions at (25.0± 0.1) °C

    NASA Astrophysics Data System (ADS)

    Abbas, Khamis A.

    2008-09-01

    The rate constants of the hydrolysis of p-substituted benzonitriles with sulfuric acid solutions (18.2 M to 10 M) have been determined spectrophotometrically at (25.1±0.1) °C. It was found that the catalytic activity of sulfuric acid was strongly inhibited by water. The logarithms of the observed rate constants were correlated with different substituent inductive (localized) and resonance (delocalized) constants. The results of the correlation studies indicated that the rate-determining step of the hydrolysis of benzonitriles in 18.2 M sulfuric acid was the addition of a nucleophile, and the hydrolysis was clearly enhanced by the electron-withdrawing inductive effect, while the rate-determining step of the hydrolysis of p-substituted benzonitriles in 10.0 M sulfuric acid was most probably the protonation of benzonitriles, and the rate constants increased by both electron-donating resonance and inductive effects. A mixture of the two mechanisms most probably occurred in 15.3 to 17.0 M sulfuric acid. HSO4 - rather thanwater most probably acted as nucleophile in the hydrolysis of benzonitriles especially at high concentrations of sulfuric acid solutions.

  1. The hydrolysis of polyimides

    NASA Technical Reports Server (NTRS)

    Hoagland, P. D.; Fox, S. W.

    1973-01-01

    Thermal polymerization of aspartic acid produces a polysuccinimide (I), a chain of aspartoyl residues. An investigation was made of the alkaline hydrolysis of the imide rings of (I) which converts the polyimide to a polypeptide. The alkaline hydrolysis of polyimides can be expected to be kinetically complex due to increasing negative charge generated by carboxylate groups. For this reason, a diimide, phthaloyl-DL-aspartoyl-beta-alanine (IIA) was synthesized for a progressive study of the hydrolysis of polyimides. In addition, this diimide (IIA) can be related to thalidomide and might be expected to exhibit similar reactivity during hydrolysis of the phthalimide ring.

  2. Acid-Labile Thermoresponsive Copolymers That Combine Fast pH-Triggered Hydrolysis and High Stability under Neutral Conditions.

    PubMed

    Zhang, Qilu; Hou, Zhanyao; Louage, Benoit; Zhou, Dingying; Vanparijs, Nane; De Geest, Bruno G; Hoogenboom, Richard

    2015-09-01

    Biodegradable polymeric materials are intensively used in biomedical applications. Of particular interest for drug-delivery applications are polymers that are stable at pH 7.4, that is, in the blood stream, but rapidly hydrolyze under acidic conditions, such as those encountered in the endo/lysosome or the tumor microenvironment. However, an increase in the acidic-degradation rate of acid-labile groups goes hand in hand with higher instability of the polymer at pH 7.4 or during storage, thus posing an intrinsic limitation on fast degradation under acidic conditions. Herein, we report that a combination of acid-labile dimethyldioxolane side chains and hydroxyethyl side chains leads to acid-degradable thermoresponsive polymers that are quickly hydrolyzed under slightly acidic conditions but stable at pH 7.4 or during storage. We ascribe these properties to high hydration of the hydroxy-containing collapsed polymer globules in conjunction with autocatalytic acceleration of the hydrolysis reactions by the hydroxy groups. PMID:26212481

  3. {beta}-Secondary and solvent deuterium kinetic isotope effects and the mechanisms of base- and acid-catalyzed hydrolysis of penicillanic acid

    SciTech Connect

    Deraniyagala, S.A.; Adediran, S.A.; Pratt, R.F.

    1995-03-24

    {beta}-Secondary and solvent deuterium kinetic isotope effects have been determined at 25 {degrees}C for the alkaline and acid-catalyzed hydrolysis of penicillanic acid. In order to determine the former isotope effect, [6,6-{sup 2}H{sub 2}]dideuteriopenicillanic acid has been synthesized. In alkaline solution, the former isotope effect was found to be 0.95 {plus_minus} 0.01. These values support the B{sub AC}2 mechanism of hydrolysis with rate-determining formation of the tetrahedral intermediate that has been proposed for other {beta}-lactams. The measured {beta}-secondary kinetic isotope for the acid-catalyzed reaction was 1.00 {plus_minus} 0.01. The data indicates that a likely pathway of acid-catalyzed hydrolysis would be that of an A{sub AC}1 mechanism with an intermediate acylium ion. If this were so, the calculated {beta}-secondary isotope effect per hydrogen coplanar with the breaking C-N bond and corrected for the inductive effect of deuterium would be 1.06 {plus_minus} 0.01. This suggests an early A{sub AC}1 transition state, which would be reasonable in this case because of destabilization of the N-protonated amide with respect to the acylium ion because of ring strain. The absence of specific participation by solvent in the transition state, as would be expected of an A{sub AC}1 but not an associative mechanism, is supported by the strongly inverse solvent deuterium kinetic isotope effect of 0.25 {plus_minus} 0.00 in 1 M HCl and 0.22 {plus_minus} 0.01 in 33.3 wt % H{sub 2}SO{sub 4}. 1 fig., 3 tabs.

  4. Effect of acid hydrolysis combined with heat moisture treatment on structure and physicochemical properties of corn starch.

    PubMed

    Sun, Qingjie; Zhu, Xiaolei; Si, Fumei; Xiong, Liu

    2015-01-01

    Modification of starch led to new products with new desirable properties. Corn starch samples modified by acid hydrolysis combined with heat moisture treatment (AH-HMT) were made by changing pH, moisture content and treated temperature. After modification, swelling power at temperature higher than 75 °C of corn starches decreased while solubility of the starches increased. After AH-HMT, pasting temperature (PT) of all treated starch samples increased. But lower peak viscosity (PKV), trough viscosity (TV) and break down (BD) of most treated starch samples were observed. AH-HMT increased the gel hardness of all treated starches. And the biggest hardness of modified starch gel was 148.419 g, improving 93.471 g compared with native starch gel. The melting temperatures (To, Tp, Tc) of modified starch increased, but the melting range and △H decreased. The X-ray pattern remained practically unchanged with or without AH-HMT. Acid hydrolysis combined with heat moisture treatment (AH-HMT) improved the functional properties of corn starch. PMID:25593372

  5. Three-step biocatalytic reaction using whole cells for efficient production of tyramine from keratin acid hydrolysis wastewater.

    PubMed

    Zhang, Hongjuan; Wei, Yu; Lu, Yang; Wu, Siping; Liu, Qian; Liu, Junzhong; Jiao, Qingcai

    2016-02-01

    Tyramine has been paid more attention in recent years as a significant metabolite of tyrosine and catecholamine drug and an intermediate of medicinal material and some drugs. In this study, an effective, green, and three-step biocatalytic synthesis method for production of tyramine starting from serine in keratin acid hydrolysis wastewater was developed and investigated. Serine deaminase from Escherichia coli was first combined with tyrosine phenol-lyase from Citrobacter koseri, to convert L-serine to L-tyrosine. L-Tyrosine can then be decarboxylated to tyramine by tyrosinede carboxylase from Lactobacillus brevis. All these enzymes originated from recombinant whole cells. Serine deaminaseand tyrosine phenol-lyase could efficiently convert L-serine in wastewater to L-tyrosine at pH 8.0, 37 °C, and Triton X-100 of 0.04% when tyrosine phenol-lyase and its corresponding substrates were sequentially added. Tyrosine conversion rate reached 98 % by L-tyrosine decarboxylase. In scale-up study, the conversion yield of L-serine in wastewater to tyrosine was up to 89 %. L-Tyrosine was decarboxylated to tyramine with a high yield 94 %. Tyramine hydrochloride was obtained with a total yield 84 %. This study has provided an efficient way of recycling keratin acid hydrolysis wastewater to produce tyramine. PMID:26476652

  6. Characterization of degradation products of ivabradine by LC-HR-MS/MS: a typical case of exhibition of different degradation behaviour in HCl and H2SO4 acid hydrolysis.

    PubMed

    Patel, Prinesh N; Borkar, Roshan M; Kalariya, Pradipbhai D; Gangwal, Rahul P; Sangamwar, Abhay T; Samanthula, Gananadhamu; Ragampeta, Srinivas

    2015-02-01

    A validated stability-indicating HPLC method was established, and comprehensive stress testing of ivabradine, a cardiotonic drug, was carried out as per ICH guidelines. Ivabradine was subjected to acidic, basic and neutral hydrolysis, oxidation, photolysis and thermal stress conditions, and the resulting degradation products were investigated by LC-PDA and LC-HR-MS/MS. The drug was found to degrade in acid and base hydrolysis. An efficient and selective stability assay method was developed on Phenomenex Luna C18 (250 × 4.6 mm, 5.0 µm) column using ammonium formate (10 mM, pH 3.0) and acetonitrile as mobile phase at 30 °C in gradient elution mode. The flow rate was 0.7 ml/min and detection wavelength was 286 nm. A total of five degradation products (I-1 to I-5) were identified and characterized by LC-HR-MS/MS in combination with accurate mass measurements. The drug exhibited different degradation behaviour in HCl and H2SO4 hydrolysis conditions. It is a unique example where two of the five degradation products in HCl hydrolysis were absent in H2SO4 acid hydrolysis. The present study provides guidance to revise the stress test for the determination of inherent stability of drugs containing lactam moiety under hydrolytic conditions. Most probable mechanisms for the formation of degradation products have been proposed on the basis of a comparison of the fragmentation pattern of the drug and its degradation products. In silico toxicity revealed that the degradation products (I-2 to I-5) were found to be severe irritants in case of ocular irritancy. The analytical assay method was validated with respect to specificity, linearity, range, precision, accuracy and robustness. PMID:25800016

  7. Forms of acid hydrolysis and gley formation and their role in the development of light-colored acid eluvial (Podzolic) horizons

    NASA Astrophysics Data System (ADS)

    Zaidel'Man, F. R.

    2010-04-01

    Nowadays, three processes, namely lessivage, acid hydrolysis, and gleying, are considered as responsible for the development of loamy and clayey podzolic soils. However, as was shown earlier, lessivage is not obligatory for their origin. In view of assessing the reasons for the formation of light-colored acid eluvial horizons, this article deals with the role of acid hydrolysis under aerobic conditions against the background of a percolative water regime and of two forms of gleying in the development of the horizons mentioned above. One form of gleying occurs under permanent anaerobic conditions against the background of a stagnant water regime; the other one is formed under pulsating anaerobic-aerobic conditions against the background of a stagnant-percolative water regime. As a result, three large genetically individual groups of soils are formed: nondifferentiated brown and gley, and differentiated podzolic soils on different parent rocks. The two latter forms of gleying are identical in their effects on the mineral substrates. They cause the iron removal from the soils. Among the three processes considered, the last one (gleying under a stagnant-percolative water regime) is the single reason for the leaching of most of the metals, the formation of light-colored acid eluvial horizons and their clay depletion, and for the differentiation of the soil profile.

  8. A rapid hydrolysis method and DABS-Cl derivatization for complete amino acid analysis of octreotide acetate by reversed phase HPLC.

    PubMed

    Akhlaghi, Yousef; Ghaffari, Solmaz; Attar, Hossein; Alamir Hoor, Amir

    2015-11-01

    Octreotide as a synthetic cyclic octapeptide is a somatostatin analog with longer half-life and more selectivity for inhibition of the growth hormone. The acetate salt of octreotide is currently used for medical treatment of somatostatin-related disorders such as endocrine and carcinoid tumors, acromegaly, and gigantism. Octreotide contains both cysteine and tryptophan residues which make the hydrolysis part of its amino acid analysis procedure very challenging. The current paper introduces a fast and additive-free method which preserves tryptophan and cysteine residues during the hydrolysis. Using only 6 M HCl, this hydrolysis process is completed in 30 min at 150 °C. This fast hydrolysis method followed by pre-column derivatization of the released amino acids with 4-N,N-dimethylaminoazobenzene-4'-sulfonyl chloride (DABS-Cl) which takes only 20 min, makes it possible to do the complete amino acid analysis of an octreotide sample in a few hours. The highly stable-colored DABS-Cl derivatives can be detected in 436 nm in a reversed phase chromatographic system, which eliminates spectral interferences to a great extent. The amino acid analysis of octreotide acetate including hydrolysis, derivatization, and reversed phase HPLC determination was validated according to International Conference of Harmonization (ICH) guidelines. PMID:26002809

  9. Further studies on the glycerol teichoic acid of walls of Staphylococcus lactis I3. Location of the phosphodiester groups and their susceptibility to hydrolysis with alkali

    PubMed Central

    Archibald, A. R.; Baddiley, J.; Heckels, J. E.; Heptinstall, S.

    1971-01-01

    1. The teichoic acid from walls of Staphylococcus lactis I3 is readily degraded in dilute alkali. 2. Degradation proceeds by selective hydrolysis of that phosphodiester group attached to an alcoholic hydroxyl group of the N-acetylglucosamine and gives a repeating unit in high yield. 3. Further studies on a different repeating unit isolated by partial acid hydrolysis have shown that the glycerol diphosphate is attached to the 4-hydroxyl group of the N-acetylglucosamine and not to the 3-hydroxyl group as was proposed earlier. 4. The susceptibility towards hydrolysis by alkali of other structural types of teichoic acid has been examined and found to vary markedly according to their structure. PMID:5158917

  10. 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

    2016-05-01

    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. PMID:26876990

  11. Effect of surfactant on hydrolysis products accumulation and short-chain fatty acids (SCFA) production during mesophilic and thermophilic fermentation of waste activated sludge: kinetic studies.

    PubMed

    Zhang, Peng; Chen, Yinguang; Zhou, Qi

    2010-09-01

    In the presence of surfactant sodium dodecylbenzene sulfonate (SDBS) the hydrolysis products accumulation and the short-chain fatty acids (SCFA) production during waste activated sludge fermentation under mesophilic and thermophilic conditions was compared with that at room temperature. In order to understand the mechanism of significant amounts of mesophilic and thermophilic hydrolysis products and SCFA observed in the presence of surfactant, the kinetic models at different SDBS dosages were developed. It was found that SDBS increased the mesophilic and thermophilic hydrolysis rate significantly, and the maximum specific utilization of hydrolysis products increased at low SDBS and decreased at high one. However, the observed maximum specific utilization of SCFA decreased seriously with SDBS increase. In the presence of SDBS the decay rate of acidogenic bacteria not only was lower than that in the absence of SDBS but decreased with the increase of SDBS under either mesophilic or thermophilic conditions. PMID:20409704

  12. An integrated process for the production of platform chemicals and diesel miscible fuels by acid-catalyzed hydrolysis and downstream upgrading of the acid hydrolysis residues with thermal and catalytic pyrolysis.

    PubMed

    Girisuta, Buana; Kalogiannis, Konstantinos G; Dussan, Karla; Leahy, James J; Hayes, Michael H B; Stefanidis, Stylianos D; Michailof, Chrysa M; Lappas, Angelos A

    2012-12-01

    This study evaluates an integrated process for the production of platform chemicals and diesel miscible biofuels. An energy crop (Miscanthus) was treated hydrothermally to produce levulinic acid (LA). Temperatures ranging between 150 and 200 °C, sulfuric acid concentrations 1-5 wt.% and treatment times 1-12 h were applied to give different combined severity factors. Temperatures of 175 and 200 °C and acid concentration of 5 wt.% were found to be necessary to achieve good yield (17 wt.%) and selectivities of LA while treatment time did not have an effect. The acid hydrolysis residues were characterized for their elemental, cellulose, hemicellulose and lignin contents, and then tested in a small-scale pyrolyzer using silica sand and a commercial ZSM-5 catalyst. Milder pretreatment yielded more oil (43 wt.%) and oil O(2) (37%) while harsher pretreatment and catalysis led to more coke production (up to 58 wt.%), less oil (12 wt.%) and less oil O(2) (18 wt.%). PMID:23073094

  13. In situ pressurized biphase acid hydrolysis, a promising approach to produce bioactive diosgenin from the tubers of Dioscorea Zingiberensis

    PubMed Central

    Yang, Huan; Yin, Hua-wu; Wang, Xue-wei; Li, Zi-hao; Shen, Yu-ping; Jia, Xiao-bin

    2015-01-01

    Background: The tubers of Dioscorea zingiberensis, is the most favorable plant material for the production of diosgenin, an important bioactive steroidal sapogenin and requisite precursor of cortin, contraceptive and sex hormone, which is the only desired product after steroidal saponins from the tubers are hydrolyzed. Objective: A novel technology, in situ pressurized biphase acid hydrolysis was constructed for the first time to simplify extraction process, increase extraction yield and decrease the consumption of mineral acids. Materials and Methods: The method developed in this study has been optimized and verified through orthogonal design for experiments, in which the effect and their significance of four factors including molarity of acid, temperature, extraction duration and sample quantity have been investigated. Then, the comparison was conducted among the newly developed method and other reported methods. The diosgenin was also isolated by column chromatography, followed by mass spectrometry and nuclear magnetic resonance analysis for structural confirmation. Results: It was found that temperature is the factor of the most influence and the highest extraction yield at 2.21% has been achieved while the hydrolysis was performed at 140°C for 1.5 h in 0.20M H2SO4 solution with petroleum ether under an uncontrolled pressurized condition. And, compared to the others, the increment in the extraction yield of new method was 20.8 ~ 74.0%, and the consumption of H2SO4 was reduced by 17 times at most. Conclusion: This method is a much cleaner and more efficient approach for extraction of diosgenin from the tubers, and is promising to be applied in pharmaceutical industry. PMID:26246743

  14. AMP kinase activation with AICAR further increases fatty acid oxidation and blunts triacylglycerol hydrolysis in contracting rat soleus muscle

    PubMed Central

    Smith, Angela C; Bruce, Clinton R; Dyck, David J

    2005-01-01

    Muscle contraction increases glucose uptake and fatty acid (FA) metabolism in isolated rat skeletal muscle, due at least in part to an increase in AMP-activated kinase activity (AMPK). However, the extent to which AMPK plays a role in the regulation of substrate utilization during contraction is not fully understood. We examined the acute effects of 5-aminoimidazole-4-carboxamide riboside (AICAR; 2 mm), a pharmacological activator of AMPK, on FA metabolism and glucose oxidation during high intensity tetanic contraction in isolated rat soleus muscle strips. Muscle strips were exposed to two different FA concentrations (low fatty acid, LFA, 0.2 mm; high fatty acid, HFA, 1 mm) to examine the role that FA availability may play in both exogenous and endogenous FA metabolism with contraction and AICAR. Synergistic increases in AMPK α2 activity (+45%; P < 0.05) were observed after 30 min of contraction with AICAR, which further increased exogenous FA oxidation (LFA: +71%, P < 0.05; HFA: +46%, P < 0.05) regardless of FA availability. While there were no changes in triacylglycerol (TAG) esterification, AICAR did increase the ratio of FA partitioned to oxidation relative to TAG esterification (LFA: +65%, P < 0.05). AICAR significantly blunted endogenous TAG hydrolysis (LFA: −294%, P < 0.001; HFA: −117%, P < 0.05), but had no effect on endogenous oxidation rates, suggesting a better matching between TAG hydrolysis and subsequent oxidative needs of the muscle. There was no effect of AICAR on the already elevated rates of glucose oxidation during contraction. These results suggest that FA metabolism is very sensitive to AMPK α2 stimulation during contraction. PMID:15774529

  15. Acid Hydrolysis and Molecular Density of Phytoglycogen and Liver Glycogen Helps Understand the Bonding in Glycogen α (Composite) Particles

    PubMed Central

    Powell, Prudence O.; Sullivan, Mitchell A.; Sheehy, Joshua J.; Schulz, Benjamin L.; Warren, Frederick J.; Gilbert, Robert G.

    2015-01-01

    Phytoglycogen (from certain mutant plants) and animal glycogen are highly branched glucose polymers with similarities in structural features and molecular size range. Both appear to form composite α particles from smaller β particles. The molecular size distribution of liver glycogen is bimodal, with distinct α and β components, while that of phytoglycogen is monomodal. This study aims to enhance our understanding of the nature of the link between liver-glycogen β particles resulting in the formation of large α particles. It examines the time evolution of the size distribution of these molecules during acid hydrolysis, and the size dependence of the molecular density of both glucans. The monomodal distribution of phytoglycogen decreases uniformly in time with hydrolysis, while with glycogen, the large particles degrade significantly more quickly. The size dependence of the molecular density shows qualitatively different shapes for these two types of molecules. The data, combined with a quantitative model for the evolution of the distribution during degradation, suggest that the bonding between β into α particles is different between phytoglycogen and liver glycogen, with the formation of a glycosidic linkage for phytoglycogen and a covalent or strong non-covalent linkage, most probably involving a protein, for glycogen as most likely. This finding is of importance for diabetes, where α-particle structure is impaired. PMID:25799321

  16. The photochemical production of organic nitrates from α-pinene and loss via acid-dependent particle phase hydrolysis

    NASA Astrophysics Data System (ADS)

    Rindelaub, Joel D.; McAvey, Kevin M.; Shepson, Paul B.

    2015-01-01

    The hydroxyl radical oxidation of α-pinene under high NOx conditions was studied in a photochemical reaction chamber to investigate organic nitrate (RONO2) production and fate between the gas and particle phases. We report an organic nitrate yield of 26 ± 7% from the oxidation of this monoterpene in the presence of nitric oxide (NO). However, the apparent organic nitrate yield was found to be highly dependent on both chamber relative humidity (RH) and seed aerosol acidity, likely as a result of particle phase hydrolysis. The particle phase loss of organic nitrates is believed to increase the gas to particle partitioning within the system, leading to decreased RONO2 yields in both the gas and particle phases at elevated RH and an apparent non-equilibrium partitioning mechanism. The hydrolysis of particle phase organic nitrates in this study, starting at low chamber relative humidity, implies that aerosol partitioning of organic nitrates may be an important sink for atmospheric NOx and may have a significant impact on regional air quality.

  17. Progressing batch hydrolysis process

    DOEpatents

    Wright, J.D.

    1985-01-10

    A progressive batch hydrolysis process is disclosed for producing sugar from a lignocellulosic feedstock. It comprises passing a stream of dilute acid serially through a plurality of percolation hydrolysis reactors charged with feed stock, at a flow rate, temperature and pressure sufficient to substantially convert all the cellulose component of the feed stock to glucose. The cooled dilute acid stream containing glucose, after exiting the last percolation hydrolysis reactor, serially fed through a plurality of pre-hydrolysis percolation reactors, charged with said feedstock, at a flow rate, temperature and pressure sufficient to substantially convert all the hemicellulose component of said feedstock to glucose. The dilute acid stream containing glucose is cooled after it exits the last prehydrolysis reactor.

  18. Response surface optimization of corn stover pretreatment using dilute phosphoric acid for enzymatic hydrolysis and ethanol production.

    PubMed

    Avci, Ayse; Saha, Badal C; Dien, Bruce S; Kennedy, Gregory J; Cotta, Michael A

    2013-02-01

    Dilute H(3)PO(4) (0.0-2.0%, v/v) was used to pretreat corn stover (10%, w/w) for conversion to ethanol. Pretreatment conditions were optimized for temperature, acid loading, and time using central composite design. Optimal pretreatment conditions were chosen to promote sugar yields following enzymatic digestion while minimizing formation of furans, which are potent inhibitors of fermentation. The maximum glucose yield (85%) was obtained after enzymatic hydrolysis of corn stover pretreated with 0.5% (v/v) acid at 180°C for 15min while highest yield for xylose (91.4%) was observed from corn stover pretreated with 1% (v/v) acid at 160°C for 10min. About 26.4±0.1g ethanol was produced per L by recombinant Escherichia coli strain FBR5 from 55.1±1.0g sugars generated from enzymatically hydrolyzed corn stover (10%, w/w) pretreated under a balanced optimized condition (161.81°C, 0.78% acid, 9.78min) where only 0.4±0.0g furfural and 0.1±0.0 hydroxylmethyl furfural were produced. PMID:23334017

  19. Dissociation and hydrolysis of ammonia-borane with solid acids and carbon dioxide: An efficient hydrogen generation system

    NASA Astrophysics Data System (ADS)

    Chandra, Manish; Xu, Qiang

    Pure hydrogen generation under mild conditions in a controllable way is important for portable devices. Recently, we have found that an aq. ammonia-borane (NH 3BH 3) solution is a potential hydrogen source with noble metal catalysts. For practical use, the development of a low-cost, efficient and safe system is desired. In this study, we found that solid acids such as cation exchange resins and zeolites, which are low-cost and safe, also exhibit high activities for the dissociation and hydrolysis of NH 3BH 3 to generate hydrogen with an H 2 to NH 3BH 3 ratio up to 3.0 at room temperature. The reaction rate depends on the type of solid acid. Especially, Dowex and Amberlyst, the two low-cost solid acids often used as catalysts in a variety of reactions, exhibit reaction kinetics higher than the noble metal catalysts. Carbon dioxide is also active as an acid for this reaction. The reaction products in solution have been identified by 11B NMR, and the evolved gases have been analyzed by mass spectrometry which indicates high purity hydrogen. This new system may have a high potential for application in fuel cells.

  20. N2O5 Hydrolysis on a Liquid Surface Coated by Long Chain Fatty Acid and Alcohol monolayers

    NASA Astrophysics Data System (ADS)

    Cosman, L. M.; Knopf, D. A.; Bertram, A. K.

    2006-12-01

    Heterogeneous reactions between aerosol particles and gas phase species play a crucial role in the atmosphere and can influence the composition of the atmosphere. Heterogeneous reactions of N2O5 with aqueous particles have a significant effect on the global NOx budget, which influences tropospheric concentrations of O3 and OH. The hydrolysis of N2O5 on sulfuric acid particles has been studied extensively and is found to be very efficient. However, surface active organic molecules are common constituents of aqueous atmospheric particles and their effect on heterogeneous reactions is still poorly understood. Previous studies have shown that organic coatings on atmospheric droplets can change the uptake of gaseous species substantially. It is important to know heterogeneous reaction rates for the hydrolysis of N2O5 on organic coated aqueous particles in order to accurately predict the oxidative capacity of the troposphere. Here we present the investigation of heterogeneous reactions of N2O5 on aqueous sulfuric acid coated by an organic monolayer. A new parallel plate flow reactor (PPFR) has been developed to allow heterogeneous reactions to be studied on a planar aqueous surface coated with an organic monolayer. In addition, a theoretical framework has been developed to derive heterogeneous kinetics taking into consideration the effects of diffusion. The PPFR coupled to a chemical ionization mass spectrometer was employed to measure reactive uptake coefficients for N2O5 on aqueous H2SO4 solutions coated with long chain fatty acids and alcohols. One key feature of this new technique is the ability to characterize the monolayer prior to kinetic measurements to determine the surface pressure and packing density of the organic molecules on the surface. Monolayers of octadecanol, hexadecanol, stearic acid, and phytanic acid were studied on 60 wt % H2SO4 solutions at 274 K. It was found that a monolayer of octadecanol reduced the uptake coefficient of N2O5 with H2SO4 by

  1. Enzymatic hydrolysis of cuttlefish (Sepia officinalis) and sardine (Sardina pilchardus) viscera using commercial proteases: effects on lipid distribution and amino acid composition.

    PubMed

    Kechaou, Emna Soufi; Dumay, Justine; Donnay-Moreno, Claire; Jaouen, Pascal; Gouygou, Jean-Paul; Bergé, Jean-Pascal; Amar, Raja Ben

    2009-02-01

    Total lipid and phospholipid recovery as well as amino acid quality and composition from cuttlefish (Sepia officinalis) and sardine (Sardina pilchardus) were compared. Enzymatic hydrolyses were performed using the three proteases Protamex, Alcalase, and Flavourzyme by the pH-stat method (24 h, pH 8, 50 degrees C). Three fractions were generated: an insoluble sludge, a soluble aqueous phase, and an oily phase. For each fraction, lipids, phospholipids, and proteins were quantified. Quantitative and qualitative analyses of the raw material and hydrolysates were performed. The degree of hydrolysis (DH) for cuttlefish viscera was 3.2% using Protamex, 6.8% using Flavourzyme, and 7% using Alcalase. DH for sardine viscera was 1.9% (using Flavourzyme), 3.1% (using Protamex) and 3.3% (using Alcalase). Dry matter yields of all hydrolysis reactions increased in the aqueous phases. Protein recovery following hydrolysis ranged from 57.2% to 64.3% for cuttlefish and 57.4% to 61.2% for sardine. Tissue disruption following protease treatment increased lipid extractability, leading to higher total lipid content after hydrolysis. At least 80% of the lipids quantified in the raw material were distributed in the liquid phases for both substrates. The hydrolysed lipids were richer in phospholipids than in the lipids extracted by classical chemical extraction, especially after Flavourzyme hydrolysis for cuttlefish and Alcalase hydrolysis for sardine. The total amino acid content differed according to the substrate and the enzyme used. However, regardless of the raw material or the protease used, hydrolysis increased the level of essential amino acids in the hydrolysates, thereby increasing their potential nutritional value for feed products. PMID:19217554

  2. Theoretical study of the hydrolysis of ethyl benzoate in acidic aqueous solution using the QM/MC/FEP method

    NASA Astrophysics Data System (ADS)

    Kaweetirawatt, Thanayuth; Kokita, Yohei; Iwai, Shiho; Sumimoto, Michinori; Hori, Kenji

    2012-09-01

    The hydrolysis of ethyl benzoate in acidic condition was theoretically studied for models with two (2W) or three water (3WA) molecules at the B3LYP/6-311++G(d,p) levels of theory. Activation free energy of solvation in aqueous solution (ΔG‡cal) was calculated using the QM/MC/FEP method. The value of the 2W model in aqueous solution was calculated to be smaller by more than 5.0 kcal mol-1 than the observed value (26.0 kcal mol-1 at 298 K). The position of the third water molecule in the 3WA model plays an essential role in producing the ΔG‡cal value (26.4 kcal mol-1) consistent with the experimental value.

  3. Effect of carboxylic acid adsorption on the hydrolysis and sintered properties of aluminum nitride powder

    SciTech Connect

    Egashira, Makoto; Shimizu, Yasuhiro; Takao, Yuji; Yamaguchi, Ryoji; Ishikawa, Yasuhiro . Dept. of Materials Science and Engineering)

    1994-07-01

    To suppress the reactivity of AlN powder with water, chemical surface modification with carboxylic acids has been investigated. It was found that the chemical stability of AlN powder increased as the number of carbon atoms in carboxylic acids used for the surface treatment increased. Among the carboxylic acids tested, stearic acid was the most promising from the viewpoint of the chemical stability of the treated powder and the thermal conductivity of the sintered ceramics prepared by cold isostatic pressing and pressureless sintering.

  4. The effect of several organic acids on phytate phosphorus hydrolysis in broiler chicks.

    PubMed

    Liem, A; Pesti, G M; Edwards, H M

    2008-04-01

    Supplementation of some organic acids to a P-deficient diet has been shown to improve phytate P utilization. Two experiments were conducted from 0 to 16 d in battery brooders to determine the effect of various organic acids supplementation on phytate P utilization. In both experiments, birds were fed P-deficient corn and soybean meal-based diets. In experiment 1, citric acid, malic acid, fumaric acid, and EDTA were supplemented. Experiment 2 had a 2 x 2 factorial design with 2 sources of Met, 2-hydroxy-4-(methylthio) butanoic acid (HMB) and dl-Met, with or without 500 U/kg of phytase. In experiment 1, the addition of citric, malic, and fumaric acids increased percentage of bone ash, but only the effect of citric acid was significant. The addition of citric and malic acids also significantly increased the retention of P and phytate P (P<0.05). In experiment 2, the addition of phytase to the diet significantly increased 16-d BW gain, feed intake, percentage of bone ash, milligrams of bone ash, phytate P disappearance, and decreased the incidence of P-deficiency rickets. Methionine source did not affect 16-d BW gain, feed intake, feed efficiency, milligrams of bone ash, or P rickets incidence. However, the birds fed HMB had a higher percentage of bone ash and phytate P disappearance compared with the groups fed dl-Met only when phytase was added to the diets. The additions of citric acid and HMB improved phytate P utilization. However, the reason why some organic acids are effective whereas others are not is not apparent. PMID:18339989

  5. Stereochemical features of the hydrolysis of 9,10-epoxystearic acid catalysed by plant and mammalian epoxide hydrolases.

    PubMed Central

    Summerer, Stephan; Hanano, Abdulsamie; Utsumi, Shigeru; Arand, Michael; Schuber, Francis; Blée, Elizabeth

    2002-01-01

    cis-9,10-epoxystearic acid was used as a tool to probe the active sites of epoxide hydrolases (EHs) of mammalian and plant origin. We have compared the stereochemical features of the hydrolysis of this substrate catalysed by soluble and membrane-bound rat liver EHs, by soluble EH (purified to apparent homogeneity) obtained from maize seedlings or celeriac roots, and by recombinant soybean EH expressed in yeast. Plant EHs were found to differ in their enantioselectivity, i.e. their ability to discriminate between the two enantiomers of 9,10-epoxystearic acid. For example, while the maize enzyme hydrated both enantiomers at the same rate, the EH from soybean exhibited very high enantioselectivity in favour of 9R,10S-epoxystearic acid. This latter enzyme also exhibited a strict stereoselectivity, i.e. it hydrolysed the racemic substrate with a very high enantioconvergence, yielding a single chiral diol product, threo-9R,10R-dihydroxystearic acid. Soybean EH shared these distinctive stereochemical features with the membrane-bound rat liver EH. The stereochemical outcome of these enzymes probably results from a stereoselective attack by the nucleophilic residue on the oxirane ring carbon having the (S)-configuration, leading to the presumed (in plant EH) covalent acyl-enzyme intermediate. In sharp contrast, the reactions catalysed by cytosolic rat liver EH exhibited a complete absence of enantioselectivity and enantioconvergence; this latter effect might be ascribed to a regioselective formation of the acyl-enzyme intermediate involving C-10 of 9,10-epoxystearic acid, independent of its configuration. Thus, compared with soybean EH, the active site of rat liver soluble EH displays a very distinct means of anchoring the oxirane ring of the fatty acid epoxides, and therefore appears to be a poor model for mapping the catalytic domain of plant EHs. PMID:12020347

  6. Stereochemical features of the hydrolysis of 9,10-epoxystearic acid catalysed by plant and mammalian epoxide hydrolases.

    PubMed

    Summerer, Stephan; Hanano, Abdulsamie; Utsumi, Shigeru; Arand, Michael; Schuber, Francis; Blée, Elizabeth

    2002-09-01

    cis-9,10-epoxystearic acid was used as a tool to probe the active sites of epoxide hydrolases (EHs) of mammalian and plant origin. We have compared the stereochemical features of the hydrolysis of this substrate catalysed by soluble and membrane-bound rat liver EHs, by soluble EH (purified to apparent homogeneity) obtained from maize seedlings or celeriac roots, and by recombinant soybean EH expressed in yeast. Plant EHs were found to differ in their enantioselectivity, i.e. their ability to discriminate between the two enantiomers of 9,10-epoxystearic acid. For example, while the maize enzyme hydrated both enantiomers at the same rate, the EH from soybean exhibited very high enantioselectivity in favour of 9R,10S-epoxystearic acid. This latter enzyme also exhibited a strict stereoselectivity, i.e. it hydrolysed the racemic substrate with a very high enantioconvergence, yielding a single chiral diol product, threo-9R,10R-dihydroxystearic acid. Soybean EH shared these distinctive stereochemical features with the membrane-bound rat liver EH. The stereochemical outcome of these enzymes probably results from a stereoselective attack by the nucleophilic residue on the oxirane ring carbon having the (S)-configuration, leading to the presumed (in plant EH) covalent acyl-enzyme intermediate. In sharp contrast, the reactions catalysed by cytosolic rat liver EH exhibited a complete absence of enantioselectivity and enantioconvergence; this latter effect might be ascribed to a regioselective formation of the acyl-enzyme intermediate involving C-10 of 9,10-epoxystearic acid, independent of its configuration. Thus, compared with soybean EH, the active site of rat liver soluble EH displays a very distinct means of anchoring the oxirane ring of the fatty acid epoxides, and therefore appears to be a poor model for mapping the catalytic domain of plant EHs. PMID:12020347

  7. Titan's Primordial Soup: Formation of Amino Acids via Low-Temperature Hydrolysis of Tholins

    NASA Astrophysics Data System (ADS)

    Neish, Catherine D.; Somogyi, Árpád; Smith, Mark A.

    2010-04-01

    Titan organic haze analogues, or "tholins," produce biomolecules when hydrolyzed at low temperature over long timescales. By using a combination of high-resolution mass spectroscopy and tandem mass spectrometry fragmentation techniques, four amino acids were identified in a tholin sample that had been hydrolyzed in a 13 wt % ammonia-water solution at 253 ± 1 K and 293 ± 1 K for 1 year. These four species have been assigned as the amino acids asparagine, aspartic acid, glutamine, and glutamic acid. This represents the first detection of biologically relevant molecules created under conditions thought to be similar to those found in impact melt pools and cryolavas on Titan, which are at a stage of chemical evolution not unlike the "primordial soup" of the early Earth. Future missions to Titan should therefore carry instrumentation capable of, but certainly not limited to, detecting amino acids and other prebiotic molecules on Titan's surface.

  8. High resolution microtubule structures reveal the structural transitions in αβ–tubulin upon GTP hydrolysis

    PubMed Central

    Alushin, Gregory M.; Lander, Gabriel C.; Kellogg, Elizabeth H.; Zhang, Rui; Baker, David; Nogales, Eva

    2014-01-01

    Summary Dynamic instability, the stochastic switching between growth and shrinkage, is essential for microtubule function. This behavior is driven by GTP hydrolysis in the microtubule lattice, and is inhibited by anticancer agents like Taxol. We provide new insight into the mechanism of dynamic instability, based on high-resolution cryo-EM structures (4.7–5.6 Å) of dynamic microtubules and microtubules stabilized by GMPCPP or Taxol. We infer that hydrolysis leads to a compaction around the E-site nucleotide at longitudinal interfaces, as well as movement of the α–tubulin intermediate domain and H7 helix. Displacement of the C-terminal helices in both α– and β–tubulin subunits suggests an effect on interactions with binding partners that contact this region. Taxol inhibits most of these conformational changes, allosterically inducing a GMPCPP-like state. Lateral interactions are similar in all conditions we examined, suggesting that microtubule lattice stability is primarily modulated at longitudinal interfaces. PMID:24855948

  9. Chemical synthesis and enzymatic, stereoselective hydrolysis of a functionalized dihydropyrimidine for the synthesis of β-amino acids.

    PubMed

    Slomka, Christin; Zhong, Sabilla; Fellinger, Anna; Engel, Ulrike; Syldatk, Christoph; Bräse, Stefan; Rudat, Jens

    2015-12-01

    A novel substrate, 6-(4-nitrophenyl)dihydropyrimidine-2,4(1H,3H)-dione (pNO2PheDU), was chemically synthesized and analytically verified for the potential biocatalytic synthesis of enantiopure β-amino acids. The hydantoinase (EC 3.5.2.2) from Arthrobacter crystallopoietes DSM20117 was chosen to prove the enzymatic hydrolysis of this substrate, since previous investigations showed activities of this enzyme toward 6-monosubstituted dihydrouracils. Whole cell biotransformations with recombinant Escherichia coli expressing the hydantoinase showed degradation of pNO2PheDU. Additionally, the corresponding N-carbamoyl-β-amino acid (NCarbpNO2 βPhe) was chemically synthesized, an HPLC-method with chiral stationary phases for detection of this product was established and thus (S)-enantioselectivity toward pNO2PheDU has been shown. Consequently this novel substrate is a potential precursor for the enantiopure β-amino acid para-nitro-β-phenylalanine (pNO2 βPhe). PMID:26705241

  10. Influence of temperature, time, liquid/solid ratio and sulfuric acid concentration on the hydrolysis of palm empty fruit bunches.

    PubMed

    Ferrer, Ana; Requejo, Ana; Rodríguez, Alejandro; Jiménez, Luis

    2013-02-01

    The influence of temperature (150-190 °C), time (0-20 min), liquid/solid ratio (6-8) and sulfuric acid concentration (0.1-0.5%), on the hydrolysis of palm empty fruit bunches (EFBs) was studied and the liquid and solid fractions were analyzed. Polynomial models were found to reproduce the experimental results with errors less than 15% in most of the cases (except for xylose concentration). Operating conditions of 190 °C for 15 min at a liquid/solid ratio of 6 and a sulfuric acid concentration of 0.1% resulted in the production of 3.12, 4.0, 2.35 and 2.28 g/L of glucose, xylose, arabinose and acetic acid, respectively, starting with 1000 g of EFBs. The yield was 67.96%. Soda-anthraquinone, ethanol and ethanolamine pulping of the solid fraction provided pulps with brightness values (63.24%, 28.78%, 48.76%), but with poor resistance properties (6.57-8.54 Nm/g for tensile index, 0.38-0.44 k N/g for burst index and 0.96-1.02 mN m2/g for tear index). Therefore it is advisable to use the pulps for speciality papers or for bioethanol-production. PMID:23266852

  11. Determination of the main impurities formed after acid hydrolysis of soybean extracts and the in vitro mutagenicity and genotoxicity studies of 5-ethoxymethyl-2-furfural.

