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Sample records for pixe-aided immobilization study

  1. [sup 31]P NMR study of immobilized artificial membrane surfaces. Structure and dynamics of immobilized phospholipids

    SciTech Connect

    Qiu, X.; Pidgeon, C. )

    1993-11-25

    Chromatography surfaces were prepared by immobilizing a single-chain ether phospholipid at approximately a monolayer density on silica particles. The chromatography particles are denoted as [sup ether]IAM.PC[sup C10/C3], and they are stable to all solvents. The structure and dynamics of the interphase created by immobilizing phospholipids on silica particles were studied by [sup 31]P NMR methods. [sup ether]IAM.PC[sup C10/C3] spontaneously wets when suspended in both aqueous and organic solvents, and [sup 31]P NMR spectra were obtained in water, methanol, chloroform, acetonitrile, and acetone. [sup 31]P NMR spectra were subjected to line-shape analysis. From line-shape analysis, the correlation times for rapid internal motion ([tau]-PLL) and wobbling ([tau]-PRP) of the phospholipid headgroup were calculated for each solvent. Immobilized phospholipid headgroups comprising the IAM interfacial region undergo rapid reorientation similar to the case of the phospholipids forming liposome membranes with [tau]-PLL approximately 1 ns. Phospholipids in liposome membranes exhibit slower wobbling motion ([tau]-PRP approximately 1 ms) in the plane of the membrane. However, the immobilized phospholipids on [sup ether]IAM.PC[sup C10/C3] surfaces wobble with correlation times [tau]-PRP that depend on the solvent bathing the [sup ether]IAM.PC[sup C10/C3] surface. 41 refs., 9 figs., 2 tabs.

  2. Preliminary studies on immobilization of lipase using chicken eggshell

    NASA Astrophysics Data System (ADS)

    Salleh, S.; Serri, N. A.; Hena, S.; Tajarudin, H. A.

    2016-06-01

    A few advantages of enzyme immobilization are reusability of expensive enzyme, improvement of stability and activity compared to crude enzyme. Various organic components can be used as carrier for enzyme immobilization such as chicken eggshell. It can be used as a carrier for immobilization as its mineral component mostly contains of calcium carbonate. In the present study, Tributyrin method was used to test enzyme activity of Rhizomucour Miehei, Candida Antarctica and Candida Rugosa. Rhizomucour Miehei shows the highest enzyme activity (360.8 mol/min/mL lipase) and was used in further experiment. Experiment was continued to study incubation time for lipase immobilization on eggshell (1-4 hours) and reaction time of esterification of sugar ester (0-72 hours). Two hours incubation time for lipase immobilization was observed and gives the highest yield of sugar ester (78.13%). Fructose and stearic acid as substrate was used for the production of sugar ester. The highest percentage of sugar ester production was shown at 36 hours of reaction time.

  3. Influence of immobilization strategies on biosensing response characteristics: A comparative study.

    PubMed

    Kaur, Gurpreet; Saha, Shibu; Tomar, Monika; Gupta, Vinay

    2016-01-01

    The immobilization technique plays an important role in fabrication of a biosensor. NiO based cholesterol biosensor has been used to study the effect of various immobilization techniques on the biosensing response characteristics. The biosensors were fabricated by immobilizing cholesterol oxidase on NiO thin films by three different immobilization techniques viz. physisorption, cross-linking and covalent binding. The study reveals a strong dependence of biosensing response on corresponding immobilization technique. The biosensor based on immobilization by covalent bonding shows superior response characteristics as compared to others owing to its zero length. The results highlight the significance of immobilization technique for biosensor fabrication.

  4. Use of immobilized enzymes in drug metabolism studies.

    PubMed

    Dulik, D M; Fenselau, C

    1988-04-01

    The immobilization of drug-metabolizing enzymes onto polymeric supports offers several advantages over use of conventional microsomal or soluble enzyme preparations. These include increased storage stability, facilitated separation of products from the incubation mixture, the ability to recover and reuse the enzyme catalyst, and in many cases, stabilization of the tertiary structure of membrane-bound enzymes. Attachment of the protein to the solid support may be accomplished by adsorption, covalent bonding, or entrapment techniques. This methodology has been successfully utilized for studies with such enzymes as cytochrome P-450, UDP-glucuronyltransferases, glutathione S-transferases, S-methyltransferases, and N-acetyltransferases. Although often employed for the synthesis of xenobiotic metabolites, immobilized enzymes have been used for mechanistic and relative reactivity studies, limited kinetic studies, and extracorporeal detoxification. Co-immobilization of multiple drug-metabolizing enzyme systems has made possible the sequential formation of metabolites arising from oxidation followed by conjugation. Immobilized enzymes may also be used in the prediction of species-dependent metabolic pathways. The potential for large-scale synthesis of drug metabolites using this methodology has been explored. PMID:3127263

  5. Study on algae removal by immobilized biosystem on sponge

    NASA Astrophysics Data System (ADS)

    Pei, Haiyan; Hu, Wenrong

    2006-10-01

    In this study, sponges were used to immobilize domesticated sludge microbes in a limited space, forming an immobilized biosystem capable of algae and microcystins removal. The removal effects on algae, microcystins and UV260 of this biosystem and the mechanism of algae removal were studied. The results showed that active sludge from sewage treatment plants was able to remove algae from a eutrophic lake’s water after 7 d of domestication. The removal efficiency for algae, organic matter and microcystins increased when the domesticated sludge was immobilized on sponges. When the hydraulic retention time (HRT) was 5h, the removal rates of algae, microcystins and UV260 were 90%, 94.17% and 84%, respectively. The immobilized biosystem consisted mostly of bacteria, the Ciliata and Sarcodina protozoans and the Rotifer metazoans. Algal decomposition by zoogloea bacteria and preying by microcreatures were the two main modes of algal removal, which occurred in two steps: first, absorption by the zoogloea; second, decomposition by the zoogloea bacteria and the predacity of the microcreatures.

  6. Platelet Immobilization on Supported Phospholipid Bilayers for Single Platelet Studies.

    PubMed

    Uhl, Eva; Donati, Alessia; Reviakine, Ilya

    2016-08-23

    The worldwide cardiovascular disease (CVD) epidemic is of grave concern. A major role in the etiology of CVDs is played by the platelets (thrombocytes). Platelets are anuclear cell fragments circulating in the blood. Their primary function is to catalyze clot formation, limiting traumatic blood loss in the case of injury. The same process leads to thrombosis in the case of CVDs, which are commonly managed with antiplatelet therapy. Platelets also have other, nonhemostatic functions in wound healing, inflammation, and tissue regeneration. They play a role in the early stages of atherosclerosis and the spread of cancer through metastases. Much remains to be learned about the regulation of these diverse platelet functions under physiological and pathological conditions. Breakthroughs in this regard are expected to come from single platelet studies and systems approaches. The immobilization of platelets at surfaces is advantageous for developing such approaches, but platelets are activated when they come in contact with foreign surfaces. In this work, we develop and validate a protocol for immobilizing platelets on supported lipid bilayers without activation due to immobilization. Our protocol can therefore be used for studying platelets with a wide variety of surface-sensitive techniques. PMID:27438059

  7. Subsurface Immobilization of Plutonium: Experimental and Model Validation Studies

    SciTech Connect

    Rittmann, Bruce E; Deo, Randhir P; Reed, Donald T

    2008-08-13

    We conducted a coordinated experimental and modeling study centered on the interaction of Shewanella alga BrY (S. alga) with plutonium species and phases. Plutonium is the key contaminant of concern at several DOE sites that are being addressed by the overall ERSP program. The over-arching goal of this research was to understand the long-term stability of bio-precipitated immobilized plutonium phases under changing redox conditions in biologically active systems. To initiate the process of plutonium immobilization, a side-by-side comparison of the bioreduction of uranyl and plutonyl species was conducted with S. alga. Uranyl was reduced in our system, consistent with literature reports, but we noted coupling between abiotic and biotic processes and observed that non-reductive pathways to precipitation typically exist. Additionally, a key role of biogenic Fe2+, which is known to reduce uranyl at low pH, is suggested. In contrast, residual organics, present in biologically active systems, reduce Pu(VI) species to Pu(V) species at near-neutral pH. The predominance of relatively weak complexes of PuO2+ is an important difference in how the uranyl and plutonyl species interacted with S. alga. Pu(V) also led to increased toxicity towards S. alga and is also more easily reduced by microbial activity. Biogenic Fe2+, produced by S. alga when Fe3+ is present as an electron acceptor, also played a key role in understanding redox controls and pathways in this system. Overall, the bioreduction of plutonyl was observed under anaerobic conditions, which favor its immobilization in the subsurface. Understanding the mechanism by which redox control is established in biologically active systems is a key aspect of remediation and immobilization strategies for actinides when they are present as subsurface contaminants.

  8. Effects of the immobilization supports on the catalytic properties of immobilized mushroom tyrosinase: a comparative study using several substrates.

    PubMed

    Marín-Zamora, María Elisa; Rojas-Melgarejo, Francisco; García-Cánovas, Francisco; García-Ruiz, Pedro Antonio

    2007-09-30

    Mushroom tyrosinase was immobilized from an extract onto glass beads covered with one of the following compounds: the crosslinked totally cinnamoylated derivatives of glycerine, D-sorbitol, D-manitol, 1,2-O-isopropylidene-alpha-D-glucofuranose, D-glucuronic acid, D-gulonic acid, sucrose, D-glucosone, D-arabinose, D-fructose, D-glucose, ethyl-D-glucopyranoside, inuline, dextrine, dextrane or starch, or the partially cinnamoylated derivative 3,5,6-tricinnamoyl-D-glucofuranose which was obtained by the acid hydrolysis of 1,2-O-isopropylidene-alpha-d-glucofuranose. The enzyme was immobilized by direct adsorption onto the support and the quantity of tyrosinase immobilized was found to increase with the hydrophobicity of the supports. The kinetic constants of immobilized tyrosinase acting on the substrates, 4-tert-butylcatechol, dopamine and DL-dopa, were studied. When immobilized tyrosinase acted on 4-tert-butylcatechol, the values of K(m)(app) were lower than these obtained for tyrosinase in solution while, when dopamine and DL-dopa were used, the K(m)(app) were higher. The order of the substrates as regards their ionizable groups, DL-dopa (two ionizable groups)>dopamine (one ionizable group)>4-tert-butylcatechol (no ionizable group) coincided with the order of the K(m)(app) values shown by tyrosinase immobilized on the hydrophobic supports, and was the inverse of that observed for tyrosinase in solution. The K(m)(app) values of immobilized tyrosinase were in all cases higher than those of soluble tyrosinase and depended on the nature of the support and the hydrophobicity of the substrate, meaning that it is possible to design supports with different degrees of selectivity towards a mixture of enzyme substrates in the reaction medium. PMID:17868943

  9. Effects of the immobilization supports on the catalytic properties of immobilized mushroom tyrosinase: a comparative study using several substrates.

    PubMed

    Marín-Zamora, María Elisa; Rojas-Melgarejo, Francisco; García-Cánovas, Francisco; García-Ruiz, Pedro Antonio

    2007-09-30

    Mushroom tyrosinase was immobilized from an extract onto glass beads covered with one of the following compounds: the crosslinked totally cinnamoylated derivatives of glycerine, D-sorbitol, D-manitol, 1,2-O-isopropylidene-alpha-D-glucofuranose, D-glucuronic acid, D-gulonic acid, sucrose, D-glucosone, D-arabinose, D-fructose, D-glucose, ethyl-D-glucopyranoside, inuline, dextrine, dextrane or starch, or the partially cinnamoylated derivative 3,5,6-tricinnamoyl-D-glucofuranose which was obtained by the acid hydrolysis of 1,2-O-isopropylidene-alpha-d-glucofuranose. The enzyme was immobilized by direct adsorption onto the support and the quantity of tyrosinase immobilized was found to increase with the hydrophobicity of the supports. The kinetic constants of immobilized tyrosinase acting on the substrates, 4-tert-butylcatechol, dopamine and DL-dopa, were studied. When immobilized tyrosinase acted on 4-tert-butylcatechol, the values of K(m)(app) were lower than these obtained for tyrosinase in solution while, when dopamine and DL-dopa were used, the K(m)(app) were higher. The order of the substrates as regards their ionizable groups, DL-dopa (two ionizable groups)>dopamine (one ionizable group)>4-tert-butylcatechol (no ionizable group) coincided with the order of the K(m)(app) values shown by tyrosinase immobilized on the hydrophobic supports, and was the inverse of that observed for tyrosinase in solution. The K(m)(app) values of immobilized tyrosinase were in all cases higher than those of soluble tyrosinase and depended on the nature of the support and the hydrophobicity of the substrate, meaning that it is possible to design supports with different degrees of selectivity towards a mixture of enzyme substrates in the reaction medium.

  10. Laccase immobilization over multi-walled carbon nanotubes: Kinetic, thermodynamic and stability studies.

    PubMed

    Tavares, Ana P M; Silva, Cláudia G; Dražić, Goran; Silva, Adrián M T; Loureiro, José M; Faria, Joaquim L

    2015-09-15

    The biocatalytic performance of immobilized enzyme systems depends mostly on the intrinsic properties of both biomolecule and support, immobilization technique and immobilization conditions. Multi-walled carbon nanotubes (MWCNTs) possess unique features for enzyme immobilization by adsorption. Enhanced catalytic activity and stability can be achieved by optimization of the immobilization conditions and by investigating the effect of operational parameters. Laccase was immobilized over MWCNTs by adsorption. The hybrid material was characterized by Fourier transformed infrared (FTIR) spectroscopy, scanning and transmission electron microscopy (SEM and TEM, respectively). The effect of different operational conditions (contact time, enzyme concentration and pH) on laccase immobilization was investigated. Optimized conditions were used for thermal stability, kinetic, and storage and operational stability studies. The optimal immobilization conditions for a laccase concentration of 3.75μL/mL were a pH of 9.0 and a contact time of 30min (522 Ulac/gcarrier). A decrease in the thermal stability of laccase was observed after immobilization. Changes in ΔS and ΔH of deactivation were found for the immobilized enzyme. The Michaelis-Menten kinetic constant was higher for laccase/MWCNT system than for free laccase. Immobilized laccase maintained (or even increased) its catalytic performance up to nine cycles of utilization and revealed long-term storage stability.

  11. Studies on the thermal inactivation of immobilized enzymes

    SciTech Connect

    Ulbrich, R.; Schellenberger, A.; Damerau, W.

    1986-04-01

    The thermal inactivation of a great number of immobilized enzymes shows a biphasic kinetics, which distinctly differs from the first-order inactivation kinetics of the corresponding soluble enzymes. As shown for ..cap alpha..-amylase, chymotrypsin, and trypsin covalently bound to silica, polystyrene, or polyacrylamide, the dependence of the remaining activities on the heating time can be well described by the sum of two exponential terms. To interpret this mathematical model function, the catalytic properties of immobilized enzymes (number of active sites in silica-bound trypsin, Km and Ea values in silica-bound ..cap alpha..-amylase and chymotrypsin) at different stages of inactivation and the influence of various factors (coupling conditions, addition of denaturants or stabilizers, etc.) on the thermal inactivation of silica-bound ..cap alpha..-amylase were studied. Furthermore, conformational alterations in the thermal denaturation of spin-labeled soluble and silica-bound ..beta..-amylase were compared by electron spin resonance (ESR) studies. The results suggest that the biphasic inactivation kinetics reflects two different pathways according to which catalytically identical enzyme molecules are predominantly inactivated. 45 references.

  12. Adhesion Peptide Immobilization on Electrospun Polymers: a Photoelectron Spectroscopy Study

    NASA Astrophysics Data System (ADS)

    Iucci, G.; Polzonetti, G.; Ghezzo, F.; Modesti, M.; Roso, M.; Dettin, M.

    2010-06-01

    The immobilization of an oligopeptide, mimicking the adhesion sequence of fibronectin, on the surface of polymer films prepared by electrospinning was investigated by XPS spectroscopy. Films of polycaprolactone (PCL) and poly(l-lactide-caprolactone) [P(LLA-CL)] were prepared by electrospinning onto aluminium substrates. A modified adhesion peptide containing a photoreactive group was immobilized on the surface of the polymer nanofibers by incubation in peptide solution and subsequent UV irradiation. XPS analysis yield evidence of successful peptide immobilization on the polymer surface; the amount of immobilized peptide increases with the concentration of the mother solution.

  13. Adhesion Peptide Immobilization on Electrospun Polymers: a Photoelectron Spectroscopy Study

    SciTech Connect

    Iucci, G.; Polzonetti, G.; Ghezzo, F.; Modesti, M.; Roso, M.; Dettin, M.

    2010-06-02

    The immobilization of an oligopeptide, mimicking the adhesion sequence of fibronectin, on the surface of polymer films prepared by electrospinning was investigated by XPS spectroscopy. Films of polycaprolactone (PCL) and poly(l-lactide-caprolactone)[P(LLA-CL)] were prepared by electrospinning onto aluminium substrates. A modified adhesion peptide containing a photoreactive group was immobilized on the surface of the polymer nanofibers by incubation in peptide solution and subsequent UV irradiation. XPS analysis yield evidence of successful peptide immobilization on the polymer surface; the amount of immobilized peptide increases with the concentration of the mother solution.

  14. Saccharides as Prospective Immobilizers of Nucleic Acids for Room-Temperature Structural EPR Studies.

    PubMed

    Kuzhelev, Andrey A; Shevelev, Georgiy Yu; Krumkacheva, Olesya A; Tormyshev, Victor M; Pyshnyi, Dmitrii V; Fedin, Matvey V; Bagryanskaya, Elena G

    2016-07-01

    Pulsed dipolar electron paramagnetic resonance (EPR) spectroscopy is a powerful tool for structural studies of biomolecules and their complexes. This method, whose applicability has been recently extended to room temperatures, requires immobilization of the studied biosystem to prevent averaging of dipolar couplings; at the same time, the modification of native conformations by immobilization must be avoided. In this work, we provide first demonstration of room-temperature EPR distance measurements in nucleic acids using saccharides trehalose, sucrose, and glucose as immobilizing media. We propose an approach that keeps structural conformation and unity of immobilized double-stranded DNA. Remarkably, room-temperature electron spin dephasing time of triarylmethyl-labeled DNA in trehalose is noticeably longer compared to previously used immobilizers, thus providing a broader range of available distances. Therefore, saccharides, and especially trehalose, can be efficiently used as immobilizers of nucleic acids, mimicking native conditions and allowing wide range of structural EPR studies at room temperatures. PMID:27320083

  15. Comperative study of catalase immobilization on chitosan, magnetic chitosan and chitosan-clay composite beads.

    PubMed

    Başak, Esra; Aydemir, Tülin; Dinçer, Ayşe; Becerik, Seda Çınar

    2013-12-01

    Catalase was immobilized on chitosan and modified chitosan. Studies were carried out on free-immobilized catalase concerning the determination of optimum temperature, pH, thermal, storage stability, reusability, and kinetic parameters. Optimum temperature and pH for free catalase and catalase immobilized were found as 35°C and 7.0, respectively. After 100 times of repeated tests, the immobilized catalases on chitosan-clay and magnetic chitosan maintain over 50% and 60% of the original activity, respectively. The ease of catalase immobilization on low-cost matrices and good stability upon immobilization in the present study make it a suitable product for further use in the food industry.

  16. [Study on gallic acid preparation by using immobilized tannase from Aspergillus niger].

    PubMed

    Guo, L H; Yang, S K

    2000-09-01

    Immobilized tannase which had higher ability of transforming tannin from Chinese gallotannin into GA was prepared by embedding tannase with calcium alginate carrier. The immobilization conditions and some properties of the immobilized tannase were studied. The results showed that: The optimal immobilized parametres were 90 mg sodium alginate, 546 u(182 u/mL) tannase and 1%-2% CaCl2. The optimum temperature and pH value of immobilized tannase were 45 degrees C and 6.5, respectively. It was stable in the temperature range of 10-50 degrees C and between pH5-7. Based on the optimum program, the laboratory gramme-grade preparation experiments of GA were done. The average yield of GA product in three preparations was upto 61%, and was about equivalent to the yield of acidic hydrolytic method which is used to produce GA industrially now. Thus, the research results possessed potential applied value.

  17. Biosorption of metal contaminants using immobilized biomass--Field studies

    SciTech Connect

    Jeffers, T.H.; Bennett, P.G.; Corwin, R.R.

    1993-01-01

    The US Bureau of Mines has developed porous beads containing immobilized biological materials such as sphagnum peat moss for extracting metal contaminants from waste waters. The beads, designated as BIO-FIX beads, have removed toxic metals from over 100 waters in laboratory tests. These waters include acid mine drainage (AMD) water from mining sites, metallurgical and chemical industry waste water, and contaminated ground water. Following the laboratory studies, cooperative field tests were conducted to evaluate the metal adsorption properties of the beads in column and low-maintenance circuits, determine bead stability in varied climatic situations, and demonstrate the beads' potential as a viable waste water treatment technique. Field results indicated that BIO-FIX beads readily adsorbed cadmium, lead, and other toxic metals from dilute waters; effluents frequently met drinking water standards and other discharge criteria. The beads exhibited excellent handling characteristics in both column and low-maintenance circuits, and continued to extract metal ions after repeated loading-elution cycles. Based on laboratory and field data, cost evaluations for using BIO-FIX technology to treat two AMD waters were prepared. Operating costs for BIO-FIX treatment, which ranged from $1.40 to $2.30 per 1,000 gal of water treated, were comparable with chemical precipitation costs.

  18. Studies of Immobilized Homogeneous Metal Catalysts on Silica Supports

    SciTech Connect

    Keith James Stanger

    2003-05-31

    The tethered, chiral, chelating diphosphine rhodium complex, which catalyzes the enantioselective hydrogenation of methyl-{alpha}-acetamidocinnamate (MAC), has the illustrated structure as established by {sup 31}P NMR and IR studies. Spectral and catalytic investigations also suggest that the mechanism of action of the tethered complex is the same as that of the untethered complex in solution. The rhodium complexes, [Rh(COD)H]{sub 4}, [Rh(COD){sub 2}]{sup +}BF{sub 4}{sup -}, [Rh(COD)Cl]{sub 2}, and RhCl{sub 3} {center_dot} 3H{sub 2}O, adsorbed on SiO{sub 2} are optimally activated for toluene hydrogenation by pretreatment with H{sub 2} at 200 C. The same complexes on Pd-SiO{sub 2} are equally active without pretreatments. The active species in all cases is rhodium metal. The catalysts were characterized by XPS, TEM, DRIFTS, and mercury poisoning experiments. Rhodium on silica catalyzes the hydrogenation of fluorobenzene to produce predominantly fluorocyclohexane in heptane and 1,2-dichloroethane solvents. In heptane/methanol and heptane/water solvents, hydrodefluorination to benzene and subsequent hydrogenation to cyclohexane occurs exclusively. Benzene inhibits the hydrodefluorination of fluorobenzene. In DCE or heptane solvents, fluorocyclohexane reacts with hydrogen fluoride to form cyclohexene. Reaction conditions can be chosen to selectively yield fluorocyclohexane, cyclohexene, benzene, or cyclohexane. The oxorhenium(V) dithiolate catalyst [-S(CH{sub 2}){sub 3}s-]Re(O)(Me)(PPh{sub 3}) was modified by linking it to a tether that could be attached to a silica support. Spectroscopic investigation and catalytic oxidation reactivity showed the heterogenized catalyst's structure and reactivity to be similar to its homogeneous analog. However, the immobilized catalyst offered additional advantages of recyclability, extended stability, and increased resistance to deactivation.

  19. Study of the efficiency of immobilized algal technology for wastewater treatment.

    PubMed

    Kanherkar, S V; Late, A M; Nalawade, P M; Bhosaleanda, B J; Dhapate, S A

    2012-01-01

    The present paper deals with the study of efficiency of immobilized algal technology in wastewater treatment. The acclimatized algal species and wastewater samples were collected from three different sampling sites such as Kham River, Waluj [MIDC], Salim Ali Lake for the study. The encapsulation of collected algal species such as Spirogyra, Cyanobacteria in mixture form and Arthospira from selected sampling sites were made in sodium alginate for wastewater treatment. The percentage efficiency of immobilized algal technology for wastewater treatment was studied with respect to physico-chemical parameters. The physicochemical parameters were analyzed before and after treatment and compared for percentage efficiency study. The results obtained from present investigation reveal that the immobilized algal technology has maximum percentage efficiency in reduction of BOD, COD, Total Hardness, Total Alkalinity, Chloride. TSS, TDS and TSS. Whereas, the minimum fluctuations were found in pH and temperature. However, the immobilized algal technology is also useful to increase the dissolved oxygen concentration in wastewater treatment process.

  20. Studies on the surface coat of Paramecium aurelia. II. Relationship to the immobilization antigen.

    PubMed

    Wyroba, E

    1977-07-11

    Correlations between the presence of surface coat and immobilization antigen of Paramecium tetraurelia were studied. Supravital, partial removal of the surface coat resulted in accelerated response of monobacterially and axenically grown cells to the homologous antiserum. Ciliates pretreated with trypsin or pronase (0.5 mg/ml for 45 min at 0-4 degrees C) were immobilized approximately twice as fast as untreated control cells. The probable localization of at least part, of the immobilization antigen within the surface coat of P. tetraurelia is discussed.

  1. Spectroscopy study of Zn, Cd, Pb and Cr ions immobilization on C-S-H phase.

    PubMed

    Żak, Renata; Deja, Jan

    2015-01-01

    Calcium silicate hydrates (C-S-H) have a large number of structural sites available for cations and anions to bind. The C-S-H phases are materials which have ability to toxic ions immobilization. Immobilization mechanisms for C-S-H include sorption, phase mixing, substitution and precipitation of insoluble compounds. This study presents the C-S-H (prepared with C/S ratios 1.0) phase as absorbent for immobilization of Zn, Cd, Pb and Cr ions. The C-S-H spectra before and after incorporation of heavy metals ions into the C-S-H structure were obtained. The effect of added heavy metals ions on the hydration phenomena was studied by means of X-ray diffractions analysis. FTIR spectra was measured. The microstructure and phase composition of C-S-H indicate that they can play an essential role in the immobilization of heavy metals. The properties of C-S-H in the presence of Zn, Cd, Pb and Cr cations were studied. The leaching ML test was used to evaluate the level of immobilization of heavy metals in C-S-H. The leached solutions are diluted and analyzed using atomic absorption spectrometry (AAS) and the activated solid particles are separated, washed, desiccated and analyzed by Fourier transform infrared (FTIR) spectroscopy. It was found that the degree of Cd, Zn, Pb and Cr cations immobilization was very high (exceeding 99.96%).

  2. Functionalized Graphene Sheets As Immobilization Matrix for Fenugreek β-Amylase: Enzyme Kinetics and Stability Studies

    PubMed Central

    Srivastava, Garima; Singh, Kritika; Talat, Mahe; Srivastava, Onkar Nath; Kayastha, Arvind M.

    2014-01-01

    β-Amylase finds application in food and pharmaceutical industries. Functionalized graphene sheets were customised as a matrix for covalent immobilization of Fenugreek β-amylase using glutaraldehyde as a cross-linker. The factors affecting the process were optimized using Response Surface Methodology based Box-Behnken design of experiment which resulted in 84% immobilization efficiency. Scanning and Transmission Electron Microscopy (SEM, TEM) and Fourier Tansform Infrared (FTIR) spectroscopy were employed for the purpose of characterization of attachment of enzyme on the graphene. The enzyme kinetic studies were carried out for obtaining best catalytic performance and enhanced reusability. Optimum temperature remained unchanged, whereas optimum pH showed shift towards acidic range for immobilized enzyme. Increase in thermal stability of immobilized enzyme and non-toxic nature of functionalized graphene can be exploited for production of maltose in food and pharmaceutical industries. PMID:25412079

  3. Immobilized low-level waste disposal options configuration study

    SciTech Connect

    Mitchell, D.E.

    1995-02-01

    This report compiles information that supports the eventual conceptual and definitive design of a disposal facility for immobilized low-level waste. The report includes the results of a joint Westinghouse/Fluor Daniel Inc. evaluation of trade-offs for glass manufacturing and product (waste form) disposal. Though recommendations for the preferred manufacturing and disposal option for low-level waste are outside the scope of this document, relative ranking as applied to facility complexity, safety, remote operation concepts and ease of retrieval are addressed.

  4. Effectiveness of pressure-immobilization first aid for snakebite requires further study.

    PubMed

    Currie, Bart J; Canale, Elizabeth; Isbister, Geoffrey K

    2008-06-01

    In the prospective Royal Darwin Hospital snakebite study, pressure-immobilization first aid (PI) was used more often than in previous studies. However, bandages were not uncommonly too loose or not applied to the whole limb and immobilization was often neglected. While PI should continue to be promoted as the standard for Australia for the present, prospective multicentre studies of snakebite with quantitative assays for blood venom concentration will hopefully better elucidate the real effectiveness of PI and define the limitations of timing of application and determine the optimum types of bandage materials to use and the pressure required to be maintained.

  5. Gamma radiation grafted polymers for immobilization of Brucella antigen in diagnostic test studies

    NASA Astrophysics Data System (ADS)

    Docters, E. H.; Smolko, E. E.; Suarez, C. E.

    The radiation grafting process has a wide field of industrial applications, and in the recent years the immobilization of biocomponents in grafted polymeric materials obtained by means of ionizing radiations is a new and important contribution to biotechnology. In the present work, gamma preirradiation grafting method was employed to produce acrylics hydrogels onto polyethylene (PE), polyvinyl chloride (PVC) and polystyrene (PS). Two monomers were used to graft the previously mentioned polymers: methacrylic acid (MAAc) and acrylamide (AAm), and several working conditions were considered as influencing the degree of grafting. All this grafted polymers were used to study the possibility of a subsequent immobilization of Brucella antigen (BAg) in diagnostic test studies (ELISA).

  6. Comparison between Free and Immobilized Ion Effects on Hydrophobic Interactions: A Molecular Dynamics Study.

    PubMed

    Huang, Kai; Gast, Sebastian; Ma, C Derek; Abbott, Nicholas L; Szlufarska, Izabela

    2015-10-15

    Fundamental studies of the effect of specific ions on hydrophobic interactions are driven by the need to understand phenomena such as hydrophobically driven self-assembly or protein folding. Using β-peptide-inspired nanorods, we investigate the effects of both free ions (dissolved salts) and proximally immobilized ions on hydrophobic interactions. We find that the free ion effect is correlated with the water density fluctuation near a nonpolar molecular surface, showing that such fluctuation can be an indicator of hydrophobic interactions in the case of solution additives. In the case of immobilized ion, our results demonstrate that hydrophobic interactions can be switched on and off by choosing different spatial arrangements of proximal ions on a nanorod. For globally amphiphilic nanorods, we find that the magnitude of the interaction can be further tuned using proximal ions with varying ionic sizes. In general, univalent proximal anions are found to weaken hydrophobic interactions. This is in contrast to the effect of free ions, which according to our simulations strengthen hydrophobic interactions. In addition, immobilized anions of increasing ionic size do not follow the same ordering (Hofmeister-like ranking) as free ions when it comes to their impact on hydrophobic interactions. The immobilized ion effect is not simply correlated with the water density fluctuation near the nonpolar side of the amphiphilic nanorod. We propose a molecular picture that explains the contrasting effects of immobilized versus free ions.

  7. Comparative study of 6-APA production by free and agar immobilized bacteria in nutrient broth culture.

    PubMed

    Dolui, A K; Das, S

    2011-04-01

    In the present study different bacterial samples were isolated from soil of different places of Dibrugarh and screened for biotransformation ability to produce 6-Aminopenicillanic acid. Among ten isolated bacterial samples, three gram positive bacterial samples designated as AKDD-2, AKDD-4 and AKDD-6 showed the production of 6-APA from penicillin G. Assessment of production of 6-APA after incubation in penicillin G (2 mg/ml) by three different samples separately in free and agar immobilization state was done by HPLC analysis. Reusability of immobilized cells was found successful up to 14 days. PMID:21614893

  8. Comparative study of 6-APA production by free and agar immobilized bacteria in nutrient broth culture.

    PubMed

    Dolui, A K; Das, S

    2011-04-01

    In the present study different bacterial samples were isolated from soil of different places of Dibrugarh and screened for biotransformation ability to produce 6-Aminopenicillanic acid. Among ten isolated bacterial samples, three gram positive bacterial samples designated as AKDD-2, AKDD-4 and AKDD-6 showed the production of 6-APA from penicillin G. Assessment of production of 6-APA after incubation in penicillin G (2 mg/ml) by three different samples separately in free and agar immobilization state was done by HPLC analysis. Reusability of immobilized cells was found successful up to 14 days.

  9. Study of bone mineral metabolism. [during body immobilization, bed rest, and space flight

    NASA Technical Reports Server (NTRS)

    Palmer, H. E.

    1975-01-01

    The use of Sr-85 as an indicator of the skeletal location and relative amount of bone demineralization which occurs during immobilization of the body or body parts, bed-rest or space flight was studied. The bone mineral replacement which occurs after immobilization was measured rather than the bone loss which occurs during immobilization. In a study with two adult beagle dogs, the Sr-85 uptake in a leg which had been immobilized for two months was 400 percent higher than the uptake in the legs in regular use. This increased uptake probably resulted from only a few percent loss in bone mineral and indicates that losses less than one percent can be easily detected and located. The sensitivity, simplicity, and low radiation dose associated with the use of this method indicates that it should receive consideration for use on humans in bed-rest and space flight studies. Methods for measuring changes in total body nitrogen and in assisting the Johnson Space Center in calibrating a whole body counter for total body potassium measurements were also investigated.

  10. An immunohistochemical study of the sciatic nerve in a rat knee immobilization model

    PubMed Central

    Yoshida, Shinya; Matsuzaki, Taro; Hoso, Masahiro

    2016-01-01

    [Purpose] This study was performed to immunohistochemically evaluate changes in the periphery of the sciatic nerve in a rat model of knee immobilization, and to assess the effects of range of motion exercise. [Subjects and Methods] Twenty-one male rats were divided randomly into three groups: control (C), immobilized (I), and exercise (E group). Rats in the I and E groups had the right knee joint immobilized for 2 weeks. In the E group, range of motion exercise was also performed. After the experimental period, the periphery of the sciatic nerve was immunohistochemically observed. [Results] Immunohistochemical staining revealed that the myelin sheath and the perineurium in all groups were laminin positive. In the C and E groups, all rats showed normal staining. In contrast, 4 rats in the I group exhibited weak labeling. [Conclusion] Our results suggest that immobilization alters the perineurium at a molecular level and the range of motion exercise is essential for maintaining the environment of the perineurium. PMID:27190437

  11. Mechanistic study for immobilization of cysteine-labeled oligopeptides on UV-activated surfaces.

    PubMed

    Ong, Lian Hao; Ding, Xiaokang; Yang, Kun-Lin

    2014-10-01

    In this study, we report immobilization of cysteine-labeled oligopeptides on UV activated surfaces decorated with N,N-dimethyl-n-octadecyl-3-aminopropyltrimethoxysilyl chloride (DMOAP). Our result shows that cysteine group, regardless of its position in the oligopeptide, is essential for successful immobilization of oligopeptide on the UV-activated surface. A possible reaction mechanism is nucleophilic addition of thiolates to surface aldehyde groups generated during UV activation. By using this technique, we are able to incorporate anchoring points into oligopeptides through cysteine residues. Furthermore, immobilized oligopeptides on the UV-activated surface is very stable even under harsh washing conditions. Finally, we show that an HPQ-containing oligopeptide can be immobilized on the UV-activated surface, but the final surface density and its ability to bind streptavidin are affected by the position of cysteine and HPQ. An oligopeptide with a cysteine at the N-terminus and a HPQ motif at the C-terminus gives the highest binding signal in the streptavidin-binding assay. This result is potentially useful for the development of functional oligopeptide microarrays for detecting target protein molecules.

  12. [Study on immobilized cells for producing alpha-amylase by using polyving alcohol as the carrier(II): The effect of fermentating conditions on the ability producing alpha-amylase of the cells immobilized with polyving alcohol as the corrier and continuous fermentation of the immobilized cells in CSTR].

    PubMed

    Liu, Z; Wang, J; Li, Z

    1998-03-01

    The effects of fermentating conditions on the ability of immobilized cells with PVA as carrier for producing alpha-amylase were studied. The continuous fermentation with the immobilized cells were tested in continuous flow stirred tank reactor (CSTR). The results showed that the adaptability of the immobilized Bacillus substilis to pH increased after immobilization. In CSTR, the immobilized cells can be fermentated continuously for 360 hrs and the activity of alpha-amylase can be kept on the level of about 170 u/ml.

  13. Kinetic modeling and docking study of immobilized lipase catalyzed synthesis of furfuryl acetate.

    PubMed

    Mathpati, Ashwini C; Badgujar, Kirtikumar C; Bhanage, Bhalchandra M

    2016-03-01

    The present work deals with the kinetic modeling and docking study for the furfuryl acetate synthesis using immobilized Burkholderia cepacia (BCL) lipase. Initially various lipases were immobilized on hydroxypropyl methyl cellulose (HPMC) and poly vinyl alcohol (PVA) base hybrid polymer matrix. After screening of various immobilized biocatalysts, HPMC:PVA:BCL was found to be a robust biocatalyst. Various reaction conditions were optimized using response surface methodology (RSM) based on a four-factor-three-level Box-Behnken design. The optimal conditions were obtained at molar ratio of 1:2 of furfuryl alcohol to acyl donor, temperature 50°C with catalyst loading of 30mg in 3mL of non-aqueous media toluene. Under these conditions 99.98% yield was obtained in 3h. The Arrhenius plot showed that the activation energy for furfuryl acetate synthesis was 10.68kcal/mol. The kinetics of reaction was studied close to optimized conditions which obey order bi-bi model. Molecular docking study was carried out to understand the active site of BCL which is responsible for the reaction. It was observed that the reaction proceeds via acylation of the active serine of BCL and demonstrating strong hydrogen bond between the substrate and histidine site. The catalyst recyclability study was carried up to five cycles. PMID:26827768

  14. Self-assembled monolayers for studying enzyme immobilization and ion recognition

    NASA Astrophysics Data System (ADS)

    Kang, Jie

    This thesis explores the use of self-assembled monolayers on gold for studying enzyme immobilization and ion recognition. Chapter 1 serves as a general introduction to biosensing, self-assembled monolayers, protein immobilization, and surface characterization techniques. Chapter 2 through Chapter 5 describe the immobilization of a redox enzyme, glucose oxidase, to a variety of functional self-assembled monolayers by either noncovalent adsorption or covalent attachment. The characteristics of different immobilization methods are investigated, and the activity of the immobilized enzyme is assessed electrochemically. Chapter 2 presents detailed procedures for measuring glucose oxidase activity by an electrochemical technique---cyclic voltammetry. Chapter 3 describes the adsorption of glucose oxidase to hydrophobic and hydrophilic self-assembled monolayers (SAMs). Significant glucose oxidase adsorption to hydrophobic, methyl-terminated SAMs was observed, while long chain, hydrophilic SAMs terminated by hydroxyl and carboxyl groups resist enzyme adsorption. Chapter 4 examines the covalent attachment of glucose oxidase to N-hydroxysuccinimide ester (NHS ester)-terminated self-assembled monolayers. The reactivity of the surface NHS ester group is found to increase as its coverage is lowered. This observation is explained by the steric effect. Chapter 5 reports the electrostatic adsorption of glucose oxidase to self-assembled monolayers of cystamine. The adsorbed enzyme shows superior activity to enzyme immobilized by other means. The rate constants of surface enzyme catalysis are determined and compared with those of the enzyme in solution. Chapter 6 is concerned with iron (III) recognition by a self-assembled monolayer terminated with a siderophore group, desferrioxamine (H3DFO). We first demonstrate that the iron coverage of the ferrioxamine (FeDFO)-terminated SAM can be successfully assayed by cyclic voltammetry. We then present results for iron (III) binding to the H3

  15. Hydrated lime for metals immobilization and explosives transformation: Treatability study.

    PubMed

    Martin, W Andy; Larson, S L; Nestler, C C; Fabian, G; O'Connor, G; Felt, D R

    2012-05-15

    Fragmentation grenades contain Composition B (RDX and TNT) within a steel shell casing. There is the potential for off-site migration of high explosives and metals from hand grenade training ranges by transport in surface water and subsurface transport in leachate. This treatability study used bench-scale columns and mesocosm-scale laboratory lysimeters to investigate the potential of hydrated lime as a soil amendment for in situ remediation of explosives and metals stabilization in hand grenade range soils. Compared to the unamended soil there was a 26-92% reduction of RDX in the leachate and runoff water from the lime treated soils and a 66-83% reduction of zinc in the leachate and runoff water samples; where the hand grenade range metals of concern were zinc, iron, and manganese. The amended soil was maintained at the target pH of greater than 10.5 for optimum explosives decomposition. The treatability study indicated a high potential of success for scale-up to an in situ field study. PMID:22445717

  16. [Applied study of the submerged macrophytes bed-immobilized bacteria in drinking water restoration].

    PubMed

    Chen, Qi-Chun; Li, Zheng-Kui; Wang, Yi-Chao; Wu, Kai; Fan, Nian-Wen

    2012-01-01

    The effect of submerged macrophytes bed-immobilized bacteria technology which applied in drinking water restoration was studied. Ammonifying bacteria, nitrobacteria, nitrosobacteria and denitrifying bacteria which isolated from Taihu Labe was immobilized to the porous carries, combined with the submerged macrophytes bed technology, we applied the new equipment in water restoration of gonghu bay, this equipment has good ability to resist storm, the denitrifying bacteria number increased from 5.4 x 10(2)-2.7 x 10(3) to 3.9 x 10(5)-9.1 x 10(5), N2O flux of experimental plot was 3-24 microg x (m2 x h)(-1), it's more than the contrast group obviously, TN concentration reduced 19% - 74%, while NO3- concentration reduced 24% -81% after the equipment running a period of time; The experimental data of 120 days showed that this technology is suitable for drinking water restoration, as it can control eutrophication.

  17. Subsurface Bio-Immobilization of Plutonium: Experiment and Model Validation Study

    SciTech Connect

    Reed, Donald; Rittmann, Bruce

    2006-06-01

    The goal of this project is to conduct a concurrent experimental and modeling study centered on the interactions of Shewanella algae BrY with plutonium and uranium species and phases. The most important objective of this research is to investigate the long-term stability of bioprecipitated immobilized actinide phases under changing redox conditions in biologically active systems. The long-term stability of bio-immobilized actinides (e.g. by bio-reduction) is a key criteria that defines the utility and effectiveness of a remediation/containment strategy for subsurface actinide contaminants. Plutonium, which is the focus of this project, is the key contaminant of concern at several DOE sites.

  18. HPLC study on the stability of bendamustine hydrochloride immobilized onto polyphosphoesters.

    PubMed

    Pencheva, Ivanka; Bogomilova, Anita; Koseva, Neli; Obreshkova, Danka; Troev, Kolio

    2008-12-01

    Novel water soluble polymer complexes of bendamustine hydrochloride, a bifunctional alkylating agent with antimetabolic and cytotoxic activity, were developed using biodegradable polymer carriers-poly(oxyethylene H-phosphonate), poly(methyloxyethylene phosphate) and poly(hydroxyoxyethylene phosphate). Bendamustine hydrochloride was immobilized onto polyphosphoesters via covalent, ionic and hydrogen bonding. The structure of the complexes formed was elucidated by (1)H, (13)C, (31)P NMR and FT-IR spectroscopy. The chemical stability of bendamustine hydrochloride in the novel complexes was studied by HPLC analysis based on a validated method with appointed analytical parameters such as specificity, repeatability, limit of quantitation, limit of detection and linearity. The results from the HPLC indicate that in neutral (pH 7) and alkaline (pH 9) media bendamustine hydrochloride in the polymer complexes is more stable than the pure bendamustine hydrochloride. The enhanced stability of the immobilized drug is explained with the drug interaction with the polymer carriers or their degradation products.

  19. Comparative study of free and immobilized lipase from Bacillus aerius and its application in synthesis of ethyl ferulate.

    PubMed

    Saun, Nitin Kumar; Narwal, Sunil Kumar; Dogra, Priyanka; Chauhan, Ghanshyam Singh; Gupta, Reena

    2014-01-01

    In the present study, a purified lipase from Bacillus aerius immobilized on celite matrix was used for synthesis of ethyl ferulate. The celite-bound lipase exposed to glutaraldehyde showed 90.02% binding efficiency. It took two hours to bind maximally onto the support. The pH and temperature optima of the immobilized lipase were same as those of free enzyme i.e 9.5 and 55°C. Among different substrates both free and immobilized lipase showed maximum affinity towards p-nitrophenyl palmitate (p-NPP). The lipase activity was found to be stimulated in the presence of Mg(2+) in case of free enzyme while Zn(2+) and Fe(3+) showed stimulatory effect on immobilized lipase whereas salt ions as well as chelating agents inhibited activity of both free and immobilized lipase. Maximum enzyme activity was observed in n-hexane as organic solvent followed by n-heptane for both free and immobilized lipase, however CCl4, acetone and benzene inhibited the enzyme activity. Moreover, all the selected detergents (SDS, Triton X-100, Tween 80 and Tween 20) had an inhibitory effect on both free and immobilized enzyme activity. The celite bound lipase (1.5%) efficiently performed maximum esterification (2.51 moles/l) of ethanol and ferulic acid (100 mM each, at a molar ratio of 1:3) when incubated at 55°C for 48 h resulting in the formation of ester ethyl ferulate. PMID:25099909

  20. Study on immobilization of yeast alcohol dehydrogenase on nanocrystalline Ni-Co ferrites as magnetic support.

    PubMed

    Shakir, Mohammad; Nasir, Zeba; Khan, Mohd Shoeb; Lutfullah; Alam, Md Fazle; Younus, Hina; Al-Resayes, Saud Ibrahim

    2015-01-01

    The covalent binding of yeast alcohol dehydrogenase (YADH) enzyme complex in a series of magnetic crystalline Ni-Co nanoferrites, synthesized via sol-gel auto combustion technique was investigated. The structural analysis, morphology and magnetic properties of Ni-Co nanoferrites were determined by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), vibrating-sample magnetometer (VSM), high resolution transmission electron microscopy (HRTEM) and Fourier transform infrared spectroscopy (FTIR). The comparative analysis of the HRTEM micrographs of bare magnetic nanoferrite particles and particles immobilized with enzyme revealed an uniform distribution of the particles in both the cases without undergoing change in the size which was found to be in the range 20-30 nm. The binding of YADH to Ni-Co nanoferrites and the possible binding mechanism have been suggested by comparing the FTIR results. The binding properties of the immobilized YADH enzyme were also studied by kinetic parameters, optimum operational pH, temperature, thermal stability and reusability. The immobilized YADH exhibits enhanced thermal stability as compared to the free enzyme over a wide range of temperature and pH, and showed good durability after recovery by magnetic separation for repeated use.

  1. Study on immobilization of yeast alcohol dehydrogenase on nanocrystalline Ni-Co ferrites as magnetic support.

    PubMed

    Shakir, Mohammad; Nasir, Zeba; Khan, Mohd Shoeb; Lutfullah; Alam, Md Fazle; Younus, Hina; Al-Resayes, Saud Ibrahim

    2015-01-01

    The covalent binding of yeast alcohol dehydrogenase (YADH) enzyme complex in a series of magnetic crystalline Ni-Co nanoferrites, synthesized via sol-gel auto combustion technique was investigated. The structural analysis, morphology and magnetic properties of Ni-Co nanoferrites were determined by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive X-ray spectroscopy (EDS), vibrating-sample magnetometer (VSM), high resolution transmission electron microscopy (HRTEM) and Fourier transform infrared spectroscopy (FTIR). The comparative analysis of the HRTEM micrographs of bare magnetic nanoferrite particles and particles immobilized with enzyme revealed an uniform distribution of the particles in both the cases without undergoing change in the size which was found to be in the range 20-30 nm. The binding of YADH to Ni-Co nanoferrites and the possible binding mechanism have been suggested by comparing the FTIR results. The binding properties of the immobilized YADH enzyme were also studied by kinetic parameters, optimum operational pH, temperature, thermal stability and reusability. The immobilized YADH exhibits enhanced thermal stability as compared to the free enzyme over a wide range of temperature and pH, and showed good durability after recovery by magnetic separation for repeated use. PMID:25450541

  2. High pressure studies on hesperitin production with hesperidinase free and immobilized in calcium alginate beads

    NASA Astrophysics Data System (ADS)

    Furtado, Andreia; Rosário, Pedro M.; Calado, António R. T.; Alfaia, António J. I.; Ribeiro, Maria H. L.

    2012-03-01

    The use of high pressure for the enzymatic synthesis of pharmacologically interesting molecules is a very important tool. Hesperidin and hesperitin exhibit anti-inflammatory, antimicrobial, antioxidant, and anticarcinogenic properties and prevent bone loss. However, hesperidin has a low bioavailability compared with hesperitin, due to the rutinoside moiety attached to the flavonoid. The aim of this work was the enzymatic production of hesperitin from hesperidin (soluble and insoluble) with hesperidinase free and immobilized in Ca-alginate beads, under high pressure conditions. The work was focused on the optimization of enzyme activity, studying the effects: pressure (50-150 MPa), temperature (35-75 °C), concentration of substrate (100-800 mg/L), and immobilization of hesperidinase. An 18-fold increase in hesperidinase residual activity was observed under high pressure conditions of 100 MPa compared to 0.1 MPa. A higher specificity of the hydrolytic reaction under high pressure (100 MPa) with a two-and three-fold increase in the ratio K cat/K M (specificity constant) at 55 °C and 75 °C was observed. A two-fold increase in the maximum activity at 100 MPa was observed with immobilized hesperinase compared to 0.1 MPa. In the second reutilization, almost a four-fold increase was obtained under high pressure conditions in comparison to atmospheric pressure.

  3. [Study on influence between activated carbon property and immobilized biological activated carbon purification effect].

    PubMed

    Wang, Guang-zhi; Li, Wei-guang; He, Wen-jie; Han, Hong-da; Ding, Chi; Ma, Xiao-na; Qu, Yan-ming

    2006-10-01

    By means of immobilizing five kinds of activated carbon, we studied the influence between the chief activated carbon property items and immobilized bioactivated carbon (IBAC) purification effect with the correlation analysis. The result shows that the activated carbon property items which the correlation coefficient is up 0.7 include molasses, abrasion number, hardness, tannin, uniform coefficient, mean particle diameter and effective particle diameter; the activated carbon property items which the correlation coefficient is up 0.5 include pH, iodine, butane and tetrachloride. In succession, the partial correlation analysis shows that activated carbon property items mostly influencing on IBAC purification effect include molasses, hardness, abrasion number, uniform coefficient, mean particle diameter and effective particle diameter. The causation of these property items bringing influence on IBAC purification is that the activated carbon holes distribution (representative activated carbon property item is molasses) provides inhabitable location and adjust food for the dominance bacteria; the mechanical resist-crash property of activated carbon (representative activated carbon property items: abrasion number and hardness) have influence on the stability of biofilm; and the particle diameter size and distribution of activated carbon (representative activated carbon property items: uniform coefficient, mean particle diameter and effective particle diameter) can directly affect the force of water in IBAC filter bed, which brings influence on the dominance bacteria immobilizing on activated carbon.

  4. Study on changes in skin extensibility during the development of joint contracture due to joint immobilization in rats

    PubMed Central

    Tasaka, Atsushi; Ono, Takeya; Oki, Sadaaki; Umei, Namiko; Ishikura, Hideki; Aihara, Kazuki; Sato, Yuta; Matsumoto, Tomohiro

    2015-01-01

    [Purpose] The purpose of this study was to elucidate whether skin extensibility decreases when a contracture develops as a result of joint immobilization. [Subjects] This study was conducted on six female Wistar rats. [Methods] The rats were divided into two experimental groups. In the immobilized group, the right ankle joints were immobilized in complete plantar flexion by plaster casts for two weeks. In the control group, the left ankle joints had no intervention. On the final day, skin extensibility was determined from a length-tension curve by collecting skin from the posterior aspect of the ankle joint and using a tensile strength tester. [Results] Compared with the control group, the immobilized group showed a significant decrease in skin extensibility. [Conclusion] The results demonstrated that the extensibility of the skin itself decreases when joint contracture develops. PMID:26504268

  5. An efficient TiO2 coated immobilized system for the degradation studies of herbicide isoproturon: durability studies.

    PubMed

    Verma, A; Prakash, N T; Toor, A P

    2014-08-01

    The investigation presents the observations on the use of cement beads for the immobilization of TiO2 for the degradation of herbicide isoproturon. The immobilized system was effective in degrading and mineralizing the herbicide for continuous thirty cycles without losing its durability. Catalyst was characterized by SEM-EDAX for checking the durability of the catalyst. The degradation rate followed first order kinetics as measured by change in absorption intensity in UV range as well as HPLC analysis. Two rounds of TiO2 coating on inert cement beads with average diameter 1.5cm at UV Intensity 25Wm(-2) calcined at 400°C were the optimized conditions for the degradation of herbicide isoproturon. More than 90% TOC and COD reduction along with ammonium ions generation (80%) confirmed the mineralization of isoproturon. Fixed bed baffled reactor studies under solar irradiations using the TiO2 immobilized beads confirmed 85% degradation after 6h. LC-MS studies confirmed the intermediates formation and their subsequent degradation using immobilized system.

  6. Kinetic studies on degradation of Reactive Red 120 dye in immobilized packed bed reactor by Bacillus cohnii RAPT1.

    PubMed

    Padmanaban, V C; Geed, Sachin RameshRao; Achary, Anant; Singh, R S

    2016-08-01

    The degradation of Reactive Red 120 using Bacillus cohnii RAPT1 immobilized on polyurethane was studied. Initial experiments indicated that the percentage removal of dye in immobilized batch was significantly higher than batch (without immobilization). The optimum process parameters such as effect of dye concentration, time of immobilization on Poly Urethane Foam, initial inoculum size, pH and temperature for removal of dye were investigated and was found as 200ppm, 36h, 300*10(6) colony forming units/ml, 8.0 and 35°C respectively. Under optimum conditions, 100% removal of dye was obtained within 4h. The kinetics of biodegradation for the batch with free cells and immobilised packed batch was found to be IInd order with kinetic constant and initial rate of reaction as 0.0408, 0.084L/(mgday) and 1632, 3360 (mg/Lday) respectively. PMID:26968121

  7. BIOINTERACTION ANALYSIS BY HIGH-PERFORMANCE AFFINITY CHROMATOGRAPHY: KINETIC STUDIES OF IMMOBILIZED ANTIBODIES

    PubMed Central

    Nelson, Mary Anne; Moser, Annette; Hage, David S.

    2009-01-01

    A system based on high-performance affinity chromatography was developed for characterizing the binding, elution and regeneration kinetics of immobilized antibodies and immunoaffinity supports. This information was provided by using a combination of frontal analysis, split-peak analysis and peak decay analysis to determine the rate constants for antibody-antigen interactions under typical sample application and elution conditions. This technique was tested using immunoaffinity supports that contained monoclonal antibodies for 2,4-dichlorophenoxyacetic acid (2,4-D). Association equilibrium constants measured by frontal analysis for 2,4-D and related compounds with the immobilized antibodies were 1.7–12 × 106 M−1 at pH 7.0 and 25°C. Split-peak analysis gave association rate constants of 1.4–12 × 105 M−1s−1 and calculated dissociation rate constants of 0.01–0.4 s−1 under the application conditions. Elution at pH 2.5 for the analytes from the antibodies was examined by peak decay analysis and gave dissociation rate constants of 0.056–0.17 s−1. A comparison of frontal analysis results after various periods of column regeneration allowed the rate of antibody regeneration to be examined, with the results giving a first-order regeneration rate constant of 2.4 × 10−4 s−1. This combined approach and the information it provides should be useful in the design and optimization of immunoaffinity chromatography and other analytical methods that employ immobilized antibodies. The methods described are not limited to the particular analytes and antibodies employed in this study but should be useful in characterizing other targets, ligands and supports. PMID:19394281

  8. Modification of silicon nitride surfaces with GOPES and APTES for antibody immobilization: computational and experimental studies

    NASA Astrophysics Data System (ADS)

    Dien To, Thien; Nguyen, Anh Tuan; Nhat Thanh Phan, Khoa; Thu Thi Truong, An; Doan, Tin Chanh Duc; Mau Dang, Chien

    2015-12-01

    Chemical modification of silicon nitride (SiN) surfaces by silanization has been widely studied especially with 3-(aminopropyl)triethoxysilane (APTES) and 3-(glycidyloxypropyl) dimethylethoxysilane (GOPES). However few reports performed the experimental and computational studies together. In this study, surface modification of SiN surfaces with GOPES and APTES covalently bound with glutaraldehyde (GTA) was investigated for antibody immobilization. The monoclonal anti-cytokeratin-FITC (MACF) antibody was immobilized on the modified SiN surfaces. The modified surfaces were characterized by water contact angle measurements, atomic force microscopy and fluorescence microscopy. The FITC-fluorescent label indicated the existence of MACF antibody on the SiN surfaces and the efficiency of the silanization reaction. Absorption of APTES and GOPES on the oxidized SiN surfaces was computationally modeled and calculated by Materials Studio software. The computational and experimental results showed that modification of the SiN surfaces with APTES and GTA was more effective than the modification with GOPES.

  9. Kinetics study of palm oil hydrolysis using immobilized lipase Candida rugosa in packed bed reactor.

    PubMed

    Min, C S; Bhatia, S; Kamaruddin, A H

    1999-01-01

    Continuous hydrolysis of palm oil triglyceride in organic solvent using immobilized Candida rugosa on the Amberlite MB-1 as a source of immobilized lipase was studied in packed bed reactor. The enzymatic kinetics of hydrolysis reaction was studied by changing the substrate concentration, reaction temperature and residence time(tau) in the reactor. At 55 degrees C, the optimum water concentration was found to be 15 % weight per volume of solution (%w/v). The Michaelis-Menten kinetic model was used to obtain the reaction parameters, Km(app) and V max(app). The activation energies were found to be quite low indicating that the lipase-catalyzed process is controlled by diffusion of substrates. The Michaelis-Menten kinetic model was found to be suitable at low water concentration 10-15 %w/v of solution. At higher water concentration, substrate inhibition model was used for data analysis. Reactor operation was found to play an important role in the palm oil hydrolysis kinetic.

  10. Study on Carrier Material of Immobilization Acetylcholinesterase For Biosensor in Detectionof Organophosphorus Pesticide Residues

    NASA Astrophysics Data System (ADS)

    Sun, Xia; Wang, Xiangyou; Jia, Chuandong

    A comparison between several immobilization materials of AChE on surface of glassy carbon electrode(GCE) was presented. The immobilization methods employed crosslinking method with glutaraldehyde as a cross-linking agent and bovine serum albumin(BSA) as a protectant, AChE was immobilized on different membranes including nylon membrane, cellulose nitrate membrane and chitosan membrane respectively. The enzyme membrane was then fixed on the surface of glassy carbon electrode(GCE) with O-ring to prepare an amperometric biosensor for the detection of organophosphorus pesticides. The activity of immobilization AChE was detected by measuring the oxidation current of thiocholine, the results showed that the activity of immobilization AChE were all different with different membrane as carrier material.Compared with nylon membrane and cellulose nitrate membrane, chitosan membrane was obviously good. So chitosan membrane can be selected as immobilized AChE carrier material.

  11. Spectroscopic study of 3-Hydroxyflavone - protein interaction in lipidic bi-layers immobilized on silver nanoparticles.

    PubMed

    Voicescu, Mariana; Ionescu, Sorana; Nistor, Cristina L

    2017-01-01

    The interaction of 3-Hydroxyflavone with serum proteins (BSA and HSA) in lecithin lipidic bi-layers (PC) immobilized on silver nanoparticles (SNPs), was studied by fluorescence and Raman spectroscopy. BSA secondary structure was quantified with a deconvolution algorithm, showing a decrease in α-helix structure when lipids were added to the solution. The effect of temperature on the rate of the excited-state intra-molecular proton transfer and on the dual fluorescence emission of 3-HF in the HSA/PC/SNPs systems was discussed. Evaluation of the antioxidant activity of 3-HF in HSA/PC/SNPs systems was also studied. The antioxidant activity of 3-HF decreased in the presence of SNPs. The results are discussed with relevance to the secondary structure of proteins and of the 3-HF based nano-systems to a topical formulation useful in the oxidative stress process. PMID:27380623

  12. Instrumentation for studying binder burnout in an immobilized plutonium ceramic wasteform

    SciTech Connect

    Mitchell, M; Pugh, D; Herman, C

    2000-04-21

    The Plutonium Immobilization Program produces a ceramic wasteform that utilizes organic binders. Several techniques and instruments were developed to study binder burnout on full size ceramic samples in a production environment. This approach provides a method for developing process parameters on production scale to optimize throughput, product quality, offgas behavior, and plant emissions. These instruments allow for offgas analysis, large-scale TGA, product quality observation, and thermal modeling. Using these tools, results from lab-scale techniques such as laser dilametry studies and traditional TGA/DTA analysis can be integrated. Often, the sintering step of a ceramification process is the limiting process step that controls the production throughput. Therefore, optimization of sintering behavior is important for overall process success. Furthermore, the capabilities of this instrumentation allows better understanding of plant emissions of key gases: volatile organic compounds (VOCs), volatile inorganics including some halide compounds, NO{sub x}, SO{sub x}, carbon dioxide, and carbon monoxide.

  13. Immobilized Kluyveromyces marxianus cells in carboxymethyl cellulose for production of ethanol from cheese whey: experimental and kinetic studies.

    PubMed

    Roohina, Fatemeh; Mohammadi, Maedeh; Najafpour, Ghasem D

    2016-09-01

    Cheese whey fermentation to ethanol using immobilized Kluyveromyces marxianus cells was investigated in batch and continuous operation. In batch fermentation, the yeast cells were immobilized in carboxymethyl cellulose (CMC) polymer and also synthesized graft copolymer of CMC with N-vinyl-2-pyrrolidone, denoted as CMC-g-PVP, and the efficiency of the two developed cell entrapped beads for lactose fermentation to ethanol was examined. The yeast cells immobilized in CMC-g-PVP performed slightly better than CMC with ethanol production yields of 0.52 and 0.49 g ethanol/g lactose, respectively. The effect of supplementation of cheese whey with lactose (42, 70, 100 and 150 g/l) on fermentative performance of K. marxianus immobilized in CMC beads was considered and the results were used for kinetic studies. The first order reaction model was suitable to describe the kinetics of substrate utilization and modified Gompertz model was quite successful to predict the ethanol production. For continuous ethanol fermentation, a packed-bed immobilized cell reactor (ICR) was operated at several hydraulic retention times; HRTs of 11, 15 and 30 h. At the HRT of 30 h, the ethanol production yield using CMC beads was 0.49 g/g which implies that 91.07 % of the theoretical yield was achieved.

  14. Immobilized Kluyveromyces marxianus cells in carboxymethyl cellulose for production of ethanol from cheese whey: experimental and kinetic studies.

    PubMed

    Roohina, Fatemeh; Mohammadi, Maedeh; Najafpour, Ghasem D

    2016-09-01

    Cheese whey fermentation to ethanol using immobilized Kluyveromyces marxianus cells was investigated in batch and continuous operation. In batch fermentation, the yeast cells were immobilized in carboxymethyl cellulose (CMC) polymer and also synthesized graft copolymer of CMC with N-vinyl-2-pyrrolidone, denoted as CMC-g-PVP, and the efficiency of the two developed cell entrapped beads for lactose fermentation to ethanol was examined. The yeast cells immobilized in CMC-g-PVP performed slightly better than CMC with ethanol production yields of 0.52 and 0.49 g ethanol/g lactose, respectively. The effect of supplementation of cheese whey with lactose (42, 70, 100 and 150 g/l) on fermentative performance of K. marxianus immobilized in CMC beads was considered and the results were used for kinetic studies. The first order reaction model was suitable to describe the kinetics of substrate utilization and modified Gompertz model was quite successful to predict the ethanol production. For continuous ethanol fermentation, a packed-bed immobilized cell reactor (ICR) was operated at several hydraulic retention times; HRTs of 11, 15 and 30 h. At the HRT of 30 h, the ethanol production yield using CMC beads was 0.49 g/g which implies that 91.07 % of the theoretical yield was achieved. PMID:27126500

  15. [Study on PVA-alginate co-immobilization of Xanthomonas ampelina TS206].

    PubMed

    Chen, Qingsen; Liu, Jian

    2003-08-01

    Ice nucleation activity and anti-leaking capability are both important technical parameters in INA (Ice nucleation active bactetia) bacteria immobilization which can be adopted on freezing concentrate. Both PVA and alginate are good medium for immobilization. They can be used on co-immobilization of ice nucleation-active bacteria (Xanthomonas ampelinaTS206). The results showed that quantity of embedding affects ice nucleation activity greatly. The order of importance to comprehensive scores of technical standard should be concentration of Sodium alginate > Boric acid > PVA > CaCl2. The optimized concentration are PVA 8%, sodium alginate 1%, CaCl2 1.1% and boric acid 5%. A conclusion can be draw that ice nucleation activity increases with the number of beads and shows little pertinency with the time of immobilization, whereas anti-leaking capability can be influenced faintly by the number of beads and the time of immobilization. PMID:16276925

  16. Kinetic study of the immobilization of galvanic sludge in clay-based matrix.

    PubMed

    Magalhães, Jorge M; Silva, João E; Castro, Fernando P; Labrincha, João A

    2005-05-20

    The viability of inertization of galvanic wastes through their incorporation in clay-based materials, such as common formulations for tiles and bricks, is here studied by determining the leaching kinetics in different media. Metals immobilization is assured by firing at reasonably high temperatures, since intimately contact and/or reaction between residue and clay particles is promoted but also due to formation of insoluble metal oxides that rest unreactive towards clay grains. For most metals, leaching rate follows a zero-order kinetic law, with values between 0.001 to 0.1 mg/(g day cm2). Leaching velocity tends to increase with rising atomic numbers: Zn < Cu < Ni < Cr. These values depend exponentially on the relative sludge content.

  17. Evaluation of capillary chromatographic supports for immobilized human purine nucleoside phosphorylase in frontal affinity chromatography studies.

    PubMed

    de Moraes, Marcela Cristina; Temporini, Caterina; Calleri, Enrica; Bruni, Giovanna; Ducati, Rodrigo Gay; Santos, Diógenes Santiago; Cardoso, Carmen Lucia; Cass, Quezia Bezerra; Massolini, Gabriella

    2014-04-18

    The aim of this work was to optimize the preparation of a capillary human purine nucleoside phosphorylase (HsPNP) immobilized enzyme reactor (IMER) for characterization and affinity screening studies of new inhibitors by frontal affinity chromatography coupled to mass spectrometry (FAC-MS). For this purpose two monolithic supports, a Chromolith Speed Rod (0.1mm I.D.×5cm) and a methacrylate-based monolithic epoxy polymeric capillary column (0.25mm I.D.×5cm) with epoxy reactive groups were considered and compared to an IMER previously developed using an open fused silica capillary. Each HsPNP-IMER was characterized in terms of catalytic activity using Inosine as standard substrate. Furthermore, they were also explored for affinity ranking experiments. Kd determination was carried out with the based fused silica HsPNP-IMER and the results are herein discussed.

  18. Biosorption of metal contaminants using immobilized biomass: A laboratory study. Rept. of Investigations/1991

    SciTech Connect

    Jeffers, T.H.; Ferguson, C.R.; Bennett, P.G.

    1991-01-01

    The U.S. Bureau of Mines has developed porous beads containing immobilized biological materials for removing metal contaminants from waste waters. The beads, designated as BIO-FIX beads, are prepared by blending biomass, such as sphagnum peat moss or algae, into a polymer solution and spraying the mixture into water. Laboratory studies were conducted to determine bead sorption and elution characteristics. Batch and continuous tests demonstrated that BIO-FIX beads sorbed arsenic, cadmium, lead, and other toxic metals from acid mine drainage waters collected from several sites. Selectivity for heavy and toxic metal ions over calcium and magnesium was demonstrated. The beads exhibited excellent metal sorption and handling characteristics in stirred tanks, column contactors, and a low-maintenance passive system. The sorption process was reversible, and metal ions were eluted from the beads using dilute mineral acids. Cyclic tests indicated that the beads continued to extract metal ions after repeated loading-elution cycles.

  19. Immobilization of DNA at Glassy Ccarbon Electrodes: A Critical Study of Adsorbed Layer

    PubMed Central

    Pedano, M. L.; Rivas, G. A.

    2005-01-01

    In this work we present a critical study of the nucleic acid layer immobilized at glassy carbon electrodes. Different studies were performed in order to assess the nature of the interaction between DNA and the electrode surface. The adsorption and electrooxidation of DNA demonstrated to be highly dependent on the surface and nature of the glassy carbon electrode. The DNA layer immobilized at a freshly polished glassy carbon electrode was very stable even after applying highly negative potentials. The electron transfer of potassium ferricyanide, catechol and dopamine at glassy carbon surfaces modified with thin (obtained by adsorption under controlled potential conditions) and thick (obtained by casting the glassy carbon surface with highly concentrated DNA solutions) DNA layers was slower than that at the bare glassy carbon electrode, although this effect was dependent on the thickness of the layer and was not charge selective. Raman experiments showed an important decrease of the vibrational modes assigned to the nucleobases residues, suggesting a strong interaction of these residues with the electrode surface. The hybridization of oligo(dG)21 and oligo(dC)21 was evaluated from the guanine oxidation signal and the reduction of the redox indicator Co(phen)33+. In both cases the chronopotentiometric response indicated that the compromise of the bases in the interaction of DNA with the electrode surface is too strong, preventing further hybridization. In summary, glassy carbon is a useful electrode material to detect DNA in a direct and very sensitive way, but not to be used for the preparation of biorecognition layers by direct adsorption of the probe sequence on the electrode surface for detecting the hybridization event.

  20. Microorganism immobilization

    DOEpatents

    Compere, Alicia L.; Griffith, William L.

    1981-01-01

    Live metabolically active microorganisms are immobilized on a solid support by contacting particles of aggregate material with a water dispersible polyelectrolyte such as gelatin, crosslinking the polyelectrolyte by reacting it with a crosslinking agent such as glutaraldehyde to provide a crosslinked coating on the particles of aggregate material, contacting the coated particles with live microorganisms and incubating the microorganisms in contact with the crosslinked coating to provide a coating of metabolically active microorganisms. The immobilized microorganisms have continued growth and reproduction functions.

  1. A noble technique a using force-sensing resistor for immobilization-device quality assurance: A feasibility study

    NASA Astrophysics Data System (ADS)

    Cho, Min-Seok; Kim, Tae-Ho; Kang, Seong-Hee; Kim, Dong-Su; Kim, Kyeong-Hyeon; Shin, Dong-Seok; Noh, Yu-Yun; Koo, Hyun-Jae; Cheon, Geum Seong; Suh, Tae Suk; Kim, Siyong

    2016-03-01

    Many studies have reported that a patient can move even when an immobilization device is used. Researchers have developed an immobilization-device quality-assurance (QA) system that evaluates the validity of immobilization devices. The QA system consists of force-sensing-resistor (FSR) sensor units, an electric circuit, a signal conditioning device, and a control personal computer (PC) with in-house software. The QA system is designed to measure the force between an immobilization device and a patient's skin by using the FSR sensor unit. This preliminary study aimed to evaluate the feasibility of using the QA system in radiation-exposure situations. When the FSR sensor unit was irradiated with a computed tomography (CT) beam and a treatment beam from a linear accelerator (LINAC), the stability of the output signal, the image artifact on the CT image, and changing the variation on the patient's dose were tested. The results of this study demonstrate that this system is promising in that it performed within the error range (signal variation on CT beam < 0.30 kPa, root-mean-square error (RMSE) of the two CT images according to presence or absence of the FSR sensor unit < 15 HU, signal variation on the treatment beam < 0.15 kPa, and dose difference between the presence and the absence of the FSR sensor unit < 0.02%). Based on the obtained results, we will volunteer tests to investigate the clinical feasibility of the QA system.

  2. Useful oriented immobilization of antibodies on chimeric magnetic particles: direct correlation of biomacromolecule orientation with biological activity by AFM studies.

    PubMed

    Marciello, Marzia; Filice, Marco; Olea, David; Velez, Marisela; Guisan, José M; Mateo, Cesar

    2014-12-16

    The preparation and performance of a suitable chimeric biosensor based on antibodies (Abs) immobilized on lipase-coated magnetic particles by means of a standing orienting strategy are presented. This novel system is based on hydrophobic magnetic particles coated with modified lipase molecules able to orient and further immobilize different Abs in a covalent way without any previous site-selective chemical modification of biomacromolecules. Different key parameters attending the process were studied and optimized. The optimal preparation was performed using a controlled loading (1 nmol Ab g(-1) chimeric support) at pH 9 and a short reaction time to recover a biological activity of about 80%. AFM microscopy was used to study and confirm the Abs-oriented immobilization on lipase-coated magnetic particles and the final achievement of a highly active and recyclable chimeric immune sensor. This direct technique was demonstrated to be a powerful alternative to the indirect immunoactivity assay methods for the study of biomacromolecule-oriented immobilizations.

  3. Study of bi-enzyme immobilization onto layered double hydroxides nanomaterials for histamine biosensor application.

    PubMed

    Baccar, Z M; Hidouri, S; Errachid, A; Sanchez, O Ruiz

    2011-10-01

    In this work, we present the development of a hybrid biomembrane based on the immobilization of diamine oxidase (DAO) into LDH thin films for histamine detection. The LDHs preselected as host matrixes are: hydrotalcites (Mg2Al(CO3)0.5(OH)6), lowaite (Mg4Fe(OH)10Cl) and hydrocalumite (Ca2Al(OH)6Cl). The immobilized probes were characterized by atomic force microscopy (AFM) and attenuated total reflection infrared spectroscopy (IR-ATR mode). The analysis of these results shows that the immobilization of DAO occurs with all type of selected LDH and is stable after a 7 day-immersion in phosphate buffer solution. The LDH incorporating magnesium or calcium divalent cations present high-quality surface topology for DAO immobilization and the ability to keep the enzyme in a well conformation for biogenic amines catabolism and histamine detection. PMID:22400262

  4. Plutonium Immobilization Project Binder Burnout and Sintering Studies (Milestone 6.6a)

    SciTech Connect

    Chandler, G.