    PubMed

    Nemitz, Marina C; Picada, Jaqueline N; da Silva, Juliana; Garcia, Ana Letícia H; Papke, Débora K M; Grivicich, Ivana; Steppe, Martin; von Poser, Gilsane L; Teixeira, Helder F

    2016-09-10

    Soybean acid hydrolyzed extracts are raw-materials widely used for manufacturing of pharmaceuticals and cosmetics products due to their high content of isoflavone aglycones. In the present study, the main sugar degradation products 5-hydroxymethyl-2-furfural (HMF) and 5-ethoxymethyl-2-furfural (EMF) were quantitatively determined after acid hydrolysis of extracts from different soybean cultivars by a validated liquid chromatography method. The furanic compounds determined in samples cover the range of 0.16-0.21mg/mL and 0.22-0.33mg/mL for HMF and EMF, respectively. Complementarily, due to the scarce literature regarding the EMF toxicology, this study also assessed the EMF mutagenicity by the Salmonella/microsome test and genotoxicity by the comet assay. The results revealed that EMF did not show mutagenicity at the range of 50-5000μg/plate in S. typhimurium strains TA98, TA97a, TA100, TA102 and TA1535, but induced DNA damage in HepG2 cells at non-cytotoxic doses of 0.1-1.3mg/mL, mainly by oxidative stress mechanisms. Based on literature of HMF genotoxicity, and considering the EMF genotoxicity results herein shown, purification procedures to remove these impurities from extracts are recommended during healthcare products development to ensure the security of the products. PMID:27475406

  12. Improving enzymatic hydrolysis of corn stover pretreated by ethylene glycol-perchloric acid-water mixture.

    PubMed

    He, Yu-Cai; Liu, Feng; Gong, Lei; Lu, Ting; Ding, Yun; Zhang, Dan-Ping; Qing, Qing; Zhang, Yue

    2015-02-01

    To improve the enzymatic saccharification of lignocellulosic biomass, a mixture of ethylene glycol-HClO4-water (88.8:1.2:10, w/w/w) was used for pretreating corn stover in this study. After the optimization in oil-bath system, the optimum pretreatment temperature and time were 130 °C and 30 min, respectively. After the saccharification of 10 g/L pretreated corn stover for 48 h, the saccharification rate was obtained in the yield of 77.4 %. To decrease pretreatment temperature and shorten pretreatment time, ethylene glycol-HClO4-water (88.8:1.2:10, w/w/w) media under microwave irradiation was employed to pretreat corn stover effectively at 100 °C and 200 W for 5 min. Finally, the recovered hydrolyzates containing glucose obtained from the enzymatic hydrolysis of pretreated corn stovers could be fermented into ethanol efficiently. These results would be helpful for developing a cost-effective pretreatment combined with enzymatic saccharification of cellulosic materials for the production of lignocellulosic ethanol. PMID:25384544

  13. Amino acid side chain induced selectivity in the hydrolysis of peptides catalyzed by a Zr(IV)-substituted Wells-Dawson type polyoxometalate.

    PubMed

    Vanhaecht, Stef; Absillis, Gregory; Parac-Vogt, Tatjana N

    2013-11-21

    In this paper the reactivity of K15H[Zr(α2-P2W17O61)2]·25H2O (1), a Zr(IV)-substituted Wells-Dawson polyoxometalate, is examined towards a series of Gly-Aa, Aa-Gly or Aa-Ser dipeptides, in which the nature and the size of the Aa amino acid side chain were varied. The rate of peptide bond hydrolysis, determined by (1)H NMR experiments, in Gly-Aa dipeptides is strongly dependent on the molecular volume and the chemical structure of the Aa side chain. When the volume of the aliphatic side chain of the Aa residue in Gly-Aa increased, a clear decrease in the hydrolysis rate was observed. Replacing one α-H in the C-terminal Gly residue of Gly-Gly by a methyl group (Gly-Ala) resulted in a 6-fold reactivity decrease, pointing towards the importance of steric factors for efficient peptide bond hydrolysis. The rate constants for peptide bond hydrolysis in Gly-Aa dipeptides at pD 5.0 and 60 °C ranged from 208.0 ± 15.6 × 10(-6) min(-1) for Gly-Ser to 5.0 ± 1.0 × 10(-6) min(-1) for Gly-Glu, reflecting the influence of the different nature of the amino acid side chains on the hydrolysis rate. Faster hydrolysis was observed for peptides containing Ser and Thr since the hydroxyl group in their side chain is able to facilitate amide bond hydrolysis by promoting an N→O acyl rearrangement. Peptides containing positively charged side chains at pD 5.0 show enhanced hydrolysis rates as a result of the secondary electrostatic interactions with the negatively charged surface of the polyoxometalate, which stabilize the peptide-polyoxometalate complex. A slow hydrolysis rate was observed for Gly-Glu, because of the preferential coordination of the carboxylate group in the side chain of Glu to Zr(IV), which prevents coordination of the peptide carbonyl group and its activation towards hydrolysis. PMID:24018583

  14. Structural basis for the hydrolysis of ATP by a nucleotide binding subunit of an amino acid ABC transporter from Thermus thermophilus.

    PubMed

    Devi, Seenivasan Karthiga; Chichili, Vishnu Priyanka Reddy; Jeyakanthan, J; Velmurugan, D; Sivaraman, J

    2015-06-01

    ATP-binding cassette (ABC) transporters are a major family of small molecule transporter proteins, and their deregulation is associated with several diseases, including cancer. Here, we report the crystal structure of the nucleotide binding domain (NBD) of an amino acid ABC transporter from Thermus thermophilus (TTHA1159) in its apo form and as a complex with ADP along with functional studies. TTHA1159 is a putative arginine ABC transporter. The apo-TTHA1159 was crystallized in dimeric form, a hitherto unreported form of an apo NBD. Structural comparison of the apo and ADP-Mg(2+) complexes revealed that Phe14 of TTHA1159 undergoes a significant conformational change to accommodate ADP, and that the bound ADP interacts with the P-loop (Gly40-Thr45). Modeling of ATP-Mg(2+):TTHA1159 complex revealed that Gln86 and Glu164 are involved in water-mediated hydrogen bonding contacts and Asp163 in Mg(2+) ion-mediated hydrogen bonding contacts with the γ-phosphate of ATP, consistent with the findings of other ABC transporters. Mutational studies confirmed the necessity of each of these residues, and a comparison of the apo/ADP Mg(2+):TTHA1159 with its ATP-complex model suggests the likelihood of a key conformational change to the Gln86 side chain for ATP hydrolysis. PMID:25916755

  15. Fuel ethanol production from corn stover under optimized dilute phosphoric acid pretreatment and enzymatic hydrolysis

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Ethanol is a renewable oxygenated fuel. Dilute acid pretreatment is a promising pretreatment technology for conversion of lignocellulosic biomass to fuel ethanol. Generation of fermentable sugars from corn stover involves pretreatment and enzymatic saccharification. Pretreatment is crucial as nat...

  16. Dilute oxalic acid pretreatment for high total sugar recovery in pretreatment and subsequent enzymatic hydrolysis.

    PubMed

    Qing, Qing; Huang, Meizi; He, Yucai; Wang, Liqun; Zhang, Yue

    2015-12-01

    Oxalic acid was evaluated as an alternative reagent to mineral inorganic acid in pretreatment of corncob to achieve high xylose yield in addition to highly digestible solid residue. A quadratic polynomial model of xylose formation was developed for optimization of pretreatment process by the response surface methodology based on the impact factors of pretreatment temperature, reaction time, acid concentration, and solid-to-liquid ratio. The highest xylose yield was 94.3 % that was obtained under the pretreatment condition of 140 °C for 40 min with 0.5 wt% oxalic acid at a solid loading of 7.5 %. Under these conditions, the xylose yield results of verification experiments were very close to the model prediction, which indicated that the model was applicable. The solid residue generated under this condition also demonstrated a satisfactory enzymatic digestibility and fermentability. PMID:26494137

  17. Hydrolysis of Np(IV) and Pu(IV) and Their Complexation by Aqueous Orthosilicic Acid Si(OH)4

    SciTech Connect

    Yusov, A B.; Fedoseev, A M.; Delegard, Calvin H.

    2004-12-10

    The hydrolysis and interaction of Np(IV) and Pu(IV) with orthosilicic acid, Si(OH)4, were studied in 0.1-1.0 M ionic strength aqueous solutions. Spectrophotometry was used to study these reactions at about 10-4 M Np(IV) and Pu(IV) concentrations. The first hydrolysis constants, Khydr, agree with the majority of earlier spectrophotometric and potentiometric data. The absorption spectra of NpOH3+ and PuOH3+ were obtained by spectral deconvolution. Reasons to explain the overestimation of Khydr obtained by other methods [by extraction of trace amounts of Np(IV) and Pu(IV) and by solubility] are discussed. Formation of the complexes NpOSi(OH)33+ and PuOSi(OH)33+ is demonstrated in the p[H+] range 1.4-2.1 and 0.7-1.4, respectively. Measured values of equilibrium constants of the reaction M4+ + Si(OH)4 ? MOSi(OH)33+ + H+ at ionic strength I=1.0 are log ?1 = 0.41?0.02 and 1.04?0.04, respectively, for Np(IV) and Pu(IV). The stability constants of the NpOSi(OH)33+ and PuOSi(OH)33+ complexes, recalculated to zero ionic strength, are log ?10 = 11.2 and 11.8, respectively. The correlation between Khydr and ?1, as observed for all earlier studied metal ions, also occurs for both Np(IV) and Pu(IV).

  18. Development of C-reactive protein certified reference material NMIJ CRM 6201-b: optimization of a hydrolysis process to improve the accuracy of amino acid analysis.

    PubMed

    Kato, Megumi; Kinumi, Tomoya; Yoshioka, Mariko; Goto, Mari; Fujii, Shin-Ichiro; Takatsu, Akiko

    2015-04-01

    To standardize C-reactive protein (CRP) assays, the National Metrology Institute of Japan (NMIJ) has developed a C-reactive protein solution certified reference material, CRM 6201-b, which is intended for use as a primary reference material to enable the SI-traceable measurement of CRP. This study describes the development process of CRM 6201-b. As a candidate material of the CRM, recombinant human CRP solution was selected because of its higher purity and homogeneity than the purified material from human serum. Gel filtration chromatography was used to examine the homogeneity and stability of the present CRM. The total protein concentration of CRP in the present CRM was determined by amino acid analysis coupled to isotope-dilution mass spectrometry (IDMS-AAA). To improve the accuracy of IDMS-AAA, we optimized the hydrolysis process by examining the effect of parameters such as the volume of protein samples taken for hydrolysis, the procedure of sample preparation prior to the hydrolysis, hydrolysis temperature, and hydrolysis time. Under optimized conditions, we conducted two independent approaches in which the following independent hydrolysis and liquid chromatography-isotope dilution mass spectrometry (LC-IDMS) were combined: one was vapor-phase acid hydrolysis (130 °C, 24 h) and hydrophilic interaction liquid chromatography-mass spectrometry (HILIC-MS) method, and the other was microwave-assisted liquid-phase acid hydrolysis (150 °C, 3 h) and pre-column derivatization liquid chromatography-tandem mass spectrometry (LC-MS/MS) method. The quantitative values of the two different amino acid analyses were in agreement within their uncertainties. The certified value was the weighted mean of the results of the two methods. Uncertainties from the value-assignment method, between-method variance, homogeneity, long-term stability, and short-term stability were taken into account in evaluating the uncertainty for a certified value. The certified value and the

  19. Progressing batch hydrolysis process

    DOEpatents

    Wright, John D.

    1986-01-01

    A progressive batch hydrolysis process for producing sugar from a lignocellulosic feedstock, comprising passing a stream of dilute acid serially through a plurality of percolation hydrolysis reactors charged with said feedstock, at a flow rate, temperature and pressure sufficient to substantially convert all the cellulose component of the feedstock to glucose; cooling said dilute acid stream containing glucose, after exiting the last percolation hydrolysis reactor, then feeding said dilute acid stream serially through a plurality of prehydrolysis percolation reactors, charged with said feedstock, at a flow rate, temperature and pressure sufficient to substantially convert all the hemicellulose component of said feedstock to glucose; and cooling the dilute acid stream containing glucose after it exits the last prehydrolysis reactor.

  20. Dependence of RIG-I Nucleic Acid-Binding and ATP Hydrolysis on Activation of Type I Interferon Response

    PubMed Central

    Baek, Yu Mi; Yoon, Soojin; Hwang, Yeo Eun

    2016-01-01

    Exogenous nucleic acids induce an innate immune response in mammalian host cells through activation of the retinoic acid-inducible gene I (RIG-I). We evaluated RIG-I protein for RNA binding and ATPase stimulation with RNA ligands to investigate the correlation with the extent of immune response through RIG-I activation in cells. RIG-I protein favored blunt-ended, double-stranded RNA (dsRNA) ligands over sticky-ended dsRNA. Moreover, the presence of the 5'-triphosphate (5'-ppp) moiety in dsRNA further enhanced binding affinity to RIG-I. Two structural motifs in RNA, blunt ends in dsRNA and 5'-ppp, stimulated the ATP hydrolysis activity of RIG-I. These structural motifs also strongly induced IFN expression as an innate immune response in cells. Therefore, we suggest that IFN induction through RIG-I activation is mainly determined by structural motifs in dsRNA that increase its affinity for RIG-I protein and stimulate ATPase activity in RIG-I. PMID:27574504

  1. Dependence of RIG-I Nucleic Acid-Binding and ATP Hydrolysis on Activation of Type I Interferon Response.

    PubMed

    Baek, Yu Mi; Yoon, Soojin; Hwang, Yeo Eun; Kim, Dong-Eun

    2016-08-01

    Exogenous nucleic acids induce an innate immune response in mammalian host cells through activation of the retinoic acid-inducible gene I (RIG-I). We evaluated RIG-I protein for RNA binding and ATPase stimulation with RNA ligands to investigate the correlation with the extent of immune response through RIG-I activation in cells. RIG-I protein favored blunt-ended, double-stranded RNA (dsRNA) ligands over sticky-ended dsRNA. Moreover, the presence of the 5'-triphosphate (5'-ppp) moiety in dsRNA further enhanced binding affinity to RIG-I. Two structural motifs in RNA, blunt ends in dsRNA and 5'-ppp, stimulated the ATP hydrolysis activity of RIG-I. These structural motifs also strongly induced IFN expression as an innate immune response in cells. Therefore, we suggest that IFN induction through RIG-I activation is mainly determined by structural motifs in dsRNA that increase its affinity for RIG-I protein and stimulate ATPase activity in RIG-I. PMID:27574504

  2. Hydrolysis of 4-Hydroxybenzoic Acid Esters (Parabens) and Their Aerobic Transformation into Phenol by the Resistant Enterobacter cloacae Strain EM

    PubMed Central

    Valkova, Nelly; Lépine, François; Valeanu, Loredana; Dupont, Maryse; Labrie, Louisette; Bisaillon, Jean-Guy; Beaudet, Réjean; Shareck, François; Villemur, Richard

    2001-01-01

    Enterobacter cloacae strain EM was isolated from a commercial dietary mineral supplement stabilized by a mixture of methylparaben and propylparaben. It harbored a high-molecular-weight plasmid and was resistant to high concentrations of parabens. Strain EM was able to grow in liquid media containing similar amounts of parabens as found in the mineral supplement (1,700 and 180 mg of methyl and propylparaben, respectively, per liter or 11.2 and 1.0 mM) and in very high concentrations of methylparaben (3,000 mg liter−1, or 19.7 mM). This strain was able to hydrolyze approximately 500 mg of methyl-, ethyl-, or propylparaben liter−1 (3 mM) in less than 2 h in liquid culture, and the supernatant of a sonicated culture, after a 30-fold dilution, was able to hydrolyze 1,000 mg of methylparaben liter−1 (6.6 mM) in 15 min. The first step of paraben degradation was the hydrolysis of the ester bond to produce 4-hydroxybenzoic acid, followed by a decarboxylation step to produce phenol under aerobic conditions. The transformation of 4-hydroxybenzoic acid into phenol was stoichiometric. The conversion of approximately 500 mg of parabens liter−1 (3 mM) to phenol in liquid culture was completed within 5 h without significant hindrance to the growth of strain EM, while higher concentrations of parabens partially inhibited its growth. PMID:11375144

  3. Theoretical study of neutral and of acid and base promoted hydrolysis of formamide

    SciTech Connect

    Krug, J.P.; Popelier, P.L.A.; Bader, R.F.W.

    1992-09-17

    This paper studies the reaction pathways for four hydrolyses of formamide. These pathways are the uncatalyzed reaction with a single water molecule, the acid catalyzed reaction with H{sub 3}O{sup +} for both oxygen and nitrogen protonation, and the reaction with the hydroxide ion.

  4. Titan's Primordial Soup: Formation of Amino Acids via Low Temperature Hydrolysis of Tholins

    NASA Astrophysics Data System (ADS)

    Neish, Catherine; Somogyi, Á.; Smith, M. A.

    2009-09-01

    Titan, Saturn's largest moon, is a world rich in the "stuff of life". Reactions occurring in its dense nitrogen-methane atmosphere produce a wide variety of organic molecules, which subsequently rain down onto its surface. Water - thought to be another important ingredient for life - is likewise abundant on Titan. Theoretical models of Titan's formation predict that its interior consists of an ice I layer several tens of kilometers thick overlying a liquid ammonia-rich water layer several hundred kilometers thick (Tobie et al., 2005). Though its surface temperature of 94K dictates that Titan is on average too cold for liquid water to persist at its surface, melting caused by impacts and/or cryovolcanism may lead to its episodic availability. Impact melt pools on Titan would likely remain liquid for 102 - 104 years before freezing (O'Brien et al., 2005). The combination of complex organic molecules and transient locales of liquid water make Titan an interesting natural laboratory for studying prebiotic chemistry. In this work, we sought to determine what biomolecules might be formed under conditions analogous to those found in transient liquid water environments on Titan. We hydrolyzed Titan organic haze analogues, or "tholins", in 13 wt. % ammonia-water at 253K and 293K for a year. Using a combination of high resolution mass spectroscopy and tandem mass spectroscopy fragmentation techniques, four amino acids were identified in the hydrolyzed tholin sample. These four species have been assigned as the amino acids asparagine, aspartic acid, glutamine, and glutamic acid. This represents the first detection of biologically relevant molecules created under conditions similar to those found in impact melt pools and cryolavas on Titan. Future missions to Titan should therefore carry instrumentation capable of detecting amino acids and other prebiotically relevant molecules on its surface This work was supported by the NASA Exobiology Program.

  5. Correction: Acid-catalyzed carboxylic acid esterification and ester hydrolysis mechanism: acylium ion as a sharing active intermediate via a spontaneous trimolecular reaction based on density functional theory calculation and supported by electrospray ionization-mass spectrometry.

    PubMed

    Shi, Hongchang; Wang, Yilei; Hua, Ruimao

    2015-12-28

    Correction for 'Acid-catalyzed carboxylic acid esterification and ester hydrolysis mechanism: acylium ion as a sharing active intermediate via a spontaneous trimolecular reaction based on density functional theory calculation and supported by electrospray ionization-mass spectrometry' by Hongchang Shi et al., Phys. Chem. Chem. Phys., 2015, DOI: 10.1039/c5cp02914g. PMID:26583937

  6. The salt stress-induced LPA response in Chlamydomonas is produced via PLA2 hydrolysis of DGK-generated phosphatidic acid[S

    PubMed Central

    Arisz, Steven A.; Munnik, Teun

    2011-01-01

    The unicellular green alga Chlamydomonas has frequently been used as a eukaryotic model system to study intracellular phospholipid signaling pathways in response to environmental stresses. Earlier, we found that hypersalinity induced a rapid increase in the putative lipid second messenger, phosphatidic acid (PA), which was suggested to be generated via activation of a phospholipase D (PLD) pathway and the combined action of a phospholipase C/diacylglycerol kinase (PLC/DGK) pathway. Lysophosphatidic acid (LPA) was also increased and was suggested to reflect a phospholipase A2 (PLA2) activity based on pharmacological evidence. The question of PA's and LPA's origin is, however, more complicated, especially as both function as precursors in the biosynthesis of phospho- and galactolipids. To address this complexity, a combination of fatty acid-molecular species analysis and in vivo 32P-radiolabeling was performed. Evidence is provided that LPA is formed from a distinct pool of PA characterized by a high α-linolenic acid (18:3n-3) content. This molecular species was highly enriched in the polyphosphoinositide fraction, which is the substrate for PLC to form diacylglycerol. Together with differential 32P-radiolabeling studies and earlier PLD-transphosphatidylation and PLA2-inhibitor assays, the data were consistent with the hypothesis that the salt-induced LPA response is primarily generated through PLA2-mediated hydrolysis of DGK-generated PA and that PLD or de novo synthesis [via endoplasmic reticulum - or plastid-localized routes] is not a major contributor. PMID:21900174

  7. Porcine liver esterase-catalyzed enantioselective hydrolysis of a prochiral diester into its optically pure (S)-ester acid, a precursor to a growth hormone secretagogue.

    PubMed

    Chartrain, M; Maligres, P; Cohen, D; Upadhyay, V; Pecore, V; Askin, D; Greasham, R

    1999-01-01

    A limited screen of several commercially-available and internally-produced lipases and esterases identified porcine liver esterase as a suitable biocatalyst for the enantioselective hydrolysis of a diester into its (S)-ester acid with high optical purity (99%). This (S)-ester acid is a precursor to an experimental growth hormone secretagogue. After identifying xanthan gum as the best emulsifier and optimizing the reaction conditions, hydrolysis rates of 1 g/l.h and final (S)-ester acid (ee > 99%) titers of about 8.5 g/l were routinely achieved. This process supported the production of preparative amounts of optically pure (S)-ester (ee > 99%) with a high reaction yield of 82%. Upon purification, the (S)-ester was successfully used in the subsequent synthetic steps to yield the growth hormone secretagogue. PMID:16232487

  8. Hydrolysis of proteins with methanesulfonic acid for improved HPLC-ICP-MS determination of seleno-methionine in yeast and nuts.

    PubMed

    Wrobel, Katarzyna; Kannamkumarath, Sasi S; Wrobel, Kazimierz; Caruso, Joseph A

    2003-01-01

    In this work, the use of methanesulfonic acid for protein hydrolysis is proposed for evaluation of Se-methionine in yeast, Brazil nuts, and possibly other selenium-rich biological samples. The hydrolysis was carried out by heating the sample with 4 mol L(-1) acid at reflux for 8 h. Two chromatographic techniques (size-exclusion and ion-pairing) coupled with ICP-MS detection were used to compare the release of Se-methionine from proteins by enzymatic (proteinase K, protease XIV) and acid hydrolyses. A more efficient liberation of Se-methionine was observed by acid hydrolysis. For quantification, the sample extracts were introduced onto a C8 Alltima column, and the separation was achieved with a mobile phase containing 5 mmol L(-1) hexanesulfonic acid in citrate buffer (pH 4.5)/methanol (95:5). The results obtained by standard addition showed 816+/-17 micro g g(-1) and 36.2+/-1.5 micro g g(-1) of selenium in the form of Se-methionine in yeast and nuts, respectively (65% and 75% of total selenium). PMID:12520449

  9. Pretreatment of Dried Distiller Grains with Solubles by Soaking in Aqueous Ammonia and Subsequent Enzymatic/Dilute Acid Hydrolysis to Produce Fermentable Sugars.

    PubMed

    Nghiem, Nhuan P; Montanti, Justin; Kim, Tae Hyun

    2016-05-01

    Dried distillers grains with solubles (DDGS), a co-product of corn ethanol production in the dry-grind process, was pretreated by soaking in aqueous ammonia (SAA) using a 15 % w/w NH4OH solution at a solid/liquid ratio of 1:10. The effect of pretreatment on subsequent enzymatic hydrolysis was studied at two temperatures (40 and 60 °C) and four reaction times (6, 12, 24, and 48 h). Highest glucose yield of 91 % theoretical was obtained for the DDGS pretreated at 60 °C and 24 h. The solubilized hemicellulose in the liquid fraction was further hydrolyzed with dilute H2SO4 to generate fermentable monomeric sugars. The conditions of acid hydrolysis included 1 and 4 wt% acid, 60 and 120 °C, and 0.5 and 1 h. Highest yields of xylose and arabinose were obtained at 4 wt% acid, 120 °C, and 1 h. The fermentability of the hydrolysate obtained by enzymatic hydrolysis of the SAA-pretreated DDGS was demonstrated in ethanol fermentation by Saccharomyces cerevisiae. The fermentability of the hydrolysate obtained by consecutive enzymatic and dilute acid hydrolysis was demonstrated using a succinic acid-producing microorganism, strain Escherichia coli AFP184. Under the fermentation conditions, complete utilization of glucose and arabinose was observed, whereas only 47 % of xylose was used. The succinic acid yield was 0.60 g/g total sugar consumed. PMID:26797927

  10. The Effect of Hydrolysis on the Mechanical Properties of Injection-Molded Poly(L-lactic acid)

    NASA Astrophysics Data System (ADS)

    Kobayashi, Satoshi; Todo, Mitsugu

    Injection molded bulk-shape Poly (L-lactide) (PLLA) specimens were hydrolyzed in the phosphate buffered solution and the mechanical properties were evaluated after the hydrolysis tests. In order to evaluated the effect of crystallinity on the hydrolysis, specimens were annealed with 70 and 130°C for 24h. Hydrolysis tests were conducted with soaking the specimens in the phosphate buffered solution (pH 7.4) in an incubator where temperature was kept as 37°C. Vickers hardness was not influenced by hydrolysis until 30 days. Tensile tests results, however, indicated the strength reduction with hydrolysis. From the appearance inspection, whitened regions were observed at the inner of the specimen. These results indicated the bulk erosion with accelerated inside erosion occurred.

  11. Difference analysis of the enzymatic hydrolysis performance of acid-catalyzed steam-exploded corn stover before and after washing with water.

    PubMed

    Zhu, Junjun; Shi, Linli; Zhang, Lingling; Xu, Yong; Yong, Qiang; Ouyang, Jia; Yu, Shiyuan

    2016-10-01

    The difference in the enzymatic hydrolysis yield of acid-catalyzed steam-exploded corn stover (ASC) before and after washing with water reached approximately 15 % under the same conditions. The reasons for the difference in the yield between ASC and washed ASC (wASC) were determined through the analysis of the composition of ASC prehydrolyzate and sugar concentration of enzymatic hydrolyzate. Salts produced by neutralization (CaSO4, Na2SO4, K2SO4, and (NH4)2SO4), sugars (polysaccharides, oligosaccharides, and monosaccharides), sugar-degradation products (weak acids and furans), and lignin-degradation products (ethyl acetate extracts and nine main lignin-degradation products) were back-added to wASC. Results showed that these products, except furans, exerted negative effect on enzymatic hydrolysis. According to the characteristics of acid-catalyzed steam explosion pretreatment, the five sugar-degradation products' mixture and salts [Na2SO4, (NH4)2SO4] showed minimal negative inhibition effect on enzymatic hydrolysis. By contrast, furans demonstrated a promotion effect. Moreover, soluble sugars, such as 13 g/L xylose (decreased by 6.38 %), 5 g/L cellobiose (5.36 %), 10 g/L glucose (3.67 %), as well as lignin-degradation products, and ethyl acetate extracts (4.87 %), exhibited evident inhibition effect on enzymatic hydrolysis. Therefore, removal of soluble sugars and lignin-degradation products could effectively promote the enzymatic hydrolysis performance. PMID:27277746

  12. High yield production of sugars from deproteinated palm kernel cake under microwave irradiation via dilute sulfuric acid hydrolysis.

    PubMed

    Fan, Suet-Pin; Jiang, Li-Qun; Chia, Chin-Hua; Fang, Zhen; Zakaria, Sarani; Chee, Kah-Leong

    2014-02-01

    Recent years, great interest has been devoted to the conversion of biomass-derived carbohydrate into sugars, such as glucose, mannose and fructose. These are important versatile intermediate products that are easily processed into high value-added biofuels. In this work, microwave-assisted dilute sulfuric acid hydrolysis of deproteinated palm kernel cake (DPKC) was systematically studied using Response Surface Methodology. The highest mannose yield (92.11%) was achieved at the optimized condition of 148°C, 0.75N H2SO4, 10min 31s and substrate to solvent (SS) ratio (w/v) of 1:49.69. Besides that, total fermentable sugars yield (77.11%), was obtained at 170°C, 0.181N H2SO4, 6min 6s and SS ratio (w/v) of 1:40. Ridge analysis was employed to further verify the optimum conditions. Thus, this work provides fundamental data of the practical use of DPKC as low cost, high yield and environmental-friendly material for the production of mannose and other sugars. PMID:24342947

  13. Evaluation of xylitol production using corncob hemicellulosic hydrolysate by combining tetrabutylammonium hydroxide extraction with dilute acid hydrolysis.

    PubMed

    Jia, Honghua; Shao, Tingting; Zhong, Chao; Li, Hengxiang; Jiang, Min; Zhou, Hua; Wei, Ping

    2016-10-20

    In this paper, we produced hemicellulosic hydrolysate from corncob by tetrabutylammonium hydroxide (TBAH) extraction and dilute acid hydrolysis combined, further evaluating the feasibility of the resultant corncob hemicellulosic hydrolysate used in xylitol production by Candida tropicalis. Optimized conditions for corncob hemicellulose extraction by TBAH was obtained via response surface methodology: time of 90min, temperature of 60°C, liquid/solid ratio of 12 (v/w), and TBAH concentration of 55%, resulting in a hemicellulose extraction of 80.07% under these conditions. The FT-IR spectrum of the extracted corncob hemicellulose is consistent with that of birchwood hemicellulose and exhibits specific absorbance of hemicelluloses at 1380, 1168, 1050, and 900cm(-1). In addition, we found that C. tropicalis can ferment the resulting corncob hemicellulosic hydrolysate with pH adjustment and activated charcoal treatment leading to a high xylitol yield and productivity of 0.77g/g and 2.45g/(Lh), respectively. PMID:27474613

  14. Different roles of protein kinase C-beta and -delta in arachidonic acid cascade, superoxide formation and phosphoinositide hydrolysis.

    PubMed Central

    Duyster, J; Schwende, H; Fitzke, E; Hidaka, H; Dieter, P

    1993-01-01

    In contrast with protein kinase C (PKC)-beta, PKC-delta is exclusively detectable in the membrane fraction of liver macrophages. After long-term treatment with phorbol 12-myristate 13-acetate (PMA) PKC-beta is depleted faster (within 3 h) than PKC-delta (> 7h). Simultaneously, pretreatment with PMA for 3 h inhibits the PMA- and zymosan-induced generation of superoxide and the PMA-induced formation of prostaglandin (PG) E2, whereas a preincubation of more than 7 h is required to affect the zymosan-induced release of PGE2 and inositol phosphates. These results support an involvement of PKC-beta in the PMA-induced activation of the arachidonic acid cascade and in superoxide formation and imply an involvement of PKC-delta in zymosan-induced phosphoinositide hydrolysis and PGE2 formation. Two phorbol ester derivates, sapintoxin A (SAPA) and 12-deoxyphorbol 13-phenylacetate 20-acetate (DOPPA), which have been previously reported to activate preferentially PLC-beta but not PKC-delta in vitro [Ryves, Evans, Olivier, Parker and Evans (1992) FEBS Lett. 288, 5-9], induce the formation of PGE2 and superoxide, down-regulate PKC-delta and potentiate inositol phosphate formation in parallel SAPA, but not DOPPA, down-regulates PKC-beta and inhibits the PMA-induced formation of eicosanoids and superoxide. Images Figure 1 Figure 2 Figure 5 PMID:8389125

  15. Evolutionary Importance of the Intramolecular Pathways of Hydrolysis of Phosphate Ester Mixed Anhydrides with Amino Acids and Peptides

    NASA Astrophysics Data System (ADS)

    Liu, Ziwei; Beaufils, Damien; Rossi, Jean-Christophe; Pascal, Robert

    2014-12-01

    Aminoacyl adenylates (aa-AMPs) constitute essential intermediates of protein biosynthesis. Their polymerization in aqueous solution has often been claimed as a potential route to abiotic peptides in spite of a highly efficient CO2-promoted pathway of hydrolysis. Here we investigate the efficiency and relevance of this frequently overlooked pathway from model amino acid phosphate mixed anhydrides including aa-AMPs. Its predominance was demonstrated at CO2 concentrations matching that of physiological fluids or that of the present-day ocean, making a direct polymerization pathway unlikely. By contrast, the occurrence of the CO2-promoted pathway was observed to increase the efficiency of peptide bond formation owing to the high reactivity of the N-carboxyanhydride (NCA) intermediate. Even considering CO2 concentrations in early Earth liquid environments equivalent to present levels, mixed anhydrides would have polymerized predominantly through NCAs. The issue of a potential involvement of NCAs as biochemical metabolites could even be raised. The formation of peptide-phosphate mixed anhydrides from 5(4H)-oxazolones (transiently formed through prebiotically relevant peptide activation pathways) was also observed as well as the occurrence of the reverse cyclization process in the reactions of these mixed anhydrides. These processes constitute the core of a reaction network that could potentially have evolved towards the emergence of translation.

  16. Enhancing bio-butanol production from biomass of Chlorella vulgaris JSC-6 with sequential alkali pretreatment and acid hydrolysis.

    PubMed

    Wang, Yue; Guo, Wanqian; Cheng, Chieh-Lun; Ho, Shih-Hsin; Chang, Jo-Shu; Ren, Nanqi

    2016-01-01

    This study presents a successful butanol production method using alkali and acid pretreated biomass of Chlorella vulgaris JSC-6. The butanol concentration, yield, and productivity were 13.1g/L, 0.58mol/mol sugar, 0.66g/L/h, respectively. Nearly 2.93L/L of biohydrogen was produced during the acidogenesis phase in ABE fermentation. The hydrogen yield and productivity were 0.39mol/mol sugar and 104.2g/L/h respectively. In addition, the high glucose consumption efficiency (97.5%) suggests that the hydrolysate pretreated with NaOH (1%) followed by H2SO4 (3%) did not contain inhibitors to the fermentation. It was also discovered that an excess amount of nitrogen sources arising from hydrolysis of highly concentrated microalgal biomass negatively affected the butanol production. This work demonstrates the technical feasibility of producing butanol from sustainable third-generation feedstock (i.e., microalgal biomass). PMID:26528906

  17. Relationship between stimulated phosphatidic acid production and inositol lipid hydrolysis in intestinal longitudinal smooth muscle from guinea pig.

    PubMed Central

    Mallows, R S; Bolton, T B

    1987-01-01

    Accumulation of [32P]phosphatidic acid (PA) and total [3H]inositol phosphates (IPs) was measured in the longitudinal smooth-muscle layer from guinea-pig small intestine. Stimulation with carbachol, histamine and substance P produced increases in accumulation of both [3H]IPs and [32P]PA over the same concentration range. The increase in [32P]PA accumulation in response to carbachol (1 microM-0.1 mM) was inhibited in the presence of atropine (0.5 microM). Buffering the external free [Ca2+] to 10 nM did not prevent the carbachol-stimulated increase in [32P]PA accumulation. Carbachol and Ca2+ appear to act synergistically to increase accumulation of [32P]PA. In contrast, although incubation with noradrenaline also increased accumulation of [3H]IPs, no increase in accumulation of [32P]PA could be detected. These results suggest that an increase in formation of IPs is not necessarily accompanied by an increase in PA formation, and imply the existence of receptor-modulated pathways regulating PA concentrations other than by phospholipase-C-catalysed inositol phospholipid hydrolysis. PMID:2451504

  18. Evolutionary Importance of the Intramolecular Pathways of Hydrolysis of Phosphate Ester Mixed Anhydrides with Amino Acids and Peptides

    PubMed Central

    Liu, Ziwei; Beaufils, Damien; Rossi, Jean-Christophe; Pascal, Robert

    2014-01-01

    Aminoacyl adenylates (aa-AMPs) constitute essential intermediates of protein biosynthesis. Their polymerization in aqueous solution has often been claimed as a potential route to abiotic peptides in spite of a highly efficient CO2-promoted pathway of hydrolysis. Here we investigate the efficiency and relevance of this frequently overlooked pathway from model amino acid phosphate mixed anhydrides including aa-AMPs. Its predominance was demonstrated at CO2 concentrations matching that of physiological fluids or that of the present-day ocean, making a direct polymerization pathway unlikely. By contrast, the occurrence of the CO2-promoted pathway was observed to increase the efficiency of peptide bond formation owing to the high reactivity of the N-carboxyanhydride (NCA) intermediate. Even considering CO2 concentrations in early Earth liquid environments equivalent to present levels, mixed anhydrides would have polymerized predominantly through NCAs. The issue of a potential involvement of NCAs as biochemical metabolites could even be raised. The formation of peptide–phosphate mixed anhydrides from 5(4H)-oxazolones (transiently formed through prebiotically relevant peptide activation pathways) was also observed as well as the occurrence of the reverse cyclization process in the reactions of these mixed anhydrides. These processes constitute the core of a reaction network that could potentially have evolved towards the emergence of translation. PMID:25501391

  19. Coordination chemistry and hydrolysis of Fe(III) in a peat humic acid studied by X-ray absorption spectroscopy

    NASA Astrophysics Data System (ADS)

    Karlsson, Torbjörn; Persson, Per

    2010-01-01

    The speciation of iron (Fe) in soils, sediments and surface waters is highly dependent on chemical interactions with natural organic matter (NOM). However, the molecular structure and hydrolysis of the Fe species formed in association with NOM is still poorly described. In this study extended X-ray absorption fine structure (EXAFS) spectroscopy was used to determine the coordination chemistry and hydrolysis of Fe(III) in solution of a peat humic acid (5010-49,200 μg Fe g -1 dry weight, pH 3.0-7.2). Data were analyzed by both conventional EXAFS data fitting and by wavelet transforms in order to facilitate the identification of the nature of backscattering atoms. Our results show that Fe occurs predominantly in the oxidized form as ferric ions and that the speciation varies with pH and Fe concentration. At low Fe concentrations (5010-9920 μg g -1; pH 3.0-7.2) mononuclear Fe(III)-NOM complexes completely dominates the speciation. The determined bond distances for the Fe(III)-NOM complexes are similar to distances obtained for Fe(III) complexed by desferrioxamine B and oxalate indicating the formation of a five-membered chelate ring structure. At higher Fe concentrations (49,200 μg g -1; pH 4.2-6.9) we detect a mixture of mononuclear Fe(III)-NOM complexes and polymeric Fe(III) (hydr)oxides with an increasing amount of Fe(III) (hydr)oxides at higher pH. However, even at pH 6.9 and a Fe concentration of 49,200 μg g -1 our data indicates that a substantial amount of the total Fe (>50%) is in the form of organic complexes. Thus, in environments with significant amounts of organic matter organic Fe complexes will be of great importance for the geochemistry of Fe. Furthermore, the formation of five-membered chelate ring structures is in line with the strong complexation and limited hydrolytic polymerization of Fe(III) in our samples and also agrees with EXAFS derived structures of Fe(III) in organic soils.

  20. Validated Method for the Characterization and Quantification of Extractable and Nonextractable Ellagitannins after Acid Hydrolysis in Pomegranate Fruits, Juices, and Extracts.

    PubMed

    García-Villalba, Rocío; Espín, Juan Carlos; Aaby, Kjersti; Alasalvar, Cesarettin; Heinonen, Marina; Jacobs, Griet; Voorspoels, Stefan; Koivumäki, Tuuli; Kroon, Paul A; Pelvan, Ebru; Saha, Shikha; Tomás-Barberán, Francisco A

    2015-07-29

    Pomegranates are one of the main highly valuable sources of ellagitannins. Despite the potential health benefits of these compounds, reliable data on their content in pomegranates and derived extracts and food products is lacking, as it is usually underestimated due to their complexity, diversity, and lack of commercially available standards. This study describes a new method for the analysis of the extractable and nonextractable ellagitannins based on the quantification of the acid hydrolysis products that include ellagic acid, gallic acid, sanguisorbic acid dilactone, valoneic acid dilactone, and gallagic acid dilactone in pomegranate samples. The study also shows the occurrence of ellagitannin C-glycosides in pomegranates. The method was optimized using a pomegranate peel extract. To quantify nonextractable ellagitannins, freeze-dried pomegranate fruit samples were directly hydrolyzed with 4 M HCl in water at 90 °C for 24 h followed by extraction of the pellet with dimethyl sulfoxide/methanol (50:50, v/v). The method was validated and reproducibility was assessed by means of an interlaboratory trial, showing high reproducibility across six laboratories with relative standard deviations below 15%. Their applicability was demonstrated in several pomegranate extracts, different parts of pomegranate fruit (husk, peels, and mesocarp), and commercial juices. A large variability has been found in the ellagitannin content (150-750 mg of hydrolysis products/g) and type (gallagic acid/ellagic acid ratios between 4 and 0.15) of the 11 pomegranate extracts studied. PMID:26158321

  1. Structural and biochemical analyses reveal how ornithine acetyl transferase binds acidic and basic amino acid substrates.

    PubMed

    Iqbal, Aman; Clifton, Ian J; Chowdhury, Rasheduzzaman; Ivison, David; Domene, Carmen; Schofield, Christopher J

    2011-09-21

    Structural and biochemical analyses reveal how ornithine acetyl-transferases catalyse the reversible transfer of an acetyl-group from a basic (ornithine) to an acidic (glutamate) amino acid by employing a common mechanism involving an acetyl-enzyme intermediate but using different side chain binding modes. PMID:21796301

  2. Gas-phase hydrolysis of triplet SO2: A possible direct route to atmospheric acid formation.

    PubMed

    Donaldson, D James; Kroll, Jay A; Vaida, Veronica

    2016-01-01

    Sulfur chemistry is of great interest to the atmospheric chemistry of several planets. In the presence of water, oxidized sulfur can lead to new particle formation, influencing climate in significant ways. Observations of sulfur compounds in planetary atmospheres when compared with model results suggest that there are missing chemical mechanisms. Here we propose a novel mechanism for the formation of sulfurous acid, which may act as a seed for new particle formation. In this proposed mechanism, the lowest triplet state of SO2 ((3)B1), which may be accessed by near-UV solar excitation of SO2 to its excited (1)B1 state followed by rapid intersystem crossing, reacts directly with water to form H2SO3 in the gas phase. For ground state SO2, this reaction is endothermic and has a very high activation barrier; our quantum chemical calculations point to a facile reaction being possible in the triplet state of SO2. This hygroscopic H2SO3 molecule may act as a condensation nucleus for water, giving rise to facile new particle formation (NPF). PMID:27417675

  3. Gas-phase hydrolysis of triplet SO2: A possible direct route to atmospheric acid formation

    NASA Astrophysics Data System (ADS)

    Donaldson, D. James; Kroll, Jay A.; Vaida, Veronica

    2016-07-01

    Sulfur chemistry is of great interest to the atmospheric chemistry of several planets. In the presence of water, oxidized sulfur can lead to new particle formation, influencing climate in significant ways. Observations of sulfur compounds in planetary atmospheres when compared with model results suggest that there are missing chemical mechanisms. Here we propose a novel mechanism for the formation of sulfurous acid, which may act as a seed for new particle formation. In this proposed mechanism, the lowest triplet state of SO2 (3B1), which may be accessed by near-UV solar excitation of SO2 to its excited 1B1 state followed by rapid intersystem crossing, reacts directly with water to form H2SO3 in the gas phase. For ground state SO2, this reaction is endothermic and has a very high activation barrier; our quantum chemical calculations point to a facile reaction being possible in the triplet state of SO2. This hygroscopic H2SO3 molecule may act as a condensation nucleus for water, giving rise to facile new particle formation (NPF).