    1999-10-28

    The Plutonium Immobilization Team has developed an integrated test program to understand and optimize the controlling variables for the sintering step of the plutonium immobilization process. Sintering is the key process step that controls the product minerology. It is expected that the sintering will be the limiting process step that controls the throughput of the production line. The goal of the current sintering test program is to better understand factors that affect the sintering process.

  5. [Study of the interaction of main potato glycoalkaloids in inhibition of immobilized butyryl cholinesterase].

    PubMed

    Arkhypova, V M; Dziadevych, S V; Jaffrezic-Renault, N; Martelet, C; Soldatkin, O P

    2006-01-01

    The interaction of main potato glycoalkaloids alpha-solanine and alpha-chaconine in inhibition of horse serum butyryl cholinesterases immobilized on the pH-sensitive field-effect transistors has been investigated. The method of isobol diagram of Loewe and Muishnek has been used for interpretation of results. It has been shown the alpha-chaconine inhibits the immobilized bytyryl cholinesterases more strongly than alpha-solanine, and their mixture has the addition effect.

  6. Luminescence study of UO[sup 2+][sub 2] binding to immobilized datura innoxia biomaterial

    SciTech Connect

    Ke, Huei-Yang D.; Rayson, G.D. )

    1993-01-01

    The spectrally and temporally resolved UO[sup 2+][sub 2] fluorescence signals have been measured for solid samples of UO[sup 2+][sub 2]-Datura, UO[sup 2+][sub 2]-(immobilized Datura), and UO[sup 2+][sub 2]-(silicate polymer) at liquid nitrogen temperature. The binding capacity of UO[sup 2+][sub 2] to Datura innoxia cell material has been enhanced significantly at pH 5 and 6 when immobilized in a polysilicate matrix. New binding sites having a greater binding strength with a lower availability have been observed for the binding of UO[sup 2+][sub 2] after immobilization of the cultured biomaterial. However, chemical alteration of the cell material resulting from this immobilization process has not been observed. The chemical environment of the binding sites responsible for the binding of UO[sup 2+][sub 2] in immobilized and free D. innoxia cell material has been demonstrated to be different. This difference results from the immobilization of the cell material within a polysilicate matrix. 19 refs., 8 figs., 4 tabs.

  7. Immobilization of flavan-3-ols onto sensor chips to study their interactions with proteins and pectins by SPR

    NASA Astrophysics Data System (ADS)

    Watrelot, Aude A.; Tran, Dong Tien; Buffeteau, Thierry; Deffieux, Denis; Le Bourvellec, Carine; Quideau, Stéphane; Renard, Catherine M. G. C.

    2016-05-01

    Interactions between plant polyphenols and biomacromolecules such as proteins and pectins have been studied by several methods in solution (e.g. isothermal titration calorimetry, dynamic light scattering, nuclear magnetic resonance and spectrophotometry). Herein, these interactions were investigated in real time by Surface Plasmon Resonance (SPR) analysis after immobilization of flavan-3-ols onto a sensor chip surface. (-)-epicatechin, (+)-catechin and flavan-3-ol oligomers with an average degree of polymerization of 2 and 8 were chemically modified using N-(2-(tritylthio)ethyl)propiolamide in order to introduce a spacer unit onto the catecholic B ring. Modified flavan-3-ols were then immobilized onto a carboxymethylated dextran surface (CM5). Immobilization was validated and further verified by evaluating flavan-3-ol interaction with bovine serum albumin (BSA), poly-L-proline or commercial pectins. BSA was found to have a stronger association with monomeric flavan-3-ols than oligomers. SPR analysis of selected flavan-3-ols immobilized onto CM5 sensor chips showed a stronger association for citrus pectins than apple pectins, regardless of flavan-3-ol degree of polymerization.

  8. Immobilization of cellulase on TiO2 nanoparticles by physical and covalent methods: a comparative study.

    PubMed

    Ahmad, Razi; Sardar, Meryam

    2014-08-01

    Immobilization of cellulase from Aspergillus niger on TiO2 nanoparticles was studied by two different approaches--physical adsorption and covalent coupling. A. niger was selected, as it is generally non-pathogenic, is found in nature in the broad range of habitats and produces cellulase extracellulary. For covalent method, TiO2 nanoparticles were modified with aminopropyltriethoxysilane (APTS). The adsorbed and covalently immobilized enzymes showed 76% and 93% activity, respectively, as compared to the free enzyme. The catalytic efficiency V(max)/K(m) increased from 0.4 to 4.0 after covalent attachment, whereas in adsorption method, it increased slightly from 0.4 to 1.2. The covalently-immobilized and adsorbed cellulase lost only 25% and 50% of their activity, respectively after 60 min of incubation at 75 degrees C. The reusability and operational stability data also showed that covalent coupling increased the stability of the enzyme. The presence of enzyme on TiO2 nanoparticles was confirmed by Fourier-transform infrared spectroscopy. The high-resolution transmission electron microscopy (HR-TEM) and atomic force microscopy (AFM) studies indicated aggregation of enzyme when adsorbed on TiO2 surface and a monolayer of enzyme in covalent attachment. In conclusion, covalently attached cellulase retained good activity and thermal stability, as compared to physically adsorbed enzyme. The lower amount of enzyme activity and thermal stability in case of physically adsorbed immobilized enzyme was due to aggregation of the enzyme after adsorption on TiO2 nanoparticles, as revealed by HR-TEM and AFM. Thus, TiO2 nanoparticles could be suitable candidates for immobilization of cellulase for industrial applications like paper, textile, detergent and food industries.

  9. Biosorption of Cu(II) by immobilized microalgae using silica: kinetic, equilibrium, and thermodynamic study.

    PubMed

    Lee, Hongkyun; Shim, Eunjung; Yun, Hyun-Shik; Park, Young-Tae; Kim, Dohyeong; Ji, Min-Kyu; Kim, Chi-Kyung; Shin, Won-Sik; Choi, Jaeyoung

    2016-01-01

    Immobilized microalgae using silica (IMS) from Micractinium reisseri KGE33 was synthesized through a sol-gel reaction. Green algal waste biomass, the residue of M. reisseri KGE33 after oil extraction, was used as the biomaterial. The adsorption of Cu(II) on IMS was tested in batch experiments with varying algal doses, pH, contact times, initial Cu(II) concentrations, and temperatures. Three types of IMSs (IMS 14, 70, and 100) were synthesized according to different algal doses. The removal efficiency of Cu(II) in the aqueous phase was in the following order: IMS 14 (77.0%) < IMS 70 (83.3%) < IMS 100 (87.1%) at pH 5. The point of zero charge (PZC) value of IMS100 was 4.5, and the optimum pH for Cu(II) adsorption was 5. Equilibrium data were described using a Langmuir isotherm model. The Langmuir model maximum Cu(II) adsorption capacity (q m) increased with the algal dose in the following order: IMS 100 (1.710 mg g(-1)) > IMS 70 (1.548 mg g(-1)) > IMS 14 (1.282 mg g(-1)). The pseudo-second-order equation fitted the kinetics data well, and the value of the second-order rate constant increased with increasing algal dose. Gibbs free energies (ΔG°) were negative within the temperature range studied, which indicates that the adsorption process was spontaneous. The negative value of enthalpy (ΔH°) again indicates the exothermic nature of the adsorption process. In addition, SEM-energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared (FT-IR), and X-ray photoelectron spectroscopy (XPS) analyses of the IMS surface reveal that the algal biomass on IMS is the main site for Cu(II) binding. This study shows that immobilized microalgae using silica, a synthesized biosorbent, can be used as a cost-effective sorbent for Cu(II) removal from the aqueous phase.

  10. Biosorption of Cu(II) by immobilized microalgae using silica: kinetic, equilibrium, and thermodynamic study.

    PubMed

    Lee, Hongkyun; Shim, Eunjung; Yun, Hyun-Shik; Park, Young-Tae; Kim, Dohyeong; Ji, Min-Kyu; Kim, Chi-Kyung; Shin, Won-Sik; Choi, Jaeyoung

    2016-01-01

    Immobilized microalgae using silica (IMS) from Micractinium reisseri KGE33 was synthesized through a sol-gel reaction. Green algal waste biomass, the residue of M. reisseri KGE33 after oil extraction, was used as the biomaterial. The adsorption of Cu(II) on IMS was tested in batch experiments with varying algal doses, pH, contact times, initial Cu(II) concentrations, and temperatures. Three types of IMSs (IMS 14, 70, and 100) were synthesized according to different algal doses. The removal efficiency of Cu(II) in the aqueous phase was in the following order: IMS 14 (77.0%) < IMS 70 (83.3%) < IMS 100 (87.1%) at pH 5. The point of zero charge (PZC) value of IMS100 was 4.5, and the optimum pH for Cu(II) adsorption was 5. Equilibrium data were described using a Langmuir isotherm model. The Langmuir model maximum Cu(II) adsorption capacity (q m) increased with the algal dose in the following order: IMS 100 (1.710 mg g(-1)) > IMS 70 (1.548 mg g(-1)) > IMS 14 (1.282 mg g(-1)). The pseudo-second-order equation fitted the kinetics data well, and the value of the second-order rate constant increased with increasing algal dose. Gibbs free energies (ΔG°) were negative within the temperature range studied, which indicates that the adsorption process was spontaneous. The negative value of enthalpy (ΔH°) again indicates the exothermic nature of the adsorption process. In addition, SEM-energy-dispersive X-ray spectroscopy (EDS), Fourier transform infrared (FT-IR), and X-ray photoelectron spectroscopy (XPS) analyses of the IMS surface reveal that the algal biomass on IMS is the main site for Cu(II) binding. This study shows that immobilized microalgae using silica, a synthesized biosorbent, can be used as a cost-effective sorbent for Cu(II) removal from the aqueous phase. PMID:25953610

  11. Study the biocatalyzing effect and mechanism of cellulose acetate immobilized redox mediators technology (CE-RM) on nitrite denitrification.

    PubMed

    Li, Haibo; Guo, Jianbo; Lian, Jing; Xi, Zhenhua; Zhao, Lijun; Liu, Xiaoyu; Zhang, Chenxiao; Yang, Jingliang

    2014-06-01

    The biocatalyzing effect of a novel cellulose acetate immobilized redox mediators technology (CE-RM) on nitrite denitrification process was studied with anthraquinone, 1,8-dichloroanthraquinone, 1,5-dichloroanthraquinone and 1,4,5,8-tetrachloroanthraquinone. The results showed that the immobilized 1,4,5,8-tetrachloroanthraquinone presented the best biocatalyzed effect which increased nitrite denitrification rate to 2.3-fold with 12 mmol/L 1,4,5,8-tetrachloroanthraquinone. The unequal biocatalyzing effect was due to the quantity and position of -Cl substituent in anthraquinone-structure. Moreover, the nitrite denitrification rate was increased with the oxidation reduction potential (ORP) values becoming more negative during the biocatalyzing process. The stabilized ORP value with 12 mmol/L immobilized 1,4,5,8-tetrachloroanthraquinone were 81 mV lower than the control. At the same time, the more OH(-) was produced with the higher nitrite removal rate achieved in the nitrite denitrification process. In addition, a positive linear correlation was found between the nitrite removal reaction constants k [gNO2(-)-N/(gVSS d)] and immobilized 1,4,5,8-tetrachloroanthraquinone concentration (C1,4,5,8-tetrachloroanthraquinone), which was k = 1.8443 C1,4,5,8-tetrachloroanthraquinone + 33.75(R(2) = 0.9411). The initial nitrite concentration of 179 mgNO2(-)-N/L resulted in the maximum nitrite removal rate, which was 6.526[gNO2(-)-N/(gVSS d)]. These results show that the application of cellulose acetate immobilized redox mediators (CE-RM) can be valuable for increasing nitrite denitrification rate.

  12. Sonoenzymatic decolourization of an azo dye employing immobilized horse radish peroxidase (HRP): a mechanistic study.

    PubMed

    Malani, Ritesh S; Khanna, Swati; Moholkar, Vijayanand S

    2013-07-15

    For degradation of biorefractory pollutants, enzymatic treatments and sonochemical treatments have shown high potential. A combined technique of sono-enzymatic treatment is of special interest as it has shown enhancement effect than the individual techniques. This work has attempted to give a mechanistic insight into the interaction of sonochemical and enzymatic treatments using immobilized horseradish peroxidase (HRP) enzyme on the decolourization of acid red dye (an azo dye). In order to segregate the effect of ultrasound and cavitation, experiments were conducted at elevated static pressure. The kinetic parameters of HRP, viz. Vmax and Km were marginally affected by immobilization. There was a minor change in pH optima and temperature optima for immobilized HRP (6.5, 25°C) from free HRP (7.0, 20-25°C). Though the specific activity of free enzyme (0.272U/mg) was found to be higher than the immobilized enzyme (0.104U/mg), immobilized enzyme exhibited higher stability (up to 3 cycles) and degradation potential than free enzyme in all experiments. The results revealed that the coupling of sonication and enzymatic treatment at high pressure in presence of polyethylene glycol (PEG) yielded the highest decolourization of acid red (61.2%). However, the total decolourization achieved with combined technique was lesser than the sum of individual techniques, indicating negative synergy between the sonochemical and enzymatic techniques. PMID:23708258

  13. Comparative study of immobilized Trametes versicolor laccase on nanoparticles and kaolinite.

    PubMed

    Hu, Xiaoke; Zhao, Xueheng; Hwang, Huey-Min

    2007-01-01

    Laccase from Trametes versicolor was immobilized on nanoparticles and kaolinite by physical adsorption or chemical covalence in which the supporters were activated by cross-linked with glutaraldehyde. Thermal and pH stabilities of immobilized laccase on these different supporters were compared. The degradation efficiencies of these immobilized laccases on oxidation of benzo[a]pyrene (BaP) were also compared. The results showed that the immobilized laccases on nanoparticles were more stable in resisting pH and thermal changes. After 48h oxidation, laccase immobilized on kaolinite using the covalent coupling method showed a higher efficiency of oxidation with the BaP residue of 23% in the presence of 1mM HBT and with BaP residue of 37% in 1mM ABTS as the mediator. The results also exhibited a significant inhibition by 1% surfactant Tween 80. According to the HPLC analysis, the oxidation products including 1,6-benzo[a]pyrene quinone, 3,6-benzo[a]pyrene quinone and 6,12-benzo[a]pyrene quinone were identified.

  14. A comparative study of enzyme immobilization strategies for multi-walled carbon nanotube glucose biosensors

    NASA Astrophysics Data System (ADS)

    Shi, Jin; Claussen, Jonathan C.; McLamore, Eric S.; Haque, Aeraj ul; Jaroch, David; Diggs, Alfred R.; Calvo-Marzal, Percy; Rickus, Jenna L.; Porterfield, D. Marshall

    2011-09-01

    This work addresses the comparison of different strategies for improving biosensor performance using nanomaterials. Glucose biosensors based on commonly applied enzyme immobilization approaches, including sol-gel encapsulation approaches and glutaraldehyde cross-linking strategies, were studied in the presence and absence of multi-walled carbon nanotubes (MWNTs). Although direct comparison of design parameters such as linear range and sensitivity is intuitive, this comparison alone is not an accurate indicator of biosensor efficacy, due to the wide range of electrodes and nanomaterials available for use in current biosensor designs. We proposed a comparative protocol which considers both the active area available for transduction following nanomaterial deposition and the sensitivity. Based on the protocol, when no nanomaterials were involved, TEOS/GOx biosensors exhibited the highest efficacy, followed by BSA/GA/GOx and TMOS/GOx biosensors. A novel biosensor containing carboxylated MWNTs modified with glucose oxidase and an overlying TMOS layer demonstrated optimum efficacy in terms of enhanced current density (18.3 ± 0.5 µA mM - 1 cm - 2), linear range (0.0037-12 mM), detection limit (3.7 µM), coefficient of variation (2%), response time (less than 8 s), and stability/selectivity/reproducibility. H2O2 response tests demonstrated that the most possible reason for the performance enhancement was an increased enzyme loading. This design is an excellent platform for versatile biosensing applications.

  15. Biosorption of metal contaminants using immobilized biomass: Field studies. Report of Investigations/1993

    SciTech Connect

    Jeffers, T.H.; Bennett, P.G.; Corwin, R.R.

    1993-01-01

    The U.S. Bureau of Mines has developed porous beads containing immobilized biological materials such as sphagnum peat moss for extracting metal contaminants from waste waters. The beads, designated as BIO-FIX beads, have removed toxic metals from over 100 waters in laboratory tests. These waters include acid mine drainage (AMD) water from mining sites, metallurgical and chemical industry waste water, and contaminated ground water. Following the laboratory studies, cooperative field tests were conducted to evaluate the metal adsorption properties of the beads in column and low-maintenance circuits, determine bead stability in varied climatic situations, and demonstrate the beads' potential as a viable waste water treatment technique. Field results indicated that BIO-FIX beads readily adsorbed cadmium, lead, and other toxic metals from dilute waters; effluents frequently met drinking water standards and other discharge criteria. The beads exhibited excellent handling characteristics in both column and low-maintenance circuits, and continued to extract metal ions after repeated loading-elution cycles.

  16. Comparative Study on the Immobilization of Lipase on Chitosan Gels Modified by Different Hydrophobic Groups

    NASA Astrophysics Data System (ADS)

    Deng, Hong-Tao; Lin, Yan; Wang, Juan-Juan; Liu, Zhong-Yang; Ma, Miao; Zheng, Fei

    The hydrophobic surface modification of chitosan gels (CS) was carried out using the amidating reaction of amido groups on a gel surface with propionic acid, stearic acid, and benzoic acid, respectively, activated by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). Lipase from Candida rugosa was immobilized by adsorption on the nascent CS, propionyl-modified gels (PCS), stearyl-modified gels (SCS), and benzoyl-modified gels (BCS), respectively. The adsorption capacity and activity of immobilized lipase were investigated. It was found that the surface modification improved the adsorption capacity of lipase, and the activity retention of immobilized lipase increased from 52.34% for CS to 57.17%, 78.26% and 69.22%, respectively, for PCS, SCS, and BCS.

  17. Study on immobilization of lipase onto magnetic microspheres with epoxy groups

    NASA Astrophysics Data System (ADS)

    Lei, Lin; Bai, Yongxiao; Li, Yanfeng; Yi, Liuxiang; Yang, Yong; Xia, Chungu

    2009-02-01

    Magnetic microspheres were synthesized by the suspension polymerization of glycidyl methacrylate (GMA), methacrylic acid (MAA) and divinyl benzene (DVB) in the presence of oleic acid-coated Fe 3O 4 nanoparticles. Triacylglycerol lipase from porcine pancreas was covalently immobilized on the magnetic microspheres via the active epoxy groups with the activity yield up to 63% (±2.3%) and enzyme loading of 39 (±0.5) mg/g supports. The resulting immobilized lipase had higher optimum temperature compared with those of free lipase and exhibited better thermal, broader pH stability and excellent reusability. Furthermore, the catalyzed capability of immobilized lipase was also investigated by catalyzing synthesis of hexyl acetate and the esterification conversion rate reached to 83% (±2.5%) after 12 h in nonaqueous solvent.

  18. Evaluation of enzyme immobilization methods for paper-based devices--A glucose oxidase study.

    PubMed

    Nery, Emilia Witkowska; Kubota, Lauro T

    2016-01-01

    Paper-based sensors gained almost explosive attention during the last few years. A large number of systems, often destined to resource limited settings is based on enzymatic reactions. Choice of an adequate immobilization method could significantly prolong the shelf-life of such sensors, especially in applications, where exposure to high temperatures during storage and transport is more than a threat. We are seeking to compare a variety of immobilization methods based on different phenomena (adsorption, entrapment in gel, microencapsulation, covalent linkage), with total of 33 methods tested. Glucose oxidase was used as a model enzyme. Enzymatic activity of immobilized samples was accompanied for a period of 24 weeks considering two sets of samples, one stored in 4 °C and other in ambient temperature.

  19. Stretching and immobilization of DNA for studies of protein-DNA interactions at the single-molecule level

    NASA Astrophysics Data System (ADS)

    Kim, Ji Hoon; Dukkipati, Venkat Ram; Pang, Stella W.; Larson, Ronald G.

    2007-04-01

    Single-molecule studies of the interactions of DNA and proteins are important in a variety of biological or biotechnology processes ranging from the protein’s search for its DNA target site, DNA replication, transcription, or repair, and genome sequencing. A critical requirement for single-molecule studies is the stretching and immobilization of otherwise randomly coiled DNA molecules. Several methods for doing so have been developed over the last two decades, including the use of forces derived from light, magnetic and electric fields, and hydrodynamic flow. Here we review the immobilization and stretching mechanisms for several of these techniques along with examples of single-molecule DNA-protein interaction assays that can be performed with each of them.

  20. Immobilization of DNA for affinity chromatography and drug-binding studies.

    PubMed Central

    Macdougall, A J; Brown, J R; Plumbridge, T W

    1980-01-01

    A method is described in which double-stranded DNA is alkylated with 4-bis-(2-chloroethyl)amino-L-phenylalanine and the product immobilized on an insoluble support via the primary amino group of the phenylalanine moiety. The DNA is hence irreversibly bound to the matrix by both strands at a limited number of points. PMID:7283977

  1. Renaturation studies of free and immobilized D-amino-acid oxidase.

    PubMed

    Carrea, G; Pasta, P; Curti, B

    1983-06-15

    The renaturation of free and Sepharose-immobilized D-amino-acid oxidase (D-amino-acid:oxygen oxidoreductase (deaminating), EC 1.4.3.3), after its denaturation with 6 M guanidine hydrochloride, was investigated. No reactivation, or extremely limited reactivation (less than or equal to 4+), was obtained with the free enzyme, is spite of various attempts including the use of dialysis or buffers containing cofactors, different types of anions, surfactants and low concentrations of denaturing agents. The main obstacle to renaturation appeared to be the interaction among denatured or partially renatured monomers giving rise to inactive aggregates. In contrast, using the immobilized enzyme approach, substantial renaturation (up to 50%) of D-amino-acid oxidase was achieved. The denaturation-renaturation process was followed by monitoring the catalytic activity as well as the intrinsic protein fluorescence. An inverse correlation was found to exist between the degree of matrix activation by CNBr and the yield of enzyme reactivation. The anions of the lyotropic series markedly influenced the reactivation, showing an effectiveness opposite to their salting-out potential (thiocyanate congruent to iodide greater than chloride greater than phosphate congruent to sulphate congruent to citrate). Instead, the anions considerably increased the activity and stability of free and immobilized enzyme, according to their salting-out potential. Immobilized monomers of D-amino-acid oxidase, which in solution undergoes self-association, showed poor capacity to interact with the free enzyme: thus they appear unsuitable for analytical and preparative purposes.

  2. Study on contraction and relaxation of experimentally denervated and immobilized muscles: Comparison with dystrophic muscles

    NASA Technical Reports Server (NTRS)

    Takamori, M.; Tsujihata, M.; Mori, M.; Hazama, R.; Ide, Y.

    1980-01-01

    The contraction-relaxation mechanism of experimentally denervated and immobilized muscles of the rabbit is examined. Results are compared with those of human dystrophic muscles, in order to elucidate the role and extent of the neurotrophic factor, and the role played by the intrinsic activity of muscle in connection with pathogenesis and pathophysiology of this disease.

  3. Physical and chemical immobilization of choline oxidase onto different porous solid supports: Adsorption studies.

    PubMed

    Passos, Marieta L C; Ribeiro, David S M; Santos, João L M; Saraiva, M Lúcia M F S

    2016-08-01

    This work carries out for the first time the comparison between the physical and chemical immobilization of choline oxidase onto aminated silica-based porous supports. The influence on the immobilization efficiency of concentration, pH, temperature and contact time between the support and choline oxidase, was evaluated. The immobilization efficiency was estimated taking into consideration the choline oxidase activity, which was assessed by using cadmium telluride (CdTe) quantum dots (QDs), obtained by hydrothermal synthesis, as photoluminescent probes. Hydrogen peroxide produced by enzyme activity was capable of quenching CdTe QDs photoluminescence. The magnitude of the PL quenching process was directly related with the enzyme activity. By comparing the chemical process with the physical adsorption, it was observed that the latter provided the highest choline oxidase immobilization. The equilibrium data were analyzed using Langmuir and Freundlich isotherms and kinetic data were fitted to the pseudo-first-order and pseudo-second-order models. Thermodynamic parameters, such as Gibbs free energy and entropy were also calculated. These results will certainly contribute to the development of new sensing schemes for choline, taking into account the growing demand for its quantification in biological samples. PMID:27241295

  4. Removal of mercury (II) from aqueous solution using papain immobilized on alginate bead: optimization of immobilization condition and modeling of removal study.

    PubMed

    Bhattacharyya, Aparupa; Dutta, Susmita; De, Parameswar; Ray, Parthasarathi; Basu, Srabanti

    2010-12-01

    Papain having the characteristics of metal binding ability is immobilized on alginate bead. Design Expert Software (Version 7.1.6) uses Response Surface Methodology (RSM) for statistical designing of operating condition for immobilization of papain on alginate bead considering concentration of papain, concentration of sodium alginate, concentration of calcium chloride and pH as numeric factors and Specific Enzymatic Activity (SEA) of immobilized papain sample as response. Immobilization using 25.96 g/L papain, 20 g/L sodium alginate and 20 g/L calcium chloride at pH 7 gives the desired product as indicated by ANOVA (Analysis of Variance). Three parameters viz., initial concentration of mercury (II), amount of AIP and pH are varied in a systematic manner. Maximum 98.88% removal of mercury (II) has been achieved within 8 min when simulated aqueous solution of mercury (II) with initial concentration of 10mg/L has been contacted with 5 g of AIP at pH 9 and at 35 degrees C in a batch contactor. A mathematical model has been developed and the value of equilibrium constant for binding of mercury (II) with AIP has been found to be 126797.3. PMID:20696575

  5. Removal of mercury (II) from aqueous solution using papain immobilized on alginate bead: optimization of immobilization condition and modeling of removal study.

    PubMed

    Bhattacharyya, Aparupa; Dutta, Susmita; De, Parameswar; Ray, Parthasarathi; Basu, Srabanti

    2010-12-01

    Papain having the characteristics of metal binding ability is immobilized on alginate bead. Design Expert Software (Version 7.1.6) uses Response Surface Methodology (RSM) for statistical designing of operating condition for immobilization of papain on alginate bead considering concentration of papain, concentration of sodium alginate, concentration of calcium chloride and pH as numeric factors and Specific Enzymatic Activity (SEA) of immobilized papain sample as response. Immobilization using 25.96 g/L papain, 20 g/L sodium alginate and 20 g/L calcium chloride at pH 7 gives the desired product as indicated by ANOVA (Analysis of Variance). Three parameters viz., initial concentration of mercury (II), amount of AIP and pH are varied in a systematic manner. Maximum 98.88% removal of mercury (II) has been achieved within 8 min when simulated aqueous solution of mercury (II) with initial concentration of 10mg/L has been contacted with 5 g of AIP at pH 9 and at 35 degrees C in a batch contactor. A mathematical model has been developed and the value of equilibrium constant for binding of mercury (II) with AIP has been found to be 126797.3.

  6. Comparative studies of Remazol Brillant Blue removal by immobilized organisms; investigation of metabolites by GC/MS and FTIR spectrometry.

    PubMed

    Akdogan, Hatice Ardag; Topuz, Merve Canpolat

    2015-01-01

    Reactive dyes are important chemical pollutants from textile industries. Treatment of effluents from dye-based industries poses a major problem, and biotreatment with white rot fungi seems to be a viable option. The biological treatment of synthetic dyes at a low cost and in the shortest possible time is used especially in dye and textile industries and leads to pollution in the wastewater dumped into the environment by these industries. For this study, decolorization of the recalcitrant dye Remazol Brilliant Blue R by immobilized Pleurotus ostreatus and Coprinus plicatilis was investigated. This dye was removed 100% (dye concentration: 10.0 mg/L) by both immobilized organisms. Extracellular ligninolytic enzyme activities were also measured during the decolorization. There was an attempt to identify metabolites with FTIR spectrometry and GC/MS at the end of the decolorization. These results indicated that the samples did not include any detectable metabolite. PMID:25905752

  7. Cervical Spine Immobilization in Sports Related Injuries: Review of Current Guidelines and a Case Study of an Injured Athlete

    PubMed Central

    Bhamra, JS; Morar, Y; Khan, WS; Deep, K; Hammer, A

    2012-01-01

    Cervical spine immobilization is an essential component of the ATLS® system. Inadequate training in the management of trauma calls and failure of early recognition can have disastrous consequences. Pre-hospital personnel are routinely involved more in the assessment and stabilization of patients in comparison to other health care professionals. This case study and review highlights the importance of early recognition, assessment and correct stabilization of cervical spine injuries both in the field and during the initial assessment in hospital. Inadequate assessment, immobilization and lack of standard guidelines on the management of suspected cervical spine trauma can result in secondary injury. Regular assessment and training of pre-hospital and medical personnel is essential to the proper management of these potentially devastating injuries. PMID:23248726

  8. Study of reaction parameters and kinetics of esterification of lauric acid with butanol by immobilized Candida antarctica lipase.

    PubMed

    Shankar, Sini; Agarwal, Madhu; Chaurasia, S P

    2013-12-01

    Esterification of lauric acid with n-butanol, catalyzed by immobilized Candida antarctica lipase (CAL) in aqueous-organic biphasic solvent system was studied. Effects of various reaction parameters on esterification were investigated, such as type and amount of solvent, amount of buffer, pH, temperature, speed of agitation, amount of enzyme, butanol and lauric acid. The most suitable reaction conditions for esterification were observed at 50 degrees C and pH 7.0 using 5000 micromoles of lauric acid, 7000 pmoles of butanol, 0.25 ml phosphate buffer, 1 ml of isooctane as the solvent and 50 mg of immobilized enzyme in the reaction medium at agitation speed of 150 rpm. Maximum esterification of 96.36% was acheived in 600 min of reaction time at n-butanol to lauric acid molar ratio of 1: 0.7. Kinetic study for the esterification of lauric acid with n-butanol using immobilized CAL was carried out and the kinetic constants were estimated by using non-linear regression method. The estimated value of Michaelis kinetic constants for butanol (KmBt) and acid (KmAc) were 451.56 (M) and 4.7 x 10(-7)(M), respectively and the value of dissociation constant (KBt) of the butanol-lipase complex was 9.41 x 10(7)(M). The estimated constants agreed fairly well with literature data.

  9. Thermodynamic study of an unusual chiral separation. Propranolol enantiomers on an immobilized cellulase

    SciTech Connect

    Fornstedt, T.; Sajonz, P.; Guiochon, G. |

    1997-02-12

    The thermodynamics of interaction of (R)- and (S)-propranolol between an acetic acid buffer (pH = 4.7 and 5.5) and the protein cellobiohydrolase I immobilized on silica gel was studied between 5 and 45{degree}C. The equilibrium data were fitted to a biLangmuir adsorption isotherm with excellent agreement. One of the two Langmuir contributions is the same for both enantiomers and accounts for the nonspecific interactions between these compounds and most sites on the surfaces (type-I, nonselective sites). It has a large saturation capacity. The second contribution accounts for the chiral selective interactions (type-II sites). It has a lower monolayer capacity than the first. The interaction enthalpy and entropy on type-I sites are -1.1 kcal/mol and +0.1 cal/(mol K), respectively. For type-II sites, they are -1.9 kcal/mol and -2.6 cal/(mol K), respectively, for (R)-propranolol and +1.6 kcal/mol and +11.6 cal/(mol K), respectively, for (S)-propranolol at pH = 5.5. This explains why at this pH the retention time of the less-retained R enantiomer decreases with increasing temperature, while the retention time of the S enantiomer increases, causing a large increase of the separation factor when the temperature is raised from 5 to 45{degree}C. The saturation capacity of the chiral contributions depends strongly on the pH, and the retention times of both enantiomers decrease with increasing temperature at pH = 4.7. 46 refs., 6 figs., 5 tabs.

  10. Solvent Stability Study with Thermodynamic Analysis and Superior Biocatalytic Activity of Burkholderia cepacia Lipase Immobilized on Biocompatible Hybrid Matrix of Poly(vinyl alcohol) and Hypromellose.

    PubMed

    Badgujar, Kirtikumar C; Bhanage, Bhalchandra M

    2014-12-26

    In the present study, we have synthesized a biocompatible hybrid carrier of hypromellose (HY) and poly(vinyl alcohol) (PVA) for immobilization of Burkholderia cepacia lipase (BCL). The immobilized biocatalyst HY:PVA:BCL was subjected to determination of half-life time (τ) and deactivation rate constant (K(D)) in various organic solvents. Biocatalyst showed higher τ-value in a nonpolar solvent like cyclohexane (822 h) as compared to that of a polar solvent such as acetone (347 h), which signifies better compatibility of biocatalyst in the nonpolar solvents. Furthermore, the K(D)-value was found to be less in cyclohexane (0.843 × 10(-3)) as compared to acetone (1.997 × 10(-3)), indicating better stability in the nonpolar solvents. Immobilized-BCL (35 mg) was sufficient to achieve 99% conversion of phenethyl butyrate (natural constituent of essential oils and has wide industrial applications) using phenethyl alcohol (2 mmol) and vinyl butyrate (6 mmol) at 44 °C in 3 h. The activation energy (E(a)) was found to be lower for immobilized-BCL than crude-BCL, indicating better catalytic efficiency of immobilized lipase BCL. The immobilized-BCL reported 6-fold superior biocatalytic activity and 8 times recyclability as compared to crude-BCL. Improved catalytic activity of immobilized enzyme in nonpolar media was also supported by thermodynamic activation parameters such as enthalpy (ΔH(⧧)), entropy (ΔS(⧧)) and Gibb's free energy (ΔG(⧧)) study, which showed that phenethyl butyrate synthesis catalyzed by immobilized-BCL was feasible as compared to crude-BCL. The present work explains a thermodynamic investigation and superior biocatalytic activity for phenethyl butyrate synthesis using biocompatible immobilized HY:PVA:BCL in nonaqueous media for the first time. PMID:25474503

  11. Differential effect of the shape of calcium alginate matrices on the physiology of immobilized neuroblastoma N2a and Vero cells: a comparative study.

    PubMed

    Kintzios, S; Yiakoumetis, I; Moschopoulou, G; Mangana, O; Nomikou, K; Simonian, A

    2007-11-30

    In order to investigate the effect of cell immobilization in calcium alginate gels on cell physiology, we immobilized Vero or N2a neuroblastoma cells in gels shaped either as spherical beads or as thin membrane layers. Throughout a culture period of 4 weeks cell viability, RNA and cytoplasmic calcium concentration and glutathione accumulation were assayed by fluorescence microscopy after provision of an appropriate dye. Non-elaborate culture conditions were applied throughout the experimental period in order to evaluate cell viability under less than optimal storage conditions. Vero cell proliferation was observed only in spherical beads, while N2a cell proliferation was observed in both configurations until the third week of culture. Increased [Ca2+]cyt could be associated with cell proliferation only when cells were immobilized in spherical beads, while a considerable decrease in the biosynthesis of reduced glutathione and RNA was observed in cells immobilized in thin membrane layers. The observed effects of the shape of the immobilization matrix may be due to differences in external mass transfer resistance. Therefore, depending on cell type, cell proliferation could have been promoted by either increased (Vero) or decreased (N2a) nutrient and oxygen flow to immobilized cells. The results of the present study could contribute to an improvement of immobilized cell sensor storability.

  12. Preparation and loading buffer study of polyvinyl alcohol-based immobilized Ti4+ affinity chromatography for phosphopeptide enrichment.

    PubMed

    Hu, Yufeng; Guo, Shuangxi; Ma, Hongbo; Ye, Ning; Ren, Xueqin

    2013-11-01

    Despite recent advances in phosphoproteomics, an efficient and simple enrichment protocol is still a challenge and of high demand aiming at large-scale plant phosphoproteomics studies. Here, we developed a novel loading buffer system for synthesized immobilized metal affinity chromatography material targeting plant samples, which was prepared by a simple one-step esterification between polyvinyl alcohol and phosphoric acid and then was subjected to immobilize Ti(4+). SEM and Fourier transform IR spectroscopy were used to assure the synthesis protocol of the polyvinyl alcohol-based Ti(4+) immobilized material, and the specific surface areas and pore volumes of the polymers were measured. The selectivity for phosphopeptide enrichment from α-casein was improved by optimizing the pH and components of the loading buffer. By using potassium hydrogen phthalate/hydrochloric acid with pH at 2.50 as the loading buffer, 19 phosphopeptides with high intensity were identified. The final optimized protocol was adapted to salt-stressed maize leaves for phosphoproteome analysis. A total of 57 phosphopeptides containing 59 phosphorylated sites from 50 phosphoproteins were identified in salt-stressed maize leaf. The research was meaningful to obtain much more information about phosphoproteins leading to the comprehension of salt resistance and salt-inducible phosphorylated processes of maize leaves.

  13. Comparative study of olive oil mill wastewater treatment using free and immobilized Coriolopsis polyzona and Pycnoporus coccineus.

    PubMed

    Neifar, Mohamed; Jaouani, Atef; Martínez, María Jesús; Penninckx, Michel J

    2012-10-01

    The efficiency of the two white-rot fungi Pycnoporus coccineus and Coriolopsis polyzona in the Olive Oil Mill Wastewater (OOMW) treatment was investigated. Both fungi were active in the decolourisation and COD removal of OOMW at 50 g/L COD, but only the first fungus remains effective on the crude effluent (COD=100 g/L). Moreover P. coccineus was less affected by oxygen supplementation and exhibited a high tolerance to agitation in comparison to C. polyzona. However, it required a nitrogen supplementation to obtain faster and higher COD removal. To overcome the negative effect of agitation on fungi growth and efficiency, immobilisation of C. polyzona and P. coccineus in polyurethane foam was applied. The immobilized system showed better COD decreases during three consecutive batches without remarkable loss of performances. The results obtained in this study suggested that immobilized C. polyzona and especially immobilized P. coccineus might be applicable to a large scale for the removal colour and COD of OOMW. PMID:23124741

  14. PRELIMINARY STUDY OF CERAMICS FOR IMMOBILIZATION OF ADVANCED FUEL CYCLE REPROCESSING WASTES

    SciTech Connect

    Fox, K.; Billings, A.; Brinkman, K.; Marra, J.

    2010-09-22

    The Savannah River National Laboratory (SRNL) developed a series of ceramic waste forms for the immobilization of Cesium/Lanthanide (CS/LN) and Cesium/Lanthanide/Transition Metal (CS/LN/TM) waste streams anticipated to result from nuclear fuel reprocessing. Simple raw materials, including Al{sub 2}O{sub 3}, CaO, and TiO{sub 2} were combined with simulated waste components to produce multiphase ceramics containing hollandite-type phases, perovskites (particularly BaTiO{sub 3}), pyrochlores, zirconolite, and other minor metal titanate phases. Identification of excess Al{sub 2}O{sub 3} via X-ray Diffraction (XRD) and Scanning Electron Microscopy with Energy Dispersive Spectroscopy (SEM/EDS) in the first series of compositions led to a Phase II study, with significantly reduced Al{sub 2}O{sub 3} concentrations and increased waste loadings. Three fabrication methodologies were used, including melting and crystallizing, pressing and sintering, and Spark Plasma Sintering (SPS), with the intent of studying phase evolution under various sintering conditions. XRD and SEM/EDS results showed that the partitioning of the waste elements in the sintered materials was very similar, despite varying stoichiometry of the phases formed. The Phase II compositions generally contained a reduced amount of unreacted Al{sub 2}O{sub 3} as identified by XRD, and had phase assemblages that were closer to the initial targets. Chemical composition measurements showed no significant issues with meeting the target compositions. However, volatilization of Cs and Mo was identified, particularly during melting, since sintering of the pressed pellets and SPS were performed at lower temperatures. Partitioning of some of the waste components was difficult to determine via XRD. SEM/EDS mapping showed that those elements, which were generally present in small concentrations, were well distributed throughout the waste forms. Initial studies of radiation damage tolerance using ion beam irradiation at Los

  15. Uranium Immobilization through Fe(II) bio-oxidation: A Column study

    SciTech Connect

    Coates, John D.

    2009-09-14

    Current research on the bioremediation of heavy metals and radionuclides is focused on the ability of reducing organisms to use these metals as alternative electron acceptors in the absence of oxygen and thus precipitate them out of solution. However, many aspects of this proposed scheme need to be resolved, not the least of which is the time frame of the treatment process. Once treatment is complete and the electron donor addition is halted, the system will ultimately revert back to an oxic state and potentially result in the abiotic reoxidation and remobilization of the immobilized metals. In addition, the possibility exists that the presence of more electropositive electron acceptors such as nitrate or oxygen will also stimulate the biological oxidation and remobilization of these contaminants. The selective nitrate-dependent biooxidation of added Fe(II) may offer an effective means of “capping off” and completing the attenuation of these contaminants in a reducing environment making the contaminants less accessible to abiotic and biotic reactions and allowing the system to naturally revert to an oxic state. Our previous DOE-NABIR funded studies demonstrated that radionuclides such as uranium and cobalt are rapidly removed from solution during the biogenic formation of Fe(III)-oxides. In the case of uranium, X-ray spectroscopy analysis indicated that the uranium was in the hexavalent form (normally soluble) and was bound to the precipitated Fe(III)-oxides thus demonstrating the bioremediative potential of this process. We also demonstrated that nitrate-dependent Fe(II)- oxidizing bacteria are prevalent in the sediment and groundwater samples collected from sites 1 and 2 and the background site of the NABIR FRC in Oakridge, TN. However, all of these studies were performed in batch experiments in the laboratory with pure cultures and although a significant amount was learned about the microbiology of nitrate-dependent bio-oxidation of Fe(II), the effects of

  16. Comparative studies on the properties of tryptophanase and tyrosine phenol-lyase immobilized directly on Sepharose or by use of Sepharose-bound pyridoxal 5'-phosphate.

    PubMed

    Fukui, S; Ikeda, S; Fujimura, M; Yamada, H; Kumagai, H

    1975-02-01

    Tryptophanase from Escherichia coli B/qt 7-A and tyrosine phenol-lyase (beta-tyrosinase) from Escherichia intermedia were immobilized on Sepharose 4B by several direct coupling reactions or through pyridoxal 5'-phosphate previously bound to Sepharose. The most active preparation of immobilized tryptophanase was obtained by coupling tetrameric apoenzyme to pyridoxal-P bound on Sepharose at the 6-position through a diazo linkage. This immobilization procedure involves the formation to Schiff base linkage between 4-formyl group of Sepharose-bound pyridoxal-P and the epsilon-amino group of the lysine residue at the active center of one subunit of tetrameric apo-tryptophanase, followed by the fixation of the Schiff base linkage by reduction with NaBH4. In the case of beta-tyrosinase having two catalytic centers, however, this method was not so suitable as the case of tryptophanase. Direct coupling of the apoenzyme to CNBr-activated Sepharose or to a bromoacetyl derivative of Sepharose gave better results. In each case, the affinity for substrate or coenzyme was scarcely influenced by the immobilization. When used repeatedly in a batch system or continuously in a flow system in the absence of added pyridoxal-P, immobilized holo-tryptophanase of holo-beta-tyrosinase gradually lost its original activity; however, supplement of pyridoxal-P to the reaction system restored its initial activity. From the kinetic analyses of these phenomena, the rate constants of coenzyme dissociation from immobilized tryptophanase and beta-tyrosinase were calculated. Upon immobilization, the pH optima of both enzymes shifted 0.5 to 1.0 pH unit to the alkaline side. Both immobilized enzymes showed higher thermal stability and resistance to a denaturing agent such as guinidine-HCl than their free counterpart. Furthermore, the reactivity of sulfhydryl group of beta-tyrosinase, in connection with its coenzyme-binding property, was conveniently studied by use of the immobilized enzyme.

  17. Study on Different Molecular Weights of Chitosan as an Immobilization Matrix for a Glucose Biosensor

    PubMed Central

    Ang, Lee Fung; Por, Lip Yee; Yam, Mun Fei

    2013-01-01

    Two chitosan samples (medium molecular weight (MMCHI) and low molecular weight (LMCHI)) were investigated as an enzyme immobilization matrix for the fabrication of a glucose biosensor. Chitosan membranes prepared from acetic acid were flexible, transparent, smooth and quick-drying. The FTIR spectra showed the existence of intermolecular interactions between chitosan and glucose oxidase (GOD). Higher catalytic activities were observed on for GOD-MMCHI than GOD-LMCHI and for those crosslinked with glutaraldehyde than using the adsorption technique. Enzyme loading greater than 0.6 mg decreased the activity. Under optimum conditions (pH 6.0, 35°C and applied potential of 0.6 V) response times of 85 s and 65 s were observed for medium molecular weight chitosan glucose biosensor (GOD-MMCHI/PT) and low molecular weight chitosan glucose biosensor (GOD-LMCHI/PT), respectively. The apparent Michaelis-Menten constant () was found to be 12.737 mM for GOD-MMCHI/PT and 17.692 mM for GOD-LMCHI/PT. This indicated that GOD-MMCHI/PT had greater affinity for the enzyme. Moreover, GOD-MMCHI/PT showed higher sensitivity (52.3666 nA/mM glucose) when compared with GOD-LMCHI/PT (9.8579 nA/mM glucose) at S/N>3. Better repeatability and reproducibility were achieved with GOD-MMCHI/PT than GOD-LMCHI/PT regarding glucose measurement. GOD-MMCHI/PT was found to give the highest enzymatic activity among the electrodes under investigation. The extent of interference encountered by GOD-MMCHI/PT and GOD-LMCHI/PT was not significantly different. Although the Nafion coated biosensor significantly reduced the signal due to the interferents under study, it also significantly reduced the response to glucose. The performance of the biosensors in the determination of glucose in rat serum was evaluated. Comparatively better accuracy and recovery results were obtained for GOD-MMCHI/PT. Hence, GOD-MMCHI/PT showed a better performance when compared with GOD-LMCHI/PT. In conclusion, chitosan membranes shave

  18. Sulfur immobilization and lithium storage on defective graphene: A first-principles study

    SciTech Connect

    Zhao, Wen; Chen, Pengcheng; Tang, Peizhe; Wu, Jian; Duan, Wenhui E-mail: dwh@phys.tsinghua.edu.cn; Li, Yuanchang E-mail: dwh@phys.tsinghua.edu.cn

    2014-01-27

    Motivated by the recent progresses and remaining technical challenges in Li-S battery, we employ defective graphene as a prototype cathode framework to illustrate how battery performance is influenced by the mesoporous carbon materials. We show that the immobilization of S unavoidably sacrifices its ability to further interact with Li, which leads to an enhanced cycle life but a decreased capacity. Based on our calculated results, we suggest a suitable S binding-energy range of ∼4–5 eV to balance the battery stability and capability under thermodynamic equilibrium conditions. Our results may promote the understanding and architecture design of Li-S battery.

  19. Studies of optimum conditions for covalent immobilization of Candida rugosa lipase on poly(gamma-glutamic acid) by RSM.

    PubMed

    Chang, S-W; Shaw, J-F; Yang, K-H; Chang, S-F; Shieh, C-J

    2008-05-01

    Poly(gamma-glutamic acid) (gamma-PGA) is a material of polymer. Immobilization of Candida rugosa lipase (Lipase AY-30) by covalent binding on gamma-PGA led to a markedly improved performance of the enzyme. Response surface methodology (RSM) and 3-level-3-factor fractional factorial design were employed to evaluate the effects of immobilization parameters, such as immobilization time (2-6h), immobilization temperature (0-26 degrees C), and enzyme/support ratio (0.1-0.5, w/w). Based on the analysis of ridge max, the optimum immobilization conditions were as follows: immobilization time 2.3h, immobilization temperature 13.3 degrees C, and enzyme/support ratio 0.41 (w/w); the highest lipase activity obtained was 1196 U/mg-protein.

  20. Mimetic biomembrane-AuNPs-graphene hybrid as matrix for enzyme immobilization and bioelectrocatalysis study.

    PubMed

    Wang, Tianshu; Liu, Jiyang; Ren, Jiangtao; Wang, Jin; Wang, Erkang

    2015-10-01

    A hybrid composite constructed of phospholipids bilayer membrane, gold nanoparticles and graphene was prepared and used as matrices for microperoxidase-11 (MP11) immobilization. The direct electrochemistry and corresponding bioelectrocatalysis of the enzyme electrode was further investigated. Phospholipid bilayer membrane protected gold nanoparticles (AuNPs) were assembled on polyelectrolyte functionalized graphene sheets through electrostatic attraction to form a hybrid bionanocomposite. Owing to the biocompatible microenvironment provided by the mimetic biomembrane, microperoxidase-11 entrapped in this matrix well retained its native structure and exhibited high bioactivity. Moreover, the AuNPs-graphene assemblies could efficiently promote the direct electron transfer between the immobilized MP11 and the substrate electrode. The as-prepared enzyme electrode presented good direct electrochemistry and electrocatalytic responses to the reduction of hydrogen peroxide (H2O2). The resulting H2O2 biosensor showed a wide linear range (2.0×10(-5)-2.8×10(-4) M), a low detection limit (2.6×10(-6) M), good reproducibility and stability. Furthermore, this sensor was used for real-time detection of H2O2 dynamically released from the tumor cells MCF-7 in response to a pro-inflammatory stimulant. PMID:26078181

  1. Si-F complexing in aqueous fluids: experimental study and implications for transport of immobile elements

    NASA Astrophysics Data System (ADS)

    Dolejš, David

    2014-05-01

    Intepretation of fluid-mineral interaction mechanisms and hydrothermal fluxes requires knowledge of predominant solubility and speciation reactions and their thermodynamic properties. Fluorine represents a hard electron donor, capable of complexing and transporting high-field strength elements, which are traditionally considered to be immobile. Reactions responsible for element mobility have general form MOx + y HF (aq) + x - y H2O = M(OH)2x-yFy (aq), and their extent and transport efficiency relies on hydrogen fluoride activity. In natural fluids, a[HF] is controlled by various fluorination equilibria including neutralization of silicates with consequent formation of silicohydroxyfluoride complexes. Quartz solubility in HF-H2O fluids was experimentally determined at 400-800 oC and 100-200 MPa using rapid-quench cold-seal pressure vessels and the mineral weight-loss method. Quartz solubility significantly increases in the presence of hydrogen fluoride: at 400 oC and 100 MPa, dissolved SiO2 ranges from 0.18 wt. % in pure H2O to 12.2 wt. % at 8.3 wt. % F in the fluid, whereas at 800 oC and 200 MPa it rises from 1.51 wt. % in pure H2O to 15.3 wt. % at 8.0 wt. % F in the fluid. The isobaric solubilities of quartz appear to be temperature-independent, i.e., effects of temperature vs. fluid density on the solubility are counteracting. The experimental data are described by the density model: log m[SiO2] = a + blog ρ + clog m[F] + dT , where a = -1.049 mol kg-1, b = 0.816 mol cm-3, c = 0.802 and d = 1.256 · 10-3 mol kg-1 K-1. Solubility isotherms have similar d(log m[SiO2])/d(log m[F]) slopes over the entire range of conditions indicating that Si(OH)2F2 is the major aqueous species. Several factors promote breakdown of silicohydroxyfluoride complexes and precipitation of silica solute: (i) decreasing temperature and pressure, i.e., fluid ascent and cooling and/or (ii) neutralization and increase in the alkali/H ratio of fluids during alteration reactions or removal of

  2. Treatment of textile effluent in a developed phytoreactor with immobilized bacterial augmentation and subsequent toxicity studies on Etheostoma olmstedi fish.

    PubMed

    Watharkar, Anuprita D; Khandare, Rahul V; Waghmare, Pankajkumar R; Jagadale, Ashwini D; Govindwar, Sanjay P; Jadhav, Jyoti P

    2015-01-01

    A static hydroponic bioreactor using nursery grown plants of Pogonatherum crinitum along with immobilized Bacillus pumilus cells was developed for the treatment of textile wastewater. Independent reactors with plants and immobilized cells were also kept for performance and efficacy evaluation. The effluent samples characterized before and after their treatment showed that the plant-bacterial consortium reactor was more efficient than those of individual plant and bacterium reactors. COD, BOD, ADMI, conductivity, turbidity, TDS and TSS of the textile effluent was found to be reduced by 78, 70, 93, 4, 90, 13 and 70% respectively within 12 d by the consortial set. HPTLC analysis revealed the transformation of the textile effluent to new products. The phytotoxicity study on Phaeseolus mungo and Sorghum vulgare seeds showed reduced toxicity of treated effluents. The animal toxicity study performed on Etheostoma olmstedi fishes showed the toxic nature of untreated effluent giving extreme stress to fishes leading to death. Histology of fish gills exposed to treated effluent was found to be less affected. The oxidative stress related enzymes like superoxide dismutase and catalase were found to show decreased activities and less lipid peroxidation in fishes exposed to treated effluent.

  3. Study of concentration of HPV DNA probe immobilization for cervical cancer detection based IDE biosensor

    NASA Astrophysics Data System (ADS)

    Roshila, M. L.; Hashim, U.; Azizah, N.

    2016-07-01

    This paper mainly illustrates regarding the detection process of Human Papillomavirus (HPV) DNA probe. HPV is the most common virus that infected to human by a sexually transmitted virus. The most common high-risk HPV are 16 and 18. Interdigitated electrode (IDE) device used as based of Titanium Dioxide (TiO2) acts as inorganic surface, where by using APTES as a linker between inorganic surface and organic surface. A strategy of rapid and sensitive for the HPV detection was proposed by integrating simple DNA extraction with a gene of DNA. The extraction of the gene of DNA will make an efficiency of the detection process. It will depend on the sequence of the capture probes and the way to support their attached. The fabrication, surface modification, immobilization and hybridization processes are characterized by current voltage (I-V) measurement by using KEITHLEY 6487. This strategy will perform a good sensitivity of HPV detection.

  4. Early dissociation of nuclear factor I from the origin during initiation of adenovirus DNA replication studied by origin immobilization.

    PubMed

    Coenjaerts, F E; van der Vliet, P C

    1994-12-11

    The DNA-binding domain of Nuclear Factor I (NFIBD) enhances initiation of adenovirus DNA replication up to 50-fold by binding to the auxiliary region of the origin and positioning the viral DNA polymerase. To study if and when NFIBD dissociates from the template, we immobilized origin DNA to glutathione-agarose beads by means of a GST-NFIBD fusion protein. This immobilized template is active in replication. By analyzing the release of prelabeled templates from the beads under different conditions, we show that NFIBD dissociates already early during initiation. During preinitiation NFIBD remains bound, but as soon as dCTP, dATP or dTTP are added, efficient dissociation occurs. A much lower dissociation level was induced by addition of dGTP. Since dCTP, dATP and dTTP are required for formation of a pTP-CAT initiation intermediate, we explain our results by conformational changes occurring in the polymerase during initiation leading to disruption of both the interaction between the polymerase and NFI as well as the interaction between NFI and the DNA.

  5. Macrocyclic receptors immobilized to monodisperse porous polymer particles by chemical grafting and physical impregnation for strontium capture: a comparative study.

    PubMed

    Song, Yang; Du, Yi; Lv, Dachao; Ye, Gang; Wang, Jianchen

    2014-06-15

    Separation of strontium is of great significance for radioactive waste treatment and environmental remediation after nuclear accidents. In this work, a novel class of adsorbent (Crown-g-MPPPs) was synthesized by chemical grafting a macrocyclic ether receptor to monodisperse porous polymer particles (MPPPs) for strontium adsorption. Meanwhile, a counterpart material (Crown@MPPPs) with the receptor molecules immobilized to the MPPPs substrate by physical impregnation was prepared. To investigate how the immobilization manner and distribution of the receptors influence the adsorption ability, a comparative study on the adsorption behaviour of the two materials towards Sr(II) in HNO3 media was accomplished. Due to the shorter diffusion path and covalently-bonded structure, Crown-g-MPPPs showed faster adsorption kinetics and better stability for cycle use. While Crown@MPPPs had the advantages of facile synthesis and higher adsorption capacity, owing to the absence of conformational constraint to form complexation with Sr(II). Kinetic functions (Lagergren pseudo-first-order/pseudo-second-order functions) and adsorption isotherm models (Langmuir/Freundlich models) were used to fit the experimental data and examine the adsorption mechanism. On this basis, a chromatographic process was proposed by using Crown@MPPPs for an effective separation of Sr(II) (91%) in simulated high level liquid waste (HLLW).

  6. Bio-functionalization of electro-synthesized polypyrrole surface by heme enzyme using a mixture of Nafion and glutaraldehyde as synergetic immobilization matrix: Conformational characterization and electrocatalytic studies

    NASA Astrophysics Data System (ADS)

    ElKaoutit, Mohammed; Naranjo-Rodriguez, Ignacio; Domínguez, Manuel; Hidalgo-Hidalgo-de-Cisneros, José Luis

    2011-10-01

    Use of a mixture of Nafion and glutaraldehyde as new immobilization matrix was described. The percentage of Nafion was optimized to prevent denaturation of horseradish peroxidase enzyme after its crosslinkage with glutaraldehyde on electro-synthesized polypyrrole surface. Topographic study by Atomic Force Microscopy (AFM) shows that the enzyme seems to have been introduced inside the ionic cluster of Nafion. The characterization of the resulting bio-interfaces by UV-vis and FT-IR shows that the intra-crosslinkage phenomena caused by the use of glutaraldehyde can be eliminated by the optimization of the concentration of Nafion additive. The secondary structure contents of native and immobilized enzyme were analyzed by a Gaussian curve fitting of the respective FT-IR spectra in the amide I region. Immobilized enzyme presented notable increasing percentages of globular and short helical structure compared with native enzyme. This indicates that immobilized enzyme was folded which is in accordance with AFM studies and supports the enzyme entrance inside ionic clutter of Nafion. Thanks to synergic effects of the polypyrrole conducting polymer and the perfluorosulfonic acid polymer Nafion, HRP enzyme was immobilized in its "native" state, the resulting biosensor was able to sense peroxide without any chemical mediator and can be categorized as third generation.

  7. Microalgal immobilization methods.

    PubMed

    Moreno-Garrido, Ignacio

    2013-01-01

    In this review, methods for the most common microalgal immobilization procedures are gathered and described. Passive (due to natural adherence of cells to surfaces) and active immobilization methods should be distinguished. Among active immobilization methods, calcium alginate entrapment is the most widely used method if living cells are intended to be immobilized, due to the chemical, optical, and mechanical characteristics of this substance. Immobilization in synthetic foams, immobilization in agar and carrageenan as well as immobilization in silica-based matrix or filters are also discussed and described. Finally, some considerations on the use of flocculation for microalgae are mentioned.

  8. Covalent immobilization of redox enzyme on electrospun nonwoven poly(acrylonitrile-co-acrylic acid) nanofiber mesh filled with carbon nanotubes: a comprehensive study.

    PubMed

    Wang, Zhen-Gang; Ke, Bei-Bei; Xu, Zhi-Kang

    2007-07-01

    In this work, novel conductive composite nanofiber mesh possessing reactive groups was electrospun from solutions containing poly(acrylonitrile-co-acrylic acid) (PANCAA) and multi-walled carbon nanotubes (MWCNTs) for redoxase immobilization, assuming that the incorporated MWCNTs could behave as electrons transferor during enzyme catalysis. The covalent immobilization of catalase from bovine liver on the neat PANCAA nanofiber mesh or the composite one was processed in the presence of EDC/NHS. Results indicated that both the amount and activity retention of bound catalase on the composite nanofiber mesh were higher than those on the neat PANCAA nanofiber mesh, and the activity increased up to 42%. Kinetic parameters, K(m) and V(max), for the catalases immobilized on the composite nanofiber mesh were lower and higher than those on the neat one, respectively. This enhanced activity might be ascribed to either promoted electron transfer through charge-transfer complexes and the pi system of carbon nanotubes or rendered biocompatibility by modified MWCNTs. Furthermore, the immobilized catalases revealed much more stability after MWCNTs were incorporated into the polymer nanofiber mesh. However, there was no significant difference in optimum pH value and temperature, thermal stability and operational stability between these two immobilized preparations, while the two ones appeared more advantageous than the free in these properties. The effect of MWCNTs incorporation on another redox enzyme, peroxidase, was also studied and it was found that the activity increased by 68% in comparison of composite one with neat preparation. PMID:17171660

  9. Study of immobilized catalysts. XXI. Synthesis and structure of titanomagnesium catalysts on polymer carriers

    SciTech Connect

    Bochkin, A.M.; Pomogailo, A.D.; Dyachkovskii, F.S.

    1987-02-01

    Methods of synthesis of polymer carriers with an organometallic functional coating were developed, and grafted titanomagnesium catalysts (GTMC) based on them were obtained. The formation of isolated Ti(III) ions was detected by the EPR method. Depending on the conditions of synthesis and type of polymer carrier, their amount reached 55% of the immobilized titanium. Experimental proofs were obtained for the formation of Ti(III) via the stage of alkylation of Ti(IV) by a magnesium-containing coating. Based on the dependence of the g-factor of the Ti(III) signals on the conditions of its formation, it was suggested that the fragments of the polymer carrier enter the coordination sphere of Ti(III) at low Ti(III) concentrations in the GTMC and that stratification reactions of Ti(III) take place at high concentrations. The existence of crystallites of the active phase with dimensions of the regions of coherent scattering of 40-75 A was found by x-ray diffraction analysis. A correlation was found between the activity of GTMC in the polymerization of ethylene and the Ti(III) content in them in the form of isolated ions.

  10. Protein immobilization techniques for microfluidic assays

    PubMed Central

    Kim, Dohyun; Herr, Amy E.

    2013-01-01

    Microfluidic systems have shown unequivocal performance improvements over conventional bench-top assays across a range of performance metrics. For example, specific advances have been made in reagent consumption, throughput, integration of multiple assay steps, assay automation, and multiplexing capability. For heterogeneous systems, controlled immobilization of reactants is essential for reliable, sensitive detection of analytes. In most cases, protein immobilization densities are maximized, while native activity and conformation are maintained. Immobilization methods and chemistries vary significantly depending on immobilization surface, protein properties, and specific assay goals. In this review, we present trade-offs considerations for common immobilization surface materials. We overview immobilization methods and chemistries, and discuss studies exemplar of key approaches—here with a specific emphasis on immunoassays and enzymatic reactors. Recent “smart immobilization” methods including the use of light, electrochemical, thermal, and chemical stimuli to attach and detach proteins on demand with precise spatial control are highlighted. Spatially encoded protein immobilization using DNA hybridization for multiplexed assays and reversible protein immobilization surfaces for repeatable assay are introduced as immobilization methods. We also describe multifunctional surface coatings that can perform tasks that were, until recently, relegated to multiple functional coatings. We consider the microfluidics literature from 1997 to present and close with a perspective on future approaches to protein immobilization. PMID:24003344

  11. Immobilizing CC chemokine receptor 4's N-terminal extracellular tail on a capillary to study its potential ligands by capillary electrophoresis.

    PubMed

    Chen, Wenjing; Li, Meina; Yakufu, Pazilaiti; Ling, Xiaomei; Qi, Hui; Xiao, Junhai; Wang, Ying

    2012-04-01

    ML40 is the equivalent peptide derived from the N terminal of CCC4 (CC chemokine receptor 4), which plays a pivotal role in allergic inflammation. A new capillary electrophoresis method was developed to study the interactions between ML40 and its potential ligands in which ML40 was immobilized on the inner wall of capillary as the stationary phase based on the covalent linking technique. The interaction between S009, a known CCR4 antagonist, and the immobilized ML40 was studied to validate the bioactivity of ML40. The electropherogram of S009 showed that the peak height was reduced and the peak width was broadened in the ML40 immobilized capillary. Otherwise, 25 computer-aided design and drafting compounds were screened out using this method. Four compounds' peak widths were broadened and their peak heights were reduced, as with S009. Meanwhile, nonlinear chromatography was used to calculate the constants for the ligand-receptor complex formation. Furthermore, the tertiary amine compounds belonging to the chiral tertiary amines of the type NRR'R″, which are optically inactive resulting from rapid pyramide inversion, were chiral separated by our protein immobilization method for the first time. In general, the methodology presented would be applicable to study compound-ML40 interactions as a reliable and robust screening method for CCR4 antagonist discovery. PMID:22245764

  12. A comparative study on decolorization of reactive azo and indigoid dyes by free/immobilized pellets of Trametes versicolor and Funalia trogii.

    PubMed

    Yildirim, Seval Cing; Yesilada, Ozfer

    2015-11-01

    The objective of the present study was to investigate decolorization of Acid Blue 74 and Reactive Blue 198 dyes by free and immobilized white rot fungal pellets in order to confirm the possibility of practical application via repeated-batch cultivation. Decolorization studies were conducted using free pellets (FP), fungal cells immobilized on activated carbon (IFCAC) and pinewood (IFCP), and also fungal cells entrapped in alginate beads (FCEAB). No additional nitrogen and carbon source was used and high decolorization rates were achieved in only dye-contained media without pH adjustment. Acid Blue 74 was decolorized 96 and 94% within 2 hr by Trametes versicolor and Funalia trogii free pellets, respectively. These values were 87 and 84% for Reactive Blue 198, in this respect. Immobilization of fungal cells on pinewood increased the usability of pellets and the average decolorization efficiency of both dyes. The micro environment changed in the presence of pinewood and increased the stability of immobilized pellets. Decolorization was performed rapidly and efficiently. Laccase activity enhanced with availability of pinewood, and high laccase production with F. trogii was obtained. After separation by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), the molecular weight of T versicolor and F. trogii laccase bands was determined 64 and 61 kDa approximately. Green bands were obtained by the activity staining process with laccase substrate (ABTS) after gel renaturation step.

  13. Application of lipase immobilized on the biocompatible ternary blend polymer matrix for synthesis of citronellyl acetate in non-aqueous media: kinetic modelling study.

    PubMed

    Badgujar, Kirtikumar C; Bhanage, Bhalchandra M

    2014-04-10

    This work reports the use of new support for immobilization of lipase Burkholderia cepacia (BCL) matrix made up of polylactic acid (PLA), chitosan (CH), and polyvinyl alcohol (PVA). Initially lipase from various microbial sources and immobilization support composition was screened to obtain a robust biocatalyst. Among various biocatalysts preparation, the PLA:PVA:CH:BCL (1:6:1:2) was worked as a robust biocatalyst for the citronellyl acetate synthesis. Various reaction parameters were studied in detail to obtain the suitable reaction conditions for model citronellyl acetate synthesis reaction. Various kinetic parameters such as r(max), K(i(citronellol)), K(m(citronellol)), K(m(vinyl acetate)) were determined using non-linear regression analysis for the ternary complex as well as bi-bi ping-pong mechanism. The experimental results and kinetic study showed that citronellyl acetate synthesis catalyzed by immobilized lipase BCL followed the ternary complex mechanism with inhibition by alcohol (citronellol). The energy of activation for citronellyl acetate synthesis was found to be lower for immobilized lipase (8.9 kcal/mol) than aggregated lipase (14.8 kcal/mol) enzyme. The developed biocatalyst showed four to fivefold higher catalytic activity and excellent recyclability (up to six cycles) than the aggregated lipase. PMID:24629263

  14. Effect of computerized cognitive training with virtual spatial navigation task during bed rest immobilization and recovery on vascular function: A pilot study

    PubMed Central

    Goswami, Nandu; Kavcic, Voyko; Marusic, Uros; Simunic, Bostjan; Rössler, Andreas; Hinghofer-Szalkay, Helmut; Pisot, Rado

    2015-01-01

    We investigated the effects of bed rest (BR) immobilization, with and without computerized cognitive training with virtual spatial navigation task (CCT), on vascular endothelium on older subjects. The effects of 14-day BR immobilization in healthy older males (n=16) of ages 53–65 years on endothelial function were studied using EndoPAT®, a noninvasive and user-independent method. From the group of 16 older men, 8 randomly received CCT during the BR, using virtual navigation tasks in a virtual environment with joystick device. In all the cases, EndoPAT assessments were done at pre- and post-BR immobilization as well as following 28 days of ambulatory recovery. The EndoPAT index increased from 1.53±0.09 (mean ± standard error of the mean) at baseline to 1.61±0.16 following immobilization (P=0.62) in the group with CCT. The EndoPAT index decreased from 2.06±0.13 (mean ± standard error of the mean) at baseline to 1.70±0.09 at the last day of BR study, day 14 (BR14) (P=0.09) in the control group. Additionally, there were no statistically significant differences between BR14 and at 28 days of follow-up (rehabilitation program) (R28). Our results show a trend of immobilization in older persons affecting the vasoconstrictory endothelial response. As the control subjects had a greater increase in EndoPAT index after R28 (+0.018) compared to subjects who had cognitive training (+0.11) (calculated from the first day of BR study), it is possible that cognitive training during BR does not improve endothelial function but rather contributes to slowing down the impairment of endothelial function. Finally, our results also show that EndoPAT may be a useful noninvasive tool to assess the vascular reactivity. PMID:25709419

  15. Effect of computerized cognitive training with virtual spatial navigation task during bed rest immobilization and recovery on vascular function: a pilot study.

    PubMed

    Goswami, Nandu; Kavcic, Voyko; Marusic, Uros; Simunic, Bostjan; Rössler, Andreas; Hinghofer-Szalkay, Helmut; Pisot, Rado

    2015-01-01

    We investigated the effects of bed rest (BR) immobilization, with and without computerized cognitive training with virtual spatial navigation task (CCT), on vascular endothelium on older subjects. The effects of 14-day BR immobilization in healthy older males (n=16) of ages 53-65 years on endothelial function were studied using EndoPAT(®), a noninvasive and user-independent method. From the group of 16 older men, 8 randomly received CCT during the BR, using virtual navigation tasks in a virtual environment with joystick device. In all the cases, EndoPAT assessments were done at pre- and post-BR immobilization as well as following 28 days of ambulatory recovery. The EndoPAT index increased from 1.53±0.09 (mean ± standard error of the mean) at baseline to 1.61±0.16 following immobilization (P=0.62) in the group with CCT. The EndoPAT index decreased from 2.06±0.13 (mean ± standard error of the mean) at baseline to 1.70±0.09 at the last day of BR study, day 14 (BR14) (P=0.09) in the control group. Additionally, there were no statistically significant differences between BR14 and at 28 days of follow-up (rehabilitation program) (R28). Our results show a trend of immobilization in older persons affecting the vasoconstrictory endothelial response. As the control subjects had a greater increase in EndoPAT index after R28 (+0.018) compared to subjects who had cognitive training (+0.11) (calculated from the first day of BR study), it is possible that cognitive training during BR does not improve endothelial function but rather contributes to slowing down the impairment of endothelial function. Finally, our results also show that EndoPAT may be a useful noninvasive tool to assess the vascular reactivity.

  16. Production of L-glutamic Acid with Corynebacterium glutamicum (NCIM 2168) and Pseudomonas reptilivora (NCIM 2598): A Study on Immobilization and Reusability

    PubMed Central

    Shyamkumar, Rajaram; Moorthy, Innasi Muthu Ganesh; Ponmurugan, Karuppiah; Baskar, Rajoo

    2014-01-01

    Background L-glutamic acid is one of the major amino acids that is present in a wide variety of foods. It is mainly used as a food additive and flavor enhancer in the form of sodium salt. Corynebacterium glutamicum (C. glutamicum) is one of the major organisms widely used for glutamic acid production. Methods The study was dealing with immobilization of C. glutamicum and mixed culture of C. glutamicum and Pseudomonas reptilivora (P. reptilivora) for L-glutamic acid production using submerged fermentation. 2, 3 and 5% sodium alginate concentrations were used for production and reusability of immobilized cells for 5 more trials. Results The results revealed that 2% sodium alginate concentration produced the highest yield (13.026±0.247 g/l by C. glutamicum and 16.026±0.475 g/l by mixed immobilized culture). Moreover, reusability of immobilized cells was evaluated in 2% concentration with 5 more trials. However, when the number of cycles increased, the production of L-glutamic acid decreased. Conclusion Production of glutamic acid using optimized medium minimizes the time needed for designing the medium composition. It also minimizes external contamination. Glutamic acid production gradually decreased due to multiple uses of beads and consequently it reduces the shelf life. PMID:25215180

  17. Pilot Study to Evaluate Hydrogen Injection for Stimulating Reduction and Immobilization of Uranium in Groundwater at an ISR Mining Site

    NASA Astrophysics Data System (ADS)

    Clapp, L. W.; Cabezas, J.; Gamboa, Y.; Fernandez, W.

    2011-12-01

    State and federal regulations require that groundwater at in-situ recovery (ISR) uranium mining operations be restored to pre-mining conditions. Reverse osmosis (RO) filtration of several pore volumes of the post-leached groundwater and reinjection of the clean permeate is the most common technology currently used for restoring groundwater at uranium ISR sites. However, this approach does not revert the formation back to its initial reducing conditions, which can potentially impede timely groundwater restoration. In-situ biostimulation of indigenous iron- and sulfate reducing bacteria by injection of organic electron donors (e.g., ethanol, acetate, and lactate) to promote soluble uranium reduction and immobilization has been the subject of previous studies. However, injection of organic substrates has been observed to cause aquifer clogging near the injection point. In addition, U(VI) solubility may be enhanced through complexation with carbonate generated by organic carbon oxidation. An alternative approach that may overcome these problems involves the use of hydrogen as a reductant to promote microbial reduction and immobilization of U(VI) in situ. To test this approach, approximately 100,000 scf of compressed hydrogen gas was injected into a leached unconsolidated sand zone over two months at an ISR mining site. During this time groundwater was recirculated between injection and extraction wells (separated by 130 ft) at a rate of about 40 gpm and bromide was coinjected as a conservative tracer. A well monitoring program has been executed since June 2009 to evaluate the performance of the hydrogen injection. Current results show that U(VI) has been reduced from 4.2 to 0.05 ppm in the area surrounding the injection well and to 2.0 ± 0.3 ppm in the area surrounding the extraction well and two intermediate monitoring wells. Other water quality changes near the injection well include significant decreases in concentrations of Mo, sulfate, Fe, Mn, bicarbonate, Ca

  18. Batch and continuous flow studies of adsorptive removal of Cr(VI) by adapted bacterial consortia immobilized in alginate beads.