  4. Gas-phase hydrolysis of triplet SO2: A possible direct route to atmospheric acid formation

    PubMed Central

    Donaldson, D. James; Kroll, Jay A.; Vaida, Veronica

    2016-01-01

    Sulfur chemistry is of great interest to the atmospheric chemistry of several planets. In the presence of water, oxidized sulfur can lead to new particle formation, influencing climate in significant ways. Observations of sulfur compounds in planetary atmospheres when compared with model results suggest that there are missing chemical mechanisms. Here we propose a novel mechanism for the formation of sulfurous acid, which may act as a seed for new particle formation. In this proposed mechanism, the lowest triplet state of SO2 (3B1), which may be accessed by near-UV solar excitation of SO2 to its excited 1B1 state followed by rapid intersystem crossing, reacts directly with water to form H2SO3 in the gas phase. For ground state SO2, this reaction is endothermic and has a very high activation barrier; our quantum chemical calculations point to a facile reaction being possible in the triplet state of SO2. This hygroscopic H2SO3 molecule may act as a condensation nucleus for water, giving rise to facile new particle formation (NPF). PMID:27417675

  5. Hydrolysis of somatostatin by human tissue kallikrein after the amino acid pair phe-Phe.

    PubMed Central

    Pimenta, D C; Juliano, M A; Juliano, L

    1997-01-01

    Somatostatin-(1-14) was hydrolysed by human tissue kallikrein at the Phe7-Trp8 bond, after a Phe-Phe pair of amino acids, with similar kinetic parameters to those described for human high- and low-molecular-mass kininogens. Substance P and human insulin, which also contain a Phe-Phe pair in their sequences, were both resistant. More details of this hydrolytic specificity of human tissue kallikrein were obtained by synthesizing and assaying internally quenched fluorescent peptides containing the sequence of somatostatin-(1-14), as well as the reactive-centre loop of human kallikrein-binding protein (kallistatin). We also observed that human tissue kallikrein hydrolysed growth hormone-releasing hormone at the Arg11-Lys12 bond, although this peptide contains in its structure a pair of leucines (Leu22-Leu23), in contrast with the Phe-Phe pair in somatostatin. We have also demonstrated the susceptibility to human tissue kallikrein of some chromogenic peptide s with the general structure of X-Phe-Phe-p-nitroanilide and D-Pro-Phe-Phe-4-methylcoumaryl-7-amide. PMID:9355730

  6. Synthesis, Chemical and Enzymatic Hydrolysis, and Aqueous Solubility of Amino Acid Ester Prodrugs of 3-Carboranyl Thymidine Analogues for Boron Neutron Capture Therapy of Brain Tumors

    PubMed Central

    Hasabelnaby, Sherifa; Goudah, Ayman; Agarwal, Hitesh K.; Abd alla, Mosaad S. M.; Tjarks, Werner

    2012-01-01

    Various water-soluble L-valine-, L-glutamate-, and glycine ester prodrugs of two 3-Carboranyl Thymidine Analogues (3-CTAs), designated N5 and N5-2OH, were synthesized for Boron Neutron Capture Therapy (BNCT) of brain tumors since the water solubilities of the parental compounds proved to be insufficient in preclinical studies. The amino acid ester prodrugs were prepared and stored as hydrochloride salts. The water solubilities of these amino acid ester prodrugs, evaluated in phosphate buffered saline (PBS) at pH 5, pH 6 and pH 7.4, improved 48 to 6600 times compared with parental N5 and N5-2OH. The stability of the amino acid ester prodrugs was evaluated in PBS at pH 7.4, Bovine serum, and Bovine cerebrospinal fluid (CSF). The rate of the hydrolysis in all three incubation media depended primarily on the amino acid promoiety and, to a lesser extend, on the site of esterification at the deoxyribose portion of the 3-CTAs. In general, 3'-amino acid ester prodrugs were less sensitive to chemical and enzymatic hydrolysis than 5'-amino acid ester prodrugs and the stabilities of the latter decreased in the following order: 5'-valine > 5'-glutamate > 5'-glycine. The rate of the hydrolysis of the 5'-amino acid ester prodrugs in Bovine CSF was overall higher than in PBS and somewhat lower than in Bovine serum. Overall, 5'-glutamate ester prodrug of N5 and the 5'-glycine ester prodrugs of N5 and N5-2OH appeared to be the most promising candidates for preclinical BNCT studies. PMID:22889558

  7. Summary Report on Gamma Radiolysis of TBP/n-dodecane in the Presence of Nitric Acid Using the Radiolysis/Hydrolysis Test Loop

    SciTech Connect

    Dean R. Peterman; Bruce J. Mincher; Catherine L. Riddle; Richard D. Tillotson

    2010-08-01

    Design and installation has been completed for a state-of-the-art radiolysis/hydrolysis test loop system. The system is used to evaluate the effects of gamma radiolysis and acid hydrolysis on the stability and performance of solvent extraction process solvents. The test loop is comprised of two main sections; the solvent irradiation and hydrolysis loop and the solvent reconditioning loop. In the solvent irradiation and hydrolysis loop, aqueous and organic phases are mixed and circulated through a gamma irradiator until the desired absorbed dose is achieved. Irradiation of the mixed phases is more representative of actual conditions in a solvent extraction process. Additionally, the contact of the organic phase with the aqueous phase will subject the solvent components to hydrolysis. This hydrolysis can be accelerated by controlling the system at an elevated temperature. At defined intervals, the organic from the irradiation/hydrolysis loop will be transferred to the solvent reconditioning loop where the solvent is contacted with scrub, strip, and solvent wash solutions which simulate process flowsheet conditions. These two processes are repeated until the total desired dose is achieved. Since all viable solvent extraction components in an advanced fuel cycle must exhibit high radiolytic and hydrolytic stability, this test loop is not limited to any one solvent system but is applicable to all systems of interest. Also, the test loop is not limited to testing of process flowsheets. It is also a valuable tool in support of fundamental research on newly identified extractants/modifiers and the impact of gamma radiation on their stability in a dynamic environment. The investigation of the radiolysis of a TBP/n-dodecane process solvent in contact with aqueous nitric acid has been performed. These studies were intended to confirm/optimize the operability of the test loop system. Additionally, these data are directly applicable to numerous other solvent extraction

  8. DFT modeling of the hydrolysis of isocyanic acid over the TiO 2 anatase (1 0 1) surface: Adsorption of HNCO species

    NASA Astrophysics Data System (ADS)

    Czekaj, Izabela; Piazzesi, Gaia; Kröcher, Oliver; Wokaun, Alexander

    2006-12-01

    The nature of the interaction of isocyanic acid (HNCO) with the active centers at the ideal anatase TiO 2 (1 0 1) surface were studied using ab initio density functional theory (DFT) method with a cluster model. Two types of adsorption of isocyanic acid are found to be likely at (1 0 1) surface - dissociative and molecular adsorption. Only molecular adsorption of HNCO leads to the direct weakening and further splitting of the N dbnd C bond, which is a necessary step for the hydrolysis of isocyanic acid. During molecular adsorption of HNCO, an energetically stable intermediate surface complex is created with an adsorption energy of -1.33 eV, in which the HNCO skeleton is changing due to new strong bonds between C-O s and N-Ti s. Based on the existence of this intermediate complex a probable reaction pathway for the hydrolysis of HNCO over the ideal anatase (1 0 1) surface was developed. A surface oxygen vacancy was formed after the decomposition of the intermediate complex and CO 2 desorption. Afterwards, water adsorbs at the oxygen vacancy site and NH 3 is successively formed. The HNCO hydrolysis over TiO 2 was found to be energetically favorable with global energy gain of about -2.08 eV.

  9. Enzymatic hydrolysis of beech wood lignocellulose at high solid contents and its utilization as substrate for the production of biobutanol and dicarboxylic acids.

    PubMed

    Tippkötter, Nils; Duwe, Anna-Maria; Wiesen, Sebastian; Sieker, Tim; Ulber, Roland

    2014-09-01

    The development of a cost-effective hydrolysis for crude cellulose is an essential part of biorefinery developments. To establish such high solid hydrolysis, a new solid state reactor with static mixing is used. However, concentrations >10% (w/w) cause a rate and yield reduction of enzymatic hydrolysis. By optimizing the synergetic activity of cellulolytic enzymes at solid concentrations of 9%, 17% and 23% (w/w) of crude Organosolv cellulose, glucose concentrations of 57, 113 and 152 g L(-1) are reached. However, the glucose yield decreases from 0.81 to 0.72 g g(-1) at 17% (w/w). Optimal conditions for hydrolysis scale-up under minimal enzyme addition are identified. As result, at 23% (w/w) crude cellulose the glucose yield increases from 0.29 to 0.49 g g(-1). As proof of its applicability, biobutanol, succinic and itaconic acid are produced with the crude hydrolysate. The potential of the substrate is proven e.g. by a high butanol yield of 0.33 g g(-1). PMID:25006020

  10. Characterization of a type I pullulanase from Anoxybacillus sp. SK3-4 reveals an unusual substrate hydrolysis.

    PubMed

    Kahar, Ummirul Mukminin; Ng, Chyan Leong; Chan, Kok-Gan; Goh, Kian Mau

    2016-07-01

    Type I pullulanases are enzymes that specifically hydrolyse α-1,6 linkages in polysaccharides. This study reports the analyses of a novel type I pullulanase (PulASK) from Anoxybacillus sp. SK3-4. Purified PulASK (molecular mass of 80 kDa) was stable at pH 5.0-6.0 and was most active at pH 6.0. The optimum temperature for PulASK was 60 °C, and the enzyme was reasonably stable at this temperature. Pullulan was the preferred substrate for PulASK, with 89.90 % adsorbance efficiency (various other starches, 56.26-72.93 % efficiency). Similar to other type I pullulanases, maltotriose was formed on digestion of pullulan by PulASK. PulASK also reacted with β-limit dextrin, a sugar rich in short branches, and formed maltotriose, maltotetraose and maltopentaose. Nevertheless, PulASK was found to preferably debranch long branches at α-1,6 glycosidic bonds of starch, producing amylose, linear or branched oligosaccharides, but was nonreactive against short branches; thus, no reducing sugars were detected. This is surprising as all currently known type I pullulanases produce reducing sugars (predominantly maltotriose) on digesting starch. The closest homologue of PulASK (95 % identity) is a type I pullulanase from Anoxybacillus sp. LM14-2 (Pul-LM14-2), which is capable of forming reducing sugars from starch. With rational design, amino acids 362-370 of PulASK were replaced with the corresponding sequence of Pul-LM14-2. The mutant enzyme formed reducing sugars on digesting starch. Thus, we identified a novel motif involved in substrate specificity in type I pullulanases. Our characterization may pave the way for the industrial application of this unique enzyme. PMID:27000839

  11. Enzymatic hydrolysis and recrystallization behavior of initially amorphous cellulose.

    PubMed

    Bertran, M S; Dale, B E

    1985-02-01

    Cellulose samples from cotton and wood pulps with varying low degrees of crystallinity (mechanically decrystallized) were studied. The influence of initial cellulose crystallinity on sugar yield after enzymatic hydrolysis was determined by two different methods. As expected, samples with low crystallinity were much more accessible to enzymatic attack and glucose yields were higher than were samples of high initial crystallinity. Hydrolysis of cellulose seems more dependent on cellulose crystallinity than on the source of cellulose. It is known that decrystallized or amorphous cellulose can recrystallize under proper conditions, e.g., during acid hydrolysis. The data reported here also reveal some recrystallization during enzymatic hydrolysis which probably occurs simulataneously with a selective enzymatic attack on the amorphous regions of cellulose. In all cases, the amorphous celluloses recrystallized in the original lattice form, that of native cellulose. PMID:18553653

  12. A study of concentrated acid hydrolysis conversion of lignocellulosic materials to sugars using a co-rotating twin-screw reactor extruder and plug flow reactor

    NASA Astrophysics Data System (ADS)

    Miller, William Scott

    Concerns about the ability of petroleum to continue supplying ever increasing global energy demands, at a price capable of generating continued economic growth, have spurred innovative research in the field of alternative energy. One alternative energy option that has the ability to provide long-term sustainable energy supplies for the global energy market is the conversion of lignocellulosic materials, via acid hydrolysis, to fermentable sugars for the production of fuel grade ethanol. This research demonstrates the ability of a co-rotating twin-screw reactor extruder and plug flow reactor to continuously convert lignocellulosic materials to fermentable sugars using high temperature concentrated acid hydrolysis. In addition to demonstrating continuous operation of the two-stage concentrated acid hydrolysis system, a number of design of experiments were conducted to model the twin-screw performance and maximize its ability to effectively solubilize lignocellulosic feedstocks in the high shear, elevated temperature, concentrated acid environment. These studies produced a base case twin-screw operating condition used to generate a standard extrudate composition for an extensive high temperature acid hydrolysis batch reactor kinetic modeling study. In this study a number of nonlinear and linear regression analyses were undertaken so that the concentration of less resistant cellulose, or the amount of solublilized extrudate cellulose, resistant cellulose, or non-solubilized extrudate cellulose, glucose, and decomposition products could be obtained as a function of time, temperature, and acid concentration. This study demonstrated that the theoretical cellulose conversion of 51% was limited by the amount of solubilized polysaccharides that could be produced in the twin-screw pretreatment. Further experimentation, showing twin-screw pretreatment lignocellulosic versatility, produced nearly identical results as the southern yellow pine sawdust experiments that were

  13. Hydrolysis of Cellulose by a Mesoporous Carbon-Fe2(SO4)3/γ-Fe2O3 Nanoparticle-Based Solid Acid Catalyst

    NASA Astrophysics Data System (ADS)

    Yamaguchi, Daizo; Watanabe, Koki; Fukumi, Shinya

    2016-02-01

    Carbon-based solid acid catalysts have shown significant potential in a wide range of applications, and they have been successfully synthesized using simple processes. Magnetically separable mesoporous carbon composites also have enormous potential, especially in separation and adsorption technology. However, existing techniques have been unable to produce a magnetically separable mesoporous solid acid catalyst because no suitable precursors have been identified. Herein we describe a magnetically separable, mesoporous solid acid catalyst synthesized from a newly developed mesoporous carbon-γ-Fe2O3 nanoparticle composite. This material exhibits an equivalent acid density and catalytic activity in the hydrolysis of microcrystalline cellulose, to that of the cellulose-derived conventional catalyst. Since it is magnetically separable, this material can be readily recovered and reused, potentially reducing the environmental impact of industrial processes to which it is applied.

  14. Hydrolysis of Cellulose by a Mesoporous Carbon-Fe2(SO4)3/γ-Fe2O3 Nanoparticle-Based Solid Acid Catalyst

    PubMed Central

    Yamaguchi, Daizo; Watanabe, Koki; Fukumi, Shinya

    2016-01-01

    Carbon-based solid acid catalysts have shown significant potential in a wide range of applications, and they have been successfully synthesized using simple processes. Magnetically separable mesoporous carbon composites also have enormous potential, especially in separation and adsorption technology. However, existing techniques have been unable to produce a magnetically separable mesoporous solid acid catalyst because no suitable precursors have been identified. Herein we describe a magnetically separable, mesoporous solid acid catalyst synthesized from a newly developed mesoporous carbon-γ-Fe2O3 nanoparticle composite. This material exhibits an equivalent acid density and catalytic activity in the hydrolysis of microcrystalline cellulose, to that of the cellulose-derived conventional catalyst. Since it is magnetically separable, this material can be readily recovered and reused, potentially reducing the environmental impact of industrial processes to which it is applied. PMID:26856604

  15. The relationship between surface tension and the industrial performance of water-soluble polymers prepared from acid hydrolysis lignin, a saccharification by-product from woody materials.

    PubMed

    Matsushita, Yasuyuki; Imai, Masanori; Iwatsuki, Ayuko; Fukushima, Kazuhiko

    2008-05-01

    In this study, water-soluble anionic and cationic polymers were prepared from sulfuric acid lignin (SAL), an acid hydrolysis lignin, and the relationship between the surface tension of these polymers and industrial performance was examined. The SAL was phenolized (P-SAL) to enhance its solubility and reactivity. Sulfonation and the Mannich reaction with aminocarboxylic acids produced water-soluble anionic polymers and high-dispersibility gypsum paste. The dispersing efficiency increased as the surface tension decreased, suggesting that the fluidity of the gypsum paste increased with the polymer adsorption on the gypsum particle surface. Water-soluble cationic polymers were prepared using the Mannich reaction with dimethylamine. The cationic polymers showed high sizing efficiency under neutral papermaking conditions; the sizing efficiency increased with the surface tension. This suggests that the polymer with high hydrophilicity spread in the water and readily adhered to the pulp surface and the rosin, showing good retention. PMID:17664066

  16. New Experimental and Theoretical Approach to the Heterogeneous Hydrolysis of NO2: Key Role of Molecular Nitric Acid and Its Complexes

    SciTech Connect

    Ramazan, K A.; Wingen, Lisa M.; Miller, Y; Chaban, Galina M.; Gerber, Robert B.; Xantheas, Sotiris S.; Finlayson-Pitts, Barbara J.

    2006-06-01

    Although heterogeneous chemistry on surfaces in the troposphere is known to be important, there are currently only a few techniques available for studying the nature of surface-adsorbed species as well as their chemistry and photochemistry under atmospheric conditions of 1 atm pressure and in the presence of water vapor. We report here a new laboratory approach using a combination of long path FTIR to identify gases and attenuated total reflectance (ATR)-FTIR that allows the simultaneous observation and quantification of gases and surface species. Theory is used to identify the surface-adsorbed intermediates and products, and to quantify their relative concentrations. At intermediate relative humidities typical of the tropospheric boundary layer, the nitric acid formed in the NO2 heterogeneous hydrolysis is shown to exist as both nitrate ions from the dissociation of nitric acid formed on the surface, and as molecular nitric acid. In both cases, the ions and HNO3 are complexed to water molecules. Upon pumping, water is selectively removed, shifting the NO3- - HNO3(H2O)y equilibria towards more dehydrated forms of HNO3. Photolysis of the nitric acid-water film using 300-400 nm radiation generates gaseous NO, while photolysis at 254 nm generates both NO and HONO, resulting in conversion of surface-adsorbed nitrogen oxides into photochemically active NOx. This suggests that the assumption that deposition or formation of nitric acid provides a permanent removal mechanism from the atmosphere may not be correct. Furthermore, a potential role of surface-adsorbed nitric acid and other species formed during the heterogeneous hydrolysis of NO2 on the oxidation of organics on surfaces, and on the generation of gas phase HONO on local to global scales, should be considered.

  17. Pretreatment by NaOH swelling and then HCl regeneration to enhance the acid hydrolysis of cellulose to glucose.

    PubMed

    Sun, Binzhe; Peng, Gege; Duan, Lian; Xu, Aihua; Li, Xiaoxia

    2015-11-01

    A simple pretreatment method, NaOH swelling at a low temperature and then HCl regeneration, was developed for depolymerization of cellulose to glucose. Cellulose was transformed into amorphous hydrogel during the treatment; and due to the easy diffusion of H(+) and Cl(-) ions into the cellulose hydrogel network as well as the strong ability of Cl(-) to disrupt the massive hydrogen bond, the hydrolysis rate was greatly enhanced. The method is effective for α-cellulose, microcrystalline cellulose, filter paper, ramie fiber and absorbent cotton. Even at a high α-cellulose concentration of 50g/L, 29.1% yield of glucose was still obtained within 10min at 160°C under microwave irradiation, where up to 16.2g/L of glucose solution was given. The influence of NaOH concentration, HCl concentration, hydrolysis temperature and time on the hydrolysis rate was investigated. The structure of cellulose hydrogel was also studied to confirm the reaction mechanism. PMID:26280097

  18. Joint action of ultrasonic and Fe³⁺ to improve selectivity of acid hydrolysis for microcrystalline cellulose.

    PubMed

    Li, Jinbao; Qiang, Dandan; Zhang, Meiyun; Xiu, Huijuan; Zhang, Xiangrong

    2015-09-20

    In this study, the combination of Fe(3+)/HCl and ultrasonic treatment was applied to selectively hydrolyze cellulose for the preparation of microcrystalline cellulose (MCC). It was found that the crystallinity and specific surface area of hydrocellulose samples were higher (78.92% and 2.23581 m(2)g(-1), respectively), compared with the method that only used Fe(3+)/HCl catalyst without ultrasonic treatment. Meanwhile, the hydrolysate can be extracted and reused for cellulose hydrolysis for three runs, which was effective in saving the dosage of chemicals and reducing the pollution of the environment without affecting the properties of hydrocellulose. Moreover, the increased concentration of total reducing sugar (TRS) after three runs may be used as a valuable source in biofuels production. The technology of cellulose hydrolysis, by retaining the crystalline region for MCC products while promoting hydrolysis of amorphous region for further utilization is of great novelty, which may prove valuable in converting biomass into chemicals and biofuels, environmentally and economically. PMID:26050886

  19. Recovery of acetic acid from pre-hydrolysis liquor of hardwood kraft-based dissolving pulp production process by reactive extraction with triisooctylamine.

    PubMed

    Yang, G; Jahan, M Sarwar; Ahsan, Laboni; Zheng, Linqiang; Ni, Yonghao

    2013-06-01

    Acetic acid was one of the main compositions of the pre-hydrolysis liquor (PHL), which was recovered by reactive extraction with triisooctylamine (TIOA) diluted with decanol. Dilution of TIOA played an important role in extracting acetic acid from the PHL. The recovery of acetic acid from the PHL by TIOA was increased from 10.34% to 66.60% with the dilution of TIOA to 20% by decanol at the HAc to TIOA molar ratio of 1, consequently, the equilibrium distribution coefficient KD increased. The effects of time, temperature and pH on the extraction process were also studied. The extraction process was very fast. The acetic acid extraction decreased from 65.13% to 57.34% with the rise of temperature to 50°C from 20°C. A higher pH increased the dissociation of acetic acid, as a result, decreased acetic acid extraction. The hemicelluloses in the PHL were unaffected on the extraction process of acetic acid. PMID:23619137

  20. Deposition of Lignin Droplets Produced During Dilute Acid Pretreatment of Maize Stems Retards Enzymatic Hydrolysis of Cellulose

    SciTech Connect

    Selig, M. J.; Viamajala, S.; Decker, S. R.; Tucker, M. P.; Himmel, M. E.; Vinzant, T. B.

    2007-01-01

    Electron microscopy of lignocellulosic biomass following high-temperature pretreatment revealed the presence of spherical formations on the surface of the residual biomass. The hypothesis that these droplet formations are composed of lignins and possible lignin carbohydrate complexes is being explored. Experiments were conducted to better understand the formation of these lignin droplets and the possible implications they might have on the enzymatic saccharification of pretreated biomass. It was demonstrated that these droplets are produced from corn stover during pretreatment under neutral and acidic pH at and above 130C, and that they can deposit back onto the surface of residual biomass. The deposition of droplets produced under certain pretreatment conditions (acidic pH; T > 150C) and captured onto pure cellulose was shown to have a negative effect (5-20%) on the enzymatic saccharification of this substrate. It was noted that droplet density (per unit area) was greater and droplet size more variable under conditions where the greatest impact on enzymatic cellulose conversion was observed. These results indicate that this phenomenon has the potential to adversely affect the efficiency of enzymatic conversion in a lignocellulosic biorefinery.

  1. Poly(adenylic acid) in small amounts, free or covalently linked to substrate, protects RNA from hydrolysis by ribonuclease.

    PubMed Central

    Karpetsky, T P; Shriver, K K; Levy, C C

    1981-01-01

    Short lengths (18 residues) of poly(A), covalently linked to the 3'-termini of Escherichia coli 5 S rRNA, induce powerful inhibitions (38-87%) of the activities of RNAases (ribonucleases) from Citrobacter sp., Enterobacter sp., bovine pancreas, human spleen and human plasma. As the polypurine chain length is extended, enzyme activity declines. Furthermore, poly(A) sequences, present only on a small subpopulation of RNA, and accounting for less than 1% of total RNA, serve to protect all RNA, polyadenylated or not, from enzyme-catalysed degradation. The quantity of 3'-terminal adenylic acid residues, relative to the amount of substrate, determines enzyme activity. The exact distribution of a fixed amount of poly(A) residues on the 3'-termini of substrate molecules is unimportant in this respect. Comparison of the efficacies of inhibition of RNAase activity, by using linked poly(A) and similar quantities of free poly(A), revealed that although the free polypurine inhibits RNAase activity, covalent linkage of poly(A) to RNA is more advantageous to the stability of an RNA substrate. However, the ratio of inhibited activities obtained by using linked or free poly(A) may change considerably with alterations in either substrate concentration or polyadenylic acid segment length. PMID:6171250

  2. Imidazole-derived 2-[N-carbamoylmethyl-alkylamino]acetic acids, substrate-dependent modulators of insulin-degrading enzyme in amyloid-β hydrolysis.

    PubMed

    Charton, Julie; Gauriot, Marion; Guo, Qing; Hennuyer, Nathalie; Marechal, Xavier; Dumont, Julie; Hamdane, Malika; Pottiez, Virginie; Landry, Valerie; Sperandio, Olivier; Flipo, Marion; Buee, Luc; Staels, Bart; Leroux, Florence; Tang, Wei-Jen; Deprez, Benoit; Deprez-Poulain, Rebecca

    2014-05-22

    Insulin degrading enzyme (IDE) is a highly conserved zinc metalloprotease that is involved in the clearance of various physiologically peptides like amyloid-beta and insulin. This enzyme has been involved in the physiopathology of diabetes and Alzheimer's disease. We describe here a series of small molecules discovered by screening. Co-crystallization of the compounds with IDE revealed a binding both at the permanent exosite and at the discontinuous, conformational catalytic site. Preliminary structure-activity relationships are described. Selective inhibition of amyloid-beta degradation over insulin hydrolysis was possible. Neuroblastoma cells treated with the optimized compound display a dose-dependent increase in amyloid-beta levels. PMID:24735644

  3. Chemometrics-assisted Spectrofluorimetric Determination of Two Co-administered Drugs of Major Interaction, Methotrexate and Aspirin, in Human Urine Following Acid-induced Hydrolysis.

    PubMed

    Maher, Hadir M; Ragab, Marwa A A; El-Kimary, Eman I

    2015-01-01

    Methotrexate (MTX) is widely used to treat rheumatoid arthritis (RA), mostly along with non-steroidal anti-inflammatory drugs (NSAIDs), the most common of which is aspirin or acetyl salicylic acid (ASA). Since NSAIDs impair MTX clearance and increase its toxicity, it was necessary to develop a simple and reliable method for the monitoring of MTX levels in urine samples, when coadministered with ASA. The method was based on the spectrofluorimetric measurement of the acid-induced hydrolysis product of MTX, 4-amino-4-deoxy-10-methylpteroic acid (AMP), along with the strongly fluorescent salicylic acid (SA), a product of acid-induced hydrolysis of aspirin and its metabolites in urine. The overlapping emission spectra were resolved using the derivative method (D method). In addition, the corresponding derivative emission spectra were convoluted using discrete Fourier functions, 8-points sin xi polynomials, (D/FF method) for better elimination of interferences. Validation of the developed methods was carried out according to the ICH guidelines. Moreover, the data obtained using derivative and convoluted derivative spectra were treated using the non-parametric Theil's method (NP), compared with the least-squares parametric regression method (LSP). The results treated with Theil's method were more accurate and precise compared with LSP since the former is less affected by the outliers. This work offers the potential of both derivative and convolution using discrete Fourier functions in addition to the effectiveness of using the NP regression analysis of data. The high sensitivity obtained by the proposed methods was promising for measuring low concentration levels of the two drugs in urine samples. These methods were efficiently used to measure the drugs in human urine samples following their co-administration. PMID:26234512

  4. Improving the enzymatic hydrolysis of dilute acid pretreated wheat straw by metal ion blocking of non-productive cellulase adsorption on lignin.

    PubMed

    Akimkulova, Ardak; Zhou, Yan; Zhao, Xuebing; Liu, Dehua

    2016-05-01

    Eleven salts were selected to screen the possible metal ions for blocking the non-productive adsorption of cellulase onto the lignin of dilute acid pretreated wheat straw. Mg(2+) was screened finally as the promising candidate. The optimal concentration of MgCl2 was 1mM, but the beneficial action was also dependent on pH, hydrolysis time and cellulase loading. Significant improvement of glucan conversion (19.3%) was observed at low cellulase loading (5FPU/g solid). Addition of isolated lignins, tannic acid and lignin model compounds to pure cellulose hydrolysis demonstrated that phenolic hydroxyl group (Ph-OH) was the main active site blocked by Mg(2+). The interaction between Mg(2+) and Ph-OH of lignin monomeric moieties followed an order of p-hydroxyphenyl (H)>guaiacyl (G)>syringyl (S). Mg(2+) blocking made the lignin surface less negatively charged, which might weaken the hydrogen bonding and electrostatically attractive interaction between lignin and cellulase enzymes. PMID:26930032

  5. Hydrolysis reactor for hydrogen production

    SciTech Connect

    Davis, Thomas A.; Matthews, Michael A.

    2012-12-04

    In accordance with certain embodiments of the present disclosure, a method for hydrolysis of a chemical hydride is provided. The method includes adding a chemical hydride to a reaction chamber and exposing the chemical hydride in the reaction chamber to a temperature of at least about 100.degree. C. in the presence of water and in the absence of an acid or a heterogeneous catalyst, wherein the chemical hydride undergoes hydrolysis to form hydrogen gas and a byproduct material.

  6. Phosphonate ester hydrolysis catalyzed by two lanthanum ions. Intramolecular nucleophilic attack of coordinated hydroxide and lewis acid activation

    SciTech Connect

    Tsubouchi, A.; Bruice, T.C.

    1995-07-19

    (8-Hydroxy-2-quinolyl)methyl (8-hydroxy-2-quinolyl)methyl phosphonate (I) has been synthesized as a model compound and investigated in terms of catalysis of hydrolysis by two metal ions in concert. Removal of one of two 8-hydroxyquinoline ligands of I to provide (8-hydroxy-2-quinolyl)methylmethylphosphonate (II) leads to the formation of the 1:1 complex (II)La, which is hydrolytically inert but subject to catalysis by free La{sup 3+}. From thermodynamic studies of metal ion complexation and comparison of the kinetics of hydrolysis of I and II in the presence of metal ions, we conclude the following. The phosphonate ester I forms a hydrolytically active 1:2 complex (I)La{sub 2} with La{sup 3+} but inert 1:1 complexes with Zn{sup 2+}, Ni{sup 2+}, Co{sup 2+}, Cu{sup 2+}, and Al{sup 3+}. The La{sup 3+} in the (I)La{sub 2} complex serve to (i) facilitate the formation of metal ligated hydroxide as an intramolecule nucleophile; (ii) stabilize the transition state of the hydrolysis by neutralization of the phosphonate negative charge; and (iii) interact with an incipient oxyanion of the leaving alcohol. The two La{sup 3+} functions operate in concert and provide nearly 10{sup 13} rate enhancement. Consequently the 1:2 complex (I)La{sub 2}(OH{sub 2}){sub n-1}(OH) may serve as a model for the 3`-5` exonuclease reaction of E. coli DNA polymerase I. 39 refs., 7 figs., 3 tabs.

  7. Comparison of the levels of 8-hydroxyguanine in DNA as measured by gas chromatography mass spectrometry following hydrolysis of DNA by Escherichia coli Fpg protein or formic acid

    PubMed Central

    Rodriguez, Henry; Jurado, Juan; Laval, Jacques; Dizdaroglu, Miral

    2000-01-01

    8-hydroxyguanine (8-OH-Gua) is one of many lesions generated in DNA by oxidative processes including free radicals. It is the most extensively investigated lesion, due to its miscoding properties and its potential role in mutagenesis, carcinogenesis and aging, and also to the existence of analytical methods using HPLC and gas chromatography mass spectrometry (GC/MS). Some studies raised the possibility of artifacts generated during sample preparation. We investigated several experimental conditions in order to eliminate possible artifacts during the measurement of 8-OH-Gua by GC/MS. Derivatization has been reported to produce artifacts by oxidation of guanine to 8-OH-Gua in acid-hydrolysates of DNA, although the extent of artifacts seems to depend on experimental conditions. For removal of 8-OH-Gua from DNA, we used either formic acid hydrolysis or specific enzymatic hydrolysis with Escherichia coli Fpg protein. Derivatization of enzyme-hydrolysates should not generate additional 8-OH-Gua because of the absence of guanine, which is not released by the enzyme, whereas guanine released by acid may be oxidized to yield 8-OH-Gua. The measurement of 8-OH-Gua in calf thymus DNA by GC/isotope-dilution MS (GC/IDMS) using these two different hydrolyses yielded similar levels of 8-OH-Gua. This indicated that no artifacts occurred during derivatization of acid-hydrolysates of DNA. Pyridine instead of acetonitrile and room temperature were used during derivatization. Pyridine reduced the level of 8-OH-Gua, when compared with acetonitrile, indicating its potential to prevent oxidation. Two different stable-isotope labeled analogs of 8-OH-Gua used as internal standards for GC/IDMS analysis yielded similar results. A comparison of the present results with the results of recent trials by the European Standards Committee for Oxidative DNA Damage (ESCODD) is also presented. PMID:10908368

  8. Real-time NMR Study of Three Small GTPases Reveals That Fluorescent 2′(3′)-O-(N-Methylanthraniloyl)-tagged Nucleotides Alter Hydrolysis and Exchange Kinetics*

    PubMed Central

    Mazhab-Jafari, Mohammad T.; Marshall, Christopher B.; Smith, Matthew; Gasmi-Seabrook, Geneviève M. C.; Stambolic, Vuk; Rottapel, Robert; Neel, Benjamin G.; Ikura, Mitsuhiko

    2010-01-01

    The Ras family of small GTPases control diverse signaling pathways through a conserved “switch” mechanism, which is turned on by binding of GTP and turned off by GTP hydrolysis to GDP. Full understanding of GTPase switch functions requires reliable, quantitative assays for nucleotide binding and hydrolysis. Fluorescently labeled guanine nucleotides, such as 2′(3′)-O-(N-methylanthraniloyl) (mant)-substituted GTP and GDP analogs, have been widely used to investigate the molecular properties of small GTPases, including Ras and Rho. Using a recently developed NMR method, we show that the kinetics of nucleotide hydrolysis and exchange by three small GTPases, alone and in the presence of their cognate GTPase-activating proteins (GAPs) and guanine nucleotide exchange factors, are affected by the presence of the fluorescent mant moiety. Intrinsic hydrolysis of mantGTP by Ras homolog enriched in brain (Rheb) is ∼10 times faster than that of GTP, whereas it is 3.4 times slower with RhoA. On the other hand, the mant tag inhibits TSC2GAP-catalyzed GTP hydrolysis by Rheb but promotes p120 RasGAP-catalyzed GTP hydrolysis by H-Ras. Guanine nucleotide exchange factor-catalyzed nucleotide exchange for both H-Ras and RhoA was inhibited by mant-substituted nucleotides, and the degree of inhibition depends highly on the GTPase and whether the assay measures association of mantGTP with, or dissociation of mantGDP from the GTPase. These results indicate that the mant moiety has significant and unpredictable effects on GTPase reaction kinetics and underscore the importance of validating its use in each assay. PMID:20018863

  9. Innovations in host and microbial sialic acid biosynthesis revealed by phylogenomic prediction of nonulosonic acid structure

    PubMed Central

    Lewis, Amanda L.; Desa, Nolan; Hansen, Elizabeth E.; Knirel, Yuriy A.; Gordon, Jeffrey I.; Gagneux, Pascal; Nizet, Victor; Varki, Ajit

    2009-01-01

    Sialic acids (Sias) are nonulosonic acid (NulO) sugars prominently displayed on vertebrate cells and occasionally mimicked by bacterial pathogens using homologous biosynthetic pathways. It has been suggested that Sias were an animal innovation and later emerged in pathogens by convergent evolution or horizontal gene transfer. To better illuminate the evolutionary processes underlying the phenomenon of Sia molecular mimicry, we performed phylogenomic analyses of biosynthetic pathways for Sias and related higher sugars derived from 5,7-diamino-3,5,7,9-tetradeoxynon-2-ulosonic acids. Examination of ≈1,000 sequenced microbial genomes indicated that such biosynthetic pathways are far more widely distributed than previously realized. Phylogenetic analysis, validated by targeted biochemistry, was used to predict NulO types (i.e., neuraminic, legionaminic, or pseudaminic acids) expressed by various organisms. This approach uncovered previously unreported occurrences of Sia pathways in pathogenic and symbiotic bacteria and identified at least one instance in which a human archaeal symbiont tentatively reported to express Sias in fact expressed the related pseudaminic acid structure. Evaluation of targeted phylogenies and protein domain organization revealed that the “unique” Sia biosynthetic pathway of animals was instead a much more ancient innovation. Pathway phylogenies suggest that bacterial pathogens may have acquired Sia expression via adaptation of pathways for legionaminic acid biosynthesis, one of at least 3 evolutionary paths for de novo Sia synthesis. Together, these data indicate that some of the long-standing paradigms in Sia biology should be reconsidered in a wider evolutionary context of the extended family of NulO sugars. PMID:19666579

  10. The Rates of Hydrolysis of Thymidyl-3', 5'-Thymidine-H-Phosphonate: The Possible Role of Nucleic Acids Linked by Diesters of Phosphorous Acid in the Origins of Life

    NASA Astrophysics Data System (ADS)

    Peyser, John R.; Ferris, James P.

    2001-08-01

    Thymidyl-3',5'-thymidine H-phosphonate undergoes acid, base, and water-catalyzed hydrolysis. The products were 3'-thymidine H-phosphonate, 5'-thymidine H-phosphonate, and thymidine in a ratio of 1:1:2. The rate constants are 1.8 × 10^-3 M^-1 sec^-1, 7.2 × 10^3 M^-1 sec^-1, and 1.5 × 10^-6 sec^-1 for acid, base and water catalysis, respectively. These values are comparable with previous reports for the rates of hydrolysis of simple dialkyl esters of phosphorous acids. The Arrhenius activation energy for the base-catalyzed reaction is 20 kcal/mol. and the enthalpy and entropy of activation are 19 kcal/mol and -14 eu., respectively. The Gibbs free energy of activation is 23 kcal/mol. The rate constants suggest that nucleic acids linked by diesters of phosphorous acid hydrolyze too rapidly in aqueous solution to have accumulated in useful amounts on the primitive Earth.

  11. Active-Site Residues of Escherichia coli DNA Gyrase Required in Coupling ATP Hydrolysis to DNA Supercoiling and Amino Acid Substitutions Leading to Novobiocin Resistance

    PubMed Central

    Gross, Christian H.; Parsons, Jonathan D.; Grossman, Trudy H.; Charifson, Paul S.; Bellon, Steven; Jernee, James; Dwyer, Maureen; Chambers, Stephen P.; Markland, William; Botfield, Martyn; Raybuck, Scott A.

    2003-01-01

    DNA gyrase is a bacterial type II topoisomerase which couples the free energy of ATP hydrolysis to the introduction of negative supercoils into DNA. Amino acids in proximity to bound nonhydrolyzable ATP analog (AMP · PNP) or novobiocin in the gyrase B (GyrB) subunit crystal structures were examined for their roles in enzyme function and novobiocin resistance by site-directed mutagenesis. Purified Escherichia coli GyrB mutant proteins were complexed with the gyrase A subunit to form the functional A2B2 gyrase enzyme. Mutant proteins with alanine substitutions at residues E42, N46, E50, D73, R76, G77, and I78 had reduced or no detectable ATPase activity, indicating a role for these residues in ATP hydrolysis. Interestingly, GyrB proteins with P79A and K103A substitutions retained significant levels of ATPase activity yet demonstrated no DNA supercoiling activity, even with 40-fold more enzyme than the wild-type enzyme, suggesting that these amino acid side chains have a role in the coupling of the two activities. All enzymes relaxed supercoiled DNA to the same extent as the wild-type enzyme did, implying that only ATP-dependent reactions were affected. Mutant genes were examined in vivo for their abilities to complement a temperature-sensitive E. coli gyrB mutant, and the activities correlated well with the in vitro activities. We show that the known R136 novobiocin resistance mutations bestow a significant loss of inhibitor potency in the ATPase assay. Four new residues (D73, G77, I78, and T165) that, when changed to the appropriate amino acid, result in both significant levels of novobiocin resistance and maintain in vivo function were identified in E. coli. PMID:12604539

  12. Contrasted enzymatic cocktails reveal the importance of cellulases and hemicellulases activity ratios for the hydrolysis of cellulose in presence of xylans.