    PubMed

    Samuel, Jastin; Pulimi, Mrudula; Paul, Madona L; Maurya, Arun; Chandrasekaran, Natarajan; Mukherjee, Amitava

    2013-01-01

    The adsorptive removal of Cr(VI) by alginate beads containing Cr(VI)-adapted Acinetobacter junii, Escherichia coli and Bacillus subtilis in batch and continuous packed bed column reactors was investigated. Under optimized conditions (pH 3.0; contact time, 180 min; 30 °C; initial Cr(VI) concentration of 100 mg/L), 65.86 mg/g adsorption capacity was recorded in the batch study. When an adsorbent dosage of 1g/L, a flow rate of 5 mL/min, a bed height of 20 cm, an initial Cr(VI) concentration of 300 mg/L was employed, a capacity of 657 mg/g was noted for the continuous column assay. The batch sorption data followed the Langmuir isotherm and pseudo second order kinetics. Five sorption/desorption cycles yielded 100%, 99.63%, 95.31%, 80.7% and 74.22% regeneration, respectively. Cr(VI) adsorption studies using spiked ground water, freshwater and domestic wastewater in a packed bed reactor demonstrated Cr(VI) removals of 64.8%, 55.08%, 56.86% respectively. Cr(VI) sorption on immobilized bacteria was confirmed with Fourier-transform infrared and Energy dispersive X-ray spectroscopy. PMID:23201524

  19. Microstructural and potential dependence studies of urease-immobilized gold nanoparticles-polypyrrole composite film for urea detection.

    PubMed

    Rajesh; Puri, Nidhi; Mishra, Sujeet K; Laskar, Mariam J; Srivastava, Avanish K

    2014-01-01

    Gold nanoparticle-polypyrrole nanocomposite film was electrochemically deposited in a single-step polymerization of pyrrole in the presence of 3-mercaptopropionic acid (MPA)-capped gold nanoparticles (GNPs) and p-toluenesulfonic acid (pTSA) on the surface of an indium tin oxide (ITO)-coated glass plate. The carboxyl functional groups surrounding the GNPs within the polymer matrix were utilized for the immobilization of urease enzyme through carbodiimide coupling reaction for the construction of a Urs/GNP(MPA)-PPy/ITO-glass bioelectrode for urea detection in Tris-HCl buffer. The resulting bioelectrode film was characterized by atomic force microscopy (AFM), high-resolution transmission electron microscopy (HRTEM), contact angle measurement, Fourier transform infrared spectroscopy (FTIR), and electrochemical techniques. The potentiometric response of the bioelectrode made of polymer nanocomposite films of two different thicknesses prepared at 100 and 250 mC cm(-2) charge densities, respectively, was studied towards the urea concentration in Tris-HCl buffer (pH 7.4). The thin polymer nanocomposite film-based bioelectrode prepared at 100 mC cm(-2) charge density exhibited a comparatively good potentiometric response than a thick 250 mC cm(-2) charge density film with a linear range of urea detection from 0.01 to 10 mM with a sensitivity of 29.7 mV per decade. PMID:24142354

  20. Solid-state NMR studies of proteins immobilized on inorganic surfaces

    DOE PAGES

    Shaw, Wendy J.

    2014-10-29

    Solid state NMR is the primary tool for studying the quantitative, site-specific structure, orientation, and dynamics of biomineralization proteins under biologically relevant conditions. Two calcium phosphate proteins, statherin and leucine rich amelogenin protein (LRAP), have been studied in depth and have different features, challenging our ability to extract design principles. More recent studies of the significantly larger full-length amelogenin represent a challenging but necessary step to ultimately investigate the full diversity of biomineralization proteins. Interactions of amino acids and silaffin peptide with silica are also being studied, along with qualitative studies of proteins interacting with calcium carbonate. Dipolar recoupling techniquesmore » have formed the core of the quantitative studies, yet, the need for isolated spin pairs makes this approach costly and time intensive. The use of multi-dimensional techniques is advancing, methodology which, despite its challenges with these difficult-to-study proteins, will continue to drive future advancements in this area.« less

  1. Solid-state NMR studies of proteins immobilized on inorganic surfaces

    SciTech Connect

    Shaw, Wendy J.

    2014-10-29

    Solid state NMR is the primary tool for studying the quantitative, site-specific structure, orientation, and dynamics of biomineralization proteins under biologically relevant conditions. Two calcium phosphate proteins, statherin and leucine rich amelogenin protein (LRAP), have been studied in depth and have different features, challenging our ability to extract design principles. More recent studies of the significantly larger full-length amelogenin represent a challenging but necessary step to ultimately investigate the full diversity of biomineralization proteins. Interactions of amino acids and silaffin peptide with silica are also being studied, along with qualitative studies of proteins interacting with calcium carbonate. Dipolar recoupling techniques have formed the core of the quantitative studies, yet, the need for isolated spin pairs makes this approach costly and time intensive. The use of multi-dimensional techniques is advancing, methodology which, despite its challenges with these difficult-to-study proteins, will continue to drive future advancements in this area.

  2. Immobilization of bovine catalase onto magnetic nanoparticles.

    PubMed

    Doğaç, Yasemin İspirli; Teke, Mustafa

    2013-01-01

    The scope of this study is to achieve carrier-bound immobilization of catalase onto magnetic particles (Fe₃O₄ and Fe₂O₃NiO₂ · H₂O) to specify the optimum conditions of immobilization. Removal of H2O2 and the properties of immobilized sets were also investigated. To that end, adsorption and then cross-linking methods onto magnetic particles were performed. The optimum immobilization conditions were found for catalase: immobilization time (15 min for Fe₃O₄; 10 min for Fe2O₃NiO₂ · H₂O), the initial enzyme concentration (1 mg/mL), amount of magnetic particles (25 mg), and glutaraldehyde concentration (3%). The activity reaction conditions (optimum temperature, optimum pH, pH stability, thermal stability, operational stability, and reusability) were characterized. Also kinetic parameters were calculated by Lineweaver-Burk plots. The optimum pH values were found to be 7.0, 7.0, and 8.0 for free enzyme, Fe₃O₄-immobilized catalases, and Fe₂O₃NiO₂ · H₂O-immobilized catalases, respectively. All immobilized catalase systems displayed the optimum temperature between 25 and 35°C. Reusability studies showed that Fe₃O₄-immobilized catalase can be used 11 times with 50% loss in original activity, while Fe2O₃NiO₂ · H₂O-immobilized catalase lost 67% of activity after the same number of uses. Furthermore, immobilized catalase systems exhibited improved thermal and pH stability. The results transparently indicate that it is possible to have binding between enzyme and magnetic nanoparticles.

  3. Ultra-small rhenium nanoparticles immobilized on DNA scaffolds: An excellent material for surface enhanced Raman scattering and catalysis studies.

    PubMed

    Anantharaj, S; Sakthikumar, K; Elangovan, Ayyapan; Ravi, G; Karthik, T; Kundu, Subrata

    2016-12-01

    Highly Sensitive and ultra-small Rhenium (Re) metal nanoparticles (NPs) were successfully stabilized in water by the staging and fencing action of the versatile biomolecule DNA that resulted in two distinct aggregated chain-like morphologies with average grain sizes of 1.1±0.1nm and 0.7±0.1nm for the very first time within a minute of reaction time. Re NPs are formed by the borohydride reduction of ammonium perrhenate (NH4ReO4) in the presence of DNA at room temperature (RT) under stirring. The morphologies were controlled by carefully monitoring the molar ratio of NH4ReO4 and DNA. The synthesized material was employed in two potential applications: as a substrate for surface enhanced Raman scattering (SERS) studies and as a catalyst for the reduction of aromatic nitro compounds. SERS study was carried out by taking methylene blue (MB) as the probe and the highest SERS enhancement factor (EF) of 2.07×10(7) was found for the aggregated chain-like having average grain size of 0.7±0.1nm. Catalytic reduction of 4-nitro phenol (4-NP), 2-nitro phenol (2-NP) and 4-nitroaniline (4-NA) with a rate constant value of 6×10(-2)min(-1), 33.83×10(-2)min(-1) and 37.4×10(-2)min(-1) have testified the excellent catalytic performance of our Re NPs immobilized on DNA. The overall study have revealed the capability of DNA in stabilizing the highly reactive Re metal at nanoscale and made them applicable in practice. The present route can also be extended to prepare one dimensional (1-D), self-assembled NPs of other reactive metals, mixed metals or even metal oxides for specific applications in water based solutions. PMID:27571687

  4. Results of an inter-laboratory study of glass formulation for the immobilization of excess plutonium

    SciTech Connect

    Peeler, D.K.

    1999-12-08

    The primary focus of the current study is to determine allowable loadings of feed streams containing different ratios of plutonium, uranium, and minor components into the LaBS glass and to evaluate thermal stability with respect to the DWPF pour.

  5. Penetrable silica microspheres for immobilization of bovine serum albumin and their application to the study of the interaction between imatinib mesylate and protein by frontal affinity chromatography.

    PubMed

    Ma, Liyun; Li, Jing; Zhao, Juan; Liao, Han; Xu, Li; Shi, Zhi-guo

    2016-01-01

    In the current study, novel featured silica, named penetrable silica, simultaneously containing macropores and mesopores, was immobilized with bovine serum albumin (BSA) via Schiff base method. The obtained BSA-SiO2 was employed as the high-performance liquid chromatographic (HPLC) stationary phase. Firstly, D- and L-tryptophan were used as probes to investigate the chiral separation ability of the BSA-SiO2 stationary phase. An excellent enantioseparation factor was obtained up to 4.3 with acceptable stability within at least 1 month. Next, the BSA-SiO2 stationary phase was applied to study the interaction between imatinib mesylate (IM) and BSA by frontal affinity chromatography. A single type of binding site was found for IM with the immobilized BSA, and the hydrogen-bonding and van der Waals interactions were expected to be contributing interactions based on the thermodynamic studies, and this was a spontaneous process. Compared to the traditional silica for HPLC stationary phase, the proposed penetrable silica microsphere possessed a larger capacity to bond more BSA, minimizing column overloading effects and enhancing enantioseparation ability. In addition, the lower running column back pressure and fast mass transfer were meaningful for the column stability and lifetime. It was a good substrate to immobilize biomolecules for fast chiral resolution and screening drug-protein interactions.

  6. Penetrable silica microspheres for immobilization of bovine serum albumin and their application to the study of the interaction between imatinib mesylate and protein by frontal affinity chromatography.

    PubMed

    Ma, Liyun; Li, Jing; Zhao, Juan; Liao, Han; Xu, Li; Shi, Zhi-guo

    2016-01-01

    In the current study, novel featured silica, named penetrable silica, simultaneously containing macropores and mesopores, was immobilized with bovine serum albumin (BSA) via Schiff base method. The obtained BSA-SiO2 was employed as the high-performance liquid chromatographic (HPLC) stationary phase. Firstly, D- and L-tryptophan were used as probes to investigate the chiral separation ability of the BSA-SiO2 stationary phase. An excellent enantioseparation factor was obtained up to 4.3 with acceptable stability within at least 1 month. Next, the BSA-SiO2 stationary phase was applied to study the interaction between imatinib mesylate (IM) and BSA by frontal affinity chromatography. A single type of binding site was found for IM with the immobilized BSA, and the hydrogen-bonding and van der Waals interactions were expected to be contributing interactions based on the thermodynamic studies, and this was a spontaneous process. Compared to the traditional silica for HPLC stationary phase, the proposed penetrable silica microsphere possessed a larger capacity to bond more BSA, minimizing column overloading effects and enhancing enantioseparation ability. In addition, the lower running column back pressure and fast mass transfer were meaningful for the column stability and lifetime. It was a good substrate to immobilize biomolecules for fast chiral resolution and screening drug-protein interactions. PMID:26573171

  7. A comparative study of the most effective amendment for Pb, Zn and Cd immobilization in contaminated soils.

    PubMed

    Szrek, Dominik; Bajda, Tomasz; Manecki, Maciej

    2011-01-01

    The problem of an extensive contamination of soils with metals can be resolved using an in situ chemical immobilization technology. Five substances (natural zeolite, bog iron ore, "Polifoska 15" fertilizer, triple superphosphate, diammonium phosphate) were tested to determine their efficiency to immobilize Zn, Pb and Cd in smelter-contaminated soil in the Upper Silesia region. Soil samples were collected at three sites located in the vincity of a Pb-Zn smelter and a sludge landfill near the town of Bukowno. Effective reduction of leachable and fitoavailable Zn, Pb and Cd concentrations in soil was observed after addition of diammonium phosphate, "Polifoska 15" fertilizer and bog iron ore amendments. Additional test proved that immobilization effect gained by these amendments sustains at low-temperature conditions. It was noticed that phosphate addition resulted in lowering pH and mobilization of As(V) in soils. Good immobilization effectiveness and lack of major adverse effects of bog iron application suggest that this is the method of choice. PMID:21961559

  8. A comparative study of immobilization techniques for urease on glass-pH-electrode and its application in urea detection in blood serum.

    PubMed

    Sahney, Rachana; Anand, S; Puri, B K; Srivastava, A K

    2006-09-25

    Different techniques have been used (physical adsorption, physically entrapped sandwich and microencapsulation) for the immobilization of urease enzyme in tetramethylorthosilicate (TMOS) derived sol-gel matrix on the sensing surface of glass-pH-electrode. No significant leaching of enzyme occurs from the microencapsulated and physically entrapped enzyme sandwich films. Potentiometric techniques have been used for the estimation of urea concentration in each instance. Various parameters of biosensor performance have been studied which indicates that microencapsulation technique is a better method of enzyme immobilization in sol-gel films derived from TMOS. The advantage of microencapsulated biosensor over others include higher sensitivity (dpH/dp(C)=2.4), lower detection limit of 52 microg mL(-1), larger linear range (0.01-30 mM) of urea determination and reasonably long-term stability of about 25 days with 80% response signal.

  9. Activation and deactivation of a robust immobilized Cp*Ir-transfer hydrogenation catalyst: a multielement in situ X-ray absorption spectroscopy study.

    PubMed

    Sherborne, Grant J; Chapman, Michael R; Blacker, A John; Bourne, Richard A; Chamberlain, Thomas W; Crossley, Benjamin D; Lucas, Stephanie J; McGowan, Patrick C; Newton, Mark A; Screen, Thomas E O; Thompson, Paul; Willans, Charlotte E; Nguyen, Bao N

    2015-04-01

    A highly robust immobilized [Cp*IrCl2]2 precatalyst on Wang resin for transfer hydrogenation, which can be recycled up to 30 times, was studied using a novel combination of X-ray absorption spectroscopy (XAS) at Ir L3-edge, Cl K-edge, and K K-edge. These culminate in in situ XAS experiments that link structural changes of the Ir complex with its catalytic activity and its deactivation. Mercury poisoning and "hot filtration" experiments ruled out leached Ir as the active catalyst. Spectroscopic evidence indicates the exchange of one chloride ligand with an alkoxide to generate the active precatalyst. The exchange of the second chloride ligand, however, leads to a potassium alkoxide-iridate species as the deactivated form of this immobilized catalyst. These findings could be widely applicable to the many homogeneous transfer hydrogenation catalysts with Cp*IrCl substructure.

  10. Direct electron transfer and electrochemical study of hemoglobin immobilized in ZnO hollow spheres.

    PubMed

    Liu, Changhua; Xu, Jing; Wu, Zongfang

    2011-10-01

    ZnO hollow spheres were firstly prepared. A new type of amperometric hydrogen peroxide biosensor was fabricated by entrapping Hemoglobin (Hb) through the ZnO hollow spheres (ZHS) nanoparticles. The composition morphology and size were studied by transmission electron microscopy. The surface topography of the prepared films was imaged by atomic force microscope (AFM). Several techniques, including UV-vis absorption spectroscopy, cyclic voltammetry, chronoamperometry were employed to characterize the performance of the biosensor. The results indicated that the ZHS nanoparticles had enhanced the performance of the hydrogen peroxide sensors. The electrochemical parameters of Hb in the ZHS were calculated by the results of the electron-transfer coefficient (α) and the apparent heterogeneous electron-transfer rate constant K (s) as 0.5 and 3.1 s(-1), respectively. The resulting biosensors showed a wide linear range from 2.1 × 10(-6) to 5.18 × 10(-3) M, with a low detection limit of 7.0 × 10(-7) M (S/N = 3) under optimized experimental conditions. The results demonstrated that the ZHS matrix may improve the protein loading with the retention of bioactivity and greatly promote the direct electron transfer, which can be attributed to its unique morphology, high specific surface area, and biocompatibility. The biosensor obtained from this study possesses high sensitivity, good reproducibility, and long-term stability. PMID:21505813

  11. Pinning of the Contact Line during Evaporation on Heterogeneous Surfaces: Slowdown or Temporary Immobilization? Insights from a Nanoscale Study.

    PubMed

    Zhang, Jianguo; Müller-Plathe, Florian; Leroy, Frédéric

    2015-07-14

    The question of the effect of surface heterogeneities on the evaporation of liquid droplets from solid surfaces is addressed through nonequilibrium molecular dynamics simulations. The mechanism behind contact line pinning which is still unclear is discussed in detail on the nanoscale. Model systems with the Lennard-Jones interaction potential were employed to study the evaporation of nanometer-sized cylindrical droplets from a flat surface. The heterogeneity of the surface was modeled through alternating stripes of equal width but two chemical types. The first type leads to a contact angle of 67°, and the other leads to a contact angle of 115°. The stripe width was varied between 2 and 20 liquid-particle diameters. On the surface with the narrowest stripes, evaporation occurred at constant contact angle as if the surface was homogeneous, with a value of the contact angle as predicted by the regular Cassie-Baxter equation. When the width was increased, the contact angle oscillated during evaporation between two boundaries whose values depend on the stripe width. The evaporation behavior was thus found to be a direct signature of the typical size of the surface heterogeneity domains. The contact angle both at equilibrium and during evaporation could be predicted from a local Cassie-Baxter equation in which the surface composition within a distance of seven fluid-particle diameters around the contact line was considered, confirming the local nature of the interactions that drive the wetting behavior of droplets. More importantly, we propose a nanoscale explanation of pinning during evaporation. Pinning should be interpreted as a drastic slowdown of the contact line dynamics rather than a complete immobilization of it during a transition between two contact angle boundaries.

  12. Immobilization routes - they're not standing still

    SciTech Connect

    Basta, N.

    1982-04-19

    This paper reviews recent developments in the applications of enzyme immobilization techniques in various industries. Following success in high-fructose corn syrup production, enzyme immobilization is now making inroads in food processing and biomass-energy conversion. New studies focus on the immobilization of whole cells.

  13. Molecular study of thermal immobilization of chromium(VI) with clay.

    PubMed

    Wei, Yu-Ling; Hsieh, Huei-Fang; Yang, Yao-Wen; Lee, Jyh-Fu; Liang, Wei-Shun

    2005-04-01

    Clay that contains kaolinite has been used extensively as a raw material for manufacturing of bricks and china at 900-1100 degrees C. This study used clay to stabilize the contaminant chromium(VI) [Cr(VI)] through a heating process at 500-1100 degrees C. X-ray absorption spectroscopic results indicated that the 500-900 degrees C heating process transformed hazardous Cr(VI) to nontoxic Cr(III); Cr2O3 was the species detected as most abundant. The 1100 degrees C heating process caused the formation of Cr2SiO5, which was not detected in the samples heated at 500-900 degrees C. Fourier transformed extended X-ray absorption fine structure spectra were fitted by use of WinXAS software. Phase shifts and backscatter(ing) amplitudes for specific atom pairs, based on the crystallographic data for CrO3 and Cr2O3, were theoretically calculated with the FEFF software. The processed XAS data show that the first shell coordination numbers were similar to each other as the temperature was increased from 500 to 900 degrees C and 1100 degrees C, implying that their Cr(III) crystallite size was relatively similar. The interatomic distance between the target center element and the first shell for the 500-1100 degrees C samples was 1.98 A. The Debye-Waller factor for the 1100 degrees C sample was increased compared with the 500 and 900 degrees C samples and probably indicates the formation of Cr2SiO5.

  14. Immobilization and characterization of a thermostable lipase.

    PubMed

    Song, Chongfu; Sheng, Liangquan; Zhang, Xiaobo

    2013-12-01

    Lipases have found a number of commercial applications. However, thermostable lipase immobilized on nanoparticle is not extensively characterized. In this study, a recombinant thermostable lipase (designated as TtL) from Thermus thermophilus WL was expressed in Escherichia coli and immobilized onto 3-APTES-modified Fe3O4@SiO2 supermagnetic nanoparticles. Based on analyses with tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis, X-ray diffraction, transmission electron microscopy, and vibrating sample magnetometer observation, the diameter of immobilized lipase nanoparticle was 18.4 (± 2.4) nm, and its saturation magnetization value was 52.3 emu/g. The immobilized lipase could be separated from the reaction medium rapidly and easily in a magnetic field. The biochemical characterizations revealed that, comparing with the free one, the immobilized lipase exhibited better resistance to temperature, pH, metal ions, enzyme inhibitors, and detergents. The K m value for the immobilized TtL (2.56 mg/mL) was found to be lower than that of the free one (3.74 mg/mL), showing that the immobilization improved the affinity of lipase for its substrate. In addition, the immobilized TtL exhibited good reusability. It retained more than 79.5 % of its initial activity after reusing for 10 cycles. Therefore, our study presented that the possibility of the efficient reuse of the thermostable lipase immobilized on supermagnetic nanoparticles made it attractive from the viewpoint of practical application. PMID:23748908

  15. Effects of yeast immobilization on bioethanol production.

    PubMed

    Borovikova, Diana; Scherbaka, Rita; Patmalnieks, Aloizijs; Rapoport, Alexander

    2014-01-01

    The current study evaluated a newer method, which includes a dehydration step, of immobilizing Saccharomyces cerevisiae L-77 and S. cerevisiae L-73 onto hydroxylapatite and chamotte ceramic supports. The efficiency of cell immobilization on chamotte was significantly higher than hydroxylapatite. Immobilized yeast preparations were investigated for their ethanol-producing capabilities. The glucose concentration in a fermentation medium was 100 mg/mL. Immobilized preparations produced the same amount of ethanol (48 ± 0.5 mg/mL) as free cells after 36 H of fermentation. During the early stages of fermentation, immobilized yeast cells produced ethanol at a higher rate than free cells. Yeast preparations immobilized on both supports (hydroxylapatite and chamotte) were successfully used in six sequential batch fermentations without any loss of activity. The chamotte support was more stable in the fermentation medium during these six cycles of ethanol production. In addition to the high level of ethanol produced by cells immobilized on chamotte, the stability of this support and its low cost make it a promising material for biotechnologies associated with ethanol production.

  16. Effectiveness of olive oil for the prevention of pressure ulcers caused in immobilized patients within the scope of primary health care: study protocol for a randomized controlled trial

    PubMed Central

    2013-01-01

    Background Pressure ulcers are considered an important issue, mainly affecting immobilized older patients. These pressure ulcers increase the care burden for the professional health service staff as well as pharmaceutical expenditure. There are a number of studies on the effectiveness of different products used for the prevention of pressure ulcers; however, most of these studies were carried out at a hospital level, basically using hyperoxygenated fatty acids (HOFA). There are no studies focused specifically on the use of olive-oil-based products and therefore this research is intended to find the most cost-effective treatment and achieve an alternative treatment. Methods/design The main objective is to assess the effectiveness of olive oil, comparing it with HOFA, to treat immobilized patients at home who are at risk of pressure ulcers. As a secondary objective, the cost-effectiveness balance of this new application with regard to the HOFA will be assessed. The study is designed as a noninferiority, triple-blinded, parallel, multi-center, randomized clinical trial. The scope of the study is the population attending primary health centers in Andalucía (Spain) in the regional areas of Malaga, Granada, Seville, and Cadiz. Immobilized patients at risk of pressure ulcers will be targeted. The target group will be treated by application of an olive-oil-based formula whereas the control group will be treated by application of HOFA to the control group. The follow-up period will be 16 weeks. The main variable will be the presence of pressure ulcers in the patient. Secondary variables include sociodemographic and clinical information, caregiver information, and whether technical support exists. Statistical analysis will include the Kolmogorov-Smirnov test, symmetry and kurtosis analysis, bivariate analysis using the Student’s t and chi-squared tests as well as the Wilcoxon and the Man-Whitney U tests, ANOVA and multivariate logistic regression analysis. Discussion The

  17. Diffusion of solutes inside bacterial colonies immobilized in model cheese depends on their physicochemical properties: a time-lapse microscopy study

    PubMed Central

    Floury, Juliane; El Mourdi, Ilham; Silva, Juliana V. C.; Lortal, Sylvie; Thierry, Anne; Jeanson, Sophie

    2015-01-01

    During cheese processing and ripening, bacteria develop as colonies. Substrates and metabolites must then diffuse either from or into the colonies. Exploring how the inner cells of the colony access the substrates or get rid of the products leads to study the diffusion of solutes inside bacterial colonies immobilized in cheese. Diffusion limitations of substrates within the bacterial colony could lead to starvation for the cells in the center of the colony. This study aimed at better understands ripening at the colony level, by investigating how diffusion phenomena inside colonies vary depending on both the physicochemical properties of the solutes and Lactococcus lactis strain. Dextrans (4, 70, and 155 kDa) and milk proteins (BSA, lactoferrin and αS1-casein) of different sizes and physicochemical properties were chosen as model of diffusing solutes, and two L. lactis strains presenting different surface properties were immobilized as colonies in a model cheese. Diffusion of solutes inside and around colonies was experimentally followed by time-lapse confocal microscopy. Dextran solutes diffused inside both lactococci colonies with a non-significantly different effective diffusion coefficient, which depended mainly on size of the solute. However, whereas flexible and neutral hydrophilic polymers such as dextran can diffuse inside colonies whatever its size, none of the three proteins investigated in this study could penetrate inside lactococci colonies. Therefore, the diffusion behavior of macromolecules through bacterial colonies immobilized in a model cheese did not only depends on the size of the diffusing solutes, but also and mainly on their physicochemical properties. Milk caseins are probably first hydrolyzed by the cell wall proteases of L. lactis and/or other proteases present in the cheese, and then the generated peptides diffuse inside colonies to be further metabolized into smaller peptides and amino acids by all the cells located inside the colonies

  18. Hyperalgesia in an immobilized rat hindlimb: effect of treadmill exercise using non-immobilized limbs.

    PubMed

    Chuganji, Sayaka; Nakano, Jiro; Sekino, Yuki; Hamaue, Yohei; Sakamoto, Junya; Okita, Minoru

    2015-01-01

    Cast immobilization of limbs causes hyperalgesia, which is a decline of the threshold of mechanical and thermal mechanical stimuli. The immobilization-induced hyperalgesia (IIH) can disturb rehabilitation and activities of daily living in patients with orthopedic disorders. However, it is unclear what therapeutic and preventive approaches can be used to alleviate IIH. Exercise that activates the descending pain modulatory system may be effective for IIH. The purpose of this study was to investigate the effects of treadmill exercise during the immobilization period, using the non-immobilized limbs, on IIH. Thirty-six 8-week-old Wistar rats were randomly divided into (1) control, (2) immobilization (Im), and (3) immobilization and treadmill exercise (Im+Ex) groups. In the Im and Im+Ex groups, the right ankle joints of each rat were immobilized in full plantar flexion with a plaster cast for an 8-week period. In the Im+Ex group, treadmill exercise (15 m/min, 30 min/day, 5 days/week) was administered during the immobilization period while the right hindlimb was kept immobilized. Mechanical hyperalgesia was measured using von Frey filaments every week. To investigate possible activation of the descending pain modulatory system, beta-endorphin expression levels in hypothalamus and midbrain periaqueductal gray were analyzed. Although IIH clearly occurred in the Im group, the hyperalgesia was partially but significantly reduced in the Im+Ex group. Beta-endorphin, which is one of the endogenous opioids, was selectively increased in the hypothalamus and midbrain periaqueductal gray of the Im+Ex group. Our data suggest that treadmill running using the non-immobilized limbs reduces the amount of hyperalgesia induced in the immobilized limb even if it is not freed. This ameliorating effect might be due to the descending pain modulatory system being activated by upregulation of beta-endorphin in the brain. PMID:25304541

  19. Plutonium Immobilization Puck Handling

    SciTech Connect

    Kriikku, E.

    1999-01-26

    The Plutonium Immobilization Project (PIP) will immobilize excess plutonium and store the plutonium in a high level waste radiation field. To accomplish these goals, the PIP will process various forms of plutonium into plutonium oxide, mix the oxide powder with ceramic precursors, press the mixture into pucks, sinter the pucks into a ceramic puck, load the pucks into metal cans, seal the cans, load the cans into magazines, and load the magazines into a Defense Waste Processing Facility (DPWF) canister. These canisters will be sent to the DWPF, an existing Savannah River Site (SRS) facility, where molten high level waste glass will be poured into the canisters encapsulating the ceramic pucks. Due to the plutonium radiation, remote equipment will perform these operations in a contained environment. The Plutonium Immobilization Project is in the early design stages and the facility will begin operation in 2005. This paper will discuss the Plutonium Immobilization puck handling conceptual design and the puck handling equipment testing.

  20. Development of immobilized cellulase

    SciTech Connect

    Luther, M.A.; Halbert, D.J.

    1982-08-01

    The immobilization of cellulase from the fungus Trichoderma reesei on the surface of calcium alginate gel spheres was investigated. The immobilized cellulase catalyzed the hydrolysis of cellulose to glucose. The linking agents, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and glutaraldehyde, decreased free-enzyme activity by 95%, and the maximum observed retention of cellulase activity after immobilization was 2%. Leakage of enzyme from the support was observed. A fivefold increase in glucose production was seen after the addition of ..beta..-glucosidase-impregnated spheres to the cellulase spheres, suggesting that cellobiose may be accumulating during the reaction. A simple economic analysis suggested that the immobilized-enzyme activity per unit volume might have to be increased by a factor of fifty to become competitive with the free-enzyme process.

  1. Principles, techniques, and applications of biocatalyst immobilization for industrial application.

    PubMed

    Eş, Ismail; Vieira, José Daniel Gonçalves; Amaral, André Corrêa

    2015-03-01

    Immobilization is one of the most effective and powerful tools used in industry, which has been studied and improved since the last century. Various immobilization techniques and support materials have been used on both laboratory and industrial scale. Each immobilization technique is applicable for a specific production mostly depending on the cost and sensibility of process. Compared to free biocatalyst systems, immobilization techniques often offer better stability, increased activity and selectivity, higher resistance, improved separation and purification, reuse of enzymes, and consequently more efficient process. Recently, many reviews have been published about immobilization systems; however, most of them have focused on a specific application or not emphasized in details. This review focuses on most commonly used techniques in industry with many recent applications including using bioreactor systems for industrial production. It is also aimed to emphasize the advantages and disadvantages of the immobilization techniques and how these systems improve process productivity compared to non-immobilized systems.

  2. Immobilization induced hypercalcemia

    PubMed Central

    Cano-Torres, Edgar Alonso; González-Cantú, Arnulfo; Hinojosa-Garza, Gabriela; Castilleja-Leal, Fernando

    2016-01-01

    Summary Immobilization hypercalcemia is an uncommon diagnosis associated with increased bone remodeling disorders and conditions associated with limited movement such as medullar lesions or vascular events. Diagnosis requires an extensive evaluation to rule out other causes of hypercalcemia. This is a report of a woman with prolonged immobilization who presented with severe hypercalcemia. This case contributes to identification of severe hypercalcemia as a result of immobility and the description of bone metabolism during this state. PMID:27252745

  3. Effect of fermentation parameters on bio-alcohols production from glycerol using immobilized Clostridium pasteurianum: an optimization study.

    PubMed

    Khanna, Swati; Goyal, Arun; Moholkar, Vijayanand S

    2013-01-01

    This article addresses the issue of effect of fermentation parameters for conversion of glycerol (in both pure and crude form) into three value-added products, namely, ethanol, butanol, and 1,3-propanediol (1,3-PDO), by immobilized Clostridium pasteurianum and thereby addresses the statistical optimization of this process. The analysis of effect of different process parameters such as agitation rate, fermentation temperature, medium pH, and initial glycerol concentration indicated that medium pH was the most critical factor for total alcohols production in case of pure glycerol as fermentation substrate. On the other hand, initial glycerol concentration was the most significant factor for fermentation with crude glycerol. An interesting observation was that the optimized set of fermentation parameters was found to be independent of the type of glycerol (either pure or crude) used. At optimum conditions of agitation rate (200 rpm), initial glycerol concentration (25 g/L), fermentation temperature (30°C), and medium pH (7.0), the total alcohols production was almost equal in anaerobic shake flasks and 2-L bioreactor. This essentially means that at optimum process parameters, the scale of operation does not affect the output of the process. The immobilized cells could be reused for multiple cycles for both pure and crude glycerol fermentation.

  4. Study of the potential of the air lift bioreactor for xylitol production in fed-batch cultures by Debaryomyces hansenii immobilized in alginate beads.

    PubMed

    Pérez-Bibbins, Belinda; de Souza Oliveira, Ricardo Pinheiro; Torrado, Ana; Aguilar-Uscanga, María Guadalupe; Domínguez, José Manuel

    2014-01-01

    Cell immobilization has shown to be especially adequate for xylitol production. This work studies the suitability of the air lift bioreactor for xylitol production by Debaryomyces hansenii immobilized in Ca-alginate operating in fed-batch cultures to avoid substrate inhibition. The results showed that the air lift bioreactor is an adequate system since the minimum air flow required for fluidization was even lower than that leading to the microaerobic conditions that trigger xylitol accumulation by this yeast, also maintaining the integrity of the alginate beads and the viability of the immobilized cells until 3 months of reuses. Maximum productivities and yields of 0.43 g/l/h and 0.71 g/g were achieved with a xylose concentration of 60 g/l after each feeding. The xylose feeding rate, the air flow, and the biomass concentration at the beginning of the fed-batch operation have shown to be critical parameters for achieving high productivities and yields. Although a maximum xylitol production of 139 g/l was obtained, product inhibition was evidenced in batch experiments, which allowed estimating at 200 and 275 g/l the IC50 for xylitol productivity and yield, respectively. The remarkable production of glycerol in the absence of glucose was noticeable, which could not only be attributed to the osmoregulatory function of this polyol in conditions of high osmotic pressure caused by high xylitol concentrations but also to the role of the glycerol synthesis pathway in the regeneration of NAD(+) in conditions of suboptimal microaeration caused by insufficient aeration or high oxygen demand when high biomass concentrations were achieved.

  5. Equilibrium isotherm and kinetic studies for the simultaneous removal of phenol and cyanide by use of S. odorifera (MTCC 5700) immobilized on coconut shell activated carbon

    NASA Astrophysics Data System (ADS)

    Singh, Neetu; Balomajumder, Chandrajit

    2016-09-01

    In this study, simultaneous removal of phenol and cyanide by a microorganism S. odorifera (MTCC 5700) immobilized onto coconut shell activated carbon surface (CSAC) was studied in batch reactor from mono and binary component aqueous solution. Activated carbon was derived from coconut shell by chemical activation method. Ferric chloride (Fecl3), used as surface modification agents was applied to biomass. Optimum biosorption conditions were obtained as a function of biosorbent dosage, pH, temperature, contact time and initial phenol and cyanide concentration. To define the equilibrium isotherms, experimental data were analyzed by five mono component isotherm and six binary component isotherm models. The higher uptake capacity of phenol and cyanide onto CSAC biosorbent surface was 450.02 and 2.58 mg/g, respectively. Nonlinear regression analysis was used for determining the best fit model on the basis of error functions and also for calculating the parameters involved in kinetic and isotherm models. The kinetic study results revealed that Fractal-like mixed first second order model and Brouser-Weron-Sototlongo models for phenol and cyanide were capable to offer accurate explanation of biosorption kinetic. According to the experimental data results, CSAC with immobilization of bacterium S. odorifera (MTCC 5700) seems to be an alternative and effective biosorbent for the elimination of phenol and cyanide from binary component aqueous solution.

  6. Immobilization-induced hypersensitivity associated with spinal cord sensitization during cast immobilization and after cast removal in rats.

    PubMed

    Hamaue, Yohei; Nakano, Jiro; Sekino, Yuki; Chuganji, Sayaka; Sakamoto, Jyunya; Yoshimura, Toshiro; Origuchi, Tomoki; Okita, Minoru

    2013-11-01

    This study examined mechanical and thermal hypersensitivity in the rat hind paw during cast immobilization of the hind limbs for 4 or 8 weeks and following cast removal. Blood flow, skin temperature, and volume of the rat hind paw were assessed in order to determine peripheral circulation of the hind limbs. Sensitization was analyzed by measuring the expression of the calcitonin gene-related peptide (CGRP) in the spinal dorsal horn following cast immobilization. Two weeks post immobilization, mechanical and thermal sensitivities increased significantly in all rats; however, peripheral circulation was not affected by immobilization. Cast immobilization for 8 weeks induced more serious hypersensitivity compared to cast immobilization for 4 weeks. Moreover, CGRP expression in the deeper lamina layer of the spinal dorsal horn increased in the rats immobilized for 8 weeks but not in those immobilized for 4 weeks. These findings suggest that immobilization-induced hypersensitivity develops during the immobilization period without affecting peripheral circulation. Our results also highlight the possibility that prolonged immobilization induces central sensitization in the spinal cord.

  7. Affinity covalent immobilization of glucoamylase onto ρ-benzoquinone-activated alginate beads: II. Enzyme immobilization and characterization.