    PubMed

    Dondelinger, Eve; Aubry, Nathalie; Ben Chaabane, Fadhel; Cohen, Céline; Tayeb, Jean; Rémond, Caroline

    2016-03-01

    Various enzymatic cocktails were produced from two Trichoderma reesei strains, a cellulase hyperproducer strain and a strain with β-glucosidase activity overexpression. By using various carbon sources (lactose, glucose, xylose, hemicellulosic hydrolysate) for strains growth, contrasted enzymatic activities were obtained. The enzymatic cocktails presented various levels of efficiency for the hydrolysis of cellulose Avicel into glucose, in presence of xylans, or not. These latter were also hydrolyzed with different extents according to cocktails. The most efficient cocktails (TR1 and TR3) on Avicel were richer in filter paper activity (FPU) and presented a low ratio FPU/β-glucosidase activity. Cocktails TR2 and TR5 which were produced on the higher amount of hemicellulosic hydrolysate, possess both high xylanase and β-xylosidase activities, and were the most efficient for xylans hydrolysis. When hydrolysis of Avicel was conducted in presence of xylans, a decrease of glucose release occurred for all cocktails compared to hydrolysis of Avicel alone. Mixing TR1 and TR5 cocktails with two different ratios of proteins (1/1 and 1/4) resulted in a gain of efficiency for glucose release during hydrolysis of Avicel in presence of xylans compared to TR5 alone. Our results demonstrate the importance of combining hemicellulase and cellulase activities to improve the yields of glucose release from Avicel in presence of xylans. In this context, strategies involving enzymes production with carbon sources comprising mixed C5 and C6 sugars or combining different cocktails produced on C5 or on C6 sugars are of interest for processes developed in the context of lignocellulosic biorefinery. PMID:27001439

  13. A study of the acid-catalyzed hydrolysis of cellulose dissolved in ionic liquids and the factors influencing the dehydration of glucose and the formation of humins.

    PubMed

    Dee, Sean J; Bell, Alexis T

    2011-08-22

    An investigation was carried out into the hydrolysis of cellulose dissolved in 1-ethyl-3-methylimidazolium chloride ([Emim][Cl]) and 1-butyl-3-methylimidazolium chloride ([Bmim][Cl]) catalyzed by mineral acids. Glucose, cellobiose, and 5-hydroxymethylfurfural (5-HMF) were observed as the primary reaction products. The initial rate of glucose formation was determined to be of first order in the concentrations of dissolved glucan and protons and of zero order in the concentration of water. The absence of a dependence on water concentration suggests that cleavage of the β-1,4-glycosidic linkages near chain ends is irreversible. The apparent activation energy for glucose formation is 96 kJ mol(-1). The absence of oligosaccharides longer than cellobiose suggests that cleavage of interior glycosidic bonds is reversible due to the slow diffusional separation of cleaved chains in the highly viscous glucan/ionic liquid solution. Progressive addition of water during the course of glucan hydrolysis inhibited the rate of glucose dehydration to 5-HMF and the formation of humins. The inhibition of glucose dehydration is attributed to stronger interaction of protons with water than the 2-OH atom of the pyranose ring of glucose, the critical step in the proposed mechanism for the formation of 5-HMF. The reduction in humin formation associated with water addition is ascribed to the lowered concentration of 5-HMF, since the formation of humins is suggested to proceed through the condensation polymerization of 5-HMF with glucose. PMID:21809450

  14. Fermentative l-lactic acid production from pretreated whole slurry of oil palm trunk treated by hydrothermolysis and subsequent enzymatic hydrolysis.

    PubMed

    Eom, In-Yong; Oh, Young-Hoon; Park, Si Jae; Lee, Seung-Hwan; Yu, Ju-Hyun

    2015-06-01

    A simple and cost-effective biochemical conversion process consisting of hydrothermal treatment, enzymatic hydrolysis and fermentation of pretreated whole slurry (PWS) was developed for producing l-lactic acid (L-LA) from oil palm trunk (OPT). When OPT was hydrothermally treated at optimal condition capable of achieving maximum yield of hemicellulosic sugars after enzymatic hydrolysis, the enzymatic digestibility of the PWS afforded a yield of 81.4% of the theoretical glucose yield (TGY). However, glucose yield from washed pretreated solid (WPS) was only 43.5% of TGY. The use of two hydrolysates from PWS and WPS for fermentation by Lactobacillus paracasei engineered to selectively produce L-LA afforded yields of 89.5% and 45.8% of the theoretical LA yield (TLY), respectively. This study confirmed the inevitable extensive sugar loss during washing of pretreated slurry due to loss of soluble starch. Alternatively, the proposed design process is considered suitable for converting OPT to L-LA without such starch loss. PMID:25768416

  15. The effect of microcrystalline cellulose crystallinity on the hydrophilic property of tablets and the hydrolysis of acetylsalicylic acid as active pharmaceutical ingredient inside tablets.

    PubMed

    Awa, Kimie; Shinzawa, Hideyuki; Ozaki, Yukihiro

    2015-08-01

    The crystal structures of active pharmaceutical ingredients and excipients should be strictly controlled because they influence pharmaceutical properties of products which cause the change in the quality or the bioavailability of the products. In this study, we investigated the effects of microcrystalline cellulose (MCC) crystallinity on the hydrophilic properties of tablets and the hydrolysis of active pharmaceutical ingredient, acetylsalicylic acid (ASA), inside tablets by using tablets containing 20% MCC as an excipient. Different levels of grinding were applied to MCC prior to tablet formulation, to intentionally cause structural variation in the MCC. The water penetration and moisture absorbability of the tablets increased with decreasing the crystallinity of MCC through higher level of grinding. More importantly, the hydrolysis of ASA inside tablets was also accelerated. These results indicate that the crystallinity of MCC has crucial effects on the pharmaceutical properties of tablets even when the tablets contain a relatively small amount of MCC. Therefore, controlling the crystal structure of excipients is important for controlling product qualities. PMID:25583304

  16. Hydrolysis of wheat flour arabinoxylan, acid-debranched wheat flour arabinoxylan and arabino-xylo-oligosaccharides by β-xylanase, α-L-arabinofuranosidase and β-xylosidase.

    PubMed

    McCleary, Barry V; McKie, Vincent A; Draga, Anna; Rooney, Edward; Mangan, David; Larkin, Jennifer

    2015-04-30

    A range of α-L-arabinofuranosyl-(1-4)-β-D-xylo-oligosaccharides (AXOS) were produced by hydrolysis of wheat flour arabinoxylan (WAX) and acid debranched arabinoxylan (ADWAX), in the presence and absence of an AXH-d3 α-L-arabinofuranosidase, by several GH10 and GH11 β-xylanases. The structures of the oligosaccharides were characterised by GC-MS and NMR and by hydrolysis by a range of α-L-arabinofuranosidases and β-xylosidase. The AXOS were purified and used to characterise the action patterns of the specific α-L-arabinofuranosidases. These enzymes, in combination with either Cellvibrio mixtus or Neocallimastix patriciarum β-xylanase, were used to produce elevated levels of specific AXOS on hydrolysis of WAX, such as 3(2)-α-L-Araf-(1-4)-β-D-xylobiose (A(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(3)XX), 2(2)-α-L-Araf-(1-4)-β-D-xylotriose (XA(2)X), 3(2)-α-L-Araf (1-4)-β-D-xylotriose (XA(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotriose (A(2+3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2+3)XX), 2(4),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(2+3)XXX) and 3(3),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(3)A(3)XX), many of which have not previously been produced in sufficient quantities to allow their use as substrates in further enzymic studies. For A(2,3)XX, yields of approximately 16% of the starting material (wheat arabinoxylan) have been achieved. Mixtures of the α-L-arabinofuranosidases, with specific action on AXOS, have been combined with β-xylosidase and β-xylanase to obtain an optimal mixture for hydrolysis of arabinoxylan to L-arabinose and D-xylose. PMID:25723624

  17. Hydrolysis-acidogenesis of food waste in solid-liquid-separating continuous stirred tank reactor (SLS-CSTR) for volatile organic acid production.

    PubMed

    Karthikeyan, Obulisamy Parthiba; Selvam, Ammaiyappan; Wong, Jonathan W C

    2016-01-01

    The use of conventional continuous stirred tank reactor (CSTR) can affect the methane (CH4) recovery in a two-stage anaerobic digestion of food waste (FW) due to carbon short circuiting in the hydrolysis-acidogenesis (Hy-Aci) stage. In this research, we have designed and tested a solid-liquid-separating CSTR (SLS-CSTR) for effective Hy-Aci of FW. The working conditions were pH 6 and 9 (SLS-CSTR-1 and -2, respectively); temperature-37°C; agitation-300rpm; and organic loading rate (OLR)-2gVSL(-1)day(-1). The volatile fatty acids (VFA), enzyme activities and bacterial population (by qPCR) were determined as test parameters. Results showed that the Hy-Aci of FW at pH 9 produced ∼35% excess VFA as compared to that at pH 6, with acetic and butyric acids as major precursors, which correlated with the high enzyme activities and low lactic acid bacteria. The design provided efficient solid-liquid separation there by improved the organic acid yields from FW. PMID:26512860

  18. Efficient hydrolysis of corncob residue through cellulolytic enzymes from Trichoderma strain G26 and L-lactic acid preparation with the hydrolysate.

    PubMed

    Xie, Lulu; Zhao, Jin; Wu, Jian; Gao, Mingfu; Zhao, Zhewei; Lei, Xiangyun; Zhao, Yi; Yang, Wei; Gao, Xiaoxue; Ma, Cuiyun; Liu, Huanfei; Wu, Fengjuan; Wang, Xingxing; Zhang, Fengwei; Guo, Pengyuan; Dai, Guifu

    2015-10-01

    To prepare fermentable hydrolysate from corncob residue (CCR), Trichoderma strain G26 was cultured on medium containing CCR for production of cellulolytic enzymes through solid-state fermentation (SSF), resulting in 71.3 IU/g (FPA), 136.2 IU/g (CMCase), 85.1 IU/g (β-glucosidase) and 11,344 IU/g (xylanase), respectively. Through a three-stage saccharification strategy, CCR was hydrolyzed by the enzymatic solution (6.5 FPU/ml) into fermentable hydrolysate containing 60.1g/l glucose (81.2% cellulose was converted at solid loading of 12.5%), 21.4% higher than that by the one-stage method. And then the hydrolysate was used to produce L-lactic acid by a previous screened strain Bacillus coagulans ZX25 in the submerged fermentation. 52.0 g/l L-lactic acid was obtained after fermentation for 44 h, with 86.5% glucose being converted to L-lactic acid. The results indicate that the strains and the hydrolysis strategy are promising for commercial production of L-lactic acid from CCR and other biomass. PMID:26143000

  19. Evaluation of soluble fraction and enzymatic residual fraction of dilute dry acid, ethylenediamine, and steam explosion pretreated corn stover on the enzymatic hydrolysis of cellulose.

    PubMed

    Qin, Lei; Liu, Li; Li, Wen-Chao; Zhu, Jia-Qing; Li, Bing-Zhi; Yuan, Ying-Jin

    2016-06-01

    This study is aimed to examine the inhibition of soluble fraction (SF) and enzymatic residual fraction (ERF) in dry dilute acid (DDA), ethylenediamine (EDA) and steam explosion (SE) pretreated corn stover (CS) on the enzymatic digestibility of cellulose. SF of DDA, EDA and SE pretreated CS has high xylose, soluble lignin and xylo-oligomer content, respectively. SF of EDA pretreated CS leads to the highest inhibition, followed by SE and DDA pretreated CS. Inhibition of ERF of DDA and SE pretreated CS is higher than that of EDA pretreated CS. The inhibition degree (A0/A) of SF is 1.76 and 1.21 times to that of ERF for EDA and SE pretreated CS, respectively. The inhibition degree of ERF is 1.05 times to that of SF in DDA pretreated CS. The quantitative analysis shows that SF of EDA pretreated CS, SF and ERF of SE pretreated CS cause significant inhibition during enzymatic hydrolysis. PMID:26970919

  20. Revealing the synergetic effects in Ni nanoparticle-carbon nanotube hybrids by scanning transmission X-ray microscopy and their application in the hydrolysis of ammonia borane.

    PubMed

    Zhao, Guanqi; Zhong, Jun; Wang, Jian; Sham, Tsun-Kong; Sun, Xuhui; Lee, Shuit-Tong

    2015-06-01

    The hybrids of carbon nanotubes (CNTs) and the supported Ni nanoparticles (NPs) have been studied by scanning transmission X-ray microscopy (STXM) and tested by the hydrolysis reaction of ammonia borane (AB, NH3BH3). Data clearly showed the existence of a strong interaction between Ni NPs and thin CNTs (C-O-Ni bonds), which favored the tunable (buffer) electronic structure of Ni NPs facilitating the catalytic process. The hydrolysis process of AB confirmed the hypothesis that the hybrids with a strong interfacial interaction would show superior catalytic performance, while the hybrids with a weak interfacial interaction show poor performance. Our results provide a wealth of detailed information regarding the electronic structure of the NP-CNT hybrids and provide guidance towards the rational design of high-performance catalysts for energy applications. PMID:25960161

  1. Quantification of organic acids in particulate matter by coupling of thermally assisted hydrolysis and methylation with thermodesorption-gas chromatography-mass spectrometry.

    PubMed

    Beiner, K; Plewka, A; Haferkorn, S; Iinuma, Y; Engewald, W; Herrmann, H

    2009-09-18

    A quantitative method for the determination of organic acids in atmospheric particles is developed. The method couples a derivatisation step (thermally assisted hydrolysis and methylation) and a Curie point pyrolyser as a thermal desorption technique and gas chromatography-mass spectrometry (CPP-GC-MS). Among the reagents tested (tetramethylammonium hydroxide (TMAH), tetramethylammonium acetate (TMAAc) and phenyltrimethylammonium hydroxide (TMPAH)), the best performance was found using TMAAc as a derivatisation reagent for the reaction time of 4s at 510 degrees C as heating temperature. Calibration was performed for a series of fatty acids (FA), dicarboxylic acids (DCA) and terpenoic acids (TA) under these conditions. Coefficients of determination (R(2)) were between 0.94 and 0.98. Limits of detection (LOD) were in the nanogram-range between 0.1 and 3.6 ng. The method is applied on atmospheric particle samples to obtain the quantification reproducibility and quantification limits. Reproducibility was determined in terms of relative standard deviations (RSD) for ambient aerosol samples collected by a high-volume-sampler (HVS, RSD=6-45%, n=10) and a Berner impactor (BI, RSD=5-34%, n=10). Based on 24h sampling time the developed method enables quantification of all three classes of acids for both sampling techniques. Calibration data and presented volume concentrations are compared with literature data. A comparison with an off-line methylation-GC-MS using BF(3) as a derivatisation reagent and capillary electrophoresis coupled mass spectrometry (CE-MS) showed a good agreement. Minimal sample preparation is the main advantage of the developed method. Depending on the sensitivity requirements the present method can be a fast and simple alternative to GC-MS techniques with conventional sample preparation steps for semi-volatile organic acids. PMID:19679312

  2. Convenient and Precise Strategy for Mapping N-Glycosylation Sites Using Microwave-Assisted Acid Hydrolysis and Characteristic Ions Recognition.

    PubMed

    Ma, Cheng; Qu, Jingyao; Meisner, Jeffrey; Zhao, Xinyuan; Li, Xu; Wu, Zhigang; Zhu, Hailiang; Yu, Zaikuan; Li, Lei; Guo, Yuxi; Song, Jing; Wang, Peng George

    2015-08-01

    N-glycosylation is one of the most prevalence protein post-translational modifications (PTM) which is involved in several biological processes. Alternation of N-glycosylation is associated with cellular malfunction and development of disease. Thus, investigation of protein N-glycosylation is crucial for diagnosis and treatment of disease. Currently, deglycosylation with peptide N-glycosidase F is the most commonly used technique in N-glycosylation analysis. Additionally, a common error in N-glycosylation site identification, resulting from protein chemical deamidation, has largely been ignored. In this study, we developed a convenient and precise approach for mapping N-glycosylation sites utilizing with optimized TFA hydrolysis, ZIC-HILIC enrichment, and characteristic ions of N-acetylglucosamine (GlcNAc) from higher-energy collisional dissociation (HCD) fragmentation. Using this method, we identified a total of 257 N-glycosylation sites and 144 N-glycoproteins from healthy human serum. Compared to deglycosylation with endoglycosidase, this strategy is more convenient and efficient for large scale N-glycosylation sites identification and provides an important alternative approach for the study of N-glycoprotein function. PMID:26161579

  3. Nutritional value of protein hydrolysis products (oligopeptides and free amino acids) as a consequence of absorption and metabolism kinetics

    NASA Technical Reports Server (NTRS)

    Rerat, A.

    1995-01-01

    When pigs were submitted to duodenal infusion of solutions containing a large percentage of small peptides (PEP) or free amino acids with the same pattern (AAL) amino acids appear in the portal blood more rapidly and more uniformly after infusion of PEP then after infusion of AAL, with the notable exception of methionine for which the opposite was true. These differences were lowered when a carbohydrate (maltose dextrin) was present in the solution, but nevertheless remained significant for the first hour after the infusion. The long-term (8-hour) uptake of free amino acids into the liver and the peripheral tissues differed in profile according to the nature of the duodenal infusion. Peripheral uptake was appreciably less well balanced after infusion of free amino acids (deficiency of threonine and phenylalanine) than after infusion of small peptides (deficiency of methionine). Accordingly, in the rat, under conditions of discontinuous enteral nutrition the mixture of small peptides was of greater nutritive value than the mixture of free amino acids. It thus appears that the absorption kinetics which results in important variations in the temporal distribution of free amino acids in the tissues may be at the origin of transitory imbalances in tissue amino acid uptake, and as a result of a lower nutritive value.

  4. Reactivity of Tannic Acid with Common Corrosion Products and Its Influence on the Hydrolysis of Iron in Alkaline Solutions

    NASA Astrophysics Data System (ADS)

    Jaén, J. A.; Araúz, E. Y.; Iglesias, J.; Delgado, Y.

    2003-06-01

    To ascertain the role of tannic acid in the anticorrosive protection of steels, the reaction between 5% tannic acid aqueous solutions with lepidocrocite, goethite, superparamagnetic goethite, akaganeite, poorly crystalline maghemite, magnetite and hematite was studied using color changes, infrared and Mössbauer spectroscopy. After three months of interaction with lepidocrocite, the formation of an iron tannate complex was detected by its dark blue color and confirmed by infrared and Mössbauer analysis. Evidence for the chemical transformation was obtained for goethite in nanoparticles and poorly crystalline maghemite after reaction for six months. The other iron compounds do not transform to another oxide or phase upon treatment with the tannic acid solution. These results showed that lepidocrocite is the most reactive phase and that the size and degree of crystallinity have strong influence on the formation of the tannate complexes. The precipitation of iron phases from alkaline solutions of iron (II) sulfate heptahydrate containing different amount of tannic acid and potassium nitrate as oxidative agent was also studied. Mössbauer and infrared results show that in the absence of tannic acid some common rust components are obtained (viz. goethite, superparamagnetic goethite, maghemite and non-stoichiometric magnetite). The presence of 0.1% tannic acid in a low alkalinity solution results in the precipitation of iron oxyhydroxides and some iron tannates. Concentrations of 1% tannic acid are required for the formation of the tannates complexes as main reaction product.

  5. Muscarinic stimulation of SK-N-BE(2) human neuroblastoma cells elicits phosphoinositide and phosphatidylcholine hydrolysis: relationship to diacylglycerol and phosphatidic acid accumulation.

    PubMed Central

    Pacini, L; Limatola, C; Frati, L; Luly, P; Spinedi, A

    1993-01-01

    Muscarinic stimulation of the human neuroblastoma cell line SK-N-BE(2) elicits hydrolysis of phosphoinositides and phosphatidylcholine (PtdCho) and produces a rapid and sustained elevation of diacylglycerol (DG) mass. PtdIns(4,5)P2 cleavage by phospholipase C (PLC) occurred immediately after carbachol (CCh) addition, and phosphoinositide hydrolysis was then sustained for at least 5 min. Cell stimulation, after extensive PtdCho labelling by long-term [3H]choline administration, resulted in an enhanced release of [3H]phosphocholine (PCho) into the external medium; enhanced [3H]PCho release, which occurred with a 15 s delay with respect to CCh addition, was particularly pronounced within the first minute of stimulation and proved to be caused by PtdCho-specific PLC activation. In fact, when cells were exposed to [3H]choline for a short period, to extensively label the intracellular PCho pool but not PtdCho, stimulation did not result in an enhanced release of [3H]PCho into the medium. PtdCho-specific phospholipase D (PLD) activation was documented by the accumulation of [3H]phosphatidylethanol in cells prelabelled with [3H]myristic acid and stimulated in the presence of 1% (v/v) ethanol; this metabolic pathway, however, proved to be a minor one leading to generation of phosphatidic acid (PtdOH) during cell stimulation, whereas DG production by the sequential action of PtdCho-specific PLD and PtdOH phosphohydrolase was not observed. Studies on cells which were double-labelled with [3H]myristic acid and [14C]arachidonic acid indicated that within 15 s of stimulation DG is uniquely derived from PtdIns(4,5)P2, whereas PtdCho is the major source at later times. Evidence is provided that rapid and selective conversion of phosphoinositide-derived DG into PtdOH may play an important role in determining the temporal accumulation profile of DG from the above-mentioned sources. PMID:8380986

  6. Reaction of ClONO{sub 2} with HCl on NAT, NAD, and frozen sulfuric acid and hydrolysis of N{sub 2}O{sub 5} and ClONO{sub 2} on frozen sulfuric acid

    SciTech Connect

    Hanson, D.R.; Ravishankara, A.R. |

    1993-12-20

    The reaction of ClONO{sub 2} with HCl on the nitric acid trihydrate (NAT) and dihydrate (NAD) and on frozen sulfuric acid solutions was studied as functions of reactant concentration and temperature representative of the stratosphere. A flow tube reactor equipped with a chemical ionization mass spectrometer detector was used. The measured reactive uptake coefficient shows a strong dependence on temperature at constant H{sub 2}O partial pressure. These results for NAT are qualitatively similar to but quantitatively different from those of Abbatt and Molina. The hydrolysis of N{sub 2}O{sub 5} and ClONO{sub 2} on frozen sulfuric acid were measured to be far less efficient than that in liquid solutions. The implications of these results to the heterogeneous chemistry of the stratosphere are discussed. 19 refs., 5 figs., 2 tabs.

  7. Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas. 3. Hydrolysis of phytic acid (IP6) and occurrence of hydrolysis products (IP5, IP4, IP3 and IP2).

    PubMed

    Rapp, C; Lantzsch, H J; Drochner, W

    2001-12-01

    Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (Aspergillus niger) were investigated with six minipigs (40-50 kg initial body weight) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5 and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2 and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet [43 U phytase/kg dry matter (DM)]; (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM); and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 07 30 h and 19 30 h, each animal was fed 350 g diet mixed with 1050 ml de-ionized water. Digesta were collected continuously and completely during a 12-h period after feeding. Mean hydrolysis rates of IP6 in the stomach as measured at the proximal duodenum of animals 1, 4, 5 and 6 were 9.0, 77.2 and 66.2% for diet 1, 2 and 3, respectively. Microbial phytase was much more effective in phytate hydrolysis than wheat phytase. Mean IP6 hydrolysis rates of the respective diets in the stomach and small intestine as measured at the distal ileum of animals 1, 2 and 3 were 19.0, 62.6 and 64.6% and were lower than treatment means of the stomach only. Differences existed between experimental animals with respect to their ability to hydrolyse IP6 in the stomach independent of the presence and source of dietary phytase. Considerable amounts of hydrolysis products occurred in both the duodenal and ileal digesta when diets 2 and 3 were fed; however, only traces were determined after ingestion of diet 1. Independent of dietary treatment, four IP5 isomers were detected, but in different amounts. PMID:11906566

  8. Synergistic Hydrolysis of Carboxymethyl Cellulose and Acid-Swollen Cellulose by Two Endoglucanases (CelZ and CelY) from Erwinia chrysanthemi†

    PubMed Central

    Zhou, Shengde; Ingram, Lonnie O.

    2000-01-01

    Erwinia chrysanthemi produces a battery of hydrolases and lyases which are very effective in the maceration of plant cell walls. Although two endoglucanases (CelZ and CelY; formerly EGZ and EGY) are produced, CelZ represents approximately 95% of the total carboxymethyl cellulase activity. In this study, we have examined the effectiveness of CelY and CelZ alone and of combinations of both enzymes using carboxymethyl cellulose (CMC) and amorphous cellulose (acid-swollen cellulose) as substrates. Synergy was observed with both substrates. Maximal synergy (1.8-fold) was observed for combinations containing primarily CelZ; the ratio of enzyme activities produced was similar to those produced by cultures of E. chrysanthemi. CelY and CelZ were quite different in substrate preference. CelY was unable to hydrolyze soluble cellooligosaccharides (cellotetraose and cellopentaose) but hydrolyzed CMC to fragments averaging 10.7 glucosyl units. In contrast, CelZ readily hydrolyzed cellotetraose, cellopentaose, and amorphous cellulose to produce cellobiose and cellotriose as dominant products. CelZ hydrolyzed CMC to fragments averaging 3.6 glucosyl units. In combination, CelZ and CelY hydrolyzed CMC to products averaging 2.3 glucosyl units. Synergy did not require the simultaneous presence of both enzymes. Enzymatic modification of the substrate by CelY increased the rate and extent of hydrolysis by CelZ. Full synergy was retained by the sequential hydrolysis of CMC, provided CelY was used as the first enzyme. A general mechanism is proposed to explain the synergy between these two enzymes based primarily on differences in substrate preference. PMID:11004164

  9. Long-Term Ritonavir Exposure Increases Fatty Acid and Glycerol Recycling in 3T3-L1 Adipocytes as Compensatory Mechanisms for Increased Triacylglycerol Hydrolysis

    PubMed Central

    Adler-Wailes, Diane C.; Guiney, Evan L.; Wolins, Nathan E.; Yanovski, Jack A.

    2010-01-01

    Lipodystrophy with high nonesterified fatty acid (FA) efflux is reported in humans receiving highly active antiretroviral therapy (HAART) to treat HIV infection. Ritonavir, a common component of HAART, alters adipocyte FA efflux, but the mechanism for this effect is not established. To investigate ritonavir-induced changes in FA flux and recycling through acylglycerols, we exposed differentiated murine 3T3-L1 adipocytes to ritonavir for 14 d. FA efflux, uptake, and incorporation into acylglycerols were measured. To identify a mediator of FA efflux, we measured adipocyte triacylglycerol lipase (ATGL) transcript and protein. To determine whether ritonavir-treated adipocytes increased glycerol backbone synthesis for FA reesterification, we measured labeled glycerol and pyruvate incorporation into triacylglycerol (TAG). Ritonavir-treated cells had increased FA efflux, uptake, and incorporation into TAG (all P < 0.01). Ritonavir increased FA efflux without consistently increasing glycerol release or changing TAG mass, suggesting increased partial TAG hydrolysis. Ritonavir-treated adipocytes expressed significantly more ATGL mRNA (P < 0.05) and protein (P < 0.05). Ritonavir increased glycerol (P < 0.01) but not pyruvate (P = 0.41), utilization for TAG backbone synthesis. Consistent with this substrate utilization, glycerol kinase transcript (required for glycerol incorporation into TAG backbone) was up-regulated (P < 0.01), whereas phosphoenolpyruvate carboxykinase transcript (required for pyruvate utilization) was down-regulated (P < 0.001). In 3T3-L1 adipocytes, long-term ritonavir exposure perturbs FA metabolism by increasing ATGL-mediated partial TAG hydrolysis, thus increasing FA efflux, and leads to compensatory increases in FA reesterification with glycerol and acylglycerols. These changes in FA metabolism may, in part, explain the increased FA efflux observed in ritonavir-associated lipodystrophy. PMID:20228169

  10. Studies of the Maltose Transport System Reveal a Mechanism for Coupling ATP Hydrolysis to Substrate Translocation without Direct Recognition of Substrate

    SciTech Connect

    Gould, Alister D.; Shilton, Brian H.

    2010-10-11

    The ATPase activity of the maltose transporter (MalFGK{sub 2}) is dependent on interactions with the maltose-binding protein (MBP). To determine whether direct interactions between the translocated sugar and MalFGK{sub 2} are important for the regulation of ATP hydrolysis, we used an MBP mutant (sMBP) that is able to bind either maltose or sucrose. We observed that maltose- and sucrose-bound sMBP stimulate equal levels of MalFGK{sub 2} ATPase activity. Therefore, the ATPase activity of MalFGK{sub 2} is coupled to translocation of maltose solely by interactions between MalFGK{sub 2} and MBP. For both maltose and sucrose, the ability of sMBP to stimulate the MalFGK{sub 2} ATPase was greatly reduced compared with wild-type MBP, indicating that the mutations in sMBP have interfered with important interactions between MBP and MalFGK{sub 2}. High resolution crystal structure analysis of sMBP shows that in the closed conformation with bound sucrose, three of four mutations are buried, and the fourth causes only a minor change in the accessible surface. In contrast, in the open form of sMBP, all of the mutations are accessible, and the main chain of Tyr{sup 62}-Gly{sup 69} is destabilized and occupies an alternative conformation due to the W62Y mutation. On this basis, the compromised ability of sMBP to stimulate ATP hydrolysis by MalFGK{sub 2} is most likely due to a disruption of interactions between MalFGK{sub 2} and the open, rather than the closed, conformation of sMBP. Modeling the open sMBP structure bound to MalFGK{sub 2} in the transition state for ATP hydrolysis points to an important site of interaction and suggests a mechanism for coupling ATP hydrolysis to substrate translocation that is independent of the exact structure of the substrate.

  11. Cyclomaltoheptaose mixed esters of anti-inflammatory drugs and short-chain fatty acids and study of their enzymatic hydrolysis in vitro.

    PubMed

    Cao, Feng; Ren, Yong; Hua, Weiyi

    2009-03-10

    In an effort to enhance the drug-loading capacity of cyclomaltoheptaose (beta-cyclodextrin, betaCD) and to combine the function of anti-inflammatory drugs with short-chain fatty acids (SCFA), ternary esters incorporating seven copies of an anti-inflammatory drug and 14 copies of a SCFA onto a beta-cyclodextrin core were designed and prepared. Acetic, propionic, or butyric esters were introduced at secondary OH groups, and ibuprofen, flurbiprofen, or felbinac was attached to primary OH groups through ester bonds. Heptakis[2,3-di-O-butanoyl-6-O-2-(biphenyl-4-yl)-ethanoyl]-cyclomaltoheptaose was very stable in aqueous and esterase solution. It was hydrolyzed by alpha-amylase (4 units/mL) with t(1/2) value of 18h. The total released amount of biphenyl acetic acid was 38% after 24h when the esterase was added after the alpha-amylase hydrolysis. The present results suggest that these nine betaCD conjugates may release the anti-inflammatory drug in the colonic contents. PMID:19185291

  12. Carbachol induces a rapid and sustained hydrolysis of polyphosphoinositide in bovine tracheal smooth muscle measurements of the mass of polyphosphoinositides, 1,2-diacylglycerol, and phosphatidic acid

    SciTech Connect

    Takuwa, Y.; Takuwa, N.; Rasmussen, H.

    1986-11-05

    The effects of carbachol on polyphosphoinositides and 1,2-diacylglycerol metabolism were investigated in bovine tracheal smooth muscle by measuring both lipid mass and the turnover of (/sup 3/H)inositol-labeled phosphoinositides. Carbachol induces a rapid reduction in the mass of phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-monophosphate and a rapid increase in the mass of 1,2-diacylglycerol and phosphatidic acid. These changes in lipid mass are sustained for at least 60 min. The level of phosphatidylinositol shows a delayed and progressive decrease during a 60-min period of carbachol stimulation. The addition of atropine reverses these responses completely. Carbachol stimulates a rapid loss in (/sup 3/H)inositol radioactivity from phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-monophosphate associated with production of (/sup 3/H)inositol trisphosphate. The carbachol-induced change in the mass of phosphoinositides and phosphatidic acid is not affected by removal of extracellular Ca/sup 2 +/ and does not appear to be secondary to an increase in intracellular Ca/sup 2 +/. These results indicate that carbachol causes phospholipase C-mediated polyphosphoinositide breakdown, resulting in the production of inositol trisphosphate and a sustained increase in the actual content of 1,2-diacylglycerol. These results strongly suggest that carbachol-induced contraction is mediated by the hydrolysis of polyphosphoinositides with the resulting generation of two messengers: inositol 1,4,5-trisphosphate and 1,2-diacylglycerol.

  13. Use of bacteria for rapid, pH-neutral, hydrolysis of the model hydrophobic carboxylic acid ester p-nitrophenyl picolinate

    PubMed Central

    Forest, Alexandra E.; Goldstine, Gordon G.; Schrodi, Yann; Murray, Sean R.

    2012-01-01

    Caulobacter crescentus, Escherichia coli and Bacillus subtilis cultures promote the hydrolysis of the model ester p-nitrophenyl picolinate (PNPP) at neutral pH with high efficiency. Hydrolysis is related to cell concentration, while the interaction of PNPP with both bacterial cells and their extracellular molecules is required for a maximum rate of PNPP hydrolysis in C. crescentus cultures. Furthermore, C. crescentus cultures hydrolyze PNPP at concentrations useful in synthetic chemistry. PMID:23144558

  14. Response surface optimization of corn stover pretreatment using dilute phosphoric acid for enzymatic hydrolysis and ethanol production

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dilute H3PO4 (0.0 - 2.0%, v/v) was used to pretreat corn stover (10%, w/w) for conversion to ethanol. Pretreatment conditions were optimized for temperature, acid loading, and time using a central composite design. Optimal pretreatment conditions were chosen to promote sugar yields following enzym...

  15. Increased yield of high purity recombinant human brain natriuretic peptide by acid hydrolysis of short fusion partner in Escherichia coli.

    PubMed

    Kanumuri, Radha Madhavi; Bajji, Chitra; Tummuru, Rajesh R; Tatireddigari, Venkat R R Arva; Mangamoori, Lakshmi Narasu; Panati, Kalpana; Narala, Venkata Ramireddy

    2015-07-01

    Recombinant human B-type natriuretic peptide (rhBNP) is a 32-amino acid peptide used to treat congestive heart failure. In this paper, we report a method for the increased production of rhBNP in Escherichia coli with high purity. hBNP was cloned with a short growth hormone fusion partner coupled with a unique acid-labile dipeptide linker to cleave the fusion protein to release the rhBNP. The recombinant fusion protein was expressed as an inclusion body (IB) and the fermentation process was optimized to produce on large scale. The IBs were recovered by cell lysis, and the pure IBs were directly treated with diluted acid to get the target peptide from the fusion protein and the resultant peptide was purified by reversed phase chromatography. The final purity of the rhBNP was more than 99% with yield of 50mg per liter of culture, which is ten times higher than the previous reports. The purified rhBNP exhibited specific biological activity similar to the standard peptide in producing cyclic-guanosine monophosphate. PMID:25823948

  16. Effect of feeding lipids recovered from fish processing waste by lactic acid fermentation and enzymatic hydrolysis on antioxidant and membrane bound enzymes in rats.

    PubMed

    Rai, Amit Kumar; Bhaskar, N; Baskaran, V

    2015-06-01

    Fish oil recovered from fresh water fish visceral waste (FVW-FO) through lactic acid fermentation (FO-LAF) and enzymatic hydrolysis (FO-EH) were fed to rats to study their influence on lipid peroxidation and activities of antioxidant and membrane bound enzyme in liver, heart and brain. Feeding of FO-LAF and FO-EH resulted in increase (P < 0.05) in lipid peroxides level in serum, liver, brain and heart tissues compared to ground nut oil (control). Activity of catalase (40-235 %) and superoxide dismutase (17-143 %) also increased (P < 0.05) with incremental level of EPA + DHA in diet. The increase was similar to cod liver oil fed rats at same concentration of EPA + DHA. FO-LAF and FO-EH increased (P < 0.05) the Na(+)K(+) ATPase activity in liver and brain microsomes, Ca(+)Mg(+) ATPase in heart microsome and acetylcholine esterase in brain microsomes when fed with 5 % EPA + DHA. There was also significant change in fatty acid composition and cholesterol/phospholipid ratio in microsomes of rat fed with FVW-FO. Feeding FVW-FO recovered by biotechnological approaches enhanced the activity of antioxidant enzymes in tissues, modulates the activities of membrane bound enzymes and improved the fatty acid composition in microsomes of tissues similar to CLO. Utilization of these processing wastes for the production of valuable biofunctional products can reduce the mounting economic values of fish oil and minimize the environmental pollution problems. PMID:26028754

  17. Regiospecific Ester Hydrolysis by Orange Peel Esterase - An Undergraduate Experiment.

    NASA Astrophysics Data System (ADS)

    Bugg, Timothy D. H.; Lewin, Andrew M.; Catlin, Eric R.

    1997-01-01

    A simple but effective experiment has been developed to demonstrate the regiospecificity of enzyme catalysis using an esterase activity easily isolated from orange peel. The experiment involves the preparation of diester derivatives of para-, meta- and ortho-hydroxybenzoic acid (e.g. methyl 4-acetoxy-benzoic acid). The derivatives are incubated with orange peel esterase, as a crude extract, and with commercially available pig liver esterase and porcine pancreatic lipase. The enzymatic hydrolysis reactions are monitored by thin layer chromatography, revealing which of the two ester groups is hydrolysed, and the rate of the enzyme-catalysed reaction. The results of a group experiment revealed that in all cases hydrolysis was observed with at least one enzyme, and in most cases the enzymatic hydrolysis was specific for production of either the hydroxy-ester or acyl-acid product. Specificity towards the ortho-substituted series was markedly different to that of the para-substituted series, which could be rationalised in the case of pig liver esterase by a published active site model.

  18. Continuous steam hydrolysis of tulip poplar

    SciTech Connect

    Fieber, C.A.; Roberts, R.S.; Faass, G.S.; Muzzy, J.D.; Colcord, A.R.; Bery, M.K.

    1982-01-01

    The continuous hydrolysis of poplar chips by steam at 300-350 psi resulted in the separation of hemicellulose (I) cellulose and lignin components. The I fraction was readily depolymerised by steam to acetic acid, furfural, methanol, and xylose.

  19. Identification of N-acylethanolamines in Dictyostelium discoideum and confirmation of their hydrolysis by fatty acid amide hydrolase[S

    PubMed Central

    Hayes, Alexander C.; Stupak, Jacek; Li, Jianjun; Cox, Andrew D.

    2013-01-01

    N-acylethanolamines (NAEs) are endogenous lipid-based signaling molecules best known for their role in the endocannabinoid system in mammals, but they are also known to play roles in signaling pathways in plants. The regulation of NAEs in vivo is partly accomplished by the enzyme fatty acid amide hydrolase (FAAH), which hydrolyses NAEs to ethanolamine and their corresponding fatty acid. Inhibition of FAAH has been shown to increase the levels of NAEs in vivo and to produce desirable phenotypes. This has led to the development of pharmaceutical-based therapies for a variety of conditions targeting FAAH. Recently, our group identified a functional FAAH homolog in Dictyostelium discoideum, leading to our hypothesis that D. discoideum also possesses NAEs. In this study, we provide a further characterization of FAAH and identify NAEs in D. discoideum for the first time. We also demonstrate the ability to modulate their levels in vivo through the use of a semispecific FAAH inhibitor and confirm that these NAEs are FAAH substrates through in vitro studies. We believe the demonstration of the in vivo modulation of NAE levels suggests that D. discoideum could be a good simple model organism in which to study NAE-mediated signaling. PMID:23187822

  20. Acid Hydrolysis of Wheat Gluten Induces Formation of New Epitopes but Does Not Enhance Sensitizing Capacity by the Oral Route: A Study in “Gluten Free” Brown Norway Rats

    PubMed Central

    Kroghsbo, Stine; Andersen, Nanna B.; Rasmussen, Tina F.; Madsen, Charlotte B.