    PubMed

    Eldin, M S Mohy; Seuror, E I; Nasr, M A; Tieama, H A

    2011-05-01

    A novel affinity covalent immobilization technique of glucoamylase enzyme onto ρ-benzoquinone-activated alginate beads was presented and compared with traditional entrapment one. Factors affecting the immobilization process such as enzyme concentration, alginate concentration, calcium chloride concentration, cross-linking time, and temperature were studied. No shift in the optimum temperature and pH of immobilized enzymes was observed. In addition, K (m) values of free and entrapped glucoamylase were found to be almost identical, while the covalently immobilized enzyme shows the lowest affinity for substrate. In accordance, V (m) value of covalently immobilized enzyme was found lowest among free and immobilized counter parts. On the other hand, the retained activity of covalently immobilized glucoamylase has been improved and was found higher than that of entrapped one. Finally, the industrial applicability of covalently immobilized glucoamylase has been investigated through monitoring both shelf and operational stability characters. The covalently immobilized enzyme kept its activity over 36 days of shelf storage and after 30 repeated use runs. Drying the catalytic beads greatly reduced its activity in the beginning but recovered its lost part during use. In general, the newly developed affinity covalent immobilization technique of glucoamylase onto ρ-benzoquinone-activated alginate carrier is simple yet effective and could be used for the immobilization of some other enzymes especially amylases.

  8. Hemopoiesis-stimulating activity of immobilized oligonucleotides and hyaluronidase during cytostatic-induced myelosuppression.

    PubMed

    Dygai, A M; Skurikhin, E G; Pershina, O V; Zhdanov, V V; Khmelevskaya, A M; Andreeva, T V; Poponina, A M; Zjuzkov, G N; Udut, E V; Khrichkova, T Ju; Simanina, E V; Miroshnichenko, L A; Stavrova, L A; Tchaikovsky, A S; Markova, T S; Gurto, R V; Brjushinina, O S; Slepichev, V A

    2011-03-01

    The hemopoiesis-stimulating effect of combined treatment with immobilized oligonucleotides and hyaluronidase preparations was studied during cytostatic-induced myelosuppression caused by cyclophosphamide administration. Immobilized hyaluronidase was shown to increase the efficiency of correction of changes in the erythroid and granulocytic hemopoietic stems with immobilized oligonucleotides. This potentiation of the effect of immobilized oligonucleotides by immobilized hyaluronidase was related to an increase in functional activity of committed hemopoietic precursors.

  9. Comparative study on functionalized SBA-15 and SBA-16 nanostructured materials used for immobilization of D-amino acid oxidase

    NASA Astrophysics Data System (ADS)

    Hy, Le Gia; Phuong, Dang Tuyet; Yen, Hoang; Hoan, Nguyen Thi Vuong; Linh, Bui Thi Hai; Thang, Hoang vinh; Hoa, Tran Thi Kim; Thang, Dinh Cao; Nguyen, Vu Thi Hanh; Thao, Phan Thi Hong; Giap, Chu Van; Tuan, Vu Anh

    2008-12-01

    SBA-15 and SBA-16 nanostucrured materials were synthesized via hydrothermal treatment and were functionalized with 3-aminopropyltriethoxysilane (APTES), and vinyltriethoxysilane (VTES). The obtained samples were characterized by different techniques such as XRD, BET, IR and TEM. After functionalization, it showed that these nanostrucrured materials were still maintained the hexagonal pore structure of the parent SBA-15 and cubic cage structure of the parent SBA-16. The non-functionalized pure silica SBA-15 and SBA-16 as well as functionalized SBA-15 and SBA-16 materials were used to immobilize DAAO, which is industrially important enzyme for the production of glutaryl 7-amino cephalosporanic acid (GL-7-ACA) from cephalosporin C (CPC). The obtained results revealed that functionalized SBA- 15 and SBA-16 materials exhibited higher enzymatic activity and stability than those of non-functionalized ones. This might be due to the enhancing of surface hydrophobicity upon functionalization. The surface functionalization of the nanostructured silicas with organic groups can enhance the interaction between enzyme and the supports and consequently increasing the operational stability of the immobilized enzyme. The loading of enzyme on SBA-15 materials was higher than that on SBA-16 samples (both functionalized and non-functionalized types). This might be explained by the difference in pore size and type (cylindrical for SBA-15 and bottle-neck for SBA-16) as well as structure shape (hexagonal for SBA-15 and cubic cage for SBA-16) of both mesoporous materials. Additionally, nature of functionalized groups significantly affected the enzymatic activity. Effects on surface binding force, nature of functional groups, pore size of supports were investigated and discussed.

  10. Ultra-low field T1 vs. T1rho at 3T and 7T: study of rotationally immobilized protein gels and animal brain tissues

    NASA Astrophysics Data System (ADS)

    Dong, Hui; Inglis, Ben; Barr, Ian; Clarke, John

    2015-03-01

    Clinical magnetic resonance imaging (MRI) machines operating in static fields of typically 1.5 T or 3 T can capture information on slow molecular dynamics utilizing the so-called T1rho technique. This technique, in which a radiofrequency (RF) spin-lock field is applied with microtesla amplitude, has been used, for example, to determine the onset time of stroke in studies on rats. The long RF pulse, however, may exceed the specific absorption rate (SAR) limit, putting subjects at risk. Ultra-low-field (ULF) MRI, based on Superconducting Quantum Interference Devices (SQUIDs), directly detects proton signals at a static magnetic field of typically 50-250 μT. Using our ULF MRI system with adjustable static field of typically 55 to 240 μT, we systematically measured the T1 and T2 dispersion profiles of rotationally immobilized protein gels (bovine serum albumin), ex vivo pig brains, and ex vivo rat brains with induced stroke. Comparing the ULF results with T1rho dispersion obtained at 3 T and 7 T, we find that the degree of protein immobilization determines the frequency-dependence of both T1 and T1rho. Furthermore, T1rho and ULF T1 show similar results for stroke, suggesting that ULF MRI may be used to image traumatic brain injury with negligible SAR. This research was supported by the Henry H. Wheeler, Jr. Brain Imaging Center and the Donaldson Trust.

  11. [The immobilization of giraffes].

    PubMed

    Wiesner, H; von Hegel, G

    1989-01-01

    The anatomical and physiological conditions of blood circulation in the giraffe are pointed out. 16 immobilizations in the giraffe of either sex are reported, of which 10 were immobilized according to the following scheme. 1. Premedication: 30 mg Xylazine 150 mg Hyaluronidase 2. 15 minutes later a halter with two long ropes is put on to hold up the animals' heads after they lay down. 3. 20 minutes after premedication the injection of 5.6-6.0 mg Etorphine (2.5-2.7 ml Immobilon) together with 150 I.U. Hyaluronidase follows. 4. We think that the most important fact is to hold the animals head and neck in an upright position during the whole time of immobilization. 5. Within 3 to 5 minutes after the intravenous application of 15 mg Diprenorphine (5.0 ml Revivon) the animals raise without any problems.

  12. A comparative study of different protein immobilization methods for the construction of an efficient nano-structured lactate oxidase-SWCNT-biosensor.

    PubMed

    Pagán, Miraida; Suazo, Dámaris; Del Toro, Nicole; Griebenow, Kai

    2015-02-15

    We constructed lactate biosensors by immobilization of lactate oxidase (LOx) onto a single-walled carbon nanotube (SWCNT) electrode. The first step of the sensor construction was the immobilization of oxidized SWCNT onto a platinum electrode modified with 4-aminothiophenol (4-ATP). Two enzyme immobilization methods were used to construct the biosensors, i.e., covalent immobilization using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and physical adsorption. Atomic force microscopy (AFM) experiments confirmed the immobilization of SWCNT during the biosensor construction and X-ray photoelectron spectroscopy (XPS) experiments confirmed covalent immobilization of LOx onto the SWCNT in the first method. The biosensor based on covalent enzyme immobilization showed a sensitivity of 5.8 μA/mM, a linearity up to 0.12 mM of L-lactate, and a detection limit of 4.0 μM. The biosensor based on protein adsorption displayed a sensitivity of 9.4 μA/mM, retaining linearity up to 0.18 mM of L-lactate with a detection limit of 3.0 μM. The difference in the biosensor response can be attributed to protein conformational or dynamical changes during covalent immobilization. The stability of the biosensors was tested at different temperatures and after different storage periods. The thermostability of the biosensors after incubation at 60 °C demonstrated that the biosensor with covalently immobilized LOx retained a higher response compared with the adsorbed protein. Long-term stability experiments show a better residual activity of 40% for the covalently immobilized protein compared to 20% of residual activity for the adsorbed protein after 25 d storage. Covalent protein immobilization was superior compared to adsorption in preserving biosensor functionality over extended time period.

  13. An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes

    PubMed Central

    Mohamad, Nur Royhaila; Marzuki, Nur Haziqah Che; Buang, Nor Aziah; Huyop, Fahrul; Wahab, Roswanira Abdul

    2015-01-01

    The current demands of sustainable green methodologies have increased the use of enzymatic technology in industrial processes. Employment of enzyme as biocatalysts offers the benefits of mild reaction conditions, biodegradability and catalytic efficiency. The harsh conditions of industrial processes, however, increase propensity of enzyme destabilization, shortening their industrial lifespan. Consequently, the technology of enzyme immobilization provides an effective means to circumvent these concerns by enhancing enzyme catalytic properties and also simplify downstream processing and improve operational stability. There are several techniques used to immobilize the enzymes onto supports which range from reversible physical adsorption and ionic linkages, to the irreversible stable covalent bonds. Such techniques produce immobilized enzymes of varying stability due to changes in the surface microenvironment and degree of multipoint attachment. Hence, it is mandatory to obtain information about the structure of the enzyme protein following interaction with the support surface as well as interactions of the enzymes with other proteins. Characterization technologies at the nanoscale level to study enzymes immobilized on surfaces are crucial to obtain valuable qualitative and quantitative information, including morphological visualization of the immobilized enzymes. These technologies are pertinent to assess efficacy of an immobilization technique and development of future enzyme immobilization strategies. PMID:26019635

  14. Stirred anaerobic sequencing batch reactor containing immobilized biomass: a behavior study when submitted to different fill times.

    PubMed

    Borges, A C; Siman, R R; Rodrigues, J A D; Ratusznei, S M; Zaiat, M; Foresti, E; Borzani, W

    2004-01-01

    The effect of the filling stage on the behavior of a mechanically stirred anaerobic sequencing batch reactor containing biomass immobilized on 1 cm polyurethane foam cubes was investigated. The reactor was made of acrylic with a capacity of 6.3 L, treating per cycle 2.5 L synthetic low-strength wastewater with a concentration of 500 mgCOD/L, at 30+/-1 degrees C. Eight-hour cycles (tC) and agitation of 500 rpm were utilized. At the beginning of each cycle 60% of the wastewater volume was treated, sufficient to completely cover the bed. The remaining volume was added at different fill times (tF) of 10, 120, 240, 260 and 480 min. The results obtained showed that ratios of tF/tC < or = 0.5 enabled organic matter removal higher than 75% and 70% for filtered and non-filtered samples, respectively. Ratios of tF/tC > 0.5, despite operation stability, resulted in loss of efficiency and formation of viscous material, similar to extra-cellular polymeric substances. PMID:15303756

  15. Feasibility study of complete nitrogen removal from domestic wastewater by consequent nitrification-denitrification using immobilized nitrifiers in gel beads

    SciTech Connect

    Libman, V.; Eliosov, B.; Argaman, Y.

    2000-02-01

    A two-stage process for complete removal of nitrogen from domestic wastewater is proposed in this paper. The two stages of this process are an aerobic stage using nitrifying microorganisms immobilized in gel beads and a conventional anoxic stage. The possible application of gel-entrapped biomass for high-rate, complete nitrogen removal from domestic wastewater was verified through theoretical and experimental work. Two assumption s were confirmed in the present work: (a) substances present in domestic wastewater do not inhibit nitrification, and (b) heterotrophic biomass that builds up on the surface of nitrifying beads is not expected to affect ammonium oxidation or remove a significant part of the biochemical oxygen demand required for denitrification. The effect of the attached heterotrophic layer on system performance was evaluated through theoretical prediction and short-term experiments. Calculations indicated that complete nitrogen removal could be achieved if the heterotrophic layer on 2-mm gel beads is less than 12 {micro}m thick. The heterotrophic biofilm observed in the experiments did not homogeneously spread over the entire bead surface. It was found that attached growth of heterotrophs is affected by mixing intensity and type of carrier material.

  16. [Nitrogen uptake and denitrification study on the joint treatment of aquatic vegetation and immobilized nitrogen cycling bacteria in Taihu Lake].

    PubMed

    Liu, Dan-Dan; Li, Zheng-Kui; Ye, Zhong-Xiang; Zhang, Wan-Guang

    2014-10-01

    Undisturbed sediment cores were collected from Meiliang Bay, Taihu Lake. Immobilized nitrogen cycling bacteria (INCB), Elodea nuttallii were added to four groups of restoration incubation chambers respectively to explore the nitrogen removal mechanism in different restoration treatments. uN tracer and isotope pairing technique were used to determine the rates of plant uptake and denitrification in different treatments. The results showed that denitrification rates were significantly different among the treatments, while cores with addition of both INCB and Elodea nuttallii achieved the highest denitrification rate of 99.35 μmol · (m2 · h)( -1) and plant uptake rate of 36.55 μg · (m2 · h) -1. Elodea nuttallii in the cores could assimilate nitrate itself and enhance coupled nitrification- denitrification. Compared with plant uptake, denitrification was the main pathway of nitrogen removal. The results also showed that the combination of Elodea nuttallii and INCB could promote benthic nitrogen removal and purification of water body.

  17. Biosensor studies of collagen and laminin binding with immobilized Escherichia coli O157:H7 and inhibition with naturally occurring food additives

    NASA Astrophysics Data System (ADS)

    Medina, Marjorie B.

    1999-01-01

    Escherichia coli O157:H7 outbreaks were mostly due to consumption of undercooked contaminated beef which resulted in severe illness and several fatalities. Recalls of contaminated meat are costly for the meat industry. Our research attempts to understand the mechanisms of bacterial adhesion on animal carcass in order to eliminate or reduce pathogens in foods. We have reported the interactions of immobilized E. coli O157:H7 cells with extracellular matrix (ECM) components using a surface plasmon resonance biosensor (BIAcore). These studies showed that immobilized bacterial cells allowed the study of real-time binding interactions of bacterial surface with the ECM compounds, collagen I, laminin and fibronectin. Collagen I and laminin bound to the E. coli sensor surface with dissociation and association rates ranging from 106 to 109. Binding of collagen I and laminin mixture resulted in synergistic binding signals. An inhibition model was derived using collagen-laminin as the ligand which binds with E. coli sensor. A select group of naturally occurring food additives was evaluated by determining their effectivity in inhibiting the collagen-laminin binding to the bacterial sensor. Bound collagen-laminin was detached from the E. coli sensor surface with the aid of an organic acid. The biosensor results were verified with cell aggregation assays which were observed with optical and electron microscopes. These biosensor studies provided understanding of bacterial adhesion to connective tissue macromolecules. It also provided a model system for the rapid assessment of potential inhibitors that can be used in carcass treatment to inhibit or reduce bacterial contamination.

  18. Matrix-Immobilized BMP-2 on Microcontact Printed Fibronectin as an in vitro Tool to Study BMP-Mediated Signaling and Cell Migration

    PubMed Central

    Hauff, Kristin; Zambarda, Chiara; Dietrich, Miriam; Halbig, Maria; Grab, Anna Luise; Medda, Rebecca; Cavalcanti-Adam, Elisabetta Ada

    2015-01-01

    During development, growth factors (GFs) such as bone morphogenetic proteins (BMPs) exert important functions in several tissues by regulating signaling for cell differentiation and migration. In vivo, the extracellular matrix (ECM) not only provides support for adherent cells, but also acts as reservoir of GFs. Several constituents of the ECM provide adhesive cues, which serve as binding sites for cell trans-membrane receptors, such as integrins. In conveying adhesion-mediated signaling to the intracellular compartment, integrins do not function alone but rather crosstalk and cooperate with other receptors, such as GF receptors. Here, we present a strategy for the immobilization of BMP-2 onto cellular fibronectin (cFN), a key protein of the ECM, to investigate GF-mediated signaling and migration. Following biotinylation, BMP-2 was linked to biotinylated cFN using NeutrAvidin as cross-linker. Characterization with quartz crystal microbalance with dissipation monitoring and enzyme-linked immunosorbent assay confirmed the efficient immobilization of BMP-2 on cFN over a period of 24 h. To validate the bioactivity of matrix-immobilized BMP-2 (iBMP-2), we investigated short- and long-term responses of C2C12 myoblasts, which are an established in vitro model for BMP-2 signaling, in comparison to soluble BMP-2 (sBMP-2) or in absence of GFs. Similarly to sBMP-2, iBMP-2 triggered Smad 1/5 phosphorylation and translocation of the complex to the nucleus, corresponding to the activation of BMP-mediated Smad-dependent pathway. Additionally, successful suppression of myotube formation was observed after 6 days in sBMP-2 and iBMP-2. We next implemented this approach in the fabrication of cFN micropatterned stripes by soft lithography. These stripes allowed cell-surface interaction only on the patterned cFN, since the surface in between was passivated, thus serving as platform for studies on directed cell migration. During a 10-h observation time, the migratory behavior

  19. Immobilizing Yarrowia lipolytica Lipase Lip2 via Improvement of Microspheres by Gelatin Modification.

    PubMed

    Xie, Rong; Cui, Caixia; Chen, Biqiang; Tan, Tianwei

    2015-10-01

    The purpose of this study was to investigate the feasibility of immobilizing Yarrowia lipolytica lipase lip2 on epoxy microspheres with or without gelatin modifications. The activity of lipase immobilized on gelatin-modified supports was twofold higher than those immobilized on native supports. There was no significant difference in the Michaelis-Menten constant (K M ) between the two immobilized lipases. However, lipase immobilized on gelatin modified supports showed an approximately fourfold higher V max than lipase immobilized on native supports. Lipase immobilization on the gelatin-modified support exhibited a significantly improved operational stability in an esterification system. After it was reused for a total of 35 batches, the ester conversion of lipase immobilized on gelatin-modified and native microspheres was 83 and 60 %, respectively. Furthermore, the immobilized lipase could be stored at 4 °C for 12 months without any loss of activity.

  20. C-Terminal-oriented Immobilization of Enzymes Using Sortase A-mediated Technique.

    PubMed

    Hata, Yuto; Matsumoto, Takuya; Tanaka, Tsutomu; Kondo, Akihiko

    2015-10-01

    In the present study, sortase A-mediated immobilization of enzymes was used for the preparation of immobilized enzymes. Thermobifida fusca YX β-glucosidase (BGL) or Streptococcus bovis 148 α-amylase (AmyA) were produced with C-terminal sortase A recognition sequences. The resulting fusion proteins were successfully immobilized on nanoparticle surfaces using sortase A. Some properties (activity, stability, and reusability) of the immobilized fusion proteins were evaluated. Both immobilized BGL and immobilized AmyA prepared by the sortase A-mediated technique retained their catalytic activity, exhibiting activities 3.0- or 1.5-fold (respectively) of those seen with the same enzymes immobilized by chemical crosslinking. Immobilized enzymes prepared by the sortase A-mediated technique did not undergo dramatic changes in stability compared with the respective free enzymes. Thus, the sortase A-mediated technique provides a promising method for immobilization of active, stable enzymes.

  1. The influence of carbon nanotubes on enzyme activity and structure: investigation of different immobilization procedures through enzyme kinetics and circular dichroism studies

    NASA Astrophysics Data System (ADS)

    Cang-Rong, Jason Teng; Pastorin, Giorgia

    2009-06-01

    In the last decade, many environmental organizations have devoted their efforts to identifying renewable biosystems, which could provide sustainable fuels and thus enhance energy security. Amidst the myriad of possibilities, some biofuels make use of different types of waste biomasses, and enzymes are often employed to hydrolyze these biomasses and produce sugars that will be subsequently converted into ethanol. In this project, we aimed to bridge nanotechnology and biofuel production: here we report on the activity and structure of the enzyme amyloglucosidase (AMG), physically adsorbed or covalently immobilized onto single-walled carbon nanotubes (SWNTs) and multi-walled carbon nanotubes (MWNTs). In fact, carbon nanotubes (CNTs) present several properties that render them ideal support systems, without the diffusion limitations displayed by porous material and with the advantage of being further functionalizable at their surface. Chemical ligation was achieved both on oxidized nanotubes (via carbodiimide chemistry), as well as on amino-functionalized nanotubes (via periodate-oxidized AMG). Results showed that AMG retained a certain percentage of its specific activity for all enzyme-carbon nanotubes complexes prepared, with the physically adsorbed samples displaying better catalytic efficiency than the covalently immobilized samples. Analysis of the enzyme's structure through circular dichroism (CD) spectroscopy revealed significant structural changes in all samples, the degree of change being consistent with the activity profiles. This study proves that AMG interacts differently with carbon nanotubes depending on the method employed. Due to the higher activity reported by the enzyme physically adsorbed onto CNTs, these samples demonstrated a vast potential for further development. At the same time, the possibility of inducing magnetic properties into CNTs offers the opportunity to easily separate them from the original solution. Hence, substances to which they

  2. Study of immobilized metal affinity chromatography sorbents for the analysis of peptides by on-line solid-phase extraction capillary electrophoresis-mass spectrometry.

    PubMed

    Ortiz-Martin, Lorena; Benavente, Fernando; Medina-Casanellas, Silvia; Giménez, Estela; Sanz-Nebot, Victoria

    2015-03-01

    Several commercial immobilized metal affinity chromatography sorbents were evaluated in this study for the analysis of two small peptide fragments of the amyloid β-protein (Aβ) (Aβ(1-15) and Aβ(10-20) peptides) by on-line immobilized metal affinity SPE-CE (IMA-SPE-CE). The performance of a nickel metal ion (Ni(II)) sorbent based on nitrilotriacetic acid as a chelating agent was significantly better than two copper metal ion (Cu(II)) sorbents based on iminodiacetic acid. A BGE of 25 mM phosphate (pH 7.4) and an eluent of 50 mM imidazole (in BGE) yielded a 25-fold and 5-fold decrease in the LODs by IMA-SPE-CE-UV for Aβ(1-15) and Aβ(10-20) peptides (0.1 and 0.5 μg/mL, respectively) with regard to CE-UV (2.5 μg/mL for both peptides). The phosphate BGE was also used in IMA-SPE-CE-MS, but the eluent needed to be substituted by a 0.5% HAc v/v solution. Under optimum preconcentration and detection conditions, reproducibility of peak areas and migration times was acceptable (23.2 and 12.0%RSD, respectively). The method was more sensitive for Aβ(10-20) peptide, which could be detected until 0.25 μg/mL. Linearity for Aβ(10-20) peptide was good in a narrow concentration range (0.25-2.5 μg/mL, R(2) = 0.93). Lastly, the potential of the optimized Ni(II)-IMA-SPE-CE-MS method for the analysis of amyloid peptides in biological fluids was evaluated by analyzing spiked plasma and serum samples. PMID:25640944

  3. [Transformation of icariin by immobilized β-glucosidase and snailase].

    PubMed

    Peng, Jing; Ma, Yi-hua; Chen, Yan; Liu, Cong-Yan; Gao, Xia; Zhou, Jing

    2015-12-01

    This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage. PMID:27169291

  4. [Transformation of icariin by immobilized β-glucosidase and snailase].

    PubMed

    Peng, Jing; Ma, Yi-hua; Chen, Yan; Liu, Cong-Yan; Gao, Xia; Zhou, Jing

    2015-12-01

    This study was performed to prepare immobilized β-glucosidase and snailase, then optimize and compare the process conditions for conversion of icariin. Immobilized β-glucosidase and snailase were prepared using crosslink-embedding method. The best conditions of the preparation process were optimized by single factor analysis and the properties of immobilized β-glucosidase and snailase were investigated. The reaction conditions including temperature, pH, substrate ratio, substrate concentration, reaction time and reusing times of the conversion of icariin using immobilized β-glucosidase or snailase were optimized. Immobilized β-glucosidase and snailase exhibited better heat stabilities and could remain about 60% activity after storage at 4 degrees C for 4 weeks. The optimized conditions for the conversion of icariin were as follows, the temperature of 50 degrees C, pH of 5.0, enzyme and substrate ratio of 1 : 1, substrate concentration of 0.1 mg x mL(-1), reaction time of 6 h for β-glucosidase and 2 h for snailase, respectively. In 5 experiments, the average conversion ratio of immobilized β-glucosidase and snailase was 70.76% and 74.97%. The results suggest an effect of promoted stabilities, prolonged lifetimes in both β-glucosidase and snailase after immobilization. The immobilized β-glucosidase and snailase exhibited a higher conversion rate and reusability compared to the free β-glucosidase and snailase. Moreover, the conversion rate of immobilized snailase was higher than that of immobilized β-glucosidase. The process of icariin conversion using immobilized β-glucosidase and snailase was moderate and feasible, which suggests that immobilized enzymes may hold a promise for industrial usage.

  5. Atrazine removal in agricultural infiltrate by bioaugmented polyvinyl alcohol immobilized and free Agrobacterium radiobacter J14a: a sand column study.

    PubMed

    Siripattanakul, Sumana; Wirojanagud, Wanpen; McEvoy, John M; Casey, Francis X M; Khan, Eakalak

    2009-01-01

    Bench-scale sand column breakthrough experiments were conducted to examine atrazine removal in agricultural infiltrate by Agrobacterium radiobacter J14a (J14a) immobilized in phosphorylated-polyvinyl alcohol compared to free J14a cells. The effects of cell loading and infiltration rate on atrazine degradation and the loss of J14a were investigated. Four sets of experiments, (i) tracers, (ii) immobilized dead cells, (iii) immobilized cells, and (iv) free cells, were performed. The atrazine biodegradation at the cell loadings of 300, 600, and 900 mg dry cells L(-1) and the infiltration rates of 1, 3, and 6 cm d(-1) were tested for 5 column pore volumes (PV). The atrazine breakthrough results indicated that the immobilized dead cells significantly retarded atrazine transport. The atrazine removal efficiencies at the infiltration rates of 1, 3, and 6 cm d(-1) were 100%, 80-97%, and 50-70%, respectively. Atrazine degradation capacity for the immobilized cells was not significantly different from the free cells. Both infiltration rate and cell loading significantly affected atrazine removal for both cell systems. The bacterial loss from the immobilized cell system was 10-100 times less than that from the free cell system. For long-term tests at 50 PV, the immobilized cell system provided consistent atrazine removal efficiency while the atrazine removal by the free cells declined gradually because of the cell loss.

  6. Industrial use of immobilized enzymes.

    PubMed

    DiCosimo, Robert; McAuliffe, Joseph; Poulose, Ayrookaran J; Bohlmann, Gregory

    2013-08-01

    Although many methods for enzyme immobilization have been described in patents and publications, relatively few processes employing immobilized enzymes have been successfully commercialized. The cost of most industrial enzymes is often only a minor component in overall process economics, and in these instances, the additional costs associated with enzyme immobilization are often not justified. More commonly the benefit realized from enzyme immobilization relates to the process advantages that an immobilized catalyst offers, for example, enabling continuous production, improved stability and the absence of the biocatalyst in the product stream. The development and attributes of several established and emerging industrial applications for immobilized enzymes, including high-fructose corn syrup production, pectin hydrolysis, debittering of fruit juices, interesterification of food fats and oils, biodiesel production, and carbon dioxide capture are reviewed herein, highlighting factors that define the advantages of enzyme immobilization. PMID:23436023

  7. Industrial use of immobilized enzymes.

    PubMed

    DiCosimo, Robert; McAuliffe, Joseph; Poulose, Ayrookaran J; Bohlmann, Gregory

    2013-08-01

    Although many methods for enzyme immobilization have been described in patents and publications, relatively few processes employing immobilized enzymes have been successfully commercialized. The cost of most industrial enzymes is often only a minor component in overall process economics, and in these instances, the additional costs associated with enzyme immobilization are often not justified. More commonly the benefit realized from enzyme immobilization relates to the process advantages that an immobilized catalyst offers, for example, enabling continuous production, improved stability and the absence of the biocatalyst in the product stream. The development and attributes of several established and emerging industrial applications for immobilized enzymes, including high-fructose corn syrup production, pectin hydrolysis, debittering of fruit juices, interesterification of food fats and oils, biodiesel production, and carbon dioxide capture are reviewed herein, highlighting factors that define the advantages of enzyme immobilization.

  8. Evaluation of two-dimensional bolus effect of immobilization/support devices on skin doses: A radiochromic EBT film dosimetry study in phantom

    SciTech Connect

    Chiu-Tsao, Sou-Tung; Chan, Maria F.

    2010-07-15

    Purpose: In this study, the authors have quantified the two-dimensional (2D) perspective of skin dose increase using EBT film dosimetry in phantom in the presence of patient immobilization devices during conventional and IMRT treatments. Methods: For 6 MV conventional photon field, the authors evaluated and quantified the 2D bolus effect on skin doses for six different common patient immobilization/support devices, including carbon fiber grid with Mylar sheet, Orfit carbon fiber base plate, balsa wood board, Styrofoam, perforated AquaPlast sheet, and alpha-cradle. For 6 and 15 MV IMRT fields, a stack of two film layers positioned above a solid phantom was exposed at the air interface or in the presence of a patient alpha-cradle. All the films were scanned and the pixel values were converted to doses based on an established calibration curve. The authors determined the 2D skin dose distributions, isodose curves, and cross-sectional profiles at the surface layers with or without the immobilization/support device. The authors also generated and compared the dose area histograms (DAHs) and dose area products from the 2D skin dose distributions. Results: In contrast with 20% relative dose [(RD) dose relative to d{sub max} on central axis] at 0.0153 cm in the film layer for 6 MV 10x10 cm{sup 2} open field, the average RDs at the same depth in the film layer were 71%, 69%, 55%, and 57% for Orfit, balsa wood, Styrofoam, and alpha-cradle, respectively. At the same depth, the RDs were 54% under a strut and 26% between neighboring struts of a carbon fiber grid with Mylar sheet, and between 34% and 56% for stretched perforated AquaPlast sheet. In the presence of the alpha-cradle for the 6 MV (15 MV) IMRT fields, the hot spot doses at the effective measurement depths of 0.0153 and 0.0459 cm were 140% and 150% (83% and 89%), respectively, of the isocenter dose. The enhancement factor was defined as the ratio of a given DAH parameter (minimum dose received in a given area) with

  9. Enzymes immobilized in mesoporous silica: a physical-chemical perspective.

    PubMed

    Carlsson, Nils; Gustafsson, Hanna; Thörn, Christian; Olsson, Lisbeth; Holmberg, Krister; Åkerman, Björn

    2014-03-01

    Mesoporous materials as support for immobilized enzymes have been explored extensively during the last two decades, primarily not only for biocatalysis applications, but also for biosensing, biofuels and enzyme-controlled drug delivery. The activity of the immobilized enzymes inside the pores is often different compared to that of the free enzymes, and an important challenge is to understand how the immobilization affects the enzymes in order to design immobilization conditions that lead to optimal enzyme activity. This review summarizes methods that can be used to understand how material properties can be linked to changes in enzyme activity. Real-time monitoring of the immobilization process and techniques that demonstrate that the enzymes are located inside the pores is discussed by contrasting them to the common practice of indirectly measuring the depletion of the protein concentration or enzyme activity in the surrounding bulk phase. We propose that pore filling (pore volume fraction occupied by proteins) is the best standard for comparing the amount of immobilized enzymes at the molecular level, and present equations to calculate pore filling from the more commonly reported immobilized mass. Methods to detect changes in enzyme structure upon immobilization and to study the microenvironment inside the pores are discussed in detail. Combining the knowledge generated from these methodologies should aid in rationally designing biocatalyst based on enzymes immobilized in mesoporous materials.

  10. Hand immobilization affects arm and shoulder postural control.

    PubMed

    Bolzoni, Francesco; Bruttini, Carlo; Esposti, Roberto; Cavallari, Paolo

    2012-07-01

    It is a common experience, immediately after the removal of a cast or a splint, to feel motor awkwardness, which is usually attributed to muscular and joint immobilization. However, the same feeling may also be perceived after a brief period of immobilization. We provide evidence that this last effect stems from changes in the cortical organization of the focal movement as well as in the associated anticipatory postural adjustments. Indeed, these two aspects of the motor act are strongly correlated, although scaled in different manners. In fact, they are both shaped in the primary motor cortex, they both undergo similar amplitude and latency modulation and, as we will show, they are both impaired by the immobilization of the lone prime mover. Neuromuscular effects of limb immobilization are well known; however, most papers focus on changes occurring in the pathways projecting to the prime mover, which acts on the immobilized joint. Conversely, this study investigates the effect of immobilization on anticipatory postural adjustments. Indeed, we show that 12 h of wrist and fingers immobilization effectively modify anticipatory postural adjustments of the elbow and the shoulder, that is, those joints not immobilized within the fixation chain. Accordingly, the motor impairment observed after short-term immobilization most likely stems from the unbalance between anticipatory postural adjustments and the focal movement.

  11. Excess Weapons Plutonium Immobilization in Russia

    SciTech Connect

    Jardine, L.; Borisov, G.B.

    2000-04-15

    The joint goal of the Russian work is to establish a full-scale plutonium immobilization facility at a Russian industrial site by 2005. To achieve this requires that the necessary engineering and technical basis be developed in these Russian projects and the needed Russian approvals be obtained to conduct industrial-scale immobilization of plutonium-containing materials at a Russian industrial site by the 2005 date. This meeting and future work will provide the basis for joint decisions. Supporting R&D projects are being carried out at Russian Institutes that directly support the technical needs of Russian industrial sites to immobilize plutonium-containing materials. Special R&D on plutonium materials is also being carried out to support excess weapons disposition in Russia and the US, including nonproliferation studies of plutonium recovery from immobilization forms and accelerated radiation damage studies of the US-specified plutonium ceramic for immobilizing plutonium. This intriguing and extraordinary cooperation on certain aspects of the weapons plutonium problem is now progressing well and much work with plutonium has been completed in the past two years. Because much excellent and unique scientific and engineering technical work has now been completed in Russia in many aspects of plutonium immobilization, this meeting in St. Petersburg was both timely and necessary to summarize, review, and discuss these efforts among those who performed the actual work. The results of this meeting will help the US and Russia jointly define the future direction of the Russian plutonium immobilization program, and make it an even stronger and more integrated Russian program. The two objectives for the meeting were to: (1) Bring together the Russian organizations, experts, and managers performing the work into one place for four days to review and discuss their work with each other; and (2) Publish a meeting summary and a proceedings to compile reports of all the excellent

  12. Disposition of surplus fissile materials via immobilization

    SciTech Connect

    Gray, L.W.; Kan, T.; Sutcliffe, W.G.; McKibben, J.M.; Danker, W.

    1995-07-23

    In the Cold War aftermath, the US and Russia have agreed to large reductions in nuclear weapons. To aid in the selection of long-term management options, the USDOE has undertaken a multifaceted study to select options for storage and disposition of surplus plutonium (Pu). One disposition alternative being considered is immobilization. Immobilization is a process in which surplus Pu would be embedded in a suitable material to produce an appropriate form for ultimate disposal. To arrive at an appropriate form, we first reviewed published information on HLW immobilization technologies to identify forms to be prescreened. Surviving forms were screened using multi-attribute utility analysis to determine promising technologies for Pu immobilization. We further evaluated the most promising immobilization families to identify and seek solutions for chemical, chemical engineering, environmental, safety, and health problems; these problems remain to be solved before we can make technical decisions about the viability of using the forms for long-term disposition of Pu. All data, analyses, and reports are being provided to the DOE Office of Fissile Materials Disposition to support the Record of Decision that is anticipated in Summer of 1996.

  13. Bacteriostatic polymer film immobilization.

    PubMed

    El-Hayek, Rami F; Dye, Kevin; Warner, John C

    2006-12-15

    Coatings of quaternary ammonium tertiary structures (QUATS) copolymerized with 4-vinylbenzylthymine (VBT) exhibited high antimicrobial activity against Escherichia coli. Immobilization of QUATS improves environmental performance by preventing release of antibacterials to the environment, helping to preclude the emergence of resistant strains. The crosslinking immobilization scheme reported herein provides a more environmentally benign and more inexpensive synthesis than previously reported, thus reducing the use of solvents, energy, and production time. Development of water soluble, thymine-based photopolymers was inspired by the UV-induced 2pi + 2pi photocyclodimerization of thymine in DNA. Copolymers of 4-vinylbenzylthymine and trimethylammonium chloride, triethylammonium chloride, or dimethyloctylammonium chloride were synthesized in different monomer ratios. The antibacterial properties were tested by coating VBT:QUATS in sterilized petri dishes, crosslinking under short UV light, spraying with aqueous suspensions of bacterial cells, air drying, and then applying agar media to promote bacterial growth. The plates were incubated for 24 h at 37 degrees C. The number of viable cells ranged from 17 to 0% growth. Immobilized VBT:QUAT copolymers are antiseptic surfaces that can be produced in an environmentally benign fashion.

  14. Immobilization of peroxidase on SPEU film via radiation grafting

    NASA Astrophysics Data System (ADS)

    Hongfei, Ha; Guanghui, Wang; Jilan, Wu

    The acrylic acid or acrylamide were grafted via radiation onto segmented polyetherurethane (SPEU) film which is a kind of biocompatible material. Then the Horse radish peroxidase was immobilized on the grafted SPEU film through chemical binding. Some quantitative relationships between the percent graft and the activity, amount of immobilized enzyme were given. The properties and application of obtained biomaterial was studied as well.