    2014-01-01

    Background Acid hydrolyzed wheat proteins (HWPs) are used in the food and cosmetic industry as emulsifiers. Cases of severe food allergic reactions caused by HWPs have been reported. Recent data suggest that these reactions are caused by HWPs produced by acid hydrolysis. Objectives To examine the sensitizing capacity of gluten proteins per se when altered by acid or enzymatic hydrolysis relative to unmodified gluten in rats naïve to gluten. Methods High IgE-responder Brown Norway (BN) rats bred on a gluten-free diet were sensitized without the use of adjuvant to three different gluten products (unmodified, acid hydrolyzed and enzymatic hydrolyzed). Rats were sensitized by intraperitoneal (i.p.) immunization three times with 200 µg gluten protein/rat or by oral dosing for 35 days with 0.2, 2 or 20 mg gluten protein/rat/day. Sera were analyzed for specific IgG and IgE and IgG-binding capacity by ELISA. IgE functionality was measured by rat basophilic leukemia (RBL) assay. Results Regardless of the route of dosing, all products had sensitizing capacity. When sensitized i.p., all three gluten products induced a strong IgG1 response in all animals. Acid hydrolyzed gluten induced the highest level of specific IgE but with a low functionality. Orally all three gluten products induced specific IgG1 and IgE but with different dose-response relations. Sensitizing rats i.p. or orally with unmodified or enzymatic hydrolyzed gluten induced specific IgG1 responses with similar binding capacity which was different from that of acid hydrolyzed gluten indicating that acid hydrolysis of gluten proteins induces formation of ‘new’ epitopes. Conclusions In rats not tolerant to gluten acid hydrolysis of gluten enhances the sensitizing capacity by the i.p. but not by the oral route. In addition, acid hydrolysis induces formation of new epitopes. This is in contrast to the enzymatic hydrolyzed gluten having an epitope pattern similar to unmodified gluten. PMID:25207551

  1. Topochemical distribution of lignin and hydroxycinnamic acids in sugar-cane cell walls and its correlation with the enzymatic hydrolysis of polysaccharides

    PubMed Central

    2011-01-01

    Background Lignin and hemicelluloses are the major components limiting enzyme infiltration into cell walls. Determination of the topochemical distribution of lignin and aromatics in sugar cane might provide important data on the recalcitrance of specific cells. We used cellular ultraviolet (UV) microspectrophotometry (UMSP) to topochemically detect lignin and hydroxycinnamic acids in individual fiber, vessel and parenchyma cell walls of untreated and chlorite-treated sugar cane. Internodes, presenting typical vascular bundles and sucrose-storing parenchyma cells, were divided into rind and pith fractions. Results Vascular bundles were more abundant in the rind, whereas parenchyma cells predominated in the pith region. UV measurements of untreated fiber cell walls gave absorbance spectra typical of grass lignin, with a band at 278 nm and a pronounced shoulder at 315 nm, assigned to the presence of hydroxycinnamic acids linked to lignin and/or to arabino-methylglucurono-xylans. The cell walls of vessels had the highest level of lignification, followed by those of fibers and parenchyma. Pith parenchyma cell walls were characterized by very low absorbance values at 278 nm; however, a distinct peak at 315 nm indicated that pith parenchyma cells are not extensively lignified, but contain significant amounts of hydroxycinnamic acids. Cellular UV image profiles scanned with an absorbance intensity maximum of 278 nm identified the pattern of lignin distribution in the individual cell walls, with the highest concentration occurring in the middle lamella and cell corners. Chlorite treatment caused a rapid removal of hydroxycinnamic acids from parenchyma cell walls, whereas the thicker fiber cell walls were delignified only after a long treatment duration (4 hours). Untreated pith samples were promptly hydrolyzed by cellulases, reaching 63% of cellulose conversion after 72 hours of hydrolysis, whereas untreated rind samples achieved only 20% hydrolyzation. Conclusion The low

  2. Effects of Cortex Peptidoglycan Structure and Cortex Hydrolysis on the Kinetics of Ca2+-Dipicolinic Acid Release during Bacillus subtilis Spore Germination

    PubMed Central

    Zhang, Pengfei; Thomas, Stacy; Li, Yong-qing

    2012-01-01

    The kinetic parameters of the release of Ca2+-dipicolinic acid (CaDPA) during germination of spore populations and multiple individual spores of Bacillus subtilis strains with major alterations in the structure of the spore peptidoglycan (PG) cortex or lacking one or both of the two redundant enzymes involved in cortex hydrolysis (cortex-lytic enzymes [CLEs]) were determined. The lack of the CLE CwlJ greatly slowed CaDPA release with a germinant receptor (GR)-dependent germinant, l-valine, or a non-GR-dependent germinant, dodecylamine. The absence of the cortex-specific PG modification muramic acid–δ-lactam also increased the time needed for full CaDPA release during germination with both types of germinants. In contrast, increased cortex PG cross-linking was associated with faster times for initiation of CaDPA release with both l-valine and dodecylamine but not with faster CaDPA release once this release had been initiated. These data suggest that the precise structure of the spore cortex plays a significant role in determining the timing and the rate of CaDPA release during B. subtilis spore germination and, further, that this effect is independent of effects of GRs. PMID:22123250

  3. Variation of S/G Ratio and Lignin Content in a Populus Family Influences the Release of Xylose by Dilute Acid Hydrolysis.

    SciTech Connect

    Davison, Brian H; Drescher, Sadie R; Tuskan, Gerald A; Davis, Dr. Mark F.; Nghiem, Nhuan Phu

    2006-01-01

    Wood samples from second generation Populus cross were shown to have different lignin contents and S/G ratios (S: syringyl-like lignin structures; G: guaiacyl-like lignin structures). The lignin contents varied from 22.7% to 25.8% and the S/G ratio from 1.8 to 2.3. Selected samples spanning these ranges were hydrolyzed with dilute (1%) sulfuric acid to release fermentable sugars. The conditions were chosen for partial hydrolysis of the hemicellulosic fraction to maximize the expression of variation among samples. The results indicated that both lignin contents and S/G ratio significantly affected the yield of xylose. For example, the xylose yield of the 25.8% lignin and 2.3 S/G (hihg lignin, high S/G) sample produced 30% of the theoretical yield, whereas the xylose yield of the 22.7% lignin and 1.8 S/G (low lignin, low S/G) was 55% of the theoretical value. These results indicate that lignin content and composition among genetic variants within a single species can influence the hydrolyzability of the biomass.

  4. The formation of isocyanic acid (HNCO) by reaction of NO, CO, and H{sub 2} over Pt/SiO{sub 2} and its hydrolysis on alumina

    SciTech Connect

    Duempelmann, R.; Cant, N.W.; Trimm, D.L.

    1996-08-01

    The reaction of mixtures of NO, CO, and H{sub 2} over Pt/SiO{sub 2} in combination with Al{sub 2}O{sub 3} has been investigated with on-line Fourier transform infrared spectroscopy as the principal analytical method. With Pt/SiO{sub 2} alone isocyanic acid (HNCO) can be observed directly as a substantial product. It comprises up to 45% of the CO converted in the temperature range from 200 to 300{degrees}C. The yield is maximized when the amounts of H{sub 2} and CO in the input stream are equal. No HNCO can be observed in experiments in which Al{sub 2}O{sub 3} is placed downstream of the Pt/SiO{sub 2} or with a Pt/Al{sub 2}O{sub 3} is placed downstream of the Pt/SiO{sub 2} or with a Pt/Al{sub 2}O{sub 3} catalyst. Hydrolysis to NH{sub 3} and CO{sub 2} is then complete. For this reason it is unlikely that HNCO will escape a catalytic converter in vehicle use but it may be a significant intermediate during the warm-up phase. The formation of HNCO can be rationalized as resulting from the hydrogenation of NCO groups present in small concentrations on the Pt surface under conditions of high CO coverage. The yield of HNCO falls steeply above 300{degrees}C when the CO coverage is falling. It is unclear if hydrolysis of HNCO occurs by direct reaction of molecular HNCO on the Al{sub 2}O{sub 3} surface or proceeds via the ubiquitious support-bound isocyanate species which have been observed in many previous studies. Previous observations of isocyanate formation in hydrogen-free systems can be rationalized as arising from HNCO formation by reverse spillover of hydrogen from OH groups on the support to generate HNCO on the platinum metal. This is then transported back to the support through the gas phase to form NCO groups. 47 refs., 7 figs.

  5. Ultrasound-assisted enzymatic hydrolysis for iodinated amino acid extraction from edible seaweed before reversed-phase high performance liquid chromatography-inductively coupled plasma-mass spectrometry.

    PubMed

    Romarís-Hortas, Vanessa; Bermejo-Barrera, Pilar; Moreda-Piñeiro, Antonio

    2013-09-27

    The combination of reverse phase high performance liquid chromatography (RP-HPLC) with inductively coupled plasma mass spectrometry (ICP-MS) was used for the determination of monoiodotyrosine (MIT) and diiodotyrosine (DIT) in edible seaweed. A sample pre-treatment based on ultrasound assisted enzymatic hydrolysis was optimized for the extraction of these iodinated amino acids. Pancreatin was selected as the most adequate type of enzyme, and parameters affecting the extraction efficiency (pH, temperature, mass of enzyme and extraction time) were evaluated by univariate approaches. In addition, extractable inorganic iodine (iodide) was also quantified by anion exchange high performance liquid chromatography (AE-HPLC) coupled with ICP-MS. The proposed procedure offered limits of detection of 1.1 and 4.3ngg(-1) for MIT and DIT, respectively. Total iodine contents in seaweed, as well as total iodine in enzymatic digests were measured by ICP-MS after microwave assisted alkaline digestion with tetramethylamonium hydroxide (TMAH) for total iodine assessment, and also by treating the pancreatin extracts (extractable total iodine assessment). The optimized procedure was successfully applied to five different types of edible seaweed. The highest total iodine content, and also the highest iodide levels, was found in the brown seaweed Kombu (6646±45μgg(-1)). Regarding iodinated amino acids, Nori (a red seaweed) was by far the one with the highest amount of both species (42±3 and 0.41±0.024μgg(-1) for MIT and DIT, respectively). In general, MIT concentrations were much higher than the amounts of DIT, which suggests that iodine from iodinated proteins in seaweed is most likely bound in the form of MIT residues. PMID:23972456

  6. Structural investigation of a uronic acid-containing polysaccharide from abalone by graded acid hydrolysis followed by PMP-HPLC-MSn and NMR analysis.

    PubMed

    Wang, Hong-xu; Zhao, Jun; Li, Dong-mei; Song, Shuang; Song, Liang; Fu, Ying-huan; Zhang, Li-peng

    2015-01-30

    A new strategy was applied to elucidate the structure of a polysaccharide from abalone gonad (AGSP). It was hydrolyzed by 0.05 M, 0.2 M, 0.5 M, and 2.0 M TFA at 100 °C for 1 h, sequentially. Every hydrolysate was ultrafiltrated (3000 Da) to collect oligo- and monosaccharides, and the final retentate was further hydrolyzed with 2.0 M TFA at 110 °C and 121 °C for 2 h, respectively. 1-Phenyl-3-methyl-5-pyrazolone (PMP) derivatization followed by HPLC-MSn analysis was applied to detect the sugar residues in these hydrolysates, which allowed proposing their location in the polysaccharide structure. The retentate after 0.5 M TFA hydrolysis was confirmed as the polysaccharide backbone, and it was further analyzed by 1D and 2D NMR spectroscopy. Thus, the structural elucidation of AGSP was accomplished, and it has a backbone of →4)-β-GlcA(1→2)-α-Man(1→ repeating unit with Fuc, Xyl and Gal in the branch. The analytical strategy demonstrated was useful to characterize the structure of polysaccharides. PMID:25497339

  7. Effects of Kraft lignin on hydrolysis/dehydration of sugars, cellulosic and lignocellulosic biomass under hot compressed water.

    PubMed

    Daorattanachai, Pornlada; Viriya-empikul, Nawin; Laosiripojana, Navadol; Faungnawakij, Kajornsak

    2013-09-01

    The effect of Kraft lignin presenting on the hydrolysis and dehydration of C5 and C6 sugars, cellulose, hemicelluloses and biomass under hot compressed water (HCW) in the presence of H3PO4 catalyst was intensively studied. The lignin strongly inhibited the acid hydrolysis of cellulose and hemicellulose to glucose and xylose, respectively. Interestingly, the admixed lignin markedly promoted the isomerization of glucose to fructose, and dehydration of fructose (except at the low catalyst loading), resulting in high 5-hydroxymethylfurfural yields. Nonetheless, lignin inhibited the hydrolysis of xylan to xylose and dehydration of xylose to furfural. Moreover, the acidity of the system significantly affects the hydrolysis/dehydration of biomass. It was revealed that the presence of lignin strongly interfered the yields of sugars and furans produced from raw corncob, while the delignified corncob provided significant improvement of product yields, confirming the observed role of lignin in the biomass conversion system via sugar platforms. PMID:23907066

  8. (19) F NMR and DFT Analysis Reveal Structural and Electronic Transition State Features for RhoA-Catalyzed GTP Hydrolysis.

    PubMed

    Jin, Yi; Molt, Robert W; Waltho, Jonathan P; Richards, Nigel G J; Blackburn, G Michael

    2016-03-01

    Molecular details for RhoA/GAP catalysis of the hydrolysis of GTP to GDP are poorly understood. We use (19) F NMR chemical shifts in the MgF3 (-) transition state analogue (TSA) complex as a spectroscopic reporter to indicate electron distribution for the γ-PO3 (-) oxygens in the corresponding TS, implying that oxygen coordinated to Mg has the greatest electron density. This was validated by QM calculations giving a picture of the electronic properties of the transition state (TS) for nucleophilic attack of water on the γ-PO3 (-) group based on the structure of a RhoA/GAP-GDP-MgF3 (-) TSA complex. The TS model displays a network of 20 hydrogen bonds, including the GAP Arg85' side chain, but neither phosphate torsional strain nor general base catalysis is evident. The nucleophilic water occupies a reactive location different from that in multiple ground state complexes, arising from reorientation of the Gln-63 carboxamide by Arg85' to preclude direct hydrogen bonding from water to the target γ-PO3 (-) group. PMID:26822702

  9. Oxidation and hydrolysis of acetic acid and methylene chloride in supercritical water as a means of remediation

    SciTech Connect

    Marrone, P.A.; Lachance, R.P.; DiNaro, J.L.

    1995-10-01

    Supercritical water oxidation (SCWO) is a promising technology proposed for the destruction of hazardous organic wastes. Unlike its well known behavior under ambient conditions, water above its critical point (374{degrees}C, 221 bar) has properties similar to that of a nonpolar solvent, primarily due to the effect of a decrease in hydrogen bonding and density that occurs near and above the critical point. The result is that nonpolar organics and oxygen exhibit complete solubility in supercritical water, while polar species such as inorganic salts are insoluble and precipitate out. In the single homogeneous phase formed, oxidation of organics with oxygen in supercritical water is rapid and complete to CO{sub 2} and H{sub 2}O. Organic heteroatoms such as halogens, sulfur, or phosphorus are converted to inorganic acids (HCl, H{sub 2}SO{sub 4}, H{sub 3}PO{sub 4}) which precipitate as salts when neutralized with added base, while nitrogen is converted to N{sub 2} and N{sub 2}O. No NO{sub x} compounds are formed due to the relatively low temperatures that exist in the SCWO process (400 - 650{degrees}C) relative to that of air incineration processes (typically 900 - 1300{degrees}C). Oxidation in supercritical water is thus an appealing means of destroying toxic organic compounds while simultaneously separating out undesired inorganics by precipitation. Applications to decontaminating soils and dilute aqueous wastes are of special interest. Earlier work has demonstrated high destruction efficiencies for various organics in SCWO.

  10. Molecular Dynamic Simulations Reveal the Structural Determinants of Fatty Acid Binding to Oxy-Myoglobin

    PubMed Central

    Chintapalli, Sree V.; Bhardwaj, Gaurav; Patel, Reema; Shah, Natasha; Patterson, Randen L.; van Rossum, Damian B.; Anishkin, Andriy; Adams, Sean H.

    2015-01-01

    The mechanism(s) by which fatty acids are sequestered and transported in muscle have not been fully elucidated. A potential key player in this process is the protein myoglobin (Mb). Indeed, there is a catalogue of empirical evidence supporting direct interaction of globins with fatty acid metabolites; however, the binding pocket and regulation of the interaction remains to be established. In this study, we employed a computational strategy to elucidate the structural determinants of fatty acids (palmitic & oleic acid) binding to Mb. Sequence analysis and docking simulations with a horse (Equus caballus) structural Mb reference reveals a fatty acid-binding site in the hydrophobic cleft near the heme region in Mb. Both palmitic acid and oleic acid attain a “U” shaped structure similar to their conformation in pockets of other fatty acid-binding proteins. Specifically, we found that the carboxyl head group of palmitic acid coordinates with the amino group of Lys45, whereas the carboxyl group of oleic acid coordinates with both the amino groups of Lys45 and Lys63. The alkyl tails of both fatty acids are supported by surrounding hydrophobic residues Leu29, Leu32, Phe33, Phe43, Phe46, Val67, Val68 and Ile107. In the saturated palmitic acid, the hydrophobic tail moves freely and occasionally penetrates deeper inside the hydrophobic cleft, making additional contacts with Val28, Leu69, Leu72 and Ile111. Our simulations reveal a dynamic and stable binding pocket in which the oxygen molecule and heme group in Mb are required for additional hydrophobic interactions. Taken together, these findings support a mechanism in which Mb acts as a muscle transporter for fatty acid when it is in the oxygenated state and releases fatty acid when Mb converts to deoxygenated state. PMID:26030763

  11. Molecular dynamic simulations reveal the structural determinants of Fatty Acid binding to oxy-myoglobin.

    PubMed

    Chintapalli, Sree V; Bhardwaj, Gaurav; Patel, Reema; Shah, Natasha; Patterson, Randen L; van Rossum, Damian B; Anishkin, Andriy; Adams, Sean H

    2015-01-01

    The mechanism(s) by which fatty acids are sequestered and transported in muscle have not been fully elucidated. A potential key player in this process is the protein myoglobin (Mb). Indeed, there is a catalogue of empirical evidence supporting direct interaction of globins with fatty acid metabolites; however, the binding pocket and regulation of the interaction remains to be established. In this study, we employed a computational strategy to elucidate the structural determinants of fatty acids (palmitic & oleic acid) binding to Mb. Sequence analysis and docking simulations with a horse (Equus caballus) structural Mb reference reveals a fatty acid-binding site in the hydrophobic cleft near the heme region in Mb. Both palmitic acid and oleic acid attain a "U" shaped structure similar to their conformation in pockets of other fatty acid-binding proteins. Specifically, we found that the carboxyl head group of palmitic acid coordinates with the amino group of Lys45, whereas the carboxyl group of oleic acid coordinates with both the amino groups of Lys45 and Lys63. The alkyl tails of both fatty acids are supported by surrounding hydrophobic residues Leu29, Leu32, Phe33, Phe43, Phe46, Val67, Val68 and Ile107. In the saturated palmitic acid, the hydrophobic tail moves freely and occasionally penetrates deeper inside the hydrophobic cleft, making additional contacts with Val28, Leu69, Leu72 and Ile111. Our simulations reveal a dynamic and stable binding pocket in which the oxygen molecule and heme group in Mb are required for additional hydrophobic interactions. Taken together, these findings support a mechanism in which Mb acts as a muscle transporter for fatty acid when it is in the oxygenated state and releases fatty acid when Mb converts to deoxygenated state. PMID:26030763

  12. A Screen for Dominant Negative Mutants of SEC18 Reveals a Role for the AAA Protein Consensus Sequence in ATP Hydrolysis

    PubMed Central

    Steel, Gregor J.; Harley, Carol; Boyd, Alan; Morgan, Alan

    2000-01-01

    An evolutionarily ancient mechanism is used for intracellular membrane fusion events ranging from endoplasmic reticulum–Golgi traffic in yeast to synaptic vesicle exocytosis in the human brain. At the heart of this mechanism is the core complex of N-ethylmaleimide-sensitive fusion protein (NSF), soluble NSF attachment proteins (SNAPs), and SNAP receptors (SNAREs). Although these proteins are accepted as key players in vesicular traffic, their molecular mechanisms of action remain unclear. To illuminate important structure–function relationships in NSF, a screen for dominant negative mutants of yeast NSF (Sec18p) was undertaken. This involved random mutagenesis of a GAL1-regulated SEC18 yeast expression plasmid. Several dominant negative alleles were identified on the basis of galactose-inducible growth arrest, of which one, sec18-109, was characterized in detail. The sec18-109 phenotype (abnormal membrane trafficking through the biosynthetic pathway, accumulation of a membranous tubular network, growth suppression, increased cell density) is due to a single A-G substitution in SEC18 resulting in a missense mutation in Sec18p (Thr394→Pro). Thr394 is conserved in most AAA proteins and indeed forms part of the minimal AAA consensus sequence that serves as a signature of this large protein family. Analysis of recombinant Sec18-109p indicates that the mutation does not prevent hexamerization or interaction with yeast α-SNAP (Sec17p), but instead results in undetectable ATPase activity that cannot be stimulated by Sec17p. This suggests a role for the AAA protein consensus sequence in regulating ATP hydrolysis. Furthermore, this approach of screening for dominant negative mutants in yeast can be applied to other conserved proteins so as to highlight important functional domains in their mammalian counterparts. PMID:10749934

  13. The double helix is dehydrated: evidence from the hydrolysis of acridinium ester-labeled probes.

    PubMed

    Becker, M; Lerum, V; Dickson, S; Nelson, N C; Matsuda, E

    1999-04-27

    A highly chemiluminescent reporter molecule, acridinium ester (AE), was tethered to single-stranded oligonucleotide probes and hybridized to complementary as well as mismatched target sequences. When tethered to single-stranded probes, AE was readily hydrolyzed by water or hydroxide ion. In contrast, when hybridized to a complementary target, hydrolysis of the AE probe was markedly inhibited. Mismatches near AE eliminated the ability of the double helix to strongly inhibit AE hydrolysis. To establish the molecular basis for these remarkable hydrolysis properties of AE-labeled probes, the binding and hydrolysis mechanisms of AE-labeled probes were examined. When tethered to single- or double-stranded nucleic acids, hydrolysis of AE was found to proceed by generalized base catalysis in which a base abstracts a proton from water and the resulting hydroxide ion then hydrolyzes AE. Analysis of the hydrolysis rates of AE bound to DNA revealed that AE binds the minor groove of DNA and that its hydrolysis is inhibited by low water activity within the minor groove of the helix. Depending upon the sequence of the DNA, the water activity of the minor groove was estimated to be at least 2-4-fold lower than bulk solution. Hydrolysis measurements of AE tethered to RNA as well as RNA/DNA hybrids argued that the grooves of these double helices are also dehydrated relative to bulk solution. Remarkably, mismatched bases, regardless of their structure or sequence context, enhanced hydrolysis of AE by inducing hydration of the double helix that spread approximately five base pairs on either side of the mismatch. PMID:10220349

  14. Comparative Study of Corn Stover Pretreated by Dilute Acid and Cellulose Solvent-Based Lignocellulose Fractionation: Enzymatic Hydrolysis, Supramolecular Structure, and Substrate Accessibility

    SciTech Connect

    Zhu, Z.; Sathitsuksanoh, N.; Vinzant, T.; Schell, D. J.; McMillian, J. D.; Zhang, Y. H. P.

    2009-07-01

    Liberation of fermentable sugars from recalcitrant biomass is among the most costly steps for emerging cellulosic ethanol production. Here we compared two pretreatment methods (dilute acid, DA, and cellulose solvent and organic solvent lignocellulose fractionation, COSLIF) for corn stover. At a high cellulase loading [15 filter paper units (FPUs) or 12.3 mg cellulase per gram of glucan], glucan digestibilities of the corn stover pretreated by DA and COSLIF were 84% at hour 72 and 97% at hour 24, respectively. At a low cellulase loading (5 FPUs per gram of glucan), digestibility remained as high as 93% at hour 24 for the COSLIF-pretreated corn stover but reached only {approx}60% for the DA-pretreated biomass. Quantitative determinations of total substrate accessibility to cellulase (TSAC), cellulose accessibility to cellulase (CAC), and non-cellulose accessibility to cellulase (NCAC) based on adsorption of a non-hydrolytic recombinant protein TGC were measured for the first time. The COSLIF-pretreated corn stover had a CAC of 11.57 m{sup 2}/g, nearly twice that of the DA-pretreated biomass (5.89 m{sup 2}/g). These results, along with scanning electron microscopy images showing dramatic structural differences between the DA- and COSLIF-pretreated samples, suggest that COSLIF treatment disrupts microfibrillar structures within biomass while DA treatment mainly removes hemicellulose. Under the tested conditions COSLIF treatment breaks down lignocellulose structure more extensively than DA treatment, producing a more enzymatically reactive material with a higher CAC accompanied by faster hydrolysis rates and higher enzymatic digestibility.

  15. Mechanism of NTP Hydrolysis by the Escherichia coli Primary Replicative Helicase DnaB Protein. 2. Nucleotide and Nucleic Acid Specificities†

    PubMed Central

    Roychowdhury, Anasuya; Szymanski, Michal R.; Jezewska, Maria J.; Bujalowski, Wlodzimierz

    2011-01-01

    The kinetic mechanism of NTP binding and hydrolysis by the Escherichia coli replicative helicase, the DnaB protein, in the absence and presence of the single-stranded DNA (ssDNA), has been quantitatively examined using the rapid quench-flow technique, under single-turnover conditions. In the case of both the free helicase and the enzyme–ssDNA complexes, the mechanism is independent of the type of base of the cofactor or the DNA; the bimolecular association is followed by the reversible chemical hydrolysis and subsequent conformational transition of the enzyme–product complex. The NTP hydrolysis step is significantly faster for the purine than for the pyrimidine cofactor, both in the absence and in the presence of the DNA. The temperature effect indicates that the nature of intermediates of the purine nucleotide, ATP, is different from the nature of the analogous intermediates of the pyrimidine nucleotide, CTP. Nevertheless, both types of cofactors seem to approach a similar “exit” state at the end of the reaction. The effect of ssDNA on the kinetics of NTP hydrolysis depends on the type of nucleotide cofactor and the base composition of the DNA and is centered at the hydrolysis step. Homoadenosine ssDNA oligomers are particularly effective in increasing the hydrolysis rate. The allosteric signal from the DNA, which activates the NTP hydrolysis, comes predominantly from the strong DNA-binding subsite. The role of the weak DNA-binding subsite is to modulate the allosteric effect of the strong subsite. The significance of these results for the mechanism of the free energy transduction by the DnaB helicase is discussed. PMID:19435286

  16. Enantioselective enzymatic hydrolysis of racemic drugs by encapsulation in sol-gel magnetic sporopollenin.

    PubMed

    Yilmaz, Elif

    2012-05-01

    Candida rugosa lipase was encapsulated within a sol-gel procedure and improved considerably by fluoride-catalyzed hydrolysis of mixtures of octyltriethoxysilane and tetraethoxysilane in the presence of magnetic sporopollenin. The catalytic properties of the immobilized lipases were evaluated into model reactions, i.e., the hydrolysis of p-nitrophenylpalmitate (p-NPP), and the enantioselective hydrolysis of racemic naproxen methyl ester, mandelic acid methyl ester or 2-phenoxypropionic acid methyl ester that were studied in aqueous buffer solution/isooctane reaction system. The encapsulated magnetic sporopollenin (Spo-M-E) was found to give 319 U/g of support with 342% activity yield. It has been observed that the percent activity yields and enantioselectivity of the magnetic sporopollenin encapsulated lipase were higher than that of the encapsulated lipase without support. The substrate specificity of the encapsulated lipase revealed more efficient hydrolysis of the racemic naproxen methyl ester and 2-phenoxypropionic acid methyl ester than racemic mandelic acid methyl ester. It was observed that excellent enantioselectivity (E > 400) was obtained for encapsulated lipase with magnetic sporopollenin with an ee value of S-Naproxen and R-2 phenoxypropionic acid about 98%. PMID:21932062

  17. Lactic Acid and Thermal Treatments Trigger the Hydrolysis of Myo-Inositol Hexakisphosphate and Modify the Abundance of Lower Myo-Inositol Phosphates in Barley (Hordeum vulgare L.)

    PubMed Central

    Metzler-Zebeli, Barbara U.; Deckardt, Kathrin; Schollenberger, Margit; Rodehutscord, Markus; Zebeli, Qendrim

    2014-01-01

    Barley is an important source of dietary minerals, but it also contains myo-inositol hexakisphosphate (InsP6) that lowers their absorption. This study evaluated the effects of increasing concentrations (0.5, 1, and 5%, vol/vol) of lactic acid (LA), without or with an additional thermal treatment at 55°C (LA-H), on InsP6 hydrolysis, formation of lower phosphorylated myo-inositol phosphates, and changes in chemical composition of barley grain. Increasing LA concentrations and thermal treatment linearly reduced (P<0.001) InsP6-phosphate (InsP6-P) by 0.5 to 1 g compared to the native barley. In particular, treating barley with 5% LA-H was the most efficient treatment to reduce the concentrations of InsP6-P, and stimulate the formation of lower phosphorylated myo-inositol phosphates such as myo-inositol tetraphosphate (InsP4) and myo-inositol pentaphosphates (InsP5). Also, LA and thermal treatment changed the abundance of InsP4 and InsP5 isomers with Ins(1,2,5,6)P4 and Ins(1,2,3,4,5)P5 as the dominating isomers with 5% LA, 1% LA-H and 5% LA-H treatment of barley, resembling to profiles found when microbial 6-phytase is applied. Treating barley with LA at room temperature (22°C) increased the concentration of resistant starch and dietary fiber but lowered those of total starch and crude ash. Interestingly, total phosphorus (P) was only reduced (P<0.05) in barley treated with LA-H but not after processing of barley with LA at room temperature. In conclusion, LA and LA-H treatment may be effective processing techniques to reduce InsP6 in cereals used in animal feeding with the highest degradation of InsP6 at 5% LA-H. Further in vivo studies are warranted to determine the actual intestinal P availability and to assess the impact of changes in nutrient composition of LA treated barley on animal performance. PMID:24967651

  18. Hydrolysis of Sequenced β-Casein Peptides Provides New Insight into Peptidase Activity from Thermophilic Lactic Acid Bacteria and Highlights Intrinsic Resistance of Phosphopeptides

    PubMed Central

    Deutsch, Stéphanie-Marie; Molle, Daniel; Gagnaire, Valérie; Piot, Michel; Atlan, Danièle; Lortal, Sylvie

    2000-01-01

    The peptidases of thermophilic lactic acid bacteria have a key role in the proteolysis of Swiss cheeses during warm room ripening. To compare their peptidase activities toward a dairy substrate, a tryptic/chymotryptic hydrolysate of purified β-casein was used. Thirty-four peptides from 3 to 35 amino acids, including three phosphorylated peptides, constitute the β-casein hydrolysate, as shown by tandem mass spectrometry. Cell extracts prepared from Lactobacillus helveticus ITG LH1, ITG LH77, and CNRZ 32, Lactobacillus delbrueckii subsp. lactis ITG LL14 and ITG LL51, L. delbrueckii subsp. bulgaricus CNRZ 397 and NCDO 1489, and Streptococcus thermophilus CNRZ 385, CIP 102303, and TA 060 were standardized in protein. The peptidase activities were assessed with the β-casein hydrolysate as the substrate at pH 5.5 and 24°C (conditions of warm room ripening) by (i) free amino acid release, (ii) reverse-phase chromatography, and (iii) identification of undigested peptides by mass spectrometry. Regardless of strain, L. helveticus was the most efficient in hydrolyzing β-casein peptides. Interestingly, cell extracts of S. thermophilus were not able to release a significant level of free proline from the β-casein hydrolysate, which was consistent with the identification of numerous dipeptides containing proline. With the three lactic acid bacteria tested, the phosphorylated peptides remained undigested or weakly hydrolyzed indicating their high intrinsic resistance to peptidase activities. Finally, several sets of peptides differing by a single amino acid in a C-terminal position revealed the presence of at least one carboxypeptidase in the cell extracts of these species. PMID:11097915

  19. Comparison of polysaccharides of Haliotis discus hannai and Volutharpa ampullacea perryi by PMP-HPLC-MS(n) analysis upon acid hydrolysis.

    PubMed

    Wang, Hongxu; Zhao, Jun; Li, Dongmei; Wen, Chengrong; Liu, Haiman; Song, Shuang; Zhu, Beiwei

    2015-10-13

    Haliotis discus hannai Ino (Haliotis) is a highly valued marine shellfish, and it is sometimes replaced by another cheaper Gastropoda mollusk, Volutharpa ampullacea perryi (Volutharpa). Polysaccharides from pleopods, viscera and gonads of these two gastropods were compared by analyzing the mono- and di-saccharides in their acid hydrolysates using high performance liquid chromatography-mass spectrometry (HPLC-MS(n)) after 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatization. Disaccharide analysis revealed the distribution of uronic acid-containing polysaccharides (UACPs) in the biological samples. GlcA-(1 → 2)-Man, GlcA-(1 → 3)-GalN, and another disaccharide consisting of a hexuronic acid linked to a hexose were found in the hydrolysates, which indicated the existence of AGSP (abalone gonad sulfated polysaccharide) with the backbone composed of → 2)-α-Man(1 → 4)-β-GlcA(1 → repeating unit, AAP (abalone glycosaminoglycan-like polysaccharide) with the backbone of → 3)-GalNAc-(1 → 2)-GlcA-(1 → 3)-GalNAc-(1 → 4)-GlcA-(1 → repeating unit, and unidentified DS1P containing a hexuronic acid linked to a hexose unit, respectively. As shown by extracted ion chromatograms (XICs), AAP was the only UACP found in pleopods of the two gastropods; gonads and viscera of Haliotis contained DS1P and AGSP, while those of Volutharpa contained DS1P, AGSP as well as AAP. Monosaccharides in the acid hydrolysates were demonstrated in XICs by extracting their corresponding PMP derivative quasi-molecular ions one by one, and the results indicated the similar conclusion to the disaccharide analysis. Therefore, it could be concluded that polysaccharides from pleopods of the two gastropods are very similar, while those from their viscera and gonads differ greatly. PMID:26279526

  20. 12(S)-hydroxyeicosatetraenoic acid and 13(S)-hydroxyoctadecadienoic acid regulation of protein kinase C-alpha in melanoma cells: role of receptor-mediated hydrolysis of inositol phospholipids.

    PubMed Central

    Liu, B; Khan, W A; Hannun, Y A; Timar, J; Taylor, J D; Lundy, S; Butovich, I; Honn, K V

    1995-01-01

    Protein kinase C (PKC) isoenzymes are essential components of cell signaling. In this study, we investigated the regulation of PKC-alpha in murine B16 amelanotic melanoma (B16a) cells by the monohydroxy fatty acids 12(S)-hydroxyeicosatetraenoic acid [12(S)-HETE] and 13(S)-hydroxyoctadecadienoic acid [13(S)-HODE]. 12(S)-HETE induced a translocation of PKC-alpha to the plasma membrane and focal adhesion plaques, leading to enhanced adhesion of B16a cells to the matrix protein fibronectin. However, 13(S)-HODE inhibited these 12(S)-HETE effects on PKC-alpha. A receptor-mediated mechanism of action for 12(S)-HETE and 13(S)-HODE is supported by the following findings. First, 12(S)-HETE triggered a rapid increase in cellular levels of diacylglycerol and inositol trisphosphate in B16a cells. 13(S)-HODE blocked the 12(S)-HETE-induced bursts of both second messengers. Second, the 12(S)-HETE-increased adhesion of B16a cells to fibronectin was sensitive to inhibition by a phospholipase C inhibitor and pertussis toxin. Finally, a high-affinity binding site (Kd = 1 nM) for 12(S)-HETE was detected in B16a cells, and binding of 12(S)-HETE to B16a cells was effectively inhibited by 13(S)-HODE (IC50 = 4 nM). In summary, our data provide evidence that regulation of PKC-alpha by 12(S)-HETE and 13(S)-HODE may be through a guanine nucleotide-binding protein-linked receptor-mediated hydrolysis of inositol phospholipids. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:7568126

  1. Pretreatment of dried distillers grains with solubles by soaking in aqueous ammonia and subsequent enzymatic/dilute acid hydrolysis to produce fermentable sugars

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Dried distillers grains with solubles (DDGS), a co-product of corn ethanol production in the dry-grind process, was pretreated by soaking in aqueous ammonia (SAA) using a 15% w/w NH4OH solution at a solid:liquid ratio of 1:10. The effect of pretreatment on subsequent enzymatic hydrolysis was studied...

  2. The Oxidatively-Induced DNA Lesions 8,5′-Cyclo-2′-Deoxyadenosine and 8-Hydroxy-2′-Deoxyadenosine Are Strongly Resistant to Acid-Induced Hydrolysis of the Glycosidic Bond

    PubMed Central

    Theruvathu, Jacob A.; Jaruga, Pawel; Dizdaroglu, Miral

    2007-01-01

    The 8,5’-cyclopurine-2’-deoxynucleosides (cPu) are unique oxidatively-induced DNA lesions in that they are specifically repaired by NER. In the absence of NER, a possible mechanism for cPu removal is spontaneous glycosidic bond hydrolysis followed by enzymic processing. Such a mechanism could be significant if the glycosidic bond in cPu were substantially destabilized, as shown for other DNA lesions. Therefore, we investigated the stability of the glycosidic bond in a cPu, 8,5’(S)-cyclo-dA (S-cdA) against acid hydrolysis. For comparison, we also studied 8-OH-dA. We found that the glycosidic bond in S-cdA is ≈40 fold more resistant to glycosidic bond hydrolysis compared to dA. Interestingly, under the same conditions, the glycosidic bond in 8-OH-dA was even more stable than in S-cdA. These studies effectively rule out any mechanism for the removal of S-cdA or 8-OH-dA from DNA that requires spontaneous glycosidic bond hydrolysis, and further support the proposed role of cPu in the neurodegeneration observed in xeroderma pigmentosum patients who lack NER. Of broader significance, since NER does not function in non-transcribed DNA sequences of terminally differentiated cells, including neurons, cPu are expected to accumulate in such sequences even in individuals with normal NER, which could be important in the ageing process. PMID:17692895

  3. Novel double prodrugs of the iron chelator N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED): Synthesis, characterization, and investigation of activation by chemical hydrolysis and oxidation.

    PubMed

    Thiele, Nikki A; Abboud, Khalil A; Sloan, Kenneth B

    2016-08-01

    The development of iron chelators suitable for the chronic treatment of diseases where iron accumulation and subsequent oxidative stress are implicated in disease pathogenesis is an active area of research. The clinical use of the strong chelator N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) and its alkyl ester prodrugs has been hindered by poor oral bioavailability and lack of conversion to the parent chelator, respectively. Here, we present novel double prodrugs of HBED that have the carboxylate and phenolate donors of HBED masked with carboxylate esters and boronic acids/esters, respectively. These double prodrugs were successfully synthesized as free bases (7a-f) or as dimesylate salts (8a-c,e), and were characterized by (1)H, (13)C, and (11)B NMR; MP; MS; and elemental analysis. The crystal structure of 8a was solved. Three of the double prodrugs (8a-c) were selected for further investigation into their abilities to convert to HBED by stepwise hydrolysis and H2O2 oxidation. The serial hydrolysis of the pinacol and methyl esters of N,N'-bis(2-boronic acid pinacol ester benzyl)ethylenediamine-N,N'-diacetic acid methyl ester dimesylate (8a) was verified by LC-MS. The macro half-lives for the hydrolyses of 8a-c, measured by UV, ranged from 3.8 to 26.3 h at 37 °C in pH 7.5 phosphate buffer containing 50% MeOH. 9, the product of hydrolysis of 8a-c and the intermediate in the conversion pathway, showed little-to-no affinity for iron or copper in UV competition experiments. 9 underwent a serial oxidative deboronation by H2O2 in N-methylmorpholine buffer to generate HBED (k = 10.3 M(-1) min(-1)). The requirement of this second step, oxidation, before conversion to the active chelator is complete may confer site specificity when only localized iron chelation is needed. Overall, these results provide proof of principle for the activation of the double prodrugs by chemical hydrolysis and H2O2 oxidation, and merit further investigation into the

  4. Ultrafast hydrolysis of a Lewis photoacid.

    PubMed

    Henrich, Joseph D; Suchyta, Scott; Kohler, Bern

    2015-02-12

    This study explores the concept that electronic excitation can dramatically enhance Lewis acidity. Specifically, it is shown that photoexcitation transforms an electron-deficient organic compound of negligible Lewis acidity in its electronic ground state into a potent excited-state Lewis acid that releases a proton from a nearby water molecule in 3.1 ps. It was shown previously (Peon et al. J. Phys. Chem. A 2001, 105, 5768) that the excited state of methyl viologen (MV(2+)) is quenched rapidly in aqueous solution with the formation of an unidentified photoproduct. In this study, the quenching mechanism and the identity of the photoproduct were investigated by the femtosecond transient absorption and fluorescence upconversion techniques. Transient absorption signals at UV probe wavelengths reveal a long-lived species with a pH-dependent lifetime due to reaction with hydronium ions at a bimolecular rate of 3.1 × 10(9) M(-1) s(-1). This species is revealed to be a charge-transfer complex consisting of a ground-state MV(2+) ion and a hydroxide ion formed when a water molecule transfers a proton to the bulk solvent. Formation of a contact ion pair between MV(2+) and hydroxide shifts the absorption spectrum of the former ion by a few nm to longer wavelengths, yielding a transient absorption spectrum with a distinctive triangle wave appearance. The slight shift of this spectrum, which is in excellent agreement with steady-state difference spectra recorded for MV(2+) at high pH, is consistent with an ion pair but not with a covalent adduct (pseudobase). The long lifetime of the ion pair at neutral pH indicates that dissociation occurs many orders of magnitude more slowly than predicted by the Smoluchowski-Debye equation. Remarkably, there is no evidence of geminate recombination, suggesting that the proton that is transferred to the solvent is conducted at least several water shells away. Although the hydrolysis mechanism has yet to be fully established, evidence suggests

  5. Weak-acid sites catalyze the hydrolysis of crystalline cellulose to glucose in water: importance of post-synthetic functionalization of the carbon surface.

    PubMed

    To, Anh The; Chung, Po-Wen; Katz, Alexander

    2015-09-14

    The direct hydrolysis of crystalline cellulose to glucose in water without prior pretreatment enables the transformation of biomass into fuels and chemicals. To understand which features of a solid catalyst are most important for this transformation, the nanoporous carbon material MSC-30 was post-synthetically functionalized by oxidation. The most active catalyst depolymerized crystalline cellulose without prior pretreatment in water, providing glucose in an unprecedented 70 % yield. In comparison, virtually no reaction was observed with MSC-30, even when the reaction was conducted in aqueous solution at pH 2. As no direct correlations between the activity of this solid-solid reaction and internal-site characteristics, such as the β-glu adsorption capacity and the rate of catalytic hydrolysis of adsorbed β-glu strands, were observed, contacts of the external surface with the cellulose crystal are thought to be key for the overall efficiency. PMID:26276901

  6. Imidazole-derived 2-[N-carbamoylmethyl-alkylamino]acetic acids, substrate-dependent modulators of insulin-degrading enzyme in amyloid-β hydrolysis

    PubMed Central

    Charton, Julie; Gauriot, Marion; Guo, Qing; Hennuyer, Nathalie; Marechal, Xavier; Dumont, Julie; Hamdane, Malika; Pottiez, Virginie; Landry, Valerie; Sperandio, Olivier; Flipo, Marion; Buee, Luc; Staels, Bart; Leroux, Florence; Tang, Wei-Jen; Deprez, Benoit; Deprez-Poulain, Rebecca

    2014-01-01

    Substrates of Insulin-Degrading Enzyme are numerous and share little homology, like amyloid-beta and insulin. Small molecules binding both at the permanent exosite and at the discontinuous, conformational catalytic site, were discovered and co-crystallized with Insulin-Degrading Enzyme. Selective inhibition of amyloid-beta degradation over insulin hydrolysis was possible. Neuroblastoma cells treated with the optimized compound display a dose-dependent increase in amyloid-beta levels. PMID:24735644

  7. Functionalised carboxylic acids in atmospheric particles: An annual cycle revealing seasonal trends and possible sources

    NASA Astrophysics Data System (ADS)

    Teich, Monique; van Pinxteren, Dominik; Herrmann, Hartmut

    2013-04-01

    acids. The high concentrations in summer could lead to the conclusion that these acids are mostly formed during photochemical processes in the atmosphere. However, the concentrations in autumn were often exceeded by the ones in winter. Therefore probably other sources beside photochemical processes have to be considered. The second group consists of aromatic compounds. Because of the high concentrations in winter it can be concluded that photochemical formation plays a minor role and primary emission sources e.g., wood combustion are likely. Further evidence in determining sources of the carboxylic acids could be obtained from the air mass origin. In general, air masses transported from East have a more anthropogenic influence than the air mass inflow from West. For all aromatic carboxylic acids higher concentrations were determined during eastern inflow, indicating anthropogenic sources. This presumption is supported by high correlations with the elemental carbon (EC). Regarding the aliphatic carboxylic there is one group with higher concentrations when the air mass is transported from West and one with higher concentrations when air mass is transported from East. In summary the findings of this study reveal a clear difference in the seasonal trends of the single target acids indicating a variety of different sources.

  8. Fundamental study of the mechanism and kinetics of cellulose hydrolysis by acids and enzymes. Final report, June 1, 1978-January 31, 1981

    SciTech Connect

    Gong, C.S.; Chang, M.

    1981-02-01

    There are three basic enzymes (e.g., endoglucanase (C/sub x/), exoglucanase (C/sub 1/) and cellobiase) comprising the majority of extracellular cellulase enzymes produced by the cellulolytic mycelial fungi, Trichoderma reesei, and other cellulolytic microorganisms. The enzymes exhibited different mode of actions in respect to the hydrolysis of cellulose and cellulose derived oligosaccharides. In combination, these enzymes complimented each other to hydrolyze cellulose to its basic constituent, glucose. The kinetics of cellobiase were developed on the basis of applying the pseudo-steady state assumption to hydrolyze cellobiose to glucose. The results indicated that cellobiase was subjected to end-product inhibition by glucose. The kinetic modeling of exoglucanase (C/sub 1/) with respect to cellodextrins was studied. Both glucose and cellobiose were found to be inhibitors of this enzyme with cellobiose being a stronger inhibitor than glucose. Similarly, endoglucanase (C/sub x/) is subject to end-product inhibition by glucose. Crystallinity of the cellulose affects the rate of hydrolysis by cellulases. Hence, the changes in crystallinity of cellulose in relation to chemical pretreatment and enzyme hydrolysis was compared. The study of cellulase biosynthesis resulted in the conclusion that exo- and endo-glucanases are co-induced while cellobiase is synthesized independent of the other two enzymes. The multiplicity of cellulase enzymes are the end results of post-translational modification during and/or after the secretion of enzymes into growth environment.

  9. Elastoviscous Transitions of Articular Cartilage Reveal a Mechanism of Synergy between Lubricin and Hyaluronic Acid

    PubMed Central

    Bonnevie, Edward D.; Galesso, Devis; Secchieri, Cynthia; Cohen, Itai; Bonassar, Lawrence J.

    2015-01-01

    When lubricated by synovial fluid, articular cartilage provides some of the lowest friction coefficients found in nature. While it is known that macromolecular constituents of synovial fluid provide it with its lubricating ability, it is not fully understood how two of the main molecules, lubricin and hyaluronic acid, lubricate and interact with one another. Here, we develop a novel framework for cartilage lubrication based on the elastoviscous transition to show that lubricin and hyaluronic acid lubricate by distinct mechanisms. Such analysis revealed nonspecific interactions between these molecules in which lubricin acts to concentrate hyaluronic acid near the tissue surface and promotes a transition to a low friction regime consistent with the theory of viscous boundary lubrication. Understanding the mechanics of synovial fluid not only provides insight into the progression of diseases such as arthritis, but also may be applicable to the development of new biomimetic lubricants. PMID:26599797

  10. Pretreatment and Enzymatic Hydrolysis

    SciTech Connect

    2006-06-01

    Activities in this project are aimed at overcoming barriers associated with high capital and operating costs and sub-optimal sugar yields resulting from pretreatment and subsequent enzymatic hydrolysis of biomass.

  11. Hydrolysis of acyloxy nitroso compounds yields nitroxyl (HNO).

    PubMed

    Sha, Xin; Isbell, T Scott; Patel, Rakesh P; Day, Cynthia S; King, S Bruce

    2006-08-01

    Nitroxyl (HNO/NO(-)), the reduced form of nitric oxide, has gained attention based on its separate chemistry and biology from nitric oxide. The inherent reactivity of HNO requires new and mechanistically unique donors for the detailed study of HNO chemistry and biology. Oxidation of cyclohexanone oxime with lead tetraacetate yields 1-nitrosocyclohexyl acetate, whereas oxidation of oximes in the presence of excess carboxylic acid gives various acyloxy nitroso compounds. These bright blue compounds exist as monomers as indicated by their infrared, proton, and carbon NMR spectra, and X-ray crystallographic analysis reveals the nitroso groups possess a "nitroxyl-like" bent configuration. Hydrolysis of these compounds produces nitrous oxide, the dimerization and dehydration product of HNO, and provides evidence for the intermediacy of HNO. Both thiols and oxidative metal complexes inhibit nitrous oxide formation. Hydrolysis of these compounds in the presence of ferric heme complexes forms ferrous nitrosyl complexes providing further evidence for the intermediacy of HNO. Kinetic analysis shows that the rate of hydrolysis depends on pH and the structure of the acyl group of the acyloxy nitroso compound. These compounds relax pre-constricted rat aortic rings similar to known HNO donors. Together, these results identify acyloxy nitroso compounds as a new class of HNO donors. PMID:16866522

  12. Re-examination of Dietary Amino Acid Sensing Reveals a GCN2-Independent Mechanism.

    PubMed

    Leib, David E; Knight, Zachary A

    2015-11-10

    Animals cannot synthesize nine essential amino acids (EAAs) and must therefore obtain them from food. Mice reportedly reject food lacking a single EAA within the first hour of feeding. This remarkable phenomenon is proposed to involve post-ingestive sensing of amino acid imbalance by the protein kinase GCN2 in the brain. Here, we systematically re-examine dietary amino acid sensing in mice. In contrast to previous results, we find that mice cannot rapidly identify threonine- or leucine-deficient food in common feeding paradigms. However, mice attain the ability to identify EAA-deficient food following 2 days of EAA deprivation, suggesting a requirement for physiologic need. In addition, we report that mice can rapidly identify lysine-deficient food without prior EAA deficit, revealing a distinct sensing mechanism for this amino acid. These behaviors are independent of the proposed amino acid sensor GCN2, pointing to the existence of an undescribed mechanism for rapid sensing of dietary EAAs. PMID:26526991

  13. Re-examination of Dietary Amino Acid Sensing Reveals a GCN2-Independent Mechanism

    PubMed Central

    Leib, David E.; Knight, Zachary A.

    2016-01-01

    SUMMARY Animals cannot synthesize nine essential amino acids (EAAs) and must therefore obtain them from food. Mice reportedly reject food lacking a single EAA within the first hour of feeding. This remarkable phenomenon is proposed to involve post-ingestive sensing of amino acid imbalance by the protein kinase GCN2 in the brain. Here, we systematically re-examine dietary amino acid sensing in mice. In contrast to previous results, we find that mice cannot rapidly identify threonine- or leucine-deficient food in common feeding paradigms. However, mice attain the ability to identify EAA-deficient food following 2 days of EAA deprivation, suggesting a requirement for physiologic need. In addition, we report that mice can rapidly identify lysine-deficient food without prior EAA deficit, revealing a distinct sensing mechanism for this amino acid. These behaviors are independent of the proposed amino acid sensor GCN2, pointing to the existence of an undescribed mechanism for rapid sensing of dietary EAAs. PMID:26526991

  14. Fatty acid profiling reveals seasonal and spatial shifts in zooplankton diet in a temperate estuary

    NASA Astrophysics Data System (ADS)

    Gonçalves, A. M. M.; Azeiteiro, U. M.; Pardal, M. A.; De Troch, M.

    2012-08-01

    Fatty acids composition of copepod and cladoceran species and their possible food sources was investigated in the Mondego estuary (southern Europe) in order to explain the seasonal variation of the small copepods Acartia clausi, Acartia tonsa, Copidodiaptomus numidicus, Temora longicornis and the freshwater cladoceran Daphnia longispina. A total of 12 zooplankton species (7 marine, 2 estuarine and 3 freshwater species) were studied. A multivariate analysis revealed a clear seasonal distribution of zooplankton species in terms of fatty acids composition and abundance, with winter and spring zooplankton species showing maximal concentrations and diversity of total fatty acids. These findings underline the role of lipids as storage during the colder seasons in a highly variable environment like an estuary. Estuarine and freshwater species showed a more diverse array of saturated and unsaturated fatty acids rather than marine species, except for Centropages typicus. Fatty acids markers of trophic position indicated the presence of two trophic levels: copepod species were primarily omnivorous, whereas cladocerans showed to be herbivorous. Our results suggest that feeding patterns of plankton change spatially and temporally, reflecting the shifts in dominance between diatoms and flagellates as well as between dinoflagellates/diatoms and small animals.

  15. Characterization of Drosophila CMP-sialic acid synthetase activity reveals unusual enzymatic properties.

    PubMed

    Mertsalov, Ilya B; Novikov, Boris N; Scott, Hilary; Dangott, Lawrence; Panin, Vladislav M

    2016-07-01

    CMP-sialic acid synthetase (CSAS) is a key enzyme of the sialylation pathway. CSAS produces the activated sugar donor, CMP-sialic acid, which serves as a substrate for sialyltransferases to modify glycan termini with sialic acid. Unlike other animal CSASs that normally localize in the nucleus, Drosophila melanogaster CSAS (DmCSAS) localizes in the cell secretory compartment, predominantly in the Golgi, which suggests that this enzyme has properties distinct from those of its vertebrate counterparts. To test this hypothesis, we purified recombinant DmCSAS and characterized its activity in vitro Our experiments revealed several unique features of this enzyme. DmCSAS displays specificity for N-acetylneuraminic acid as a substrate, shows preference for lower pH and can function with a broad range of metal cofactors. When tested at a pH corresponding to the Golgi compartment, the enzyme showed significant activity with several metal cations, including Zn(2+), Fe(2+), Co(2+) and Mn(2+), whereas the activity with Mg(2+) was found to be low. Protein sequence analysis and site-specific mutagenesis identified an aspartic acid residue that is necessary for enzymatic activity and predicted to be involved in co-ordinating a metal cofactor. DmCSAS enzymatic activity was found to be essential in vivo for rescuing the phenotype of DmCSAS mutants. Finally, our experiments revealed a steep dependence of the enzymatic activity on temperature. Taken together, our results indicate that DmCSAS underwent evolutionary adaptation to pH and ionic environment different from that of counterpart synthetases in vertebrates. Our data also suggest that environmental temperatures can regulate Drosophila sialylation, thus modulating neural transmission. PMID:27114558

  16. Analysis of twenty phenolic compounds in human urine: hydrochloric acid hydrolysis, solid-phase extraction based on K2CO 3-treated silica, and gas chromatography tandem mass spectrometry.

    PubMed

    Lu, Dasheng; Feng, Chao; Wang, Dongli; Lin, Yuanjie; Ip, Ho Sai Simon; She, Jianwen; Xu, Qian; Wu, Chunhua; Wang, Guoquan; Zhou, Zhijun

    2015-05-01

    This study developed a new method for the analysis of 20 phenolic compounds in human urine. The urine samples were prepared by hydrochloric acid (HCl) hydrolysis, liquid-liquid extraction (LLE), and solid-phase extraction (SPE) cleanup. We found that HCl hydrolysis is of similar effectiveness to, and much cheaper than, the traditional enzymatic method. Vanillic acid was co-eluted with butyl paraben and interfered with the determination of butyl paraben in urine. K2CO3-treated-silica-gel SPE was designed to efficiently eliminate interference from the endogenous organic acids (especially vanillic acid) in urine. After derivatization, the samples were analyzed by large-volume-injection gas chromatography-tandem mass spectrometry (LVI-GC-MS-MS). Good linearity (R (2) ≥ 0.996) was established in the range 0.1-100 ng mL(-1) for all analytes. Method detection limits (MDLs) were 0.7-9.8 pg mL(-1). Intraday (n = 5) and interday (n = 5 days) validation was performed, with satisfactory accuracy (recovery: 70-126 % and 73-107 %, respectively) and precision (RSD ≤ 19 %) at two levels (low: 0.1 and 0.5 ng mL(-1); high: 5 and 10 ng mL(-1)). The method was used in a population study and achieved more than 85 % detection for most analytes; mean analyte concentrations were in the range 0.01-185 ng mL(-1). The method is suitable for the analysis of multiple phenolic metabolites in human urine. PMID:25903021

  17. Water Stress Responses of Tomato Mutants Impaired in Hormone Biosynthesis Reveal Abscisic Acid, Jasmonic Acid and Salicylic Acid Interactions.

    PubMed

    Muñoz-Espinoza, Valeria A; López-Climent, María F; Casaretto, José A; Gómez-Cadenas, Aurelio

    2015-01-01

    To investigate the putative crosstalk between JA and ABA in Solanum lycopersicum plants in response to drought, suppressor of prosystemin-mediated responses2 (spr2, JA-deficient) and flacca (flc, ABA-deficient) mutants together with the naphthalene/salicylate hydroxylase (NahG) transgenic (SA-deficient) line were used. Hormone profiling and gene expression of key enzymes in ABA, JA and SA biosynthesis were analyzed during early stages of drought. ABA accumulation was comparable in spr2 and wild type (WT) plants whereas expression of 9-cis-epoxycarotenoid dioxygenase 1 (NCED1) and NCED2 was different, implying a compensation mechanism between NCED genes and an organ-specific regulation of NCED1 expression. JA levels and 12-oxo-phytodienoic acid reductase 3 (OPR3) expression in flc plants suggest that ABA regulates the induction of the OPR3 gene in roots. By contrast, ABA treatment to flc plants leads to a reduction of JA and SA contents. Furthermore, different pattern of SA accumulation (and expression of isochorismate synthase and phenylalanine ammonia lyase 1) was observed between WT seedlings and mutants, suggesting that SA plays an important role on the early response of tomato plants to drought and also that JA and ABA modulate its biosynthesis. Finally, hormone profiling in spr2 and NahG plants indicate a crosstalk between JA and SA that could enhance tolerance of tomato to water stress. PMID:26635826

  18. Water Stress Responses of Tomato Mutants Impaired in Hormone Biosynthesis Reveal Abscisic Acid, Jasmonic Acid and Salicylic Acid Interactions

    PubMed Central

    Muñoz-Espinoza, Valeria A.; López-Climent, María F.; Casaretto, José A.; Gómez-Cadenas, Aurelio

    2015-01-01

    To investigate the putative crosstalk between JA and ABA in Solanum lycopersicum plants in response to drought, suppressor of prosystemin-mediated responses2 (spr2, JA-deficient) and flacca (flc, ABA-deficient) mutants together with the naphthalene/salicylate hydroxylase (NahG) transgenic (SA-deficient) line were used. Hormone profiling and gene expression of key enzymes in ABA, JA and SA biosynthesis were analyzed during early stages of drought. ABA accumulation was comparable in spr2 and wild type (WT) plants whereas expression of 9-cis-epoxycarotenoid dioxygenase 1 (NCED1) and NCED2 was different, implying a compensation mechanism between NCED genes and an organ-specific regulation of NCED1 expression. JA levels and 12-oxo-phytodienoic acid reductase 3 (OPR3) expression in flc plants suggest that ABA regulates the induction of the OPR3 gene in roots. By contrast, ABA treatment to flc plants leads to a reduction of JA and SA contents. Furthermore, different pattern of SA accumulation (and expression of isochorismate synthase and phenylalanine ammonia lyase 1) was observed between WT seedlings and mutants, suggesting that SA plays an important role on the early response of tomato plants to drought and also that JA and ABA modulate its biosynthesis. Finally, hormone profiling in spr2 and NahG plants indicate a crosstalk between JA and SA that could enhance tolerance of tomato to water stress. PMID:26635826

  19. Lipidomic profiling reveals protective function of fatty acid oxidation in cocaine-induced hepatotoxicity[S

    PubMed Central

    Shi, Xiaolei; Yao, Dan; Gosnell, Blake A.; Chen, Chi

    2012-01-01

    During cocaine-induced hepatotoxicity, lipid accumulation occurs prior to necrotic cell death in the liver. However, the exact influences of cocaine on the homeostasis of lipid metabolism remain largely unknown. In this study, the progression of subacute hepatotoxicity, including centrilobular necrosis in the liver and elevation of transaminase activity in serum, was observed in a three-day cocaine treatment, accompanying the disruption of triacylglycerol (TAG) turnover. Serum TAG level increased on day 1 of cocaine treatment but remained unchanged afterwards. In contrast, hepatic TAG level was elevated continuously during three days of cocaine treatment and was better correlated with the development of hepatotoxicity. Lipidomic analyses of serum and liver samples revealed time-dependent separation of the control and cocaine-treated mice in multivariate models, which was due to the accumulation of long-chain acylcarnitines together with the disturbances of many bioactive phospholipid species in the cocaine-treated mice. An in vitro function assay confirmed the progressive inhibition of mitochondrial fatty acid oxidation after the cocaine treatment. Cotreatment of fenofibrate significantly increased the expression of peroxisome proliferator-activated receptor α (PPARα)-targeted genes and the mitochondrial fatty acid oxidation activity in the cocaine-treated mice, resulting in the inhibition of cocaine-induced acylcarnitine accumulation and other hepatotoxic effects. Overall, the results from this lipidomics-guided study revealed that the inhibition of fatty acid oxidation plays an important role in cocaine-induced liver injury. PMID:22904346

  20. Metabolomic Analyses of Leishmania Reveal Multiple Species Differences and Large Differences in Amino Acid Metabolism

    PubMed Central

    Wang, Lijie; Zhang, Tong; Watson, David G.; Silva, Ana Marta; Coombs, Graham H.

    2015-01-01

    Comparative genomic analyses of Leishmania species have revealed relatively minor heterogeneity amongst recognised housekeeping genes and yet the species cause distinct infections and pathogenesis in their mammalian hosts. To gain greater information on the biochemical variation between species, and insights into possible metabolic mechanisms underpinning visceral and cutaneous leishmaniasis, we have undertaken in this study a comparative analysis of the metabolomes of promastigotes of L. donovani, L. major and L. mexicana. The analysis revealed 64 metabolites with confirmed identity differing 3-fold or more between the cell extracts of species, with 161 putatively identified metabolites differing similarly. Analysis of the media from cultures revealed an at least 3-fold difference in use or excretion of 43 metabolites of confirmed identity and 87 putatively identified metabolites that differed to a similar extent. Strikingly large differences were detected in their extent of amino acid use and metabolism, especially for tryptophan, aspartate, arginine and proline. Major pathways of tryptophan and arginine catabolism were shown to be to indole-3-lactate and arginic acid, respectively, which were excreted. The data presented provide clear evidence on the value of global metabolomic analyses in detecting species-specific metabolic features, thus application of this technology should be a major contributor to gaining greater understanding of how pathogens are adapted to infecting their hosts. PMID:26368322

  1. Escherichia coli DnaB Helicase–DnaC Protein Complex: Allosteric Effects of the Nucleotides on the Nucleic Acid Binding and the Kinetic Mechanism of NTP Hydrolysis. 3†

    PubMed Central

    Roychowdhury, Anasuya; Szymanski, Michal R.; Jezewska, Maria J.; Bujalowski, Wlodzimierz

    2011-01-01

    Allosteric interactions between the DNA- and NTP-binding sites of the Escherichia coli DnaB helicase engaged in the DnaB–DnaC complex and the mechanism of NTP hydrolysis by the complex have been examined using the fluorescence titration, analytical ultracentrifugation, and rapid quench-flow technique. Surprisingly, the ssDNA affinity of the DnaB–DnaC complex is independent of the structure of the phosphate group of the cofactor bound to the helicase. Thus, the DnaC protein eliminates the antagonistic allosteric effect of NTP and NDP on the ssDNA affinity of the enzyme. The protein changes the engagement of the DNA-binding subsites of the helicase in interactions with the nucleic acid, depending on the structure of the phosphate group of the present nucleotide cofactor and profoundly affects the structure of the bound DNA. Moreover, the ssDNA affinity of the helicase in the DnaB–DnaC complex is under the control of the nucleotide-binding site of the DnaC protein. The protein does not affect the NTP hydrolysis mechanism of the helicase. Nevertheless, the rate of the chemical step is diminished in the DnaB–DnaC complex. In the tertiary DnaB–DnaC–ssDNA complex, the ssDNA changes the internal dynamics between intermediates of the pyrimidine cofactor, in a manner independent of the base composition of the DNA, while the hydrolysis step of the purine cofactor is specifically stimulated by the homoadenosine ssDNA. The significance of these results for functional activities of the DnaB–DnaC complex is discussed. PMID:19432487

  2. Linear polyethylenimine produced by partial acid hydrolysis of poly(2-ethyl-2-oxazoline) for DNA and siRNA delivery in vitro

    PubMed Central

    Fernandes, Julio C; Qiu, Xingping; Winnik, Françoise M; Benderdour, Mohamed; Zhang, Xiaoling; Dai, Kerong; Shi, Qin

    2013-01-01

    Polyethylenimines (PEIs) are the most efficient synthetic vectors for gene delivery available to date. With its high charge density and strong proton-buffering effect, PEI has an ability to condense DNA and small interfering RNA at physiologic pH. However, the polymer suffers from the disadvantage of high cellular toxicity. To reduce its cellular toxicity, we synthesized linear PEIs by partial hydrolysis of poly(2-ethyl-2-oxazoline). Three linear PEIs with different hydrolysis percentages (30%, 70%, and 96%, respectively) were produced as PEI30, PEI70, and PEI96. PEI30 and PEI96 cannot be considered as suitable transfection agents because of low transfection efficiency (PEI30) or high cellular toxicity (PEI96). PEI70 displayed very weak cell toxicity. The charge density of this polymer (PEI70) was strong enough to condense DNA and small interfering RNA at a physiologic pH of 7.4. Our results also show that PEI70 was highly efficient in DNA delivery and small interfering RNA-mediated knockdown of target genes. Thus, polymers such as PEI70 appear to be very promising vectors for gene delivery. PMID:24204139

  3. 5-hydroxymethyl-2-furfural and derivatives formed during acid hydrolysis of conjugated and bound phenolics in plant foods and the effects on phenolic content and antioxidant capacity.

    PubMed

    Chen, Peter X; Tang, Yao; Zhang, Bing; Liu, Ronghua; Marcone, Massimo F; Li, Xihong; Tsao, Rong

    2014-05-21

    A common protocol for the extraction of phenolic aglycons or bound phenolics in plants generally involves hydrothermal hydrolysis in an aqueous methanol or ethanol solution containing 2-4 N HCl. However, as shown in the present study, this process also forms 5-(hydroxymethyl)furan-2-carbaldehyde (HMF) and its derivative products 5-(methoxymethyl)furan-2-carbaldehyde (MMF) and 5-(ethoxymethyl)furan-2-carbaldehyde (EMF), as identified by HPLC-DAD-ESI-MS/MS and NMR. These compounds are commonly misidentified as phenolics due to similar UV absorption at 280 nm. In this study, production of HMF, MMF, and EMF was shown to be dependent on the solvent condition and duration and temperature of hydrolysis. Fruits and vegetables produced HMF more readily than grains. HMF and its derivatives were subjected to various spectrophotometric antioxidant assays [2-diphenyl-1-picryhydrazyl radical scavenging activity (DPPH), ferric-reducing antioxidant power (FRAP), and oxygen radical absorbing capacity (ORAC)] and displayed antioxidant activity mainly in the ORAC assay. Results of this study help avoid overestimation of phenolic content and antioxidant activities of plant foods. PMID:24796380

  4. Antiparasitic evaluation of betulinic acid derivatives reveals effective and selective anti-Trypanosoma cruzi inhibitors.

    PubMed

    Meira, Cássio Santana; Barbosa-Filho, José Maria; Lanfredi-Rangel, Adriana; Guimarães, Elisalva Teixeira; Moreira, Diogo Rodrigo Magalhães; Soares, Milena Botelho Pereira

    2016-07-01

    Betulinic acid is a pentacyclic triterpenoid with several biological properties already described, including antiparasitic activity. Here, the anti-Trypanosoma cruzi activity of betulinic acid and its semi-synthetic amide derivatives (BA1-BA8) was investigated. The anti-Trypanosoma cruzi activity and selectivity were enhanced in semi-synthetic derivatives, specially on derivatives BA5, BA6 and BA8. To understand the mechanism of action underlying betulinic acid anti-T. cruzi activity, we investigated ultrastructural changes by electron microscopy. Ultrastructural studies showed that trypomastigotes incubated with BA5 had membrane blebling, flagella retraction, atypical cytoplasmic vacuoles and Golgi cisternae dilatation. Flow cytometry analysis showed that parasite death is mainly caused by necrosis. Treatment with derivatives BA5, BA6 or BA8 reduced the invasion process, as well as intracellular parasite development in host cells, with a potency and selectivity similar to that observed in benznidazole-treated cells. More importantly, the combination of BA5 and benznidazole revealed synergistic effects on trypomastigote and amastigote forms of T. cruzi. In conclusion, we demonstrated that BA5 compound is an effective and selective anti-T. cruzi agent. PMID:27080160

  5. Towards the design of organocatalysts for nerve agents remediation: The case of the active hydrolysis of DCNP (a Tabun mimic) catalyzed by simple amine-containing derivatives.

    PubMed

    Barba-Bon, Andrea; Martínez-Máñez, Ramón; Sancenón, Félix; Costero, Ana M; Gil, Salvador; Pérez-Pla, Francisco; Llopis, Elisa

    2015-11-15

    We report herein a study of the hydrolysis of Tabun mimic DCNP in the presence of different amines, aminoalcohols and glycols as potential suitable organocatalysts for DCNP degradation. Experiments were performed in CD3CN in the presence of 5% D2O, which is a suitable solvent mixture to follow the DCNP hydrolysis. These studies allowed the definition of different DCNP depletion paths, resulting in the formation of diethylphosphoric acid, tetraethylpyrophosphate and phosphoramide species as final products. Without organocatalysts, DCNP hydrolysis occurred mainly via an autocatalysis path. Addition of tertiary amines in sub-stoichiometric amounts largely enhanced DCNP depletion whereas non-tertiary polyamines reacted even faster. Glycols induced very slight increment in the DCNP hydrolysis, whereas DCNP hydrolysis increased sharply in the presence of certain aminoalcohols especially, 2-(2-aminoethylamino)ethanol. For the latter compound, DCNP depletion occurred ca. 80-fold faster than in the absence of organocatalysts. The kinetic studies revealed that DCNP hydrolysis in the presence of 2-(2-aminoethylamino)ethanol occurred via a catalytic process, in which the aminoalcohol was involved. DCNP hydrolysis generally depended strongly on the structure of the amine, and it was found that the presence of the OHCH2CH2N moiety in the organocatalyst structure seems important to induce a fast degradation of DCNP. PMID:26005922

  6. Cation–Anion Interactions within the Nucleic Acid Ion Atmosphere Revealed by Ion Counting

    PubMed Central

    Gebala, Magdalena; Giambasu, George M.; Lipfert, Jan; Bisaria, Namita; Bonilla, Steve; Li, Guangchao; York, Darrin M.; Herschlag, Daniel

    2016-01-01

    The ion atmosphere is a critical structural, dynamic, and energetic component of nucleic acids that profoundly affects their interactions with proteins and ligands. Experimental methods that “count” the number of ions thermodynamically associated with the ion atmosphere allow dissection of energetic properties of the ion atmosphere, and thus provide direct comparison to theoretical results. Previous experiments have focused primarily on the cations that are attracted to nucleic acid polyanions, but have also showed that anions are excluded from the ion atmosphere. Herein, we have systematically explored the properties of anion exclusion, testing the zeroth-order model that anions of different identity are equally excluded due to electrostatic repulsion. Using a series of monovalent salts, we find, surprisingly, that the extent of anion exclusion and cation inclusion significantly depends on salt identity. The differences are prominent at higher concentrations and mirror trends in mean activity coefficients of the electrolyte solutions. Salts with lower activity coefficients exhibit greater accumulation of both cations and anions within the ion atmosphere, strongly suggesting that cation–anion correlation effects are present in the ion atmosphere and need to be accounted for to understand electrostatic interactions of nucleic acids. To test whether the effects of cation–anion correlations extend to nucleic acid kinetics and thermodynamics, we followed the folding of P4–P6, a domain of the Tetrahymena group I ribozyme, via single-molecule fluorescence resonance energy transfer in solutions with different salts. Solutions of identical concentration but lower activity gave slower and less favorable folding. Our results reveal hitherto unknown properties of the ion atmosphere and suggest possible roles of oriented ion pairs or anion-bridged cations in the ion atmosphere for electrolyte solutions of salts with reduced activity. Consideration of these new

  7. Regulatory link between steryl ester formation and hydrolysis in the yeast Saccharomyces cerevisiae.

    PubMed

    Ploier, Birgit; Korber, Martina; Schmidt, Claudia; Koch, Barbara; Leitner, Erich; Daum, Günther

    2015-07-01

    Steryl esters and triacylglycerols are the major storage lipids of the yeast Saccharomyces cerevisiae. Steryl esters are formed in the endoplasmic reticulum by the two acyl-CoA:sterol acyltransferases Are1p and Are2p, whereas steryl ester hydrolysis is catalyzed by the three steryl ester hydrolases Yeh1p, Yeh2p and Tgl1p. To shed light on the regulatory link between steryl ester formation and hydrolysis in the maintenance of cellular sterol and free fatty acid levels we employed yeast mutants which lacked the enzymes catalyzing the degradation of steryl esters. These studies revealed feedback regulation of steryl ester formation by steryl ester hydrolysis although in a Δtgl1Δyeh1Δyeh2 triple mutant the gene expression levels of ARE1 and ARE2 as well as protein levels and stability of Are1p and Are2p were not altered. Nevertheless, the capacity of the triple mutant to synthesize steryl esters was significantly reduced as shown by in vitro and in vivo labeling of lipids with [(14)C]oleic acid and [(14)C]acetate. Enzymatic analysis revealed that inhibition of steryl ester formation occurred at the enzyme level. As the amounts and the formation of sterols and fatty acids were also decreased in the triple mutant we concluded that defects in steryl ester hydrolysis also caused feedback inhibition on the formation of sterols and fatty acids which serve as precursors for steryl ester formation. In summary, this study demonstrates a regulatory link within the steryl ester metabolic network which contributes to non-polar lipid homeostasis in yeast cells. PMID:25720564

  8. {sup 27}Al and {sup 23}Na MAS NMR and powder x-ray diffraction studies of sodium aluminate speciation and the mechanistics of aluminum hydroxide precipitation upon acid hydrolysis

    SciTech Connect

    Bradley, S.M.; Hanna, J.V.

    1994-08-24

    {sup 27}Al and {sup 23}Na MAS NMR, powder X-ray diffraction, and infrared spectroscopic investigations of freeze-dried sodium aluminates and aluminum hydroxides formed through acid hydrolysis have been undertaken, with OH/Al hydrolysis ratios between 5.3 and 2.8 being analyzed. Numerous {sup 27}AlNMR resonances were observed, the intensities of which vary as a function of OH/Al ratio, and these have been assigned to four-, five-, and six-coordinate aluminum species constituting a variety of structural moieties. The dominant species at an OH/Al ratio above 4.4 appears to be a Q{sup o}Na[Al(OH);{sub 4}] salt, as indicated by a {sup 27}Al resonance at 86.6 ppm. In addition, a second, broader resonance at 71.3 ppm demonstrates the simultaneous existence of further four-coordinate aluminum species linked thorough oxo bonds to other four-coordinate aluminums (e.g., Q{sup 2} [Al(OH);{sub 2}(OAl){sub 2}];{sup x-}). At an OH/Al ratio between 4.4 and 4.1, a water-soluble phase forms that contains both four- and six-coordinate aluminum. At OH/Al ratios fo 4.0 and below, a water-soluble phase forms that contains both four-and six-coordinate aluminum. AT OH/Al ratios of 4.0 and below, a water-insoluble phase exists possessing four-, five-, and six-coordinate aluminum. At OH/Al{le}3.9 range exhibits {sup 27}Al chemical shifts similar to those reported for transitional aluminas such as {gamma}-, {eta}-, and 0-Al{sub 2}O{sub 3} and an infrared spectrum similar to pseudo-spinel gels, suggesting that a pseudo-spinel intermediate is the first phase involved in the crystallization of gibbsite. The resonance assigned to five-coordinate aluminum probably results from species involved in the transformation of the pseudo-spinal phase to pseudo-boehmite. The formation of gibbssite on the acid hydrolysis of alkaline sodium aluminate solutions thus appears to follow the pathway pseudo-spinel {r_arrow} pseudo-boehmite {r_arrow} bayerite {r_arrow} gibbsite. 82 refs., 7 figs., 3 tabs.

  9. Analysis of TP53 mutation spectra reveals the fingerprint of the potent environmental carcinogen, aristolochic acid

    PubMed Central

    Hollstein, M; Moriya, M.; Grollman, AP; Olivier, M

    2013-01-01

    Genetic alterations in cancer tissues may reflect the mutational fingerprint of environmental carcinogens. Here we review the evidence that support the role of aristolochic acid (AA) in inducing a mutational fingerprint in the tumor suppressor gene TP53 in urothelial carcinomas of the upper urinary tract (UUT). Exposure to AA, a nitrophenathrene carboxylic acid present in certain herbal remedies and in flour prepared from wheat grain contaminated with seeds of Aristolochia clematitis, has been linked to chronic nephropathy and UUT. TP53 mutations in UUT of individuals exposed to AA reveal a unique pattern of mutations characterised by A to T transversions on the non-transcribed strand, which cluster at hotspots rarely mutated in other cancers. This unusual pattern, originally discovered in UUTs from two different populations, one in Taiwan, and one in the Balkans, has been reproduced experimentally by treating mouse cells that harbour human TP53 sequences with AA. The convergence of molecular epidemiological and experimental data establishes a clear causal association between exposure to the human carcinogen AA and UUT cancer. Despite bans on the sale of herbs containing AA, their use continues, raising global public health concern and an urgent need to identify populations at risk. PMID:23422071

  10. NREL 2012 Achievement of Ethanol Cost Targets: Biochemical Ethanol Fermentation via Dilute-Acid Pretreatment and Enzymatic Hydrolysis of Corn Stover

    SciTech Connect

    Tao, L.; Schell, D.; Davis, R.; Tan, E.; Elander, R.; Bratis, A.

    2014-04-01

    For the DOE Bioenergy Technologies Office, the annual State of Technology (SOT) assessment is an essential activity for quantifying the benefits of biochemical platform research. This assessment has historically allowed the impact of research progress achieved through targeted Bioenergy Technologies Office funding to be quantified in terms of economic improvements within the context of a fully integrated cellulosic ethanol production process. As such, progress toward the ultimate 2012 goal of demonstrating cost-competitive cellulosic ethanol technology can be tracked. With an assumed feedstock cost for corn stover of $58.50/ton this target has historically been set at $1.41/gal ethanol for conversion costs only (exclusive of feedstock) and $2.15/gal total production cost (inclusive of feedstock) or minimum ethanol selling price (MESP). This year, fully integrated cellulosic ethanol production data generated by National Renewable Energy Laboratory (NREL) researchers in their Integrated Biorefinery Research Facility (IBRF) successfully demonstrated performance commensurate with both the FY 2012 SOT MESP target of $2.15/gal (2007$, $58.50/ton feedstock cost) and the conversion target of $1.41/gal through core research and process improvements in pretreatment, enzymatic hydrolysis, and fermentation.

  11. Acid-labile mPEG-Vinyl Ether-1,2-Dioleylglycerol Lipids with Tunable pH Sensitivity: Synthesis and Structural Effects on Hydrolysis Rates, DOPE Liposome Release Performance and Pharmacokinetics

    PubMed Central

    Shin, Junhwa; Shum, Pochi; Grey, Jessica; Fujiwara, Shin-ichi; Malhotra, Guarov S.; González-Bonet, Andres; Hyun, Seok-Hee; Moase, Elaine; Allen, Theresa M.; Thompson, David H.

    2012-01-01

    A family of 3-methoxypoly(ethylene glycol)-vinyl ether-1,2-dioleylglycerol (mPEG-VE-DOG) lipopolymer conjugates, designed on the basis of DFT calculations to possess a wide range of proton affinities, was synthesized and tested for their hydrolysis kinetics in neutral and acidic buffers. Extruded ~100 nm liposomes containing these constructs in ≥90 mol% 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) produced dispersions that retained their calcein cargo for more than 2 days at pH 7.5, but released the encapsulated contents over a wide range of timescales as a function of the electronic properties of the vinyl ether linkage, the solution pH and the mPEG-VE-DOG composition in the membrane. The in vivo performance of two different 90:10 DOPE:mPEG-VE-DOG compositions was also evaluated for blood circulation time and biodistribution in mice, using 125I-tyraminylinulin as a label. The pharmacokinetic profiles gave a T1/2 of 7 h and 3 h for 90:10 DOPE:ST302 and 90:10 DOPE:ST502, respectively, with the liposomes being cleared predominantly by liver and spleen uptake. The behavior of these DOPE:mPEG-VE-DOG formulations is consistent with their relative rates of vinyl ether hydrolysis, i.e., the more acid-sensitive mPEG-VE-DOG derivatives produce faster leakage rates from DOPE:mPEG-VE-DOG liposomes, but decreased the blood circulation times in mice. These findings suggest that the vinyl ether-based PEG-lipid derivatives are promising agents for stabilizing acid-sensitive DOPE liposomes to produce formulations with a priori control over their pH-responsiveness in vitro. Our data also suggest, however, that the same factors that contribute to enhanced acid-sensitivity of the DOPE:mPEG-VE-DOG dispersions are also likely responsible for their reduced pharmacokinetic profiles. PMID:23030381

  12. Acid-labile mPEG-vinyl ether-1,2-dioleylglycerol lipids with tunable pH sensitivity: synthesis and structural effects on hydrolysis rates, DOPE liposome release performance, and pharmacokinetics.

    PubMed

    Shin, Junhwa; Shum, Pochi; Grey, Jessica; Fujiwara, Shin-ichi; Malhotra, Guarov S; González-Bonet, Andres; Hyun, Seok-Hee; Moase, Elaine; Allen, Theresa M; Thompson, David H

    2012-11-01

    A family of 3-methoxypoly(ethylene glycol)-vinyl ether-1,2-dioleylglycerol (mPEG-VE-DOG) lipopolymer conjugates, designed on the basis of DFT calculations to possess a wide range of proton affinities, was synthesized and tested for their hydrolysis kinetics in neutral and acidic buffers. Extruded ∼100 nm liposomes containing these constructs in ≥90 mol % 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) produced dispersions that retained their calcein cargo for more than 2 days at pH 7.5, but released the encapsulated contents over a wide range of time scales as a function of the electronic properties of the vinyl ether linkage, the solution pH, and the mPEG-VE-DOG composition in the membrane. The in vivo performance of two different 90:10 DOPE:mPEG-VE-DOG compositions was also evaluated for blood circulation time and biodistribution in mice, using (125)I-tyraminylinulin as a label. The pharmacokinetic profiles gave a t(1/2) of 7 and 3 h for 90:10 DOPE:ST302 and 90:10 DOPE:ST502, respectively, with the liposomes being cleared predominantly by liver and spleen uptake. The behavior of these DOPE:mPEG-VE-DOG formulations is consistent with their relative rates of vinyl ether hydrolysis, i.e., the more acid-sensitive mPEG-VE-DOG derivatives produced faster leakage rates from DOPE:mPEG-VE-DOG liposomes, but decreased the blood circulation times in mice. These findings suggest that the vinyl ether-based PEG-lipid derivatives are promising agents for stabilizing acid-sensitive DOPE liposomes to produce formulations with a priori control over their pH responsiveness in vitro. Our data also suggest, however, that the same factors that contribute to enhanced acid sensitivity of the DOPE:mPEG-VE-DOG dispersions are also likely responsible for their reduced pharmacokinetic profiles. PMID:23030381

  13. Comparative genomics of lactic acid bacteria reveals a niche-specific gene set

    PubMed Central

    2009-01-01

    Background The recently sequenced genome of Lactobacillus helveticus DPC4571 [1] revealed a dairy organism with significant homology (75% of genes are homologous) to a probiotic bacteria Lb. acidophilus NCFM [2]. This led us to hypothesise that a group of genes could be determined which could define an organism's niche. Results Taking 11 fully sequenced lactic acid bacteria (LAB) as our target, (3 dairy LAB, 5 gut LAB and 3 multi-niche LAB), we demonstrated that the presence or absence of certain genes involved in sugar metabolism, the proteolytic system, and restriction modification enzymes were pivotal in suggesting the niche of a strain. We identified 9 niche specific genes, of which 6 are dairy specific and 3 are gut specific. The dairy specific genes identified in Lactobacillus helveticus DPC4571 were lhv_1161 and lhv_1171, encoding components of the proteolytic system, lhv_1031 lhv_1152, lhv_1978 and lhv_0028 encoding restriction endonuclease genes, while bile salt hydrolase genes lba_0892 and lba_1078, and the sugar metabolism gene lba_1689 from Lb. acidophilus NCFM were identified as gut specific genes. Conclusion Comparative analysis revealed that if an organism had homologs to the dairy specific geneset, it probably came from a dairy environment, whilst if it had homologs to gut specific genes, it was highly likely to be of intestinal origin. We propose that this "barcode" of 9 genes will be a useful initial guide to researchers in the LAB field to indicate an organism's ability to occupy a specific niche. PMID:19265535

  14. Formation of hydroxyapatite by hydrolysis of alpha-tricalcium phosphate

    NASA Astrophysics Data System (ADS)

    Durucan, Caner

    Low-temperature cement-type formation of hydroxyapatite [Ca10(PO4)6(OH)2 or HAp) has value in terms of developing synthetic compounds similar in compositions to those formed by natural mineralization of bone. Understanding the in vitro kinetics of formation of the synthetic composition could produce insights into developing hard tissue analogs. The kinetics and chemistry of cement-type formation of HAp by hydrolysis of particulate alpha-tricalcium phosphate (alpha-Ca 3(PO4)2 or alpha-TCP) were examined. In particular, the effects of reaction temperature, synthesis route, inorganic salt additives and presence of biodegradable polymers (poly(alpha-hydroxyl acids) on the hydrolysis rate and microstructural/mechanical properties of HAp were determined using the following analytical techniques: isothermal calorimetry, x-ray diffraction, scanning electron microscsopy (SEM), fourier transform infrared spectroscopy (FTIR), solution chemistry, diametrical compression and 3-point bending tests. For the phase-pure alpha-TCP/water system the complete reaction times and morphologies of the resultant HAp were found to be strongly dependent on reaction temperature over a range of 37°C to 56°C. Isothermal calorimetry analyses revealed a thermally activated hydrolysis mechanism, leading to higher reaction rates with an increase in hydrolysis temperature. The microstructure of the resultant HAp typically had entangled, flake-like morphology, with HAp formed at 37°C having a smaller crystalline size than that formed at 45°C and 56°C. The cement hardening contributed to entanglement at the microstructural level. In all cases the hydrated product was phase pure calcium-deficient hydroxyapatite [Ca10-x(HPO4) x(PO4)6-x(OH)2-x], and no other intermediates or by-products were formed through the complete transformation. According to the proposed kinetic model, a two-step mechanism was found to control the overall hydrolysis reaction and thereby HAp formation at 37°C. During the first

  15. Serotonin stimulates phospholipase A sub 2 and the release of arachidonic acid in hippocampal neurons by a type 2 serotonin receptor that is independent of inositolphospholipid hydrolysis

    SciTech Connect

    Felder, C.C.; Ma, A.L.; Axelrod, J.; Kanterman, R.Y. )

    1990-03-01

    Serotonin (5-HT) stimulated the release of arachidonic acid in hippocampal neurons cocultured with glial cells but not in glial cultures alone. Similar results were observed for the 5-HT-stimulated release of inositol phosphates. These results suggest a neural but not glial origin of both responses. Pharmacological studies suggested that release of arachidonic acid and inositol phosphates was mediated by a type 2 5-HTT (5-HT{sub 2}) receptor. 5-HT-stimulated release of arachidonic acid was also detected in cortical neurons, which contain high levels of 5-HT{sub 2} receptors, but not striatum, spinal cord, or cerebellar granule cells, which have very low levels or are devoid of 5-HT{sub 2} receptors. The phorbol ester phorbol 12-myristate 13-acetate augmented the 5-HT-stimulated release of arachidonic acid but inhibited the 5-HT-stimulated release of inositol phosphates. 5-HT-stimulated release of arachidonic acid, but not inositol phosphates, was dependent on extracellular calcium. 5-HT stimulated the release of ({sup 3}H)lysophosphatidylcholine from ({sup 3}H)choline-labeled cells with no increase in the release of ({sup 3}H)choline or phospho({sup 3}H)choline. These data suggest that 5-HT stimulated the release of arachidonic acid in hippocampal neurons through the activation of phospholipase A{sub 2}, independent of the activation of phospholipase C.