  15. Study of Molecular Conformation and Activity-Related Properties of Lipase Immobilized onto Core-Shell Structured Polyacrylic Acid-Coated Magnetic Silica Nanocomposite Particles.

    PubMed

    Esmaeilnejad-Ahranjani, Parvaneh; Kazemeini, Mohammad; Singh, Gurvinder; Arpanaei, Ayyoob

    2016-04-01

    A facile approach for the preparation of core-shell structured poly(acrylic acid) (PAA)-coated Fe3O4 cluster@SiO2 nanocomposite particles as the support materials for the lipase immobilization is reported. Low- or high-molecular-weight (1800 and 100,000, respectively) PAA molecules were covalently attached onto the surface of amine-functionalized magnetic silica nanoacomposite particles. The successful preparation of particles were verified by scanning transmission electron microscopy (STEM), X-ray diffraction (XRD), vibrating sample magnetometer (VSM), thermogravimetric analysis (TGA), zeta potential measurement, and Fourier-transform infrared (FTIR) techniques. Once lipase is covalently immobilized onto the particles with an average diameter of 210 ± 50 nm, resulting from high binding sites concentrations on the low- and high-molecular-weight PAA-coated particles, high lipase immobilization efficiencies (86.2% and 89.9%, respectively), and loading capacities (786 and 816 mg g(-1), respectively) are obtained. Results from circular dichroism (CD) analysis and catalytic activity tests reveal an increase in the β-sheet content of lipase molecules upon immobilization, along with an enhancement in their activities and stabilities. The lipases immobilized onto the low- and high-molecular-weight PAA-coated particles show maximum activities at 55 and 50 °C, respectively, which are ∼28% and ∼15% higher than that of the free lipase at its own optimum temperature (40 °C), respectively. The immobilized lipases exhibit excellent performance at broader temperature and pH ranges and high thermal and storage stabilities, as well as superior reusability. These prepared magnetic nanocomposite particles can be offered as suitable support materials for efficient immobilization of enzymes and improvement of the immobilized enzymes properties.

  16. The biosorption capacity of biochar for 4-bromodiphengl ether: study of its kinetics, mechanism, and use as a carrier for immobilized bacteria.

    PubMed

    Du, Jingting; Sun, Pengfei; Feng, Zhuo; Zhang, Xin; Zhao, Yuhua

    2016-02-01

    Polybrominated diphenyl ethers (PBDEs) are known as ubiquitous pollutants in ecological systems and thus pose a great threat to the health of humans and other organisms due to their bioamplification and bioaccumulation along the food chain. The present study was designed to investigate the biosorption capacity of biochar for the removal of 4-monobromodiphengl ether and its synergistic effect when used as a carrier to immobilize the 4-monobromodiphengl ether-degrading strain Sphingomonas sp. DZ3. The raw biochar material was prepared by pyrolyzing maize straw at 350 °C under oxygen-limited conditions. The maximum biosorption capacity of biochar for 4-bromodiphengl ether was determined to be 50.23 mg/L under an initial concentration of 800 mg/L at pH 7.0 and 40 °C. The data obtained from the biosorption studies were fitted successfully with the pseudo-first-order kinetic and Freundlich isotherm models. The Weber-Morris model analysis indicated that intraparticle diffusion was the limiting step in the biosorption of 4-bromodiphengl ether onto the biosorbent. The values of thermodynamic parameters △G0 were calculated as -24.61 kJ/mol (20 °C), -24.35 kJ/mol (30 °C), and -23.98 kJ/mol (40 °C), △S(0) was -8.45 kJ/mol/K, and △H(0) was 21.36 kJ/mol. The artificial neural network analysis indicated that the initial concentration appeared to be the most influential parameter on the biosorption processes. The removal rate of 4-bromodiphengl ether achieved using the biochar-microorganism system was increased by 63 and 83% compared with the rates obtained with biochar and the strain individually, respectively. The morphology of the biochar and immobilized strain was determined using a scanning electron microscope, and information of the surface functional groups of biochar was obtained through an infrared spectra study. PMID:26498816

  17. The biosorption capacity of biochar for 4-bromodiphengl ether: study of its kinetics, mechanism, and use as a carrier for immobilized bacteria.

    PubMed

    Du, Jingting; Sun, Pengfei; Feng, Zhuo; Zhang, Xin; Zhao, Yuhua

    2016-02-01

    Polybrominated diphenyl ethers (PBDEs) are known as ubiquitous pollutants in ecological systems and thus pose a great threat to the health of humans and other organisms due to their bioamplification and bioaccumulation along the food chain. The present study was designed to investigate the biosorption capacity of biochar for the removal of 4-monobromodiphengl ether and its synergistic effect when used as a carrier to immobilize the 4-monobromodiphengl ether-degrading strain Sphingomonas sp. DZ3. The raw biochar material was prepared by pyrolyzing maize straw at 350 °C under oxygen-limited conditions. The maximum biosorption capacity of biochar for 4-bromodiphengl ether was determined to be 50.23 mg/L under an initial concentration of 800 mg/L at pH 7.0 and 40 °C. The data obtained from the biosorption studies were fitted successfully with the pseudo-first-order kinetic and Freundlich isotherm models. The Weber-Morris model analysis indicated that intraparticle diffusion was the limiting step in the biosorption of 4-bromodiphengl ether onto the biosorbent. The values of thermodynamic parameters △G0 were calculated as -24.61 kJ/mol (20 °C), -24.35 kJ/mol (30 °C), and -23.98 kJ/mol (40 °C), △S(0) was -8.45 kJ/mol/K, and △H(0) was 21.36 kJ/mol. The artificial neural network analysis indicated that the initial concentration appeared to be the most influential parameter on the biosorption processes. The removal rate of 4-bromodiphengl ether achieved using the biochar-microorganism system was increased by 63 and 83% compared with the rates obtained with biochar and the strain individually, respectively. The morphology of the biochar and immobilized strain was determined using a scanning electron microscope, and information of the surface functional groups of biochar was obtained through an infrared spectra study.

  18. Uranium immobilization and nuclear waste

    SciTech Connect

    Duffy, C.J.; Ogard, A.E.

    1982-02-01

    Considerable information useful in nuclear waste storage can be gained by studying the conditions of uranium ore deposit formation. Further information can be gained by comparing the chemistry of uranium to nuclear fission products and other radionuclides of concern to nuclear waste disposal. Redox state appears to be the most important variable in controlling uranium solubility, especially at near neutral pH, which is characteristic of most ground water. This is probably also true of neptunium, plutonium, and technetium. Further, redox conditions that immobilize uranium should immobilize these elements. The mechanisms that have produced uranium ore bodies in the Earth's crust are somewhat less clear. At the temperatures of hydrothermal uranium deposits, equilibrium models are probably adequate, aqueous uranium (VI) being reduced and precipitated by interaction with ferrous-iron-bearing oxides and silicates. In lower temperature roll-type uranium deposits, overall equilibrium may not have been achieved. The involvement of sulfate-reducing bacteria in ore-body formation has been postulated, but is uncertain. Reduced sulfur species do, however, appear to be involved in much of the low temperature uranium precipitation. Assessment of the possibility of uranium transport in natural ground water is complicated because the system is generally not in overall equilibrium. For this reason, Eh measurements are of limited value. If a ground water is to be capable of reducing uranium, it must contain ions capable of reducing uranium both thermodynamically and kinetically. At present, the best candidates are reduced sulfur species.

  19. Immobilization of horseradish peroxidase on nonwoven polyester fabric coated with chitosan.

    PubMed

    Mohamed, Saleh A; Aly, A S; Mohamed, Tarek M; Salah, Hala A

    2008-02-01

    The immobilization of horseradish peroxidase (HRP) on composite membrane has been investigated. This membrane was prepared by coating nonwoven polyester fabric with chitosan glutamate in the presence of glutraldehyde as a crosslinking agent. The physicochemical properties of soluble and immobilized HRP were evaluated. The soluble HRP lost 90% of its activity after 4 weeks of storage at 4 degrees C, whereas the immobilized enzyme retained 85% of its original activity at the same time. A reusability study of immobilized HRP showed that the enzyme retained 54% of its activity after 10 cycles of reuse. Soluble and immobilized HRP showed the same pH optima at pH 5.5. The immobilized enzyme had significant stability at different pH values, where it had maximum stability at pH 3.0 and 6.0. The kinetic properties indicated that the immobilized enzyme had more affinity toward substrates than soluble enzyme. The soluble and immobilized enzymes had temperature optima at 30 and 40 degrees C and were stable up to 40 and 50 degrees C, respectively. The stability of HRP against metal ion inactivation was improved after immobilization. Immobilized HRP exhibited high resistance to proteolysis by trypsin. The immobilized HRP was more resistant to inactivation induced by urea, Triton X-100, and organic solvents compared to its soluble counterpart. The immobilized HRP showed very high yield of immobilization and markedly high stabilization against several forms of denaturants that offer potential for several applications. PMID:18456948

  20. Stability improvement of immobilized lactoperoxidase using polyaniline polymer.

    PubMed

    Jafary, Fariba; Kashanian, Soheila; Sharieat, Ziadin Samsam; Jafary, Farzaneh; Omidfar, Kobra; Paknejad, Maliheh

    2012-12-01

    Enzyme engineering via immobilization techniques is perfectly compatible against the other chemical or biological approximate to improve enzyme functions and stability. In this study lactoperoxidase was immobilized onto polyaniline polymer activated with glutaraldehyde as a bifunctional agent, to improve enzyme properties. Polyaniline polymer was used due its unique physical and chemical properties to immobilize lactoperoxidase (LPO). The optimum activity of immobilized LPO was observed at pH 6 and 55 °C, which has been increased about 10 °C for the immobilized enzyme. The immobilized enzyme maintained absolutely active for 60 days whereas the native enzyme lost 80 % of its initial activity within this period of time. Moreover, the immobilized enzyme can be reused for several times without loss of activity. The kinetic parameter studies showed slight differences between free and immobilized enzymes. The K(m) and K(m.app) were calculated to be 0.6 and 0.4; also V(max) and V(max.app) were 1.3 and 0.9 respectively.

  1. Modified chitosan microspheres in non-aggregated amylase immobilization.

    PubMed

    Rana, Medha; Kumari, Amita; Chauhan, Ghanshyam S; Chauhan, Kalpana

    2014-05-01

    Immobilized enzymes are useful as reusable catalysts in industrial processes. In this study, α-amylase was used as a model enzyme to evaluate the propensity of synthesized porous chitosan microspheres as immobilization matrix. Chitosan microspheres were synthesized by grafting and covalent gelation technique using acrylamide (AAm) and glutaraldehyde (GA) as chemical agents, respectively. The synthesized chitosan-cl-poly(AAm) demonstrated amylase immobilization capacity of 350 mg/g. Furthermore, SEM results supported the porous microsphere structure for chitosan-cl-poly(AAm) with non-aggregated amylase immobilization, which accounts for comparable activity of immobilized amylase (3.28 μmol/ml/min) in contrast to free amylase (3.46 μmol/ml/min). The immobilized α-amylase was characterized for optimal pH and temperature activity and showed better resistance to temperature and pH inactivation in contrast to free amylase. The immobilized amylase retained more than 60% of its initial activity when stored at 4°C for 30 days and retained 50% of its initial activity after seven successive repeated-use cycles. In conclusion, the study can be used as base for the immobilization of competent industrial biocatalysts in non-aggregated active structure.

  2. Effects of immobilization on spermiogenesis

    NASA Technical Reports Server (NTRS)

    Meitner, E. R.

    1980-01-01

    The influence of immobilization stress on spermiogenesis in rats was investigated. After 96 hour immobilization, histological changes began to manifest themselves in the form of practically complete disappearance of cell population of the wall of seminiferous tubule as well as a markedly increased number of cells with pathologic mitoses. Enzymological investigations showed various changes of activity (of acid and alkaline phosphatase and nonspecific esterase) in the 24, 48, and 96 hour immobilization groups.

  3. Removal and recovery of nickel(II) from aqueous solution by loofa sponge-immobilized biomass of Chlorella sorokiniana: characterization studies.

    PubMed

    Akhtar, N; Iqbal, J; Iqbal, M

    2004-04-30

    The biosorption process for the removal of nickel(II) by loofa sponge-immobilized biomass of Chlorella sorokiniana (LIBCS), a newly developed immobilized biosorbent, was characterized. Effects of environmental factors on metal uptake capacity of LIBCS were studied and compared with free biomass of C. sorokiniana (FBCS). Nickel(II) removal by LIBCS was found to be influenced by pH of the solution, initial metal concentration, and biomass concentration. The biosorption of nickel(II) ions by both LIBCS and FBCS increased as the initial concentration of nickel(II) ions increased in the medium. No loss to biosorption capacity of LIBCS for nickel(II) was found due to the presence of loofa sponge, indeed as compared to FBCS an increase of 25.3% was noted in the biosorption capacity of LIBCS. Maximum biosorption capacities for FBCS and LIBCS were found as 48.08 and 60.38 mg nickel(II)/g, respectively, whereas the amount of nickel(II) ions adsorbed on the plain loofa sponge was 6.1mg/g. During these biosorption studies, LIBCS exhibited excellent physical and chemical stability without any significant release/loss of microalgal biomass from loofa sponge matrix. The kinetics of nickel(II) removal was extremely fast reaching at equilibrium in about 15 min for LIBCS and 20 min for FBCS. The biosorption equilibrium was well described by the Langmuir and Freundlich adsorption isotherms. The biosorption capacities were found to be solution pH dependent and the maximum adsorption was found at a solution pH 4-5. The LIBCS could be regenerated using 75 mM HCl, with up to 98% recovery. The LIBCS were shown to be robust and stable with little decrease in the nickel(II) uptake capacity when used in consecutive seven biosorption-desorption cycles. Continuous removal of nickel(II) from electroplating effluent by LIBCS packed in fixed bed column bioreactor confirm the possibility of developing a biological treatment process for the removal of toxic metals from authentic wastewater. PMID

  4. Cervical spine immobilization in the elderly population

    PubMed Central

    Phan, Kevin; Mobbs, Ralph J.; Wilson, David; Ball, Jonathon

    2016-01-01

    Background Immobilization of the cervical spine is a cornerstone of spinal injury management. In the context of suspected cervical spine injury, patients are immobilized in a ‘neutral position’ based on the head and trunk resting on a flat surface. It is hypothesized that the increased thoracic kyphosis and loss of cervical lordosis seen in elderly patients may require alternative cervical immobilization, compared with the ‘neutral position’. Methods To investigate this, an audit of pan-scan CT performed on consecutive major trauma patients aged over 65 years was carried out over a 6-month period. Utilizing the pan-CT’s localizing scout film, a novel measurement, the ‘chin-brow horizontal’ angle was independently measured by a senior spine surgeon (RJM) and a neurosurgeon (PJR) with the gantry used as a horizontal zero- degree reference. The benefit of the ‘chin-brow horizontal’ angle in the trauma setting is it can be assessed from the bedside whilst the patient is immobilized against a flat surface. Results During the 6-month study period, 58 patients were identified (30 male, 28 female), with an average age of 77.6 years (minimum 65, maximum 97). Results showed that ‘chin-brow horizontal’ angles varied widely, between +15.8 degrees in flexion to −30.5 degrees in extension (mean −12.4 degrees in extension, standard deviation 9.31 degrees. The interobserver correlation was 0.997 (95% CI: 0.995–0.998). Conclusions These findings suggest that, due to degenerative changes commonly seen in elderly patients, the routine use of the ‘neutral position’ adopted for cervical spine immobilization may not be appropriate in this population. We suggest that consideration be taken in cervical spine immobilization, with patients assessed on an individual basis including the fracture morphology, to minimize the risk of fracture displacement and worsened neurological deficit.

  5. Characterization and study of the orientation of immobilized enzymes by tryptic digestion and HPLC-MS: design of an efficient catalyst for the synthesis of cephalosporins.

    PubMed

    Temporini, Caterina; Bonomi, Paolo; Serra, Immacolata; Tagliani, Auro; Bavaro, Teodora; Ubiali, Daniela; Massolini, Gabriella; Terreni, Marco

    2010-06-14

    An innovative approach to determine the orientation of penicillin G acylase (PGA) from Escherichia coli covalently immobilized onto solid supports has been developed. This method is based on tryptic digestion of immobilized PGA followed by HPLC-MS analysis of the released peptides which are supposed to be only those exposed toward the reaction medium and not directly bound to the solid support. To this purpose, PGA was immobilized on Eupergit C (acrylic hydrophobic resin) and glyoxyl-agarose (hydrophilic resin) functionalized with epoxy and aldehyde groups, respectively, both involving the Lys residues of the protein. The peptide maps obtained were analyzed to derive the orientation of immobilized PGA, as the position of the detected Lys gave indication concerning the accessibility of the different areas of the protein. The results indicate that PGA immobilization on both supports involves mainly Lys located near the binding pocket (70%). Some differences in the enzyme orientation on the two supports can be deduced by the presence of different unbound Lys residues in the released peptides, specific to each support (Lys 117alpha for PGA-Eupergit C; Lys 163alpha and Lys 165alpha for PGA-glyoxyl-agarose). These results have been correlated with the data obtained in the kinetically controlled synthesis and indicate that the orientation of PGA on both supports is partially unfavorable, driving the active site near the support surface. This type of orientation of the enzyme enhances the effect of the nature of the support and of the binding chemistry on the catalytic properties. The information obtained indicated the most suitable support and activation strategy to design an immobilized acylase with good synthetic properties for preparative processes. The glyoxyl-Eupergit C support with enhanced porosity synergically combines the mechanical stability and synthetic performances of immobilized PGA and was successfully used in the synthesis of several cephalosporins.

  6. A study on the immobilization of selenium oxyanions by H2/Pd(s) in aqueous solution: confirmation of the one-electron reduction barrier of selenate.

    PubMed

    Puranen, Anders; Jansson, Mats; Jonsson, Mats

    2010-07-30

    Selenium is a trace element of concern in several geochemical contexts, due to the potentially high mobility of the selenium oxyanions and the narrow range between deficiency and toxicity of the element. For high level nuclear waste repositories the long-lived fission product 79Se has been identified as a potential key dose contributor for the long-term safety. This paper deals with the catalytic effect of Pd(s) on the H2 reduction of selenium oxyanions which was studied experimentally in aqueous solutions containing bicarbonate and chloride. Pd-catalysts and hydrogen have been proposed for the remediation of various groundwater contaminants and can also serve as a model substance for catalytic noble metal inclusions present in spent nuclear fuel. In this study selenite (SeO3(2)--) was found to adsorb on Pd. In the presence of hydrogen the rate of selenite removal increased yielding elemental Se. However, no adsorption or reduction of selenate (SeO4(2)--) was observed. A simple radiation chemical experiment revealed a notable barrier towards stepwise one-electron reduction of selenate to selenite. This provides an explanation for the lower reactivity of selenate in systems where reductive immobilization of selenite as well as selenate is thermodynamically favorable.

  7. Preparation and characterization of. beta. -D-glucosidase immobilized in calcium alginate

    SciTech Connect

    Woodward, J.; Krasniak, S.R.; Smith, R.D.; Spielberg, F.; Zachry, G.S.

    1982-01-01

    The immobilization of enzymes, organelles, and microbial cells within calcium alginate gel spheres has been reported. Few reports exist, however, on the use of these spheres for enzyme immobilization. The study reported here describes the immobilization of ..beta..-D-glucosidase within calcium alginate gel spheres and addresses the problems of loss of enzyme activity in the spheres via inhibition and leakage.

  8. IN SITU LEAD IMMOBILIZATION BY APATITE

    EPA Science Inventory

    Lead contamination is of environmental concern due to its effect on human health. The purpose of this study was to develop a technology to immobilize Pb in situ in contaminated soils and wastes using apatite. Hydroxyapatite [Ca10(PO4)6(O...

  9. Electron Tomography of Cryo-Immobilized Plant Tissue: A Novel Approach to Studying 3D Macromolecular Architecture of Mature Plant Cell Walls In Situ

    PubMed Central

    Sarkar, Purbasha; Bosneaga, Elena; Yap, Edgar G.; Das, Jyotirmoy; Tsai, Wen-Ting; Cabal, Angelo; Neuhaus, Erica; Maji, Dolonchampa; Kumar, Shailabh; Joo, Michael; Yakovlev, Sergey; Csencsits, Roseann; Yu, Zeyun; Bajaj, Chandrajit; Downing, Kenneth H.; Auer, Manfred

    2014-01-01

    Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D) organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT) electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼2 nm), and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF), cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin) samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we characterized the

  10. Electron tomography of cryo-immobilized plant tissue: a novel approach to studying 3D macromolecular architecture of mature plant cell walls in situ.

    PubMed

    Sarkar, Purbasha; Bosneaga, Elena; Yap, Edgar G; Das, Jyotirmoy; Tsai, Wen-Ting; Cabal, Angelo; Neuhaus, Erica; Maji, Dolonchampa; Kumar, Shailabh; Joo, Michael; Yakovlev, Sergey; Csencsits, Roseann; Yu, Zeyun; Bajaj, Chandrajit; Downing, Kenneth H; Auer, Manfred

    2014-01-01

    Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of plant cell wall composition is therefore essential for improving the fuel production process. The precise spatial three-dimensional (3D) organization of cellulose, hemicellulose, pectin and lignin within plant cell walls remains unclear to date since the microscopy techniques used so far have been limited to two-dimensional, topographic or low-resolution imaging, or required isolation or chemical extraction of the cell walls. In this paper we demonstrate that by cryo-immobilizing fresh tissue, then either cryo-sectioning or freeze-substituting and resin embedding, followed by cryo- or room temperature (RT) electron tomography, respectively, we can visualize previously unseen details of plant cell wall architecture in 3D, at macromolecular resolution (∼ 2 nm), and in near-native state. Qualitative and quantitative analyses showed that wall organization of cryo-immobilized samples were preserved remarkably better than conventionally prepared samples that suffer substantial extraction. Lignin-less primary cell walls were well preserved in both self-pressurized rapidly frozen (SPRF), cryo-sectioned samples as well as high-pressure frozen, freeze-substituted and resin embedded (HPF-FS-resin) samples. Lignin-rich secondary cell walls appeared featureless in HPF-FS-resin sections presumably due to poor stain penetration, but their macromolecular features could be visualized in unprecedented details in our cryo-sections. While cryo-tomography of vitreous tissue sections is currently proving to be instrumental in developing 3D models of lignin-rich secondary cell walls, here we confirm that the technically easier method of RT-tomography of HPF-FS-resin sections could be used immediately for routine study of low-lignin cell walls. As a proof of principle, we characterized the

  11. Metabolic alkalosis during immobilization in monkeys (M. nemestrina)

    NASA Technical Reports Server (NTRS)

    Young, D. R.; Yeh, I.; Swenson, R. S.

    1983-01-01

    The systemic and renal acid-base response of monkeys during ten weeks of immobilization was studied. By three weeks of immobilization, arterial pH and bicarbonate concentrations were elevated (chronic metabolic alkalosis). Net urinary acid excretion increased in immobilized animals. Urinary bicarbonate excretion decreased during the first three weeks of immobilization, and then returned to control levels. Sustained increases in urinary ammonium excretion were seen throughout the time duration of immobilization. Neither potassium depletion nor hypokalemia was observed. Most parameters returned promptly to the normal range during the first week of recovery. Factors tentatively associated with changes in acid-base status of monkeys include contraction of extracellular fluid volume, retention of bicarbonate, increased acid excretion, and possible participation of extrarenal buffers.

  12. Refolding of firefly luciferase immobilized on agarose beads.

    PubMed

    Zako, T; Deguchi, H; Kitayama, A; Ueda, H; Nagamune, T

    2000-03-01

    The renaturation yield of the denatured firefly luciferase decreased strongly with increasing protein concentration in a renaturation buffer, because of aggregation. In this study, firefly luciferase was immobilized on agarose beads at a high concentration. Although the protein concentration was extremely high (about 100-fold) compared to that of soluble luciferase, the renaturation yield was comparable with that for the soluble one. Thus, immobilization was shown to be effective for avoiding aggregation of firefly luciferase. It was also shown that the optimum buffer conditions for renaturation of the immobilized luciferase were the same as those for the renaturation in solution. Also, it was indicated that electrostatic interactions between a protein and the matrix have a negative effect on renaturation of the immobilized luciferase since the renaturation yield decreased at acidic pH only for the immobilized luciferase. These novel observations are described in detail in this paper.

  13. Hydroxyl Radical Generation and DNA Nuclease Activity: A Mechanistic Study Based on a Surface-Immobilized Copper Thioether Clip-Phen Derivative.

    PubMed

    Romo, Adolfo I B; Abreu, Dieric S; de F Paulo, Tércio; Carepo, Marta S P; Sousa, Eduardo H S; Lemus, Luis; Aliaga, Carolina; Batista, Alzir A; Nascimento, Otaciro R; Abruña, Héctor D; Diógenes, Izaura C N

    2016-07-11

    Coordination compounds of copper have been invoked as major actors in processes involving the reduction of molecular oxygen, mostly with the generation of radical species the assignment for which has, so far, not been fully addressed. In the present work, we have carried out studies in solution and on surfaces to gain insights into the nature of the radical oxygen species (ROS) generated by a copper(II) coordination compound containing a thioether clip-phen derivative, 1,3-bis(1,10-phenanthrolin-2-yloxy)-N-(4-(methylthio)benzylidene)propan-2-amine (2CP-Bz-SMe), enabling its adsorption/immobilization to gold surfaces. Whereas surface plasmon resonance (SPR) and electrochemistry of the adsorbed complex indicated the formation of a dimeric Cu(I) intermediate containing molecular oxygen as a bridging ligand, scanning electrochemical microscopy (SECM) and nuclease assays pointed to the generation of a ROS species. Electron paramagnetic resonance (EPR) data reinforced such conclusions, indicating that radical production was dependent on the amount of oxygen and H2 O2 , thus pointing to a mechanism involving a Fenton-like reaction that results in the production of OH(.) .

  14. Hydroxyl Radical Generation and DNA Nuclease Activity: A Mechanistic Study Based on a Surface-Immobilized Copper Thioether Clip-Phen Derivative.

    PubMed

    Romo, Adolfo I B; Abreu, Dieric S; de F Paulo, Tércio; Carepo, Marta S P; Sousa, Eduardo H S; Lemus, Luis; Aliaga, Carolina; Batista, Alzir A; Nascimento, Otaciro R; Abruña, Héctor D; Diógenes, Izaura C N

    2016-07-11

    Coordination compounds of copper have been invoked as major actors in processes involving the reduction of molecular oxygen, mostly with the generation of radical species the assignment for which has, so far, not been fully addressed. In the present work, we have carried out studies in solution and on surfaces to gain insights into the nature of the radical oxygen species (ROS) generated by a copper(II) coordination compound containing a thioether clip-phen derivative, 1,3-bis(1,10-phenanthrolin-2-yloxy)-N-(4-(methylthio)benzylidene)propan-2-amine (2CP-Bz-SMe), enabling its adsorption/immobilization to gold surfaces. Whereas surface plasmon resonance (SPR) and electrochemistry of the adsorbed complex indicated the formation of a dimeric Cu(I) intermediate containing molecular oxygen as a bridging ligand, scanning electrochemical microscopy (SECM) and nuclease assays pointed to the generation of a ROS species. Electron paramagnetic resonance (EPR) data reinforced such conclusions, indicating that radical production was dependent on the amount of oxygen and H2 O2 , thus pointing to a mechanism involving a Fenton-like reaction that results in the production of OH(.) . PMID:27310653

  15. Immobilization of Pb, Cd, and Zn in a contaminated soil using eggshell and banana stem amendments: metal leachability and a sequential extraction study.

    PubMed

    Ashrafi, Mehrnaz; Mohamad, Sharifah; Yusoff, Ismail; Shahul Hamid, Fauziah

    2015-01-01

    Heavy-metal-contaminated soil is one of the major environmental pollution issues all over the world. In this study, two low-cost amendments, inorganic eggshell and organic banana stem, were applied to slightly alkaline soil for the purpose of in situ immobilization of Pb, Cd, and Zn. The artificially metal-contaminated soil was treated with 5% eggshell or 10% banana stem. To simulate the rainfall conditions, a metal leaching experiment for a period of 12 weeks was designed, and the total concentrations of the metals in the leachates were determined every 2 weeks. The results from the metal leaching analysis revealed that eggshell amendment generally reduced the concentrations of Pb, Cd, and Zn in the leachates, whereas banana stem amendment was effective only on the reduction of Cd concentration in the leachates. A sequential extraction analysis was carried out at the end of the experiment to find out the speciation of the heavy metals in the amended soils. Eggshell amendment notably decreased mobility of Pb, Cd, and Zn in the soil by transforming their readily available forms to less accessible fractions. Banana stem amendment also reduced exchangeable form of Cd and increased its residual form in the soil.

  16. Two-dimensional (2D) infrared correlation study of the structural characterization of a surface immobilized polypeptide film stimulated by pH

    NASA Astrophysics Data System (ADS)

    Chae, Boknam; Son, Seok Ho; Kwak, Young Jun; Jung, Young Mee; Lee, Seung Woo

    2016-11-01

    The pH-induced structural changes to surface immobilized poly (L-glutamic acid) (PLGA) films were examined by Fourier transform infrared (FTIR) spectroscopy and two-dimensional (2D) correlation analysis. Significant spectral changes were observed in the FTIR spectra of the surface immobilized PLGA film between pH 6 and 7. The 2D correlation spectra constructed from the pH-dependent FTIR spectra of the surface immobilized PLGA films revealed the spectral changes induced by the alternations of the protonation state of the carboxylic acid group in the PLGA side chain. When the pH was increased from 6 to 8, weak spectral changes in the secondary structure of the PLGA main chain were induced by deprotonation of the carboxylic acid side group.

  17. Optimization of adsorptive immobilization of alcohol dehydrogenases.

    PubMed

    Trivedi, Archana; Heinemann, Matthias; Spiess, Antje C; Daussmann, Thomas; Büchs, Jochen

    2005-04-01

    In this work, a systematic examination of various parameters of adsorptive immobilization of alcohol dehydrogenases (ADHs) on solid support is performed and the impact of these parameters on immobilization efficiency is studied. Depending on the source of the enzymes, these parameters differently influence the immobilization efficiency, expressed in terms of residual activity and protein loading. Residual activity of 79% was achieved with ADH from bakers' yeast (YADH) after optimizing the immobilization parameters. A step-wise drying process has been found to be more effective than one-step drying. A hypothesis of deactivation through bubble nucleation during drying of the enzyme/glass bead suspension at low drying pressure (<45 kPa) is experimentally verified. In the case of ADH from Lactobacillus brevis (LBADH), >300% residual activity was found after drying. Hyperactivation of the enzyme is probably caused by structural changes in the enzyme molecule during the drying process. ADH from Thermoanaerobacter species (ADH T) is found to be stable under drying conditions (>15 kPa) in contrast to LBADH and YADH.

  18. Silanization and antibody immobilization on SU-8

    NASA Astrophysics Data System (ADS)

    Joshi, Manoj; Pinto, Richard; Rao, V. Ramgopal; Mukherji, Soumyo

    2007-01-01

    SU-8, an epoxy based negative photoresist, has emerged as a structural material for microfabricated sensors due to its attractive mechanical properties like low Young's modulus and chemical properties like inertness to various chemicals used in microfabrication. It can be used to fabricate MEMS structures of high aspect ratio. However, the use of SU-8 in BioMEMS application has been limited by the fact that immobilization of biomolecules on SU-8 surfaces has not been reported. In this study, the epoxy groups on the SU-8 surface were hydrolyzed in the presence of sulphochromic solution. Following this, the surface was treated with [3-(2-aminoethyl) aminopropyl]-trimethoxysilane (AEAPS). The silanized SU-8 surface was used to incubate human immunoglobulin (HIgG). The immobilization of HIgG was proved by allowing FITC tagged goat anti-human IgG to react with HIgG. This process of antibody immobilization was used to immobilize HIgG on microfabricated SU-8 cantilevers.

  19. Affinity chromatography of immobilized actin and myosin.

    PubMed Central

    Bottomley, R C; Trayer, I P

    1975-01-01

    Actin and myosin were immobilized by coupling them to agarose matrices. Both immobilized G-actin and immobilized myosin retain most of the properties of the proteins in free solution and are reliable over long periods of time. Sepharose-F-actin, under the conditions used in this study, has proved unstable and variable in its properties. Sepharose-G-actin columns were used to bind heavy meromyosin and myosin subfragment 1 specifically and reversibly. The interaction involved is sensitive to variation in ionic strength, such that myosin itself is not retained by the columns at the high salt concentration required for its complete solubilization. Myosin, rendered soluble at low ionic strength by polyalanylation, will interact successfully with the immobilized actin. The latter can distinguish between active and inactive fractions of the proteolytic and polyalanyl myosin derivatives, and was used in the preparation of these molecules. The complexes formed between the myosin derivatives and Sepharose-G-actin can be dissociated by low concentrations of ATP, ADP and pyrophosphate in both the presence and the absence of Mg2+. The G-actin columns were used to evaluate the results of chemical modifications of myosin subfragments on their interactions with actin. F-Actin in free solution is bound specifically and reversibly to columns of insolubilized myosin. Thus, with elution by either ATP or pyrophosphate, actin has been purified in one step from extracts of acetone-dried muscle powder. PMID:241335

  20. Plutonium Immobilization Canister Loading

    SciTech Connect

    Hamilton, E.L.

    1999-01-26

    This disposition of excess plutonium is determined by the Surplus Plutonium Disposition Environmental Impact Statement (SPD-EIS) being prepared by the Department of Energy. The disposition method (Known as ''can in canister'') combines cans of immobilized plutonium-ceramic disks (pucks) with vitrified high-level waste produced at the SRS Defense Waste Processing Facility (DWPF). This is intended to deter proliferation by making the plutonium unattractive for recovery or theft. The envisioned process remotely installs cans containing plutonium-ceramic pucks into storage magazines. Magazines are then remotely loaded into the DWPF canister through the canister neck with a robotic arm and locked into a storage rack inside the canister, which holds seven magazines. Finally, the canister is processed through DWPF and filled with high-level waste glass, thereby surrounding the product cans. This paper covers magazine and rack development and canister loading concepts.

  1. Comparison and evaluation of immobilization methods for preparing bacterial probes using acidophilic bioleaching bacteria Acidithiobacillus thiooxidans for AFM studies.

    PubMed

    Diao, Mengxue; Taran, Elena; Mahler, Stephen M; Nguyen, Anh V

    2014-07-01

    We evaluated different strategies for constructing bacterial probes for atomic force microscopy studies of bioleaching Acidithiobacillus thiooxidans interacting with pyrite mineral surfaces. Of three available techniques, the bacterial colloidal probe technique is the most reliable and provides a versatile platform for quantifying true interactive forces between bioleaching microorganisms and mineral surfaces.

  2. Student Collaboration in a Series of Integrated Experiments to Study Enzyme Reactor Modeling with Immobilized Cell-Based Invertase

    ERIC Educational Resources Information Center

    Taipa, M. A^ngela; Azevedo, Ana M.; Grilo, Anto´nio L.; Couto, Pedro T.; Ferreira, Filipe A. G.; Fortuna, Ana R. M.; Pinto, Ine^s F.; Santos, Rafael M.; Santos, Susana B.

    2015-01-01

    An integrative laboratory study addressing fundamentals of enzyme catalysis and their application to reactors operation and modeling is presented. Invertase, a ß-fructofuranosidase that catalyses the hydrolysis of sucrose, is used as the model enzyme at optimal conditions (pH 4.5 and 45 °C). The experimental work involves 3 h of laboratory time…

  3. Remote handling in the Plutonium Immobilization Project -- Second stage immobilization

    SciTech Connect

    Kriikku, E.

    1999-12-21

    The Savannah River Site (SRS) will immobilize excess plutonium in ceramic pucks and seal the pucks inside welded cans. Automated equipment will place these cans in magazines and the magazines in a Defense Waste Processing Facility (DWPF) canister. The DWPF will fill the canister with glass for permanent storage. Due to the radiation, remote equipment will perform these operations in a contained environment. The Plutonium Immobilization Project is in the conceptual design stage and the facility will begin operation in 2008. This paper discusses the Plutonium Immobilization Project phase 2 automation equipment conceptual design, equipment design, and work completed.

  4. SERR Spectroelectrochemical Study of Cytochrome cd1 Nitrite Reductase Co-Immobilized with Physiological Redox Partner Cytochrome c552 on Biocompatible Metal Electrodes.