  16. Serotonin stimulates phospholipase A2 and the release of arachidonic acid in hippocampal neurons by a type 2 serotonin receptor that is independent of inositolphospholipid hydrolysis.

    PubMed Central

    Felder, C C; Kanterman, R Y; Ma, A L; Axelrod, J

    1990-01-01

    Serotonin (5-HT) stimulated the release of arachidonic acid in hippocampal neurons cocultured with glial cells but not in glial cultures alone. Similar results were observed for the 5-HT-stimulated release of inositol phosphates. These results suggest a neural but not glial origin of both responses. Pharmacological studies suggested that release of arachidonic acid and inositol phosphates was mediated by a type 2 5-HT (5-HT2) receptor. 5-HT-stimulated release of arachidonic acid was also detected in cortical neurons, which contain high levels of 5-HT2 receptors, but not striatum, spinal cord, or cerebellar granule cells, which have very low levels or are devoid of 5-HT2 receptors. The phorbol ester phorbol 12-myristate 13-acetate augmented the 5-HT-stimulated release of arachidonic acid but inhibited the 5-HT-stimulated release of inositol phosphates. 5-HT-stimulated release of arachidonic acid, but not inositol phosphates, was dependent on extracellular calcium. 5-HT stimulated the release of [3H]lysophosphatidylcholine from [3H]choline-labeled cells with no increase in the release of [3H]choline or phospho[3H]choline. These data suggest that 5-HT stimulated the release of arachidonic acid in hippocampal neurons through the activation of phospholipase A2, independent of the activation of phospholipase C. PMID:2315313

  17. Polyploid genome of Camelina sativa revealed by isolation of fatty acid synthesis genes

    PubMed Central

    2010-01-01

    Background Camelina sativa, an oilseed crop in the Brassicaceae family, has inspired renewed interest due to its potential for biofuels applications. Little is understood of the nature of the C. sativa genome, however. A study was undertaken to characterize two genes in the fatty acid biosynthesis pathway, fatty acid desaturase (FAD) 2 and fatty acid elongase (FAE) 1, which revealed unexpected complexity in the C. sativa genome. Results In C. sativa, Southern analysis indicates the presence of three copies of both FAD2 and FAE1 as well as LFY, a known single copy gene in other species. All three copies of both CsFAD2 and CsFAE1 are expressed in developing seeds, and sequence alignments show that previously described conserved sites are present, suggesting that all three copies of both genes could be functional. The regions downstream of CsFAD2 and upstream of CsFAE1 demonstrate co-linearity with the Arabidopsis genome. In addition, three expressed haplotypes were observed for six predicted single-copy genes in 454 sequencing analysis and results from flow cytometry indicate that the DNA content of C. sativa is approximately three-fold that of diploid Camelina relatives. Phylogenetic analyses further support a history of duplication and indicate that C. sativa and C. microcarpa might share a parental genome. Conclusions There is compelling evidence for triplication of the C. sativa genome, including a larger chromosome number and three-fold larger measured genome size than other Camelina relatives, three isolated copies of FAD2, FAE1, and the KCS17-FAE1 intergenic region, and three expressed haplotypes observed for six predicted single-copy genes. Based on these results, we propose that C. sativa be considered an allohexaploid. The characterization of fatty acid synthesis pathway genes will allow for the future manipulation of oil composition of this emerging biofuel crop; however, targeted manipulations of oil composition and general development of C. sativa should

  18. Acidic Properties and Structure-Activity Correlations of Solid Acid Catalysts Revealed by Solid-State NMR Spectroscopy.

    PubMed

    Zheng, Anmin; Li, Shenhui; Liu, Shang-Bin; Deng, Feng

    2016-04-19

    Solid acid materials with tunable structural and acidic properties are promising heterogeneous catalysts for manipulating and/or emulating the activity and selectivity of industrially important catalytic reactions. On the other hand, the performances of acid-catalyzed reactions are mostly dictated by the acidic features, namely, type (Brønsted vs Lewis acidity), amount, strength, and local environment of acid sites. The latter is relevant to their location (intra- vs extracrystalline), and possible confinement and Brønsted-Lewis acid synergy effects that may strongly affect the host-guest interactions, reaction mechanism, and shape selectivity of the catalytic system. This account aims to highlight some important applications of state-of-the-art solid-state NMR (SSNMR) techniques for exploring the structural and acidic properties of solid acid catalysts as well as their catalytic performances and relevant reaction pathway invoked. In addition, density functional theory (DFT) calculations may be exploited in conjunction with experimental SSNMR studies to verify the structure-activity correlations of the catalytic system at a microscopic scale. We describe in this Account the developments and applications of advanced ex situ and/or in situ SSNMR techniques, such as two-dimensional (2D) double-quantum magic-angle spinning (DQ MAS) homonuclear correlation spectroscopy for structural investigation of solid acids as well as study of their acidic properties. Moreover, the energies and electronic structures of the catalysts and detailed catalytic reaction processes, including the identification of reaction species, elucidation of reaction mechanism, and verification of structure-activity correlations, made available by DFT theoretical calculations were also discussed. Relevant discussions will focus primarily on results obtained from our laboratories in the past decade, including (i) quantitative and qualitative acidity characterization utilizing assorted probe molecules

  19. Effect of pretreatment and enzymatic hydrolysis on the physical-chemical composition and morphologic structure of sugarcane bagasse and sugarcane straw.

    PubMed

    Moretti, Marcia Maria de Souza; Perrone, Olavo Micali; Nunes, Christiane da Costa Carreira; Taboga, Sebastião; Boscolo, Maurício; da Silva, Roberto; Gomes, Eleni

    2016-11-01

    The present work aimed to study the effect of the pretreatment of sugarcane bagasse and straw with microwave irradiation in aqueous and acid glycerol solutions on their chemical composition, fiber structure and the efficiency of subsequent enzymatic hydrolysis. Thermogravimetric analysis showed that the pretreatment acted mainly on the lignin and hemicellulose fractions of the bagasse, whereas, in the straw, lesser structural and chemical changes were observed. The images from transmission electron microscopy (TEM) revealed that treating bagasse and straw with acid glycerol solution loosened the cell walls and there was a breakdown in the pit membrane. The treated material was submitted to hydrolysis for 72h and higher yields of reducing sugars were observed compared to the untreated material (250.9mg/g from straw and 197.4mg/g from bagasse). TEM images after hydrolysis confirmed the possible points of access of the enzymes to the secondary cell wall region of the pretreated biomass. PMID:27578061

  20. Continuous steam hydrolysis of tulip poplar

    SciTech Connect

    Fieber, C.; Colcord, A.R.; Faass, S.; Muzzy, J.D.; Roberts, R.S.

    1982-08-01

    To produce ethanol from hardwood it is desirable to fractionate the hardwood in order to produce a relatively pure cellulosic pulp for dilute acid hydrolysis. An experimental investigation of continuous steam hydrolysis of tulip poplar wood chips indicates that over 90% of the lignin present can be extracted by 0.1N sodium hydroxide, resulting in a cellulose pulp containing over 90% hexosan. The study was performed using a Stake Technology, Ltd., continuous digester rated at one oven dry ton per hour of wood chips. The yields of hexosans, hexoses, xylan, xylose, lignin, furfural, acetic acid and methanol were determined as a function of residence time and steam pressure in the digester. The information provides a basis for establishing a material and energy balance for a hardwood to ethanol plant.

  1. Non-catalytic steam hydrolysis of fats

    SciTech Connect

    Deibert, M.C.

    1992-08-28

    Hydrolysis of fats and oils produces fatty acid and glycerol. The catalyzed, liquid phase Colgate-Emry process, state-of-the-art, produces impure products that require extensive energy investment for their purification to commercial grade. Non-catalytic steam hydrolysis may produce products more easily purified. A bench-scale hydrolyzer was designed and constructed to contact descending liquid fat or oil with rising superheated steam. Each of the five stages in the reactor was designed similar to a distillation column stage to promote intimate liquid-gas contact. Degree of hydrolysis achieved in continuous tests using tallow feed were 15% at 280C and 35% at 300C at a tallow-to-steam mass feed ratio of 4.2. At a feed ratio of 9.2, the degree of hydrolysis was 21% at 300C. Decomposition was strongly evident at 325C but not at lower temperatures. Soybean oil rapidly polymerized under reaction conditions. Batch tests at 320C produced degrees of hydrolyses of between 44% and 63% using tallow and palm oil feeds. Over 95% fatty acids were present in a clean, readily separated organic portion of the overhead product from most tests. The test reactor had serious hydraulic resistance to liquid down-flow which limited operation to very long liquid residence times. These times are in excess of those that tallow and palm oil are stable at the reaction temperature. Little glycerol and extensive light organics were produced indicating that unexplained competing reactions to hydrolysis occurred in the experimental system. Further tests using an improved reactor will be required.

  2. Comparative Transcriptomics Reveals Jasmonic Acid-Associated Metabolism Related to Cotton Fiber Initiation.

    PubMed

    Wang, Liman; Zhu, Youmin; Hu, Wenjing; Zhang, Xueying; Cai, Caiping; Guo, Wangzhen

    2015-01-01

    Analysis of mutants and gene expression patterns provides a powerful approach for investigating genes involved in key stages of plant fiber development. In this study, lintless-fuzzless XinWX and linted-fuzzless XinFLM with a single genetic locus difference for lint were used to identify differentially expressed genes. Scanning electron microscopy showed fiber initiation in XinFLM at 0 days post anthesis (DPA). Fiber transcriptional profiling of the lines at three initiation developmental stages (-1, 0, 1 DPA) was performed using an oligonucleotide microarray. Loop comparisons of the differentially expressed genes within and between the lines was carried out, and functional classification and enrichment analysis showed that gene expression patterns during fiber initiation were heavily associated with hormone metabolism, transcription factor regulation, lipid transport, and asparagine biosynthetic processes, as previously reported. Further, four members of the allene-oxide cyclase (AOC) family that function in jasmonate biosynthesis were parallel up-regulation in fiber initiation, especially at -1 DPA, compared to other tissues and organs in linted-fuzzed TM-1. Real time-quantitative PCR (RT-qPCR) analysis in different fiber mutant lines revealed that AOCs were up-regulated higher at -1 DPA in lintless-fuzzless than that in linted-fuzzless and linted-fuzzed materials, and transcription of the AOCs was increased under jasmonic acid (JA) treatment. Expression analysis of JA biosynthesis-associated genes between XinWX and XinFLM showed that they were up-regulated during fiber initiation in the fuzzless-lintless mutant. Taken together, jasmonic acid-associated metabolism was related to cotton fiber initiation. Parallel up-regulation of AOCs expression may be important for normal fiber initiation development, while overproduction of AOCs might disrupt normal fiber development. PMID:26079621

  3. Amino Acid Proximities in Two Sup35 Prion Strains Revealed by Chemical Cross-linking.

    PubMed

    Wong, Shenq-Huey; King, Chih-Yen

    2015-10-01

    Strains of the yeast prion [PSI] are different folding patterns of the same Sup35 protein, which stacks up periodically to form a prion fiber. Chemical cross-linking is employed here to probe different fiber structures assembled with a mutant Sup35 fragment. The photo-reactive cross-linker, p-benzoyl-l-phenylalanine (pBpa), was biosynthetically incorporated into bacterially prepared recombinant Sup(1-61)-GFP, containing the first 61 residues of Sup35, followed by the green fluorescent protein. Four methionine substitutions and two alanine substitutions were introduced at fixed positions in Sup(1-61) to allow cyanogen bromide cleavage to facilitate subsequent mass spectrometry analysis. Amyloid fibers of pBpa and Met/Ala-substituted Sup(1-61)-GFP were nucleated from purified yeast prion particles of two different strains, namely VK and VL, and shown to faithfully transmit specific strain characteristics to yeast expressing the wild type Sup35 protein. Intra- and intermolecular cross-linking were distinguished by tandem mass spectrometry analysis on fibers seeded from solutions containing equal amounts of (14)N- and (15)N-labeled protein. Fibers propagating the VL strain type exhibited intra- and intermolecular cross-linking between amino acid residues 3 and 28, as well as intra- and intermolecular linking between 32 and 55. Inter- and intramolecular cross-linking between residues 32 and 55 were detected in fibers propagating the VK strain type. Adjacencies of amino acid residues in space revealed by cross-linking were used to constrain possible chain folds of different [PSI] strains. PMID:26265470

  4. Comparative Transcriptomics Reveals Jasmonic Acid-Associated Metabolism Related to Cotton Fiber Initiation

    PubMed Central

    Wang, Liman; Zhu, Youmin; Hu, Wenjing; Zhang, Xueying; Cai, Caiping; Guo, Wangzhen

    2015-01-01

    Analysis of mutants and gene expression patterns provides a powerful approach for investigating genes involved in key stages of plant fiber development. In this study, lintless-fuzzless XinWX and linted-fuzzless XinFLM with a single genetic locus difference for lint were used to identify differentially expressed genes. Scanning electron microscopy showed fiber initiation in XinFLM at 0 days post anthesis (DPA). Fiber transcriptional profiling of the lines at three initiation developmental stages (-1, 0, 1 DPA) was performed using an oligonucleotide microarray. Loop comparisons of the differentially expressed genes within and between the lines was carried out, and functional classification and enrichment analysis showed that gene expression patterns during fiber initiation were heavily associated with hormone metabolism, transcription factor regulation, lipid transport, and asparagine biosynthetic processes, as previously reported. Further, four members of the allene-oxide cyclase (AOC) family that function in jasmonate biosynthesis were parallel up-regulation in fiber initiation, especially at -1 DPA, compared to other tissues and organs in linted-fuzzed TM-1. Real time-quantitative PCR (RT-qPCR) analysis in different fiber mutant lines revealed that AOCs were up-regulated higher at -1 DPA in lintless-fuzzless than that in linted-fuzzless and linted-fuzzed materials, and transcription of the AOCs was increased under jasmonic acid (JA) treatment. Expression analysis of JA biosynthesis-associated genes between XinWX and XinFLM showed that they were up-regulated during fiber initiation in the fuzzless-lintless mutant. Taken together, jasmonic acid-associated metabolism was related to cotton fiber initiation. Parallel up-regulation of AOCs expression may be important for normal fiber initiation development, while overproduction of AOCs might disrupt normal fiber development. PMID:26079621

  5. One Step Continuous Flow Synthesis of Highly Substituted Pyrrole-3-Carboxylic Acid Derivatives via in situ Hydrolysis of tert-Butyl Esters

    PubMed Central

    Herath, Ananda; Cosford, Nicholas D. P.

    2010-01-01

    The first one-step, continuous flow synthesis of pyrrole-3-carboxylic acids directly from tert-butyl acetoacetates, amines and 2-bromoketones is reported. The HBr generated as a by-product in the Hantzsch reaction was utilized in the flow method to saponify the t-butyl esters in situ to provide the corresponding acids in a single microreactor. The protocol was used in the multistep synthesis of pyrrole-3-carboxamides, including two CB1 inverse agonists, directly from commercially available starting materials in a single continuous process. PMID:20964284

  6. PLA recycling by hydrolysis at high temperature

    NASA Astrophysics Data System (ADS)

    Cristina, Annesini Maria; Rosaria, Augelletti; Sara, Frattari; Fausto, Gironi

    2016-05-01

    In this work the process of PLA hydrolysis at high temperature was studied, in order to evaluate the possibility of chemical recycling of this polymer bio-based. In particular, the possibility to obtain the monomer of lactic acid from PLA degradation was investigated. The results of some preliminary tests, performed in a laboratory batch reactor at high temperature, are presented: the experimental results show that the complete degradation of PLA can be obtained in relatively low reaction times.

  7. Hydrolysis of phytic acid by intrinsic plant or supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas.

    PubMed

    Rapp, C; Lantzsch, H J; Drochner, W

    2001-12-01

    Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant-cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet (43 U Phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. Duodenal recovery of dry matter and total phosphorus were 100% in the period between two feedings, irrespective of dietary treatment. In animals fed the wheat-based diet, dry matter left the stomach faster (p < 0.05) during the first hour after feeding than in animals fed the corn-based diets (41.3 vs. 31.0 and 25.8% of intake, respectively). Supplemented microbial phytase did not affect ileal dry matter digestibility of the corn-based diet. In the first hour after feeding, phosphorus concentration of the duodenal digesta of animals fed corn-based diets with or without supplemented microbial phytase (5.86, 6.19 mg total P/g DM) exceeded the dietary level considerably (4.30 and 4.21 mg total P/g DM) indicating a higher solubility of corn than wheat phosphorus in the stomach. Apparent ileal P absorption was higher (p < 0.05) in the wheat-based diet (37.6%) and corn-based diet supplemented with microbial phytase (34.3%) than in the unsupplemented corn-based diet (17.6%). PMID:11906564

  8. Assay of phenolic compounds from four species of Ber (Ziziphus mauritiana L.) Fruits: Comparision of three base hydrolysis procedure for quantification of total phenolic acids

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The present study was undertaken to investigate the flavonoids profile in four species of ber (Ziziphus mauritiana Lamk) fruit and to compare various techniques for the analysis of total phenolic acids. The 12 flavonoids identified were quercetin 3-O-robinobioside, quercetin 3-O-rutinoside, querceti...

  9. Thioglycoside hydrolysis catalyzed by {beta}-glucosidase

    SciTech Connect

    Shen Hong; Byers, Larry D.

    2007-10-26

    Sweet almond {beta}-glucosidase (EC 3.2.1.21) has been shown to have significant thioglycohydrolase activity. While the K{sub m} values for the S- and O-glycosides are similar, the k{sub cat} values are about 1000-times lower for the S-glycosides. Remarkably, the pH-profile for k{sub cat}/K{sub m} for hydrolysis of p-nitrophenyl thioglucoside (pNPSG) shows the identical dependence on a deprotonated carboxylate (pK{sub a} 4.5) and a protonated group (pK{sub a} 6.7) as does the pH-profile for hydrolysis of the corresponding O-glycoside. Not surprisingly, in spite of the requirement for the presence of this protonated group in catalytically active {beta}-glucosidase, thioglucoside hydrolysis does not involve general acid catalysis. There is no solvent kinetic isotope effect on the enzyme-catalyzed hydrolysis of pNPSG.

  10. Phenolic Profiling of Caffeic Acid O-Methyltransferase-Deficient Poplar Reveals Novel Benzodioxane Oligolignols1

    PubMed Central

    Morreel, Kris; Ralph, John; Lu, Fachuang; Goeminne, Geert; Busson, Roger; Herdewijn, Piet; Goeman, Jan L.; Van der Eycken, Johan; Boerjan, Wout; Messens, Eric

    2004-01-01

    Caffeic acid O-methyltransferase (COMT) catalyzes preferentially the methylation of 5-hydroxyconiferaldehyde to sinapaldehyde in monolignol biosynthesis. Here, we have compared HPLC profiles of the methanol-soluble phenolics fraction of xylem tissue from COMT-deficient and control poplars (Populus spp.), using statistical analysis of the peak heights. COMT down-regulation results in significant concentration differences for 25 of the 91 analyzed peaks. Eight peaks were exclusively detected in COMT-deficient poplar, of which four could be purified for further identification using mass spectrometry/mass spectrometry, nuclear magnetic resonance, and spiking of synthesized reference compounds. These new compounds were derived from 5-hydroxyconiferyl alcohol or 5-hydroxyconiferaldehyde and were characterized by benzodioxane moieties, a structural type that is also increased in the lignins of COMT-deficient plants. One of these four benzodioxanes amounted to the most abundant oligolignol in the HPLC profile. Furthermore, all of the differentially accumulating oligolignols involving sinapyl units were either reduced in abundance or undetectable. The concentration levels of all identified oligolignols were in agreement with the relative supply of monolignols and with their chemical coupling propensities, which supports the random coupling hypothesis. Chiral HPLC analysis of the most abundant benzodioxane dimer revealed the presence of both enantiomers in equal amounts, indicating that they were formed by radical coupling reactions under simple chemical control rather than guided by dirigent proteins. PMID:15563622

  11. Neuroinformatics analyses reveal GABAt and SSADH as major proteins involved in anticonvulsant activity of valproic acid.

    PubMed

    Piplani, Sakshi; Verma, Prabhakar Kumar; Kumar, Ajit

    2016-07-01

    The unequivocal hypotheses about anticonvulsant activity of valproic acid (VPA) have always been a basic hurdle in designing next generation neurotherapeutics, particularly the anti-epileptic drugs. The present study reports about a comprehensive in-silico investigation into qualitative and quantitative binding of VPA and corresponding natural ligands of four major enzymes involved in neurotransmissions, namely-GABA transaminase (GABAt), α-keto glutarate dehydrogenase (α-KGDH), Succinate Semialdehyde dehydrogenase (SSADH) and Glutamate Decarboxylase (GAD), respectively. The molecular docking analyses revealed that VPA inhibits GABAt and α-KGDH through allosteric while SSADH through competitive mode of binding. There is an observed elevation in binding of glutamate over GAD in the presence of VPA. The docking inhibition constant (Ki) of VPA to all the studied enzymatic receptors were observed to be well below the therapeutic concentration of VPA in blood, except for α-KGDH, thus favouring GABAergic over glutamatergic mode of anticonvulsant activity of VPA. The report is probably the first comprehensive in-silico molecular study about VPA action. PMID:27261619

  12. Evolution and Functional Implications of the Tricarboxylic Acid Cycle as Revealed by Phylogenetic Analysis

    PubMed Central

    Cavalcanti, João Henrique Frota; Esteves-Ferreira, Alberto A.; Quinhones, Carla G.S.; Pereira-Lima, Italo A.; Nunes-Nesi, Adriano; Fernie, Alisdair R.; Araújo, Wagner L.

    2014-01-01

    The tricarboxylic acid (TCA) cycle, a crucial component of respiratory metabolism, is composed of a set of eight enzymes present in the mitochondrial matrix. However, most of the TCA cycle enzymes are encoded in the nucleus in higher eukaryotes. In addition, evidence has accumulated demonstrating that nuclear genes were acquired from the mitochondrial genome during the course of evolution. For this reason, we here analyzed the evolutionary history of all TCA cycle enzymes in attempt to better understand the origin of these nuclear-encoded proteins. Our results indicate that prior to endosymbiotic events the TCA cycle seemed to operate only as isolated steps in both the host (eubacterial cell) and mitochondria (alphaproteobacteria). The origin of isoforms present in different cell compartments might be associated either with gene-transfer events which did not result in proper targeting of the protein to mitochondrion or with duplication events. Further in silico analyses allow us to suggest new insights into the possible roles of TCA cycle enzymes in different tissues. Finally, we performed coexpression analysis using mitochondrial TCA cycle genes revealing close connections among these genes most likely related to the higher efficiency of oxidative phosphorylation in this specialized organelle. Moreover, these analyses allowed us to identify further candidate genes which might be used for metabolic engineering purposes given the importance of the TCA cycle during development and/or stress situations. PMID:25274566

  13. Enzymatic hydrolysis of steryl glycosides for their analysis in foods.

    PubMed

    Münger, Linda H; Nyström, Laura

    2014-11-15

    Steryl glycosides (SG) contribute significantly to the total intake of phytosterols. The standard analytical procedure involving acid hydrolysis fails to reflect the correct sterol profile of SG due to isomerization of some of the labile sterols. Therefore, various glycosylases were evaluated for their ability to hydrolyse SG under milder conditions. Using a pure SG mixture in aqueous solution, the highest glycolytic activity, as demonstrated by the decrease in SG and increase in free sterols was achieved using inulinase preparations (decrease of >95%). High glycolytic activity was also demonstrated using hemicellulase (63%). The applicability of enzymatic hydrolysis using inulinase preparations was further verified on SG extracted from foods. For example in potato peel Δ(5)-avenasteryl glucoside, a labile SG, was well preserved and contributed 26.9% of the total SG. Therefore, enzymatic hydrolysis is suitable for replacing acid hydrolysis of SG in food lipid extracts to accurately determine the sterol profile of SG. PMID:24912717

  14. Process Design Report for Wood Feedstock: Lignocellulosic Biomass to Ethanol Process Desing and Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic Hydrolysis Current and Futuristic Scenarios

    SciTech Connect

    Wooley, Robert; Ruth, Mark; Sheehan, John; Ibsen, Kelly; Majdeski, Henry; Galves, Adrian

    1999-07-01

    The National Renewable Energy Laboratory (NREL) has undertaken a complete review and update of the process design and economic model for the biomass-to-ethanol process based on co-current dilute acid prehydrolysis, along with simultaneous saccharification (enzymatic) and co-fermentation. The process design includes the core technologies being researched by the U.S. Department of Energy (DOE): prehydrolysis, simultaneous saccharification and co-fermentation, and cellulase enzyme production.

  15. A Motif Unique to the Human Dead-Box Protein DDX3 Is Important for Nucleic Acid Binding, ATP Hydrolysis, RNA/DNA Unwinding and HIV-1 Replication

    PubMed Central

    Di Cicco, Giulia; Dietrich, Ursula; Maga, Giovanni

    2011-01-01

    DEAD-box proteins are enzymes endowed with nucleic acid-dependent ATPase, RNA translocase and unwinding activities. The human DEAD-box protein DDX3 has been shown to play important roles in tumor proliferation and viral infections. In particular, DDX3 has been identified as an essential cofactor for HIV-1 replication. Here we characterized a set of DDX3 mutants biochemically with respect to nucleic acid binding, ATPase and helicase activity. In particular, we addressed the functional role of a unique insertion between motifs I and Ia of DDX3 and provide evidence for its implication in nucleic acid binding and HIV-1 replication. We show that human DDX3 lacking this domain binds HIV-1 RNA with lower affinity. Furthermore, a specific peptide ligand for this insertion selected by phage display interferes with HIV-1 replication after transduction into HelaP4 cells. Besides broadening our understanding of the structure-function relationships of this important protein, our results identify a specific domain of DDX3 which may be suited as target for antiviral drugs designed to inhibit cellular cofactors for HIV-1 replication. PMID:21589879

  16. Kinetics of the alkaline hydrolysis of nitrocellulose.

    PubMed

    Christodoulatos, C; Su, T L; Koutsospyros, A

    2001-01-01

    Cellulose nitrate (nitrocellulose) is an explosive solid substance used in large quantities in various formulations of rocket and gun propellants. Safe destruction of nitrocellulose can be achieved by alkaline hydrolysis, which converts it to biodegradable products that can then be treated by conventional biological processes. The kinetics of the alkaline hydrolysis of munitions-grade nitrocellulose in sodium hydroxide solutions were investigated in completely mixed batch reactors. Experiments were conducted using solutions of alkaline strength ranging from 0.1 to 15% by mass and temperatures in the range of 30 to 90 degrees C. Regression analysis of the kinetic data revealed that alkaline hydrolysis of nitrocellulose is of the order 1.0 and 1.5 with respect to nitrocellulose and hydroxide concentration, respectively. The activation energy of the hydrolysis reaction was found to be 100.9 kJ/mol with a preexponential Arrhenius constant of 4.73 x 10(13). Nitrite and nitrate, in a 3:1 ratio, were the primary nitrogen species present in the posthydrolysis solution. The kinetic information is pertinent to the development and optimization of nitrocellulose chemical-biological treatment systems. PMID:11563378

  17. 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.

  18. Use of an algal hydrolysate to improve enzymatic hydrolysis of anaerobically digested fiber

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study investigated the use of acid hydrolyzed algae to enhance the enzymatic hydrolysis of cellulosic biomass. We first characterized wastewater-grown algal samples and determined the optimal conditions (acid concentration, reaction temperature, and reaction time) for algal hydrolysis using di...

  19. A Series of α-Amino Acid Ester Prodrugs of Camptothecin: In vitro Hydrolysis and A549 Human Lung Carcinoma Cell Cytotoxicity

    PubMed Central

    Deshmukh, Manjeet; Chao, Piyun; Kutscher, Hilliard L.; Gao, Dayuan; Sinko, Patrick J.

    2013-01-01

    The objective of the present study was to identify a camptothecin (CPT) prodrug with optimal release and cytotoxicity properties for immobilization on a passively targeted microparticle delivery system. A series of α-amino acid ester prodrugs of CPT were synthesized, characterized and evaluated. Four CPT prodrugs were synthesized with increasing aliphatic chain length (glycine (Gly) (2a), alanine (Ala) (2b), aminobutyric acid (Abu) (2c) and norvaline (Nva) (2d)). Prodrug reconversion was studied at pH 6.6, 7.0 and 7.4 corresponding to tumor, lung and extracellular/physiological pH, respectively. Cytotoxicity was evaluated in A549 human lung carcinoma cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The hydrolytic reconversion rate to parent CPT increased with decreasing side chain length as well as increasing pH. The Hill slope of 2d was significantly less than CPT and the other prodrugs tested, indicating a higher cell death rate at lower concentrations. These results suggest that 2d is the best candidate for a passively targeted sustained release lung delivery system. PMID:20063889

  20. Ecological roles of dominant and rare prokaryotes in acid mine drainage revealed by metagenomics and metatranscriptomics.

    PubMed

    Hua, Zheng-Shuang; Han, Yu-Jiao; Chen, Lin-Xing; Liu, Jun; Hu, Min; Li, Sheng-Jin; Kuang, Jia-Liang; Chain, Patrick S G; Huang, Li-Nan; Shu, Wen-Sheng

    2015-06-01

    High-throughput sequencing is expanding our knowledge of microbial diversity in the environment. Still, understanding the metabolic potentials and ecological roles of rare and uncultured microbes in natural communities remains a major challenge. To this end, we applied a 'divide and conquer' strategy that partitioned a massive metagenomic data set (>100 Gbp) into subsets based on K-mer frequency in sequence assembly to a low-diversity acid mine drainage (AMD) microbial community and, by integrating with an additional metatranscriptomic assembly, successfully obtained 11 draft genomes most of which represent yet uncultured and/or rare taxa (relative abundance <1%). We report the first genome of a naturally occurring Ferrovum population (relative abundance >90%) and its metabolic potentials and gene expression profile, providing initial molecular insights into the ecological role of these lesser known, but potentially important, microorganisms in the AMD environment. Gene transcriptional analysis of the active taxa revealed major metabolic capabilities executed in situ, including carbon- and nitrogen-related metabolisms associated with syntrophic interactions, iron and sulfur oxidation, which are key in energy conservation and AMD generation, and the mechanisms of adaptation and response to the environmental stresses (heavy metals, low pH and oxidative stress). Remarkably, nitrogen fixation and sulfur oxidation were performed by the rare taxa, indicating their critical roles in the overall functioning and assembly of the AMD community. Our study demonstrates the potential of the 'divide and conquer' strategy in high-throughput sequencing data assembly for genome reconstruction and functional partitioning analysis of both dominant and rare species in natural microbial assemblages. PMID:25361395

  1. Ecological roles of dominant and rare prokaryotes in acid mine drainage revealed by metagenomics and metatranscriptomics

    DOE PAGESBeta

    Hua, Zheng-Shuang; Han, Yu-Jiao; Chen, Lin-Xing; Liu, Jun; Hu, Min; Li, Sheng-Jin; Kuang, Jia-Liang; Chain, Patrick SG; Huang, Li-Nan; Shu, Wen-Sheng

    2014-11-07

    Here we report that high-throughput sequencing is expanding our knowledge of microbial diversity in the environment. Still, understanding the metabolic potentials and ecological roles of rare and uncultured microbes in natural communities remains a major challenge. To this end, we applied a ‘divide and conquer’ strategy that partitioned a massive metagenomic data set (>100 Gbp) into subsets based on K-mer frequency in sequence assembly to a low-diversity acid mine drainage (AMD) microbial community and, by integrating with an additional metatranscriptomic assembly, successfully obtained 11 draft genomes most of which represent yet uncultured and/or rare taxa (relative abundance <1%). We reportmore » the first genome of a naturally occurring Ferrovum population (relative abundance >90%) and its metabolic potentials and gene expression profile, providing initial molecular insights into the ecological role of these lesser known, but potentially important, microorganisms in the AMD environment. Gene transcriptional analysis of the active taxa revealed major metabolic capabilities executed in situ, including carbon- and nitrogen-related metabolisms associated with syntrophic interactions, iron and sulfur oxidation, which are key in energy conservation and AMD generation, and the mechanisms of adaptation and response to the environmental stresses (heavy metals, low pH and oxidative stress). Remarkably, nitrogen fixation and sulfur oxidation were performed by the rare taxa, indicating their critical roles in the overall functioning and assembly of the AMD community. Finally, our study demonstrates the potential of the ‘divide and conquer’ strategy in high-throughput sequencing data assembly for genome reconstruction and functional partitioning analysis of both dominant and rare species in natural microbial assemblages.« less

  2. Ecological roles of dominant and rare prokaryotes in acid mine drainage revealed by metagenomics and metatranscriptomics

    PubMed Central

    Hua, Zheng-Shuang; Han, Yu-Jiao; Chen, Lin-Xing; Liu, Jun; Hu, Min; Li, Sheng-Jin; Kuang, Jia-Liang; Chain, Patrick SG; Huang, Li-Nan; Shu, Wen-Sheng

    2015-01-01

    High-throughput sequencing is expanding our knowledge of microbial diversity in the environment. Still, understanding the metabolic potentials and ecological roles of rare and uncultured microbes in natural communities remains a major challenge. To this end, we applied a ‘divide and conquer' strategy that partitioned a massive metagenomic data set (>100 Gbp) into subsets based on K-mer frequency in sequence assembly to a low-diversity acid mine drainage (AMD) microbial community and, by integrating with an additional metatranscriptomic assembly, successfully obtained 11 draft genomes most of which represent yet uncultured and/or rare taxa (relative abundance <1%). We report the first genome of a naturally occurring Ferrovum population (relative abundance >90%) and its metabolic potentials and gene expression profile, providing initial molecular insights into the ecological role of these lesser known, but potentially important, microorganisms in the AMD environment. Gene transcriptional analysis of the active taxa revealed major metabolic capabilities executed in situ, including carbon- and nitrogen-related metabolisms associated with syntrophic interactions, iron and sulfur oxidation, which are key in energy conservation and AMD generation, and the mechanisms of adaptation and response to the environmental stresses (heavy metals, low pH and oxidative stress). Remarkably, nitrogen fixation and sulfur oxidation were performed by the rare taxa, indicating their critical roles in the overall functioning and assembly of the AMD community. Our study demonstrates the potential of the ‘divide and conquer' strategy in high-throughput sequencing data assembly for genome reconstruction and functional partitioning analysis of both dominant and rare species in natural microbial assemblages. PMID:25361395

  3. Ecological roles of dominant and rare prokaryotes in acid mine drainage revealed by metagenomics and metatranscriptomics

    SciTech Connect

    Hua, Zheng-Shuang; Han, Yu-Jiao; Chen, Lin-Xing; Liu, Jun; Hu, Min; Li, Sheng-Jin; Kuang, Jia-Liang; Chain, Patrick SG; Huang, Li-Nan; Shu, Wen-Sheng

    2014-11-07

    Here we report that high-throughput sequencing is expanding our knowledge of microbial diversity in the environment. Still, understanding the metabolic potentials and ecological roles of rare and uncultured microbes in natural communities remains a major challenge. To this end, we applied a ‘divide and conquer’ strategy that partitioned a massive metagenomic data set (>100 Gbp) into subsets based on K-mer frequency in sequence assembly to a low-diversity acid mine drainage (AMD) microbial community and, by integrating with an additional metatranscriptomic assembly, successfully obtained 11 draft genomes most of which represent yet uncultured and/or rare taxa (relative abundance <1%). We report the first genome of a naturally occurring Ferrovum population (relative abundance >90%) and its metabolic potentials and gene expression profile, providing initial molecular insights into the ecological role of these lesser known, but potentially important, microorganisms in the AMD environment. Gene transcriptional analysis of the active taxa revealed major metabolic capabilities executed in situ, including carbon- and nitrogen-related metabolisms associated with syntrophic interactions, iron and sulfur oxidation, which are key in energy conservation and AMD generation, and the mechanisms of adaptation and response to the environmental stresses (heavy metals, low pH and oxidative stress). Remarkably, nitrogen fixation and sulfur oxidation were performed by the rare taxa, indicating their critical roles in the overall functioning and assembly of the AMD community. Finally, our study demonstrates the potential of the ‘divide and conquer’ strategy in high-throughput sequencing data assembly for genome reconstruction and functional partitioning analysis of both dominant and rare species in natural microbial assemblages.

  4. Effect of initial pH on mesophilic hydrolysis and acidification of swine manure.

    PubMed

    Lin, Lin; Wan, Chunli; Liu, Xiang; Lee, Duu-Jong; Lei, Zhongfang; Zhang, Yi; Tay, Joo Hwa

    2013-05-01

    Effects of initial pH (3-12) on mesophilic hydrolysis and acidification reactions of swine manure was studied. The initial pH changed the microbial community in the suspension so as to affect hydrolysis and acidification reactions on swine manure. At pH 10-12 the Clostridium alkalicellum and/or Corynebacterium humireducens were enriched and the soluble chemical oxygen demand (SCOD), total volatile fatty acids (VFAs), proteins and carbohydrates from manure were increased in quantities. In particular, at pH 10 the VFA concentration peaked at 13,600 mg-COD/L, with acetate and propionate accounting for 71.8% of the total VFAs. Acidic environment facilitates release of ammonium from manure. The Butyricimonas sp. was found existing at initial pH 5 which led to accumulated quantities of butyrate. Initial pH adjustment was revealed to be an effective way to manipulate rates and end products of hydrolysis and acidification of swine manure. PMID:23567695

  5. Rapid quantitative analysis of diosgenin in the tubers of Dioscorea zingiberensis C.H. Wright by coupling cellulose enzymolysis and two-phase acid hydrolysis in tandem with HPLC-UV.

    PubMed

    Yang, Huan; Chen, Bin; Wang, Xu-bo; Chue, Phen-wei; Shen, Yu-ping; Xia, Guo-hua; Jia, Xiao-bin

    2013-10-01

    A rapid method was developed to quantify diosgenin in Rhizoma Dioscoreae Zingiberensis. For the first time, sample solution was prepared by coupling pretreatment of raw material in cellulase and two-phase acid hydrolysis. After reconstitution, analysis was carried out on a C18 column, at 30°C, with acetonitrile and water (70:30, v/v) as mobile phase with flow rate of 1.0 mL min(- 1). Detection was carried out at 202 nm. Good linearity (r(2) = 0.9998) was established between concentration of analyte and peak area. The precision was >99% and the RSD of diosgenin contents for repeatability was 1.81%. The accuracy was supported with recoveries at 98.8%, 101.6% and 101.2%. The sample solution prepared using the proposed method contained higher content of diosgenin and was stable for 48 h. Due to the high efficiency of sample preparation and high reliability of the HPLC method, it is feasible to use this method for routine analysis of diosgenin in the herb. PMID:23672189

  6. The nature of amino acid 482 of human ABCG2 affects substrate transport and ATP hydrolysis but not substrate binding

    PubMed Central

    Ejendal, Karin F.K.; Diop, Ndeye Khady; Schweiger, Linda C.; Hrycyna, Christine A.

    2006-01-01

    Several members of the ATP-binding cassette (ABC) transporter superfamily, including P-glycoprotein and the half-transporter ABCG2, can confer multidrug resistance to cancer cells in culture by functioning as ATP-dependent efflux pumps. ABCG2 variants harboring a mutation at arginine 482 have been cloned from several drug-resistant cell lines, and these variants differ in their substrate transport phenotype. In this study, we changed the wild-type arginine 482 in human ABCG2 to each one of the 19 other standard amino acids and expressed each one transiently in HeLa cells. Using the 5D3 antibody that recognizes a cell surface epitope of ABCG2, we observed that all the mutants were expressed at the cell surface. However, the mutant ABCG2 proteins differed markedly in transport activity. All of the variants were capable of transporting one or more of the substrates used in this study, with the exception of the R482K mutant, which is completely devoid of transport ability. Six of the mutants (R482G, R482H, R482K, R482P, R482T, and R482Y) and the wild-type protein (R482wt) were selected for studies of basal and stimulated ATPase activity and photoaffinity labeling with the substrate analog [125I]iodoarylazidoprazosin. Whereas these seven ABCG2 variants differed markedly in ATPase activity, all were able to specifically bind the substrate analog [125I]iodoarylazidoprazosin. These data suggest that residue 482 plays an important role in substrate transport and ATP turnover, but that the nature of this amino acid may not be important for substrate recognition and binding. PMID:16815914

  7. Biphenylsulfonacetic Acid Inhibitors of the Human Papillomavirus Type 6 E1 Helicase Inhibit ATP Hydrolysis by an Allosteric Mechanism Involving Tyrosine 486

    PubMed Central

    White, Peter W.; Faucher, Anne-Marie; Massariol, Marie-Josée; Welchner, Ewald; Rancourt, Jean; Cartier, Mireille; Archambault, Jacques

    2005-01-01

    Human papillomaviruses (HPVs) are the causative agents of benign and malignant lesions of the epithelium. Despite their high prevalence, there is currently no antiviral drug for the treatment of HPV-induced lesions. The ATPase and helicase activities of the highly conserved E1 protein of HPV are essential for viral DNA replication and pathogenesis and hence are considered valid antiviral targets. We recently described novel biphenylsulfonacetic acid inhibitors of the ATPase activity of E1 from HPV type 6 (HPV6). Based on kinetics and mutagenesis studies, we now report that these compounds act by an allosteric mechanism. They are hyperbolic competitive inhibitors of the ATPase activity of HPV6 E1 and also inhibit its helicase activity. Compounds in this series can also inhibit the ATPase activity of the closely related enzyme from HPV11; however, the most potent inhibitors of HPV6 E1 are significantly less active against the type 11 protein. We identified a single critical residue in HPV6 E1, Tyr-486, substituted by a cysteine in HPV11, which is primarily responsible for this difference in inhibitor potency. Interestingly, HPV18 E1, which also has a tyrosine at this position, could be inhibited by biphenylsulfonacetic acid derivatives, thereby raising the possibility that this class of inhibitors could be optimized as antiviral agents against multiple HPV types. These studies implicate Tyr-486 as a key residue for inhibitor binding and define an allosteric pocket on HPV E1 that can be exploited for future drug discovery efforts. PMID:16304143

  8. Lipidomic profiling of Saccharomyces cerevisiae and Zygosaccharomyces bailii reveals critical changes in lipid composition in response to acetic acid stress.

    PubMed

    Lindberg, Lina; Santos, Aline Xs; Riezman, Howard; Olsson, Lisbeth; Bettiga, Maurizio

    2013-01-01

    When using microorganisms as cell factories in the production of bio-based fuels or chemicals from lignocellulosic hydrolysate, inhibitory concentrations of acetic acid, released from the biomass, reduce the production rate. The undissociated form of acetic acid enters the cell by passive diffusion across the lipid bilayer, mediating toxic effects inside the cell. In order to elucidate a possible link between lipid composition and acetic acid stress, the present study presents detailed lipidomic profiling of the major lipid species found in the plasma membrane, including glycerophospholipids, sphingolipids and sterols, in Saccharomyces cerevisiae (CEN.PK 113_7D) and Zygosaccharomyces bailii (CBS7555) cultured with acetic acid. Detailed physiological characterization of the response of the two yeasts to acetic acid has also been performed in aerobic batch cultivations using bioreactors. Physiological characterization revealed, as expected, that Z. bailii is more tolerant to acetic acid than S. cerevisiae. Z. bailii grew at acetic acid concentrations above 24 g L(-1), while limited growth of S. cerevisiae was observed after 11 h when cultured with only 12 g L(-1) acetic acid. Detailed lipidomic profiling using electrospray ionization, multiple-reaction-monitoring mass spectrometry (ESI-MRM-MS) showed remarkable changes in the glycerophospholipid composition of Z. bailii, including an increase in saturated glycerophospholipids and considerable increases in complex sphingolipids in both S. cerevisiae (IPC 6.2×, MIPC 9.1×, M(IP)2C 2.2×) and Z. bailii (IPC 4.9×, MIPC 2.7×, M(IP)2C 2.7×), when cultured with acetic acid. In addition, the basal level of complex sphingolipids was significantly higher in Z. bailii than in S. cerevisiae, further emphasizing the proposed link between lipid saturation, high sphingolipid levels and acetic acid tolerance. The results also suggest that acetic acid tolerance is associated with the ability of a given strain to generate large

  9. Competing addition and hydrolysis of the cytidylylcytidylyladenosine terminal residues of transfer ribonucleic acid isolated from the non-lactating bovine mammary gland

    PubMed Central

    Herrington, M. D.; Hawtrey, A. O.

    1970-01-01

    1. The enzyme fraction obtained from the pH5 enzyme of non-lactating bovine mammary gland between 40 and 100% ammonium sulphate saturation markedly inhibited the AMP-incorporating activity of rat liver nucleotide-incorporating enzyme. This inhibitory effect has been attributed to high nuclease activity which can be partially removed by adsorption of the enzyme fraction on to calcium phosphate gel. 2. The degradation action of the calcium phosphate-purified enzyme is confined mainly to the terminal trinucleotide sequence -pCpCpA of tRNA, its effect being analogous to that of venom phosphodiesterase. This enzyme is heat labile and very readily loses its degradative activity. 3. Treatment of the enzyme fraction with Macaloid results in complete removal of the phosphodiesterase, leaving an enzyme capable of incorporating AMP into tRNA. 4. Transfer RNA extracted from non-lactating bovine mammary gland in the presence of polyvinyl sulphate and Macaloid is able to accept amino acids with an efficiency 30% of that shown by lactating bovine mammary-gland tRNA isolated under identical conditions. PMID:4321270

  10. Anandamide transport is independent of fatty-acid amide hydrolase activity and is blocked by the hydrolysis-resistant inhibitor AM1172

    PubMed Central

    Fegley, D.; Kathuria, S.; Mercier, R.; Li, C.; Goutopoulos, A.; Makriyannis, A.; Piomelli, D.

    2004-01-01

    The endogenous cannabinoid anandamide is removed from the synaptic space by a high-affinity transport system present in neurons and astrocytes, which is inhibited by N-(4-hydroxyphenyl)-arachidonamide (AM404). After internalization, anandamide is hydrolyzed by fatty-acid amide hydrolase (FAAH), an intracellular membrane-bound enzyme that also cleaves AM404. Based on kinetic evidence, it has recently been suggested that anandamide internalization may be mediated by passive diffusion driven by FAAH activity. To test this possibility, in the present study, we have investigated anandamide internalization in wild-type and FAAH-deficient (FAAH–/–) mice. Cortical neurons from either mouse strain internalized [3H]anandamide through a similar mechanism, i.e., via a rapid temperature-sensitive and saturable process, which was blocked by AM404. Moreover, systemic administration of AM404 to either wild-type or FAAH–/– mice enhanced the hypothermic effects of exogenous anandamide, a response that was prevented by the CB1 cannabinoid antagonist rimonabant (SR141716A). The results indicate that anandamide internalization in mouse brain neurons is independent of FAAH activity. In further support of this conclusion, the compound N-(5Z, 8Z, 11Z, 14Z eicosatetraenyl)-4-hydroxybenzamide (AM1172) blocked [3H]anandamide internalization in rodent cortical neurons and human astrocytoma cells without acting as a FAAH substrate or inhibitor. AM1172 may serve as a prototype for novel anandamide transport inhibitors with increased metabolic stability. PMID:15138300

  11. Process Design and Economics for Biochemical Conversion of Lignocellulosic Biomass to Ethanol: Dilute-Acid Pretreatment and Enzymatic Hydrolysis of Corn Stover

    SciTech Connect

    Humbird, D.; Davis, R.; Tao, L.; Kinchin, C.; Hsu, D.; Aden, A.; Schoen, P.; Lukas, J.; Olthof, B.; Worley, M.; Sexton, D.; Dudgeon, D.

    2011-03-01

    This report describes one potential biochemical ethanol conversion process, conceptually based upon core conversion and process integration research at NREL. The overarching process design converts corn stover to ethanol by dilute-acid pretreatment, enzymatic saccharification, and co-fermentation. Building on design reports published in 2002 and 1999, NREL, together with the subcontractor Harris Group Inc., performed a complete review of the process design and economic model for the biomass-to-ethanol process. This update reflects NREL's current vision of the biochemical ethanol process and includes the latest research in the conversion areas (pretreatment, conditioning, saccharification, and fermentation), optimizations in product recovery, and our latest understanding of the ethanol plant's back end (wastewater and utilities). The conceptual design presented here reports ethanol production economics as determined by 2012 conversion targets and 'nth-plant' project costs and financing. For the biorefinery described here, processing 2,205 dry ton/day at 76% theoretical ethanol yield (79 gal/dry ton), the ethanol selling price is $2.15/gal in 2007$.

  12. Molecular dynamic simulations reveal the structural determinants of fatty acid binding to oxy-myoglobin

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The mechanism(s) by which fatty acids are sequestered and transported in muscle have not been fully elucidated. A potential key player in this process is the protein myoglobin (Mb). Indeed, there is a catalogue of empirical evidence supporting direct interaction of globins with fatty acid metabolite...

  13. Flavour formation from hydrolysis of pork sarcoplasmic protein extract by a unique LAB culture isolated from Harbin dry sausage.

    PubMed

    Chen, Qian; Liu, Qian; Sun, Qinxiu; Kong, Baohua; Xiong, Youling

    2015-02-01

    The lactic acid bacteria Pediococcus pentosaceus, Lactobacillus brevis, Lactobacillus curvatus, and Lactobacillus fermentum isolated from Harbin dry sausage were assessed for their protein hydrolysis and flavour development in pork muscle sarcoplasmic protein extracts. Gel electrophoresis indicated that sarcoplasmic proteins were degraded by all of the strains, especially by P. pentosaceus and L. curvatus. Trichloroacetic acid-soluble peptides increased in all of the samples (P < 0.05), especially samples inoculated with P. pentosaceus. Samples inoculated with P. pentosaceus and L. curvatus had higher free amino acid contents than did the other two strains(P < 0.05), and glutamic acid and alanine appeared to be the predominant free amino acids. The volatile compound analysis indicated that the highest aldehydes, alcohols and acid contents were found in the sample with P. pentosaceus followed by L. curvatus. The results revealed that P. pentosaceus could be appropriate for use as a meat starter culture. PMID:25460113

  14. Revealing Nucleic Acid Mutations Using Förster Resonance Energy Transfer-Based Probes.

    PubMed

    Junager, Nina P L; Kongsted, Jacob; Astakhova, Kira

    2016-01-01

    Nucleic acid mutations are of tremendous importance in modern clinical work, biotechnology and in fundamental studies of nucleic acids. Therefore, rapid, cost-effective and reliable detection of mutations is an object of extensive research. Today, Förster resonance energy transfer (FRET) probes are among the most often used tools for the detection of nucleic acids and in particular, for the detection of mutations. However, multiple parameters must be taken into account in order to create efficient FRET probes that are sensitive to nucleic acid mutations. In this review; we focus on the design principles for such probes and available computational methods that allow for their rational design. Applications of advanced, rationally designed FRET probes range from new insights into cellular heterogeneity to gaining new knowledge of nucleic acid structures directly in living cells. PMID:27472344

  15. Nicosulfuron: alcoholysis, chemical hydrolysis, and degradation on various minerals.

    PubMed

    Sabadie, Jean

    2002-01-30

    Alcoholysis (methanol or ethanol) and hydrolysis (pH ranging from 4 to 11) of the herbicide nicosulfuron at 30 degrees C principally involves the breakdown of the urea part of the molecule. A high yield of the corresponding carbamate was obtained along with aminopyrimidine during alcoholysis. Hydrolysis led to both aminopyrimidine and pyridylsulfonamide. The latter compound may be easily cyclized (pH > or = 7). First-order kinetics describe the rates of alcoholysis and hydrolysis well. The rate constants (0.44 days(-1) for methanolysis) decreased from 0.50 to 0.002 days(-1) as pH increased from 4 to 8, then remained stable under alkaline conditions. In acidic or neutral solution, the hydrolysis path appeared prevalent (> or =70%), whereas in an alkaline medium it decreased when pH increased. The chemical degradation of nicosulfuron on various dry minerals (calcium bentonite, kaolinite, silica gel, H(+) bentonite, montmorillonite K10, and alumina) was investigated at 30 degrees C. The best conditions for the degradation are obtained on acidic minerals after herbicide deposition using the liquid method. Under these conditions an acceptable correlation with pseudo-first-order kinetics was observed, and the major degradation path is similar to that proposed for chemical hydrolysis. Conversely, alumina seemed to favor other unknown degradation processes. The hydrolysis paths of nicosulfuron and rimsulfuron appeared to be different. PMID:11804524

  16. Lactobacillus reuteri 100-23 Modulates Urea Hydrolysis in the Murine Stomach

    PubMed Central

    Wilson, Charlotte M.; Loach, Diane; Lawley, Blair; Bell, Tracey; Sims, Ian M.; O'Toole, Paul W.; Zomer, Aldert

    2014-01-01

    Comparisons of in vivo (mouse stomach) and in vitro (laboratory culture) transcriptomes of Lactobacillus reuteri strain 100-23 were made by microarray analysis. These comparisons revealed the upregulation of genes associated with acid tolerance, including urease production, in the mouse stomach. Inactivation of the ureC gene reduced the acid tolerance of strain 100-23 in vitro, and the mutant was outcompeted by the wild type in the gut of ex-Lactobacillus-free mice. Urine analysis showed that stable isotope-labeled urea, administered by gavage, was metabolized to a greater extent in Lactobacillus-free mice than animals colonized by strain 100-23. This surprising observation was associated with higher levels of urease activity and fecal-type bacteria in the stomach digesta of Lactobacillus-free mice. Despite the modulation of urea hydrolysis in the stomach, recycling of urea nitrogen in the murine host was not affected since the essential amino acid isoleucine, labeled with a stable isotope, was detected in the livers of both Lactobacillus-free and 100-23-colonized animals. Therefore, our experiments reveal a new and unexpected impact of Lactobacillus colonization on urea hydrolysis in the murine gut. PMID:25063664

  17. Effect of Pre-Stressing on the Acid-Stress Response in Bifidobacterium Revealed Using Proteomic and Physiological Approaches.

    PubMed

    Jin, Junhua; Qin, Qian; Guo, Huiyuan; Liu, Songling; Ge, Shaoyang; Zhang, Hongxing; Cui, Jianyun; Ren, Fazheng

    2015-01-01

    Weak acid resistance limits the application of Bifidobacteria as a probiotic in food. The acid tolerance response (ATR), caused by pre-stressing cells at a sublethal pH, could improve the acid resistance of Bifidobacteria to subsequent acid stress. In this study, we used Bifidobacterium longum sub. longum BBMN68 to investigate the effect of the ATR on the acid stress response (ASR), and compared the difference between the ATR and the ASR by analyzing the two-dimensional-PAGE protein profiles and performing physiological tests. The results revealed that a greater abundance of proteins involved in carbohydrate metabolism and protein protection was present after the ASR than after the ATR in Bifidobacterium. Pre-stressing cells increased the abundance of proteins involved in energy production, amino acid metabolism, and peptidoglycan synthesis during the ASR of Bifidobacterium. Moreover, after the ASR, the content of ATP, NH3, thiols, and peptidoglycan, the activity of H+-ATPase, and the maintenance of the intracellular pH in the pre-stressed Bifidobacterium cells was significantly higher than in the uninduced cells. These results provide the first explanation as to why the resistance of Bifidobacterium to acid stress improved after pre-stressing. PMID:25689631

  18. Hydrolysis of Al3+ from constrained molecular dynamics.

    PubMed

    Ikeda, Takashi; Hirata, Masaru; Kimura, Takaumi

    2006-02-21

    We investigated the hydrolysis reactions of Al(3+) in AlCl(3) aqueous solution using the constrained molecular dynamics based on the Car-Parrinello molecular-dynamics method. By employing the proton-aluminum coordination number as a reaction coordinate in the constrained molecular dynamics the deprotonation as well as dehydration processes are successfully realized. From our free-energy difference of DeltaG(0) approximately 8.0 kcal mol(-1) the hydrolysis constant pK(a1) is roughly estimated as 5.8, comparable to the literature value of 5.07. We show that the free-energy difference for the hydrolysis of Al(3+) in acidic conditions is at least 4 kcal mol(-1) higher than that in neutral condition, indicating that the hydrolysis reaction is inhibited by the presence of excess protons located around the hydrated ion, in agreement with the change of the predominant species by pH. PMID:16497053

  19. Molecular annotation of ketol-acid reductoisomerases from Streptomyces reveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity.

    PubMed

    Verdel-Aranda, Karina; López-Cortina, Susana T; Hodgson, David A; Barona-Gómez, Francisco

    2015-03-01

    The 6-phosphogluconate dehydrogenase superfamily oxidize and reduce a wide range of substrates, making their functional annotation challenging. Ketol-acid reductoisomerase (KARI), encoded by the ilvC gene in branched-chain amino acids biosynthesis, is a promiscuous reductase enzyme within this superfamily. Here, we obtain steady-state enzyme kinetic parameters for 10 IlvC homologues from the genera Streptomyces and Corynebacterium, upon eight selected chemically diverse substrates, including some not normally recognized by enzymes of this superfamily. This biochemical data suggested a Streptomyces biosynthetic interlock between proline and the branched-chain amino acids, mediated by enzyme substrate promiscuity, which was confirmed via mutagenesis and complementation analyses of the proC, ilvC1 and ilvC2 genes in Streptomyces coelicolor. Moreover, both ilvC orthologues and paralogues were analysed, such that the relationship between gene duplication and functional diversification could be explored. The KARI paralogues present in S. coelicolor and Streptomyces lividans, despite their conserved high sequence identity (97%), were shown to be more promiscuous, suggesting a recent functional diversification. In contrast, the KARI paralogue from Streptomyces viridifaciens showed selectivity towards the synthesis of valine precursors, explaining its recruitment within the biosynthetic gene cluster of valanimycin. These results allowed us to assess substrate promiscuity indices as a tool to annotate new molecular functions with metabolic implications. PMID:25296650

  20. Molecular annotation of ketol-acid reductoisomerases from Streptomyces reveals a novel amino acid biosynthesis interlock mediated by enzyme promiscuity

    PubMed Central

    Verdel-Aranda, Karina; López-Cortina, Susana T; Hodgson, David A; Barona-Gómez, Francisco

    2015-01-01

    The 6-phosphogluconate dehydrogenase superfamily oxidize and reduce a wide range of substrates, making their functional annotation challenging. Ketol-acid reductoisomerase (KARI), encoded by the ilvC gene in branched-chain amino acids biosynthesis, is a promiscuous reductase enzyme within this superfamily. Here, we obtain steady-state enzyme kinetic parameters for 10 IlvC homologues from the genera Streptomyces and Corynebacterium, upon eight selected chemically diverse substrates, including some not normally recognized by enzymes of this superfamily. This biochemical data suggested a Streptomyces biosynthetic interlock between proline and the branched-chain amino acids, mediated by enzyme substrate promiscuity, which was confirmed via mutagenesis and complementation analyses of the proC, ilvC1 and ilvC2 genes in Streptomyces coelicolor. Moreover, both ilvC orthologues and paralogues were analysed, such that the relationship between gene duplication and functional diversification could be explored. The KARI paralogues present in S. coelicolor and Streptomyces lividans, despite their conserved high sequence identity (97%), were shown to be more promiscuous, suggesting a recent functional diversification. In contrast, the KARI paralogue from Streptomyces viridifaciens showed selectivity towards the synthesis of valine precursors, explaining its recruitment within the biosynthetic gene cluster of valanimycin. These results allowed us to assess substrate promiscuity indices as a tool to annotate new molecular functions with metabolic implications. PMID:25296650

  1. Stereoconfiguration of bisphosphatidic and semilysobisphosphatidic acids from cultured hamster fibroblasts (BHK cells).

    PubMed

    Somerharju, P; Brotherus, J; Kahma, K; Renkonen, O

    1977-04-26

    Monolayers of hamster fibroblasts (BHK cells) were incubated in Eagle's minimal essential medium under conditions where an increase in the levels of all cellular bisphosphatidic acids takes place. Bisphosphatidic acid and semilysobisphosphatidic were isolated from these cells and subjected to strong alkaline hydrolysis. Stereochemical analysis of the hydrolysis products revealed that the majority of the molecules of both lipids are derivatives of sn-1-glycerophospho-sn-1'-glycerol, the structure previously found to be the "backbone" of lysobisphosphatidic acid, (bis(monoacylglycerol)phosphate) from BHK cells and other sources. This finding suggests a close metabolic relationship between the three bisphosphatidic acid derivatives of BHK cells. PMID:857898

  2. Lignocellulosic Biomass to Ethanol Process Design and Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic Hydrolysis for Corn Stover

    SciTech Connect

    Aden, A.; Ruth, M.; Ibsen, K.; Jechura, J.; Neeves, K.; Sheehan, J.; Wallace, B.; Montague, L.; Slayton, A.; Lukas, J.

    2002-06-01

    This report is an update of NREL's ongoing process design and economic analyses of processes related to developing ethanol from lignocellulosic feedstocks. The U.S. Department of Energy (DOE) is promoting the development of ethanol from lignocellulosic feedstocks as an alternative to conventional petroleum-based transportation fuels. DOE funds both fundamental and applied research in this area and needs a method for predicting cost benefits of many research proposals. To that end, the National Renewable Energy Laboratory (NREL) has modeled many potential process designs and estimated the economics of each process during the last 20 years. This report is an update of the ongoing process design and economic analyses at NREL. We envision updating this process design report at regular intervals; the purpose being to ensure that the process design incorporates all new data from NREL research, DOE funded research and other sources, and that the equipment costs are reasonable and consistent with good engineering practice for plants of this type. For the non-research areas this means using equipment and process approaches as they are currently used in industrial applications. For the last report, published in 1999, NREL performed a complete review and update of the process design and economic model for the biomass-to-ethanol process utilizing co-current dilute acid prehydrolysis with simultaneous saccharification (enzymatic) and co-fermentation. The process design included the core technologies being researched by the DOE: prehydrolysis, simultaneous saccharification and co-fermentation, and cellulase enzyme production. In addition, all ancillary areas--feed handling, product recovery and purification, wastewater treatment (WWT), lignin combustor and boiler-turbogenerator, and utilities--were included. NREL engaged Delta-T Corporation (Delta-T) to assist in the process design evaluation, the process equipment costing, and overall plant integration. The process design and

  3. Determination of free and total myo-inositol in infant formula and adult/pediatric nutritional formula by high- performance anion exchange chromatography with pulsed amperometric detection, including a novel total extraction using microwave-assisted acid hydrolysis and enzymatic treatment: first action 2012.12.

    PubMed

    Ellingson, David; Pritchard, Ted; Foy, Pamela; King, Kathryn; Mitchell, Barbara; Austad, John; Winters, Doug; Sullivan, Darryl; Dowell, Dawn

    2013-01-01

    After an assessment of data generated from a single-laboratory validation study published in J. AOAC Int. 95, 1469-1478 (2012), a method for determining total myo-inositol in infant formula and adult/ pediatric nutritional formula by high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), including extraction by using microwave-assisted acid hydrolysis and enzymatic treatment was presented for consideration by AOAC during the AOAC Annual Meeting held in Las Vegas, NV, from September 30 to October 3, 2012. The Expert Review Panel on Infant Formula and Adult Nutritionals concluded that the method met the criteria set by the standard method performance requirements (SMPRs) for the determination of free myo-inositol and approved the method as AOAC Official First Action. The method also determines total myo-inositol, but includes bound sources that the SMPRs exclude. The method involves using HPAEC-PAD for free myo-inositol and a total myo-inositol determination by two different techniques. The first technique uses the conventional acid hydrolysis with 6 h incubation in an autoclave. The second uses a microwave-assisted acid hydrolysis with enzymatic treatment that decreases the extraction time. PMID:24282949

  4. Pretreatment and enzymatic hydrolysis of lignocellulosic biomass

    NASA Astrophysics Data System (ADS)

    Corredor, Deisy Y.

    The performance of soybean hulls and forage sorghum as feedstocks for ethanol production was studied. The main goal of this research was to increase fermentable sugars' yield through high-efficiency pretreatment technology. Soybean hulls are a potential feedstock for production of bio-ethanol due to their high carbohydrate content (≈50%) of nearly 37% cellulose. Soybean hulls could be the ideal feedstock for fuel ethanol production, because they are abundant and require no special harvesting and additional transportation costs as they are already in the plant. Dilute acid and modified steam-explosion were used as pretreatment technologies to increase fermentable sugars yields. Effects of reaction time, temperature, acid concentration and type of acid on hydrolysis of hemicellulose in soybean hulls and total sugar yields were studied. Optimum pretreatment parameters and enzymatic hydrolysis conditions for converting soybean hulls into fermentable sugars were identified. The combination of acid (H2SO4, 2% w/v) and steam (140°C, 30 min) efficiently solubilized the hemicellulose, giving a pentose yield of 96%. Sorghum is a tropical grass grown primarily in semiarid and dry parts of the world, especially in areas too dry for corn. The production of sorghum results in about 30 million tons of byproducts mainly composed of cellulose, hemicellulose, and lignin. Forage sorghum such as brown midrib (BMR) sorghum for ethanol production has generated much interest since this trait is characterized genetically by lower lignin concentrations in the plant compared with conventional types. Three varieties of forage sorghum and one variety of regular sorghum were characterized and evaluated as feedstock for fermentable sugar production. Fourier transform infrared spectroscopy (FTIR), scanning electron microscope (SEM) and X-Ray diffraction were used to determine changes in structure and chemical composition of forage sorghum before and after pretreatment and enzymatic hydrolysis

  5. The Crystal Structure of the Adenylation Enzyme VinN Reveals a Unique β-Amino Acid Recognition Mechanism*

    PubMed Central

    Miyanaga, Akimasa; Cieślak, Jolanta; Shinohara, Yuji; Kudo, Fumitaka; Eguchi, Tadashi

    2014-01-01

    Adenylation enzymes play important roles in the biosynthesis and degradation of primary and secondary metabolites. Mechanistic insights into the recognition of α-amino acid substrates have been obtained for α-amino acid adenylation enzymes. The Asp residue is invariant and is essential for the stabilization of the α-amino group of the substrate. In contrast, the β-amino acid recognition mechanism of adenylation enzymes is still unclear despite the importance of β-amino acid activation for the biosynthesis of various natural products. Herein, we report the crystal structure of the stand-alone adenylation enzyme VinN, which specifically activates (2S,3S)-3-methylaspartate (3-MeAsp) in vicenistatin biosynthesis. VinN has an overall structure similar to that of other adenylation enzymes. The structure of the complex with 3-MeAsp revealed that a conserved Asp230 residue is used in the recognition of the β-amino group of 3-MeAsp similar to α-amino acid adenylation enzymes. A mutational analysis and structural comparison with α-amino acid adenylation enzymes showed that the substrate-binding pocket of VinN has a unique architecture to accommodate 3-MeAsp as a β-amino acid substrate. Thus, the VinN structure allows the first visualization of the interaction of an adenylation enzyme with a β-amino acid and provides new mechanistic insights into the selective recognition of β-amino acids in this family of enzymes. PMID:25246523

  6. The crystal structure of the adenylation enzyme VinN reveals a unique β-amino acid recognition mechanism.

    PubMed

    Miyanaga, Akimasa; Cieślak, Jolanta; Shinohara, Yuji; Kudo, Fumitaka; Eguchi, Tadashi

    2014-11-01

    Adenylation enzymes play important roles in the biosynthesis and degradation of primary and secondary metabolites. Mechanistic insights into the recognition of α-amino acid substrates have been obtained for α-amino acid adenylation enzymes. The Asp residue is invariant and is essential for the stabilization of the α-amino group of the substrate. In contrast, the β-amino acid recognition mechanism of adenylation enzymes is still unclear despite the importance of β-amino acid activation for the biosynthesis of various natural products. Herein, we report the crystal structure of the stand-alone adenylation enzyme VinN, which specifically activates (2S,3S)-3-methylaspartate (3-MeAsp) in vicenistatin biosynthesis. VinN has an overall structure similar to that of other adenylation enzymes. The structure of the complex with 3-MeAsp revealed that a conserved Asp(230) residue is used in the recognition of the β-amino group of 3-MeAsp similar to α-amino acid adenylation enzymes. A mutational analysis and structural comparison with α-amino acid adenylation enzymes showed that the substrate-binding pocket of VinN has a unique architecture to accommodate 3-MeAsp as a β-amino acid substrate. Thus, the VinN structure allows the first visualization of the interaction of an adenylation enzyme with a β-amino acid and provides new mechanistic insights into the selective recognition of β-amino acids in this family of enzymes. PMID:25246523

  7. ESTIMATION OF PHOSPHATE ESTER HYDROLYSIS RATE CONSTANTS. I. ALKALINE HYDROLYSIS

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to allow the calculation of alkaline hydrolysis rate constants of phosphate esters in water. The rate is calculated from the energy difference between the initial and transition state...

  8. ESTIMATION OF PHOSPHATE ESTER HYDROLYSIS RATE CONSTANTS - ALKALINE HYDROLYSIS

    EPA Science Inventory

    SPARC (SPARC Performs Automated Reasoning in Chemistry) chemical reactivity models were extended to allow the calculation of alkaline hydrolysis rate constants of phosphate esters in water. The rate is calculated from the energy difference between the initial and transition state...

  9. Proteomics-Based Metabolic Modeling Reveals That Fatty Acid Oxidation (FAO) Controls Endothelial Cell (EC) Permeability*

    PubMed Central

    Patella, Francesca; Schug, Zachary T.; Persi, Erez; Neilson, Lisa J.; Erami, Zahra; Avanzato, Daniele; Maione, Federica; Hernandez-Fernaud, Juan R.; Mackay, Gillian; Zheng, Liang; Reid, Steven; Frezza, Christian; Giraudo, Enrico; Fiorio Pla, Alessandra; Anderson, Kurt; Ruppin, Eytan; Gottlieb, Eyal; Zanivan, Sara

    2015-01-01

    Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model that predicts the metabolic fluxes in ECs when cultured on a tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. Remarkably, global phosphoproteomic changes measured upon acute CPT1A inhibition pinpointed altered calcium signaling. Indeed, CPT1A inhibition increases intracellular calcium oscillations. Finally, inhibiting CPT1A induces hyperpermeability in vitro and leakage of blood vessel in vivo, which were restored blocking calcium influx or replenishing the tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability. PMID:25573745

  10. Proteomics-based metabolic modeling reveals that fatty acid oxidation (FAO) controls endothelial cell (EC) permeability.

    PubMed

    Patella, Francesca; Schug, Zachary T; Persi, Erez; Neilson, Lisa J; Erami, Zahra; Avanzato, Daniele; Maione, Federica; Hernandez-Fernaud, Juan R; Mackay, Gillian; Zheng, Liang; Reid, Steven; Frezza, Christian; Giraudo, Enrico; Fiorio Pla, Alessandra; Anderson, Kurt; Ruppin, Eytan; Gottlieb, Eyal; Zanivan, Sara

    2015-03-01

    Endothelial cells (ECs) play a key role to maintain the functionality of blood vessels. Altered EC permeability causes severe impairment in vessel stability and is a hallmark of pathologies such as cancer and thrombosis. Integrating label-free quantitative proteomics data into genome-wide metabolic modeling, we built up a model that predicts the metabolic fluxes in ECs when cultured on a tridimensional matrix and organize into a vascular-like network. We discovered how fatty acid oxidation increases when ECs are assembled into a fully formed network that can be disrupted by inhibiting CPT1A, the fatty acid oxidation rate-limiting enzyme. Acute CPT1A inhibition reduces cellular ATP levels and oxygen consumption, which are restored by replenishing the tricarboxylic acid cycle. Remarkably, global phosphoproteomic changes measured upon acute CPT1A inhibition pinpointed altered calcium signaling. Indeed, CPT1A inhibition increases intracellular calcium oscillations. Finally, inhibiting CPT1A induces hyperpermeability in vitro and leakage of blood vessel in vivo, which were restored blocking calcium influx or replenishing the tricarboxylic acid cycle. Fatty acid oxidation emerges as central regulator of endothelial functions and blood vessel stability and druggable pathway to control pathological vascular permeability. PMID:25573745

  11. Deciphering Carbamoylpolyoxamic Acid Biosynthesis Reveals Unusual Acetylation Cycle Associated with Tandem Reduction and Sequential Hydroxylation.

    PubMed

    Qi, Jianzhao; Wan, Dan; Ma, Hongmin; Liu, Yuanzhen; Gong, Rong; Qu, Xudong; Sun, Yuhui; Deng, Zixin; Chen, Wenqing

    2016-08-18

    Polyoxin, produced by Streptomcyes cacaoi var. asoensis and Streptomyces aureochromogenes, contains two non-proteinogenic amino acids, carbamoylpolyoxamic acid (CPOAA) and polyoximic acid. Although the CPOAA moiety is highly unusual, its biosynthetic logic has remained enigmatic for decades. Here, we address CPOAA biosynthesis by reconstitution of its pathway. We demonstrated that its biosynthesis is initiated by a versatile N-acetyltransferase, PolN, catalyzing L-glutamate (1) to N-acetyl glutamate (2). Remarkably, we verified that PolM, a previously annotated dehydrogenase, catalyzes an unprecedented tandem reduction of acyl-phosphate to aldehyde, and subsequently to alcohol. We also unveiled a distinctive acetylation cycle catalyzed by PolN to synthesize α-amino-δ-hydroxyvaleric acid (6). Finally, we report that PolL is capable of converting a rare sequential hydroxylation of α-amino-δ-carbamoylhydroxyvaleric acid (7) to CPOAA. PolL represents an intriguing family of Fe(II)-dependent α-ketoglutarate dioxygenase with a cupin fold. These data illustrate several novel enzymatic reactions, and also set a foundation for rational pathway engineering for polyoxin production. PMID:27541195

  12. Hydrolysis of phytic acid by intrinsic plant and supplemented microbial phytase (Aspergillus niger) in the stomach and small intestine of minipigs fitted with re-entrant cannulas. 2. Phytase activity.

    PubMed

    Rapp, C; Lantzsch, H J; Drochner, W

    2001-12-01

    Hydrolysis of phytate in the stomach and the small intestine as influenced by intrinsic plant (wheat) and supplemented microbial phytase (A. niger) were investigated with six minipigs (40-50 kg initial BW) fitted with re-entrant cannulas in the duodenum, 30 cm posterior to the pylorus (animals 1, 4, 5, and 6) and ileocecal re-entrant cannulas, 5 cm prior the ileocecal junction (animals 1, 2, and 3), respectively. Dietary treatments were as follows: (1) diet 1, a corn-based diet (43 U phytase/kg DM); (2) diet 2, diet 1 supplemented with microbial phytase (818 U/kg DM) and (3) diet 3, a wheat-based diet (1192 U/kg DM). At 0730 and 1930 per animal 350 g diet mixed with 1050 ml de-ionized water were fed. Digesta were collected continuously and completely during 12 h after feeding. In the duodenal digesta, 70% of the microbial phytase (diet 2) and 45% of the wheat phytase (diet 3) were recovered within 12 h after ingestion of the phytases, whereas only negligible amounts were detected in the digesta of pigs fed the phytase-poor corn-based diet 1. Most phytase activity passed through the stomach within the first hour after feeding. Microbial phytase activity at pH 2.8 was less sensitive to acidic pHs, such as those found in the stomach, than phytase activity at pH 5.3. Phytase activities in the digesta of the distal ileum did not depend either on source or amount of dietary phytase activity. PMID:11906565

  13. A kinetic study of Trichoderma reesei Cel7B catalyzed cellulose hydrolysis.

    PubMed

    Song, Xiangfei; Zhang, Shujun; Wang, Yefei; Li, Jingwen; He, Chunyan; Yao, Lishan

    2016-06-01

    One prominent feature of Trichoderma reesei (Tr) endoglucanases catalyzed cellulose hydrolysis is that the reaction slows down quickly after it starts (within minutes). But the mechanism of the slowdown is not well understood. A structural model of Tr- Cel7B catalytic domain bound to cellulose was built computationally and the potentially important binding residues were identified and tested experimentally. The 13 tested mutants show different binding properties in the adsorption to phosphoric acid swollen cellulose and filter paper. Though the partitioning parameter to filter paper is about 10 times smaller than that to phosphoric acid swollen cellulose, a positive correlation is shown for two substrates. The kinetic studies show that the reactions slow down quickly for both substrates. This slowdown is not correlated to the binding constant but anticorrelated to the enzyme initial activity. The amount of reducing sugars released after 24h by Cel7B in phosphoric acid swollen cellulose, Avicel and filter paper cellulose hydrolysis is correlated with the enzyme activity against a soluble substrate p-nitrophenyl lactoside. Six of the 13 tested mutants, including N47A, N52D, S99A, N323D, S324A, and S346A, yield ∼15-35% more reducing sugars than the wild type (WT) Cel7B in phosphoric acid swollen cellulose and filter paper hydrolysis. This study reveals that the slowdown of the reaction is not due to the binding of the enzyme to cellulose. The activity of Tr- Cel7B against the insoluble substrate cellulose is determined by the enzyme's capability in hydrolyzing the soluble substrate. PMID:27178789

  14. Mechanistic Modeling Reveals the Critical Knowledge Gaps in Bile Acid-Mediated DILI.

    PubMed

    Woodhead, J L; Yang, K; Brouwer, K L R; Siler, S Q; Stahl, S H; Ambroso, J L; Baker, D; Watkins, P B; Howell, B A

    2014-01-01

    Bile salt export pump (BSEP) inhibition has been proposed to be an important mechanism for drug-induced liver injury (DILI). Modeling can prioritize knowledge gaps concerning bile acid (BA) homeostasis and thus help guide experimentation. A submodel of BA homeostasis in rats and humans was constructed within DILIsym, a mechanistic model of DILI. In vivo experiments in rats with glibenclamide were conducted, and data from these experiments were used to validate the model. The behavior of DILIsym was analyzed in the presence of a simulated theoretical BSEP inhibitor. BSEP inhibition in humans is predicted to increase liver concentrations of conjugated chenodeoxycholic acid (CDCA) and sulfate-conjugated lithocholic acid (LCA) while the concentration of other liver BAs remains constant or decreases. On the basis of a sensitivity analysis, the most important unknowns are the level of BSEP expression, the amount of intestinal synthesis of LCA, and the magnitude of farnesoid-X nuclear receptor (FXR)-mediated regulation. PMID:25006780

  15. Amino acid racemization reveals differential protein turnover in osteoarthritic articular and meniscal cartilages

    PubMed Central

    Stabler, Thomas V; Byers, Samuel S; Zura, Robert D; Kraus, Virginia Byers

    2009-01-01

    Introduction Certain amino acids within proteins have been reported to change from the L form to the D form over time. This process is known as racemization and is most likely to occur in long-lived low-turnover tissues such as normal cartilage. We hypothesized that diseased tissue, as found in an osteoarthritic (OA) joint, would have increased turnover reflected by a decrease in the racemized amino acid content. Methods Using high-performance liquid chromatography methods, we quantified the L and D forms of amino acids reported to racemize in vivo on a biological timescale: alanine, aspartate (Asp), asparagine (Asn), glutamate, glutamine, isoleucine, leucine (Leu), and serine (Ser). Furthermore, using a metabolically inactive control material (tooth dentin) and a control material with normal metabolism (normal articular cartilage), we developed an age adjustment in order to make inferences about the state of protein turnover in cartilage and meniscus. Results In the metabolically inactive control material (n = 25, ages 13 to 80 years) and the normal metabolizing control material (n = 19, ages 17 to 83 years), only Asp + Asn (Asx), Ser, and Leu showed a significant change (increase) in racemization with age (P < 0.01). The age-adjusted proportions of racemized to total amino acid (D/D+L expressed as a percentage of the control material) for Asx, Ser, and Leu when compared with the normal articular cartilage control were 97%, 74%, and 73% in OA meniscal cartilage and 97%, 70%, and 78% in OA articular cartilage. We also observed lower amino acid content in OA articular and meniscal cartilages compared with normal articular cartilage as well as a loss of total amino acids with age in the OA meniscal but not the OA articular cartilage. Conclusions These data demonstrate comparable anabolic responses for non-lesioned OA articular cartilage and OA meniscal cartilage but an excess of catabolism over anabolism for the meniscal cartilage. PMID:19267899

  16. Integration of computational modeling with membrane transport studies reveals new insights into amino acid exchange transport mechanisms