    PubMed

    Silveira, Célia M; Quintas, Pedro O; Moura, Isabel; Moura, José J G; Hildebrandt, Peter; Almeida, M Gabriela; Todorovic, Smilja

    2015-01-01

    Cytochrome cd1 nitrite reductases (cd1NiRs) catalyze the one-electron reduction of nitrite to nitric oxide. Due to their catalytic reaction, cd1NiRs are regarded as promising components for biosensing, bioremediation and biotechnological applications. Motivated by earlier findings that catalytic activity of cd1NiR from Marinobacter hydrocarbonoclasticus (Mhcd1) depends on the presence of its physiological redox partner, cytochrome c552 (cyt c552), we show here a detailed surface enhanced resonance Raman characterization of Mhcd1 and cyt c552 attached to biocompatible electrodes in conditions which allow direct electron transfer between the conducting support and immobilized proteins. Mhcd1 and cyt c552 are co-immobilized on silver electrodes coated with self-assembled monolayers (SAMs) and the electrocatalytic activity of Ag // SAM // Mhcd1 // cyt c552 and Ag // SAM // cyt c552 // Mhcd1 constructs is tested in the presence of nitrite. Simultaneous evaluation of structural and thermodynamic properties of the immobilized proteins reveals that cyt c552 retains its native properties, while the redox potential of apparently intact Mhcd1 undergoes a ~150 mV negative shift upon adsorption. Neither of the immobilization strategies results in an active Mhcd1, reinforcing the idea that subtle and very specific interactions between Mhcd1 and cyt c552 govern efficient intermolecular electron transfer and catalytic activity of Mhcd1.

  5. Advances in ethanol production using immobilized cell systems

    SciTech Connect

    Margaritis, A.; Merchant, F.J.A.

    1984-01-01

    The application of immobilized cell systems for the production of ethanol has resulted in substantial improvements in the efficiency of the process when compared to the traditional free cell system. In this review, the various methods of cell immobilization employed in ethanol production systems have been described in detail. Their salient features, performance characteristics, advantages and limitations have been critically assessed. More recently, these immobilized cell systems have also been employed for the production of ethanol from non-conventional feedstocks such as Jerusalem artichoke extracts, cheese whey, cellulose, cellobiose and xylose. Ethanol production by immobilized yeast and bacterial cells has been attempted in various bioreactor types. Although most of these studies have been carried out using laboratory scale prototype bioreactors, it appears that only fluidized bed, horizontally packed bed bioreactors and tower fermenters may find application on scale-up. Several studies have indicated that upon immobilization, yeast cells performing ethanol fermentation exhibit more favourable physiological and metabolic properties. This, in addition to substantial improvements in ethanol productivities by immobilized cell systems, is indicative of the fact that future developments in the production of ethanol and alcoholic beverages will be directed towards the use of immobilized cell systems. 291 references.

  6. Immobilization of amyloglucosidase onto macroporous cryogels for continuous glucose production from starch.

    PubMed

    Uygun, Murat; Akduman, Begüm; Ergönül, Bülent; Aktaş Uygun, Deniz; Akgöl, Sinan; Denizli, Adil

    2015-01-01

    Poly(methyl methacrylate-glycidyl methacrylate) [Poly(MMA-GMA)] cryogels were synthesized using monomers of methylmethacrylic acid and epoxy group bearing GMA via radical cryopolymerization technique. Synthesized cryogels were used for the immobilization of amyloglucosidase to the cryogel surface using epoxy chemistry. Characterizations of the free and immobilized amyloglucosidase were carried out by comparing the optimum and kinetic parameters of enzymes. For this, pH and temperature profiles of free and immobilized preparation were studied and, it was found that, optimum pH of enzyme was not change upon immobilization (pH 5.0), while optimum temperature of the enzyme shifted 10 °C to warmer region after immobilization (optimum temperatures for free and immobilized enzyme were 55 and 65 °C, respectively). Kinetic parameters of free and immobilized enzyme were also investigated and Km values of free and immobilized amyloglucosidase were found to be 2.743 and 0.865 mg/mL, respectively. Vmax of immobilized amyloglucosidase was found to be (0.496 µmol/min) about four times less than that of free enzyme (2.020 µmol/min). Storage and operational stabilities of immobilized amyloglucosidase were also studied and it was showed that immobilized preparation had much more stability than free preparation. In the present work, amyloglucosidase immobilized poly(MMA-GMA) cryogels were used for continuous glucose syrup production from starch for the first time. Efficiency of immobilized enzyme was investigated and released amount of glucose was found to be 2.54 mg/mL at the end of the 5 min of hydrolysis. The results indicate that the epoxy functionalized cryogels offer a good alternative for amyloglucosidase immobilization applications with increased operational and thermal stability, and reusability. Also, these cryogels can be used for immobilization of other industrially valuable enzymes beyond amyloglucosidase. PMID:26235358

  7. Immobilization of amyloglucosidase onto macroporous cryogels for continuous glucose production from starch.

    PubMed

    Uygun, Murat; Akduman, Begüm; Ergönül, Bülent; Aktaş Uygun, Deniz; Akgöl, Sinan; Denizli, Adil

    2015-01-01

    Poly(methyl methacrylate-glycidyl methacrylate) [Poly(MMA-GMA)] cryogels were synthesized using monomers of methylmethacrylic acid and epoxy group bearing GMA via radical cryopolymerization technique. Synthesized cryogels were used for the immobilization of amyloglucosidase to the cryogel surface using epoxy chemistry. Characterizations of the free and immobilized amyloglucosidase were carried out by comparing the optimum and kinetic parameters of enzymes. For this, pH and temperature profiles of free and immobilized preparation were studied and, it was found that, optimum pH of enzyme was not change upon immobilization (pH 5.0), while optimum temperature of the enzyme shifted 10 °C to warmer region after immobilization (optimum temperatures for free and immobilized enzyme were 55 and 65 °C, respectively). Kinetic parameters of free and immobilized enzyme were also investigated and Km values of free and immobilized amyloglucosidase were found to be 2.743 and 0.865 mg/mL, respectively. Vmax of immobilized amyloglucosidase was found to be (0.496 µmol/min) about four times less than that of free enzyme (2.020 µmol/min). Storage and operational stabilities of immobilized amyloglucosidase were also studied and it was showed that immobilized preparation had much more stability than free preparation. In the present work, amyloglucosidase immobilized poly(MMA-GMA) cryogels were used for continuous glucose syrup production from starch for the first time. Efficiency of immobilized enzyme was investigated and released amount of glucose was found to be 2.54 mg/mL at the end of the 5 min of hydrolysis. The results indicate that the epoxy functionalized cryogels offer a good alternative for amyloglucosidase immobilization applications with increased operational and thermal stability, and reusability. Also, these cryogels can be used for immobilization of other industrially valuable enzymes beyond amyloglucosidase.

  8. Immobilization of a Plant Lipase from Pachira aquatica in Alginate and Alginate/PVA Beads.

    PubMed

    Bonine, Bárbara M; Polizelli, Patricia Peres; Bonilla-Rodriguez, Gustavo O

    2014-01-01

    This study reports the immobilization of a new lipase isolated from oleaginous seeds of Pachira aquatica, using beads of calcium alginate (Alg) and poly(vinyl alcohol) (PVA). We evaluated the morphology, number of cycles of reuse, optimum temperature, and temperature stability of both immobilization methods compared to the free enzyme. The immobilized enzymes were more stable than the free enzyme, keeping 60% of the original activity after 4 h at 50°C. The immobilized lipase was reused several times, with activity decreasing to approximately 50% after 5 cycles. Both the free and immobilized enzymes were found to be optimally active between 30 and 40°C.

  9. Uranium Immobilization in Wetland Soils

    NASA Astrophysics Data System (ADS)

    Jaffe, Peter R.; Koster van Groos, Paul G.; Li, Dien; Chang, Hyun-Shik; Seaman, John C.; Kaplan, Daniel I.; Peacock, Aaron D.; Scheckel, Kirk

    2014-05-01

    In wetlands, which are a major feature at the groundwater-surface water interface, plants deliver oxygen to the subsurface to keep root tissue aerobic. Some of this oxygen leaches into the rhizosphere where it will oxidize iron that typically precipitates on or near roots. Furthermore, plans provide carbon via root exudates and turnover, which in the presence of the iron oxides drives the activity of heterotrophic iron reducers in wetland soils. Oxidized iron is an important electron acceptor for many microbially-driven transformations, which can affect the fate and transport of several pollutants. It has been shown that heterotrophic iron reducing organisms, such as Geobacter sp., can reduce water soluble U(VI) to insoluble U(IV). The goal of this study was to determine if and how iron cycling in the wetland rhizosphere affects uranium dynamics. For this purpose, we operated a series of small-scale wetland mesocosms in a greenhouse to simulate the discharge of uranium-contaminated groundwater to surface waters. The mesocosms were operated with two different Fe(II) loading rates, two plant types, and unplanted controls. The mesocosms contained zones of root exclusion to differentiate between the direct presence and absence of roots in the planted mesocosms. The mesocosms were operated for several month to get fully established, after which a U(VI) solution was fed for 80 days. The mesocosms were then sacrificed and analyzed for solid-associated chemical species, microbiological characterization, micro-X-ray florescence (µ-XRF) mapping of Fe and U on the root surface, and U speciation via X-ray Absorption Near Edge Structure (XANES). Results showed that bacterial numbers including Geobacter sp., Fe(III), as well as total uranium, were highest on roots, followed by sediments near roots, and lowest in zones without much root influence. Results from the µ-XRF mapping on root surfaces indicated a strong spatial correlation between Fe and U. This correlation was

  10. Nanoporous Gold as a Solid Support for Protein Immobilization for the Development of Immunoassays, and for Biomolecular Interaction Studies

    NASA Astrophysics Data System (ADS)

    Pandey, Binod Prasad

    Nanoporous gold (NPG) is a versatile material of high surface area to volume ratio that can be readily modified with self-assembled monolayers of alkanethiols to which biomolecules can be linked. NPG presents new opportunities for the development of immunoassays, and for the development of carbohydrate based assays. This thesis explores the use of NPG as a support for self-assembled monolayers, their linkage to antibody-enzyme conjugates for immunoassay development, and for the study and application of carbohydrate-protein interactions. Direct kinetic electrochemical immunoassays were developed on NPG for prostate specific antigen (PSA) and carcinoembryonic antigen (CEA). The decrease in enzymatic conversion of p-aminophenylphosphate to p-aminophenol, by alkaline phosphatase conjugated to an antibody, due to steric hindrance caused by the presence of antigen on antibody, was observed as a drop in peak current in square-wave voltammetry. Detection limit of these assays was 0.075 ng mL -1 and 0.015 ng mL-1 for PSA and CEA, respectively. Similarly, the linear range of determination of these biomarkers extended up to 30 ng mL-1 and 10 ng mL-1 for PSA and CEA, respectively. Minimal interference was observed using newborn calf serum as a substitute for the human serum matrix. A rapid and sensitive enzyme linked lectinsorbant assay was also developed for the study of glycoprotein-lectin interactions on the NPG surface. Self-assembled monolayers of alkanethiols on NPG were characterized by cyclic voltammetry and electrochemical impedance spectroscopy. Similarly, the applicability of this surface for the formation of carbohydrate monolayers and its application for lectin carbohydrate interactions was also studied. Pure and mixed SAMs of 8-mercaptooctyl β-D-mannopyranoside (αMan-C8-SH) and α-D-Gal-(1→4)-β-D-Gal-(1α)-D-Glc-1-O-mercaptooctane (Gb3-C8-SH) with alkanethiols having varying tail groups were prepared. Binding affinity and binding kinetics of concanavalin A

  11. Antimony (Sb) and lead (Pb) in contaminated shooting range soils: Sb and Pb mobility and immobilization by iron based sorbents, a field study.

    PubMed

    Okkenhaug, Gudny; Grasshorn Gebhardt, Karl-Alexander; Amstaetter, Katja; Bue, Helga Lassen; Herzel, Hannes; Mariussen, Espen; Rossebø Almås, Åsgeir; Cornelissen, Gerard; Breedveld, Gijs D; Rasmussen, Grete; Mulder, Jan

    2016-04-15

    Small-arm shooting ranges often receive a significant input of lead (Pb), copper (Cu) and antimony (Sb) from ammunition. The goal of the present study was to investigate the mobility, distribution and speciation of Pb and Sb pollution under field conditions in both untreated and sorbent-amended shooting range soil. Elevated Sb (19-349μgL(-1)) and Pb (7-1495μgPbL(-1)) concentrations in the porewater of untreated soil over the four-year test period indicated a long-term Sb and Pb source to the adjacent environment in the absence of remedial measures. Mixing ferric oxyhydroxide powder (CFH-12) (2%) together with limestone (1%) into the soil resulted in an average decrease of Sb and Pb porewater concentrations of 66% and 97%, respectively. A similar reduction was achieved by adding 2% zerovalent iron (Fe°) to the soil. The remediation effect was stable over the four-year experimental period indicating no remobilization. Water- and 1M NH4NO3-extractable levels of Sb and Pb in field soil samples indicated significant immobilization by both treatments (89-90% for Sb and 89-99% for Pb). Results from sequential extraction analysis indicate fixation of Sb and Pb in less accessible fractions like amorphous iron oxides or even more crystalline and residual mineral phases, respectively. This work shows that amendment with Fe-based sorbents can be an effective method to reduce the mobility of metals both in cationic and anionic form in polluted shooting range soil.

  12. Antimony (Sb) and lead (Pb) in contaminated shooting range soils: Sb and Pb mobility and immobilization by iron based sorbents, a field study.

    PubMed

    Okkenhaug, Gudny; Grasshorn Gebhardt, Karl-Alexander; Amstaetter, Katja; Bue, Helga Lassen; Herzel, Hannes; Mariussen, Espen; Rossebø Almås, Åsgeir; Cornelissen, Gerard; Breedveld, Gijs D; Rasmussen, Grete; Mulder, Jan

    2016-04-15

    Small-arm shooting ranges often receive a significant input of lead (Pb), copper (Cu) and antimony (Sb) from ammunition. The goal of the present study was to investigate the mobility, distribution and speciation of Pb and Sb pollution under field conditions in both untreated and sorbent-amended shooting range soil. Elevated Sb (19-349μgL(-1)) and Pb (7-1495μgPbL(-1)) concentrations in the porewater of untreated soil over the four-year test period indicated a long-term Sb and Pb source to the adjacent environment in the absence of remedial measures. Mixing ferric oxyhydroxide powder (CFH-12) (2%) together with limestone (1%) into the soil resulted in an average decrease of Sb and Pb porewater concentrations of 66% and 97%, respectively. A similar reduction was achieved by adding 2% zerovalent iron (Fe°) to the soil. The remediation effect was stable over the four-year experimental period indicating no remobilization. Water- and 1M NH4NO3-extractable levels of Sb and Pb in field soil samples indicated significant immobilization by both treatments (89-90% for Sb and 89-99% for Pb). Results from sequential extraction analysis indicate fixation of Sb and Pb in less accessible fractions like amorphous iron oxides or even more crystalline and residual mineral phases, respectively. This work shows that amendment with Fe-based sorbents can be an effective method to reduce the mobility of metals both in cationic and anionic form in polluted shooting range soil. PMID:26799225

  13. Waste immobilization process development at the Savannah River Plant

    SciTech Connect

    Charlesworth, D L

    1986-01-01

    Processes to immobilize various wasteforms, including waste salt solution, transuranic waste, and low-level incinerator ash, are being developed. Wasteform characteristics, process and equipment details, and results from field/pilot tests and mathematical modeling studies are discussed.

  14. Immobilized soybean hull peroxidase for the oxidation of phenolic compounds in coffee processing wastewater.

    PubMed

    Chagas, Pricila Maria Batista; Torres, Juliana Arriel; Silva, Maria Cristina; Corrêa, Angelita Duarte

    2015-11-01

    Chitosan beads were prepared, using glutaraldehyde as a crosslinking agent for the immobilization of soybean hull peroxidase (SBP). The activity of free and immobilized SBP was studied. The optimum pH was 6.0 for both the free and immobilized enzyme; however, enzyme activity became more dependent on the temperature after immobilization. This study evaluated the potential use of immobilized and free enzyme in the oxidation of caffeic acid, of synthetic phenolic solution (SPS) and of total phenolic compounds in coffee processing wastewater (CPW). Some factors, such as reaction time, amount of H2O2 and caffeic acid were evaluated, in order to determine the optimum conditions for enzyme performance. Both enzymes showed a potential in the removal of caffeic acid, SPS and CPW, and immobilized SBP had the highest oxidation performance. The immobilized enzyme showed a potential of 50% in the oxidation of caffeic acid after 4 consecutive cycles.

  15. FTIR and Mössbauer spectroscopic study of sodium-aluminum-iron phosphate glassy materials for high level waste immobilization

    NASA Astrophysics Data System (ADS)

    Stefanovsky, S. V.; Stefanovsky, O. I.; Remizov, M. B.; Belanova, E. A.; Kozlov, P. V.; Glazkova, Ya. S.; Sobolev, A. V.; Presniakov, I. A.; Kalmykov, S. N.; Myasoedov, B. F.

    2015-11-01

    Complex sodium-aluminum-iron phosphate glassy materials with various Al2O3 to Fe2O3 ratio containing high level waste (HLW) surrogate were characterized by X-ray diffraction and scanning electron microscopy and studied in details by Fourier transform infrared (FTIR) spectroscopy. The samples with high Al2O3 content and not containing Fe2O3 were predominantly amorphous but subjected to devitrification under annealing. Addition of B2O3 and partial Fe2O3 substitution for Al2O3 in the materials increases their resistance to devitrification whereas further substitution and NiO incorporation significantly increase the tendency to devitrification. FTIR spectra demonstrate changes in the structure of glassy materials caused by both structural variations in the anionic motif and occurrence of crystalline phases in the materials. According to Mössbauer spectroscopy data, iron in the glassy samples is present as octahedrally coordinated Fe3+ ions while in the partly devitrified samples iron is partitioned among vitreous and crystalline phases entering the vitreous phase mainly as Fe3+O6 units and crystalline phases as major Fe3+ and minor Fe2+ ions in a magnetically ordered state and participating in a "fast" electronic exchange.

  16. Evaluation of cohesive and elastic support bandages for joint immobilization.

    PubMed

    Wilder, R P; Doctor, A; Paley, R J; Saunders, T J; Edlich, R F

    1989-01-01

    The purpose of this clinical study was to compare the performance of a new cohesive bandage to that of elastic bandages for joint immobilization. The magnitude of joint immobilization by these bandages was quantitated during isokinetic exercise using a computerized dynamometer. The degree to which the cohesive and elastic bandages reduced range of motion and peak torque of plantar and dorsiflexion was not significantly different. After exercising for 1 hour, the elastic bandage loosens, reducing its ability to immobilize the joint. In contrast, the cohesive bandage maintains its configuration, despite active exercise for 1 hour.

  17. Surface immobilization of antibody on silk fibroin through conformational transition.

    PubMed

    Lu, Qiang; Wang, Xiaoqin; Zhu, Hesun; Kaplan, David L

    2011-07-01

    In recent studies silk fibroin has been explored as a new material platform for biosensors. Based on these developments, a procedure for the immobilization of antibodies on silk fibroin substrates was developed as a route to functionalizing these biosensor systems. By controlling the conformational transition of the silk fibroin, a primary antibody was immobilized and enriched at the surface of silk fibroin substrates under mild reaction conditions to maintain antibody function. Compared to chemical crosslinking, the immobilization efficiency in the present approach was increased significantly. This method, achieving high loading of antibody while retaining function, improves the feasibility of silk fibroin as a platform material for biosensor applications.

  18. N-15 NMR study of the immobilization of 2,4- and 2,6-dinitrotoluene in aerobic compost

    USGS Publications Warehouse

    Thorn, K.A.; Pennington, J.C.; Kennedy, K.R.; Cox, L.G.; Hayes, C.A.; Porter, B.E.

    2008-01-01

    Large-scale aerobic windrow composting has been used to bioremediate washout lagoon soils contaminated with the explosives TNT (2,4,6- trinitrotoluene) and RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) at several sites within the United States. We previously used 15N NMR to investigate the reduction and binding of T15NT in aerobic bench -scale reactors simulating the conditions of windrow composting. These studies have been extended to 2,4-dinitrotoluene (2,4DNT) and 2,6-dinitrotoluene (2,6DNT), which, as impurities in TNT, are usually present wherever soils have been contaminated with TNT. Liquid-state 15N NMR analyses of laboratory reactions between 4-methyl-3-nitroaniline-15N, the major monoamine reduction product of 2,4DNT, and the Elliot soil humic acid, both in the presence and absence of horseradish peroxidase, indicated that the amine underwent covalent binding with quinone and other carbonyl groups in the soil humic acid to form both heterocyclic and non-heterocyclic condensation products. Liquid-state 15N NMR analyses of the methanol extracts of 20 day aerobic bench-scale composts of 2,4-di-15N-nitrotoluene and 2,6-di-15N-nitrotoluene revealed the presence of nitrite and monoamine, but not diamine, reduction products, indicating the occurrence of both dioxygenase enzyme and reductive degradation pathways. Solid-state CP/MAS 15N NMR analyses of the whole composts, however, suggested that reduction to monoamines followed by covalent binding of the amines to organic matter was the predominant pathway. ?? 2008 American Chemical Society.

  19. Polymer-assisted iron oxide magnetic nanoparticle immobilized keratinase

    NASA Astrophysics Data System (ADS)

    Konwarh, Rocktotpal; Karak, Niranjan; Rai, Sudhir Kumar; Mukherjee, Ashis Kumar

    2009-06-01

    Nanotechnology holds the prospect for avant-garde changes to improve the performance of materials in various sectors. The domain of enzyme biotechnology is no exception. Immobilization of industrially important enzymes onto nanomaterials, with improved performance, would pave the way to myriad application-based commercialization. Keratinase produced by Bacillus subtilis was immobilized onto poly(ethylene glycol)-supported Fe3O4 superparamagnetic nanoparticles. The optimization process showed that the highest enzyme activity was noted when immobilized onto cyanamide-activated PEG-assisted MNP prepared under conditions of 25 °C and pH 7.2 of the reaction mixture before addition of H2O2 (3% w/w), 2% (w/v) PEG6000 and 0.062:1 molar ratio of PEG to FeCl2·4H2O. Further statistical optimization using response surface methodology yielded an R2 value that could explain more than 94% of the sample variations. Along with the magnetization studies, the immobilization of the enzyme onto the PEG-assisted MNP was characterized by UV, XRD, FTIR and TEM. The immobilization process had resulted in an almost fourfold increase in the enzyme activity over the free enzyme. Furthermore, the immobilized enzyme exhibited a significant thermostability, storage stability and recyclability. The leather-industry-oriented application of the immobilized enzyme was tested for the dehairing of goat-skin.

  20. Enhanced growth of lactobacilli in soymilk upon immobilization on agrowastes.

    PubMed

    Teh, Sue-Siang; Ahmad, Rosma; Wan-Abdullah, Wan-Nadiah; Liong, Min-Tze

    2010-04-01

    Cell immobilization is an alternative to microencapsulation for the maintenance of cells in a liquid medium. The objective of this study was to evaluate the effects of agrowastes from durian (Durio zibethinus), cempedak (Artocarpus champeden), and mangosteen (Garcinia mangostana) as immobilizers for lactobacilli grown in soymilk. Rinds from the agrowastes were separated from the skin, dried, and ground (150 microm) to form powders and used as immobilizers. Scanning electron microscopy revealed that lactobacilli cells were attached and bound to the surface of the immobilizers. Immobilized cells of Lactobacillus acidophilus FTDC 1331, L. acidophilus FTDC 2631, L. acidophilus FTDC 2333, L. acidophilus FTDC 1733, and L. bulgaricus FTCC 0411 were inoculated into soymilk, stored at room temperature (25 degrees C) and growth properties were evaluated over 168 h. Soymilk inoculated with nonimmobilized cells was used as the control. Utilization of substrates, concentrations of lactic and acetic acids, and changes in pH were evaluated in soymilk over 186 h. Immobilized lactobacilli showed significantly better growth (P < 0.05) compared to the control, accompanied by higher production of lactic and acetic acids in soymilk. Soymilk containing immobilized cells showed greater reduction of soy sugars such as stachyose, raffinose, sucrose, fructose, and glucose compared to the control (P < 0.05).

  1. Acetylcholinesterase immobilized onto PEI-coated silica nanoparticles.

    PubMed

    Tumturk, Hayrettin; Yüksekdag, Hazer

    2016-01-01

    Polyethyleneimine (PEI) coated-silica nanoparticles were prepared by the Stöber method. The formation and the structure of the nanoparticles were characterized by ATR-FT-IR spectroscopy and transmission electron microscopy (TEM). TEM images of the silica and PEI-coated nanoparticles revealed that they were well dispersed and that there was no agglomeration. The acetylcholineesterase enzyme was immobilized onto these nanoparticles. The effects of pH and temperature on the storage stability of the free and immobilized enzyme were investigated. The optimum pHs for free and immobilized enzymes were determined as 7.0 and 8.0, respectively. The optimum temperatures for free and immobilized enzymes were found to be 30.0 and 35.0°C, respectively. The maximum reaction rate (Vmax) and the Michaelis-Menten constant (Km) were investigated for the free and immobilized enzyme. The storage stability of acetylcholinesterase was increased when immobilized onto the novel PEI-coated silica nanoparticles. The reuse numbers of immobilized enzyme were also studied. These hybrid nanoparticles are desirable as carriers for biomedical applications.

  2. Immunosuppressive agent leflunomide: a SWNTs-immobilized dihydroortate dehydrogenase inhibitory effect and computational study of its adsorption properties on zigzag single walled (6,0) carbon and boron nitride nanotubes as controlled drug delivery devices.

    PubMed

    Raissi, Heidar; Mollania, Fariba

    2014-06-01

    Leflunomide [HWA 486 or RS-34821, 5-methyl-N-(4trifluoromethylphenyl)-4-isoxazole carboximide] is an immunosuppressive agent effective in the treatment of rheumatoid arthritis. Dihydroortate dehydrogenase (DHODH, EC 1.3.3.1) immobilization on the nanotubes was carried out and biochemical characterization of free and immobilized enzyme was determined. In comparison with free enzyme, the immobilized DHODH showed improved stability and reusability for investigation of inhibition pattern of drugs such as leflunomide. The experimental data showed that, DHODH was inhibited by the active metabolite of leflunomide (RS-61980) with a Ki and KI of 0.82 and 0.06 mM, respectively. Results exhibited mixed-type inhibition kinetics towards dihydroorotate as a substrate in the free and immobilized enzyme. Furthermore, the behavior of anticancer drug leflunomide adsorbed on the external surface of zigzag single walled (6,0) carbon and boron nitride nanotubes (SWCNT and SWBNNT) was studied by means of DFT calculations at the B3LYP/6-31G(*) level of theory. The larger adsorption energies and charges transfer showed that the adsorption of leflunomide onto SWBNNT is more stable than that the adsorption of leflunomide onto SWCNT. Frontier molecular orbitals (HOMO and LUMO) suggest that adsorption of leflunomide onto SWBNNT behave as charge transfer compounds with leflunomide as an electron donor and SWBNNT as an electron acceptor. Thus, nanotubes (NTs) have been proposed and actively explored as multipurpose innovative carriers for drug delivery and diagnostic application. The AIM theory has been also applied to analyze the properties of the bond critical points: their electron densities and their laplacians. Also, the natural bond orbital (NBO) calculations were performed to derive natural atomic orbital occupancies, and partial charges of the interacting atoms in the equilibrium tube-molecule distance. PMID:24566615

  3. Immunosuppressive agent leflunomide: a SWNTs-immobilized dihydroortate dehydrogenase inhibitory effect and computational study of its adsorption properties on zigzag single walled (6,0) carbon and boron nitride nanotubes as controlled drug delivery devices.

    PubMed

    Raissi, Heidar; Mollania, Fariba

    2014-06-01

    Leflunomide [HWA 486 or RS-34821, 5-methyl-N-(4trifluoromethylphenyl)-4-isoxazole carboximide] is an immunosuppressive agent effective in the treatment of rheumatoid arthritis. Dihydroortate dehydrogenase (DHODH, EC 1.3.3.1) immobilization on the nanotubes was carried out and biochemical characterization of free and immobilized enzyme was determined. In comparison with free enzyme, the immobilized DHODH showed improved stability and reusability for investigation of inhibition pattern of drugs such as leflunomide. The experimental data showed that, DHODH was inhibited by the active metabolite of leflunomide (RS-61980) with a Ki and KI of 0.82 and 0.06 mM, respectively. Results exhibited mixed-type inhibition kinetics towards dihydroorotate as a substrate in the free and immobilized enzyme. Furthermore, the behavior of anticancer drug leflunomide adsorbed on the external surface of zigzag single walled (6,0) carbon and boron nitride nanotubes (SWCNT and SWBNNT) was studied by means of DFT calculations at the B3LYP/6-31G(*) level of theory. The larger adsorption energies and charges transfer showed that the adsorption of leflunomide onto SWBNNT is more stable than that the adsorption of leflunomide onto SWCNT. Frontier molecular orbitals (HOMO and LUMO) suggest that adsorption of leflunomide onto SWBNNT behave as charge transfer compounds with leflunomide as an electron donor and SWBNNT as an electron acceptor. Thus, nanotubes (NTs) have been proposed and actively explored as multipurpose innovative carriers for drug delivery and diagnostic application. The AIM theory has been also applied to analyze the properties of the bond critical points: their electron densities and their laplacians. Also, the natural bond orbital (NBO) calculations were performed to derive natural atomic orbital occupancies, and partial charges of the interacting atoms in the equilibrium tube-molecule distance.

  4. Optimizing immobilization of avidin on surface-modified magnetic nanoparticles: characterization and application of protein-immobilized nanoparticles.

    PubMed

    Yang, Tao; Sun, Shuguo; Ma, Meihu; Lin, Qinlu; Zhang, Lin; Li, Yan; Luo, Feijun

    2015-10-01

    A simple optimization method of immobilization of avidin on magnetic nanoparticles (MNPs)' surface was proposed in this study. The avidin-immobilized MNPs were then developed and used to immobilize a model enzyme [Horseradish peroxidase (HRP)]. The loading capacity (LC) and activity of avidin-immobilized MNPs were optimized through selecting the most appropriate nanoparticle's size and shape, glutaraldehyde concentration, cross-linking reaction time, ultrasonic processing time, and initial concentration of avidin. The LC under optimized conditions was 63.37 ± 1.29 mg avidin/g MNPs, and the immobilized protein was still able to maintain its high biological activity of 10.86 ± 0.13 U/mg (biotin-binding activity of nature avidin was 14.1 U/mg) and better thermal stability compared to free avidin. A highly reusable, stable, and easily recovered immobilized HRP was obtained using MNPs as carriers. The immobilized HRP was reused repeatedly more than 9 times and retained more than 65 % of its original activity. PMID:26224655

  5. Treating Wastewater With Immobilized Enzymes

    NASA Technical Reports Server (NTRS)

    Jolly, Clifford D.

    1991-01-01

    Experiments show enzymes are immobilized on supporting materials to make biocatalyst beds for treatment of wastewater. With suitable combination of enzymes, concentrations of various inorganic and organic contaminants, including ammonia and urea, reduced significantly.

  6. Potentiometric biosensor for determination of urea in milk using immobilized Arthrobacter creatinolyticus urease.

    PubMed

    Ramesh, Rajendran; Puhazhendi, Puhazhselvan; Kumar, Jitendra; Gowthaman, Marichetti Kuppuswami; D'Souza, Stanislaus Francis; Kamini, Numbi Ramudu

    2015-04-01

    The extracellular urease from Arthrobacter creatinolyticus was partially purified by ammonium sulfate precipitation and immobilized on PAN [poly(acrylonitrile-methylmethacrylate-sodium vinylsulfonate)] membrane. The urease immobilized PAN membrane exhibited an activity of 97.92U/cm(2) under the optimum conditions of 1.0% enzyme concentration, 15% glutaraldehyde, 24h immobilization time and temperature of 4°C. The changes in surface morphology of the membrane after immobilization were studied by SEM and ATR-FTIR analysis. Immobilized membrane was associated with potentiometric electrode for calibration of biosensor and the results showed a linear response for wide range of urea concentration from 1 to 100mM. The immobilized urease had good storage stability for a period of 70days at 4°C and could be effectively reused for 13cycles. Urease immobilized membrane was also employed in analysis of urea spiked milk samples.

  7. Biomimetic CO₂ sequestration using purified carbonic anhydrase from indigenous bacterial strains immobilized on biopolymeric materials.

    PubMed

    Sharma, Anjana; Bhattacharya, Abhishek; Shrivastava, Ankita

    2011-04-01

    The present study deals with immobilization of purified CA and whole cell of Pseudomonas fragi, Micrococcus lylae, and Micrococcus luteus 2 on different biopolymer matrices. Highest enzyme immobilization was achieved with P. fragi CA (89%) on chitosan-KOH beads, while maximum cell immobilization was achieved with M. lylae (75%) on chitosan-NH(4)OH beads. A maximum increase of 1.08-1.18 fold stability between 35 and 55°C was observed for M. lylae immobilized CA. The storage stability was improved by 2.02 folds after immobilization. FTIR spectra confirmed the adsorption of CA on chitosan-KOH beads following hydrophilic interactions. Calcium carbonate precipitation was achieved using chitosan-KOH immobilized P. fragi CA. More than 2 fold increase in sequestration potential was observed for immobilized system as compared to free enzyme. XRD spectra revealed calcite as the dominant phase in biomimetically produced calcium carbonate. PMID:22112959

  8. Enhanced starch hydrolysis using α-amylase immobilized on cellulose ultrafiltration affinity membrane.

    PubMed

    Konovalova, Viktoriia; Guzikevich, Kateryna; Burban, Anatoliy; Kujawski, Wojciech; Jarzynka, Karolina; Kujawa, Joanna

    2016-11-01

    In order to prepare ultrafiltration membranes possessing biocatalytic properties, α-amylase has been immobilized on cellulose membranes. Enzyme immobilization was based on a covalent bonding between chitosan and a surface of cellulose membrane, followed by an attachment of Cibacron Blue F3G-A dye as affinity ligand. Various factors affecting the immobilization process, such as enzyme concentration, pH of modifying solution, zeta-potential of membrane surface, and stability of immobilized enzyme were studied. The applicability of immobilized α-amylase has been investigated in ultrafiltration processes. The immobilization of α-amylase on membrane surface allows to increase the value of mass transfer coefficient and to decrease the concentration polarization effect during ultrafiltration of starch solutions. The enzyme layer on the membrane surface prevents a rapid increase of starch concentration due to the amylase hydrolysis of starch in the boundary layer. The presented affinity immobilization technique allows also for the regeneration of membranes from inactivated enzyme.

  9. Immobilization of Polymeric Luminophor on Nanoparticles Surface

    NASA Astrophysics Data System (ADS)

    Bolbukh, Yuliia; Podkoscielna, Beata; Lipke, Agnieszka; Bartnicki, Andrzej; Gawdzik, Barbara; Tertykh, Valentin

    2016-04-01

    Polymeric luminophors with reduced toxicity are of the priorities in the production of lighting devices, sensors, detectors, bioassays or diagnostic systems. The aim of this study was to develop a method of immobilization of the new luminophor on a surface of nanoparticles and investigation of the structure of the grafted layer. Monomer 2,7-(2-hydroxy-3-methacryloyloxypropoxy)naphthalene (2,7-NAF.DM) with luminophoric properties was immobilized on silica and carbon nanotubes in two ways: mechanical mixing with previously obtained polymer and by in situ oligomerization with chemisorption after carrier's modification with vinyl groups. The attached polymeric (or oligomeric) surface layer was studied using thermal and spectral techniques. Obtained results confirm the chemisorption of luminophor on the nanotubes and silica nanoparticles at the elaborated synthesis techniques. The microstructure of 2,7-NAF.DM molecules after chemisorption was found to be not changed. The elaborated modification approach allows one to obtain nanoparticles uniformly covered with polymeric luminophor.

  10. Summary report on geochemical barrier special study. [Geochemically modify tailings to immobilize contaminants with modifiers such as peat, limestone, and hydrated lime

    SciTech Connect

    Not Available

    1988-12-01

    Long-term management of uranium mill tailings must provide assurance that soluble contaminants will not migrate beyond the Point of Compliance. Conventional management alternatives provide containment through the use of physical barriers which are designed to prevent migration of water through the tailings pile. An alternative is to geochemically modify the tailings to immobilize the contaminants. This investigation examined three potential geochemical modifiers to determine their ability to immobilize inorganic groundwater contaminants found in uranium mill tailings. These modifiers were hydrated lime (Ca(OH)[sub 2]), limestone (CaCO[sub 3]), and a sphaegnum peat moss. This investigation focused on both the geochemical interactions between the tailings and the modifiers, and the effects the modifiers had on the physical strength of the tailings. The geochemical investigations began with characterization of the tailings by X-ray diffraction and scanning electron microscopy. This was followed by batch leaching experiments in which various concentrations of each modifier were added to tailings in shaker flasks and allowed to come to equilibrium. Finally, column experiments were conducted to simulate flow through a tailings pile. The results show that all of the modifiers were at least moderately effective at immobilizing most of the groundwater contaminants of concern at uranium mill tailings sites. Hydrated lime was able to achieve 90 percent concentration reduction of arsenic, cadmium, selenium, uranium, and sulfate when added at a two percent concentration. Limestone was somewhat less effective and peat removed greater than 90 percent of arsenic, lead, uranium, and sulfate at a one percent concentration. The column tests showed that kinetic and/or mass transfer limitations are important and that sufficient time must be allowed for the immobilization reactions to occur.

  11. The Collaborative Study on the Enzymatic Analysis of Positional Distribution of Short- and Medium-chain Fatty Acids in Milk Fat Using Immobilized Candida antarctica Lipase B.

    PubMed

    Yoshinaga, Kazuaki; Sato, Shinichi; Sasaki, Ryo; Asada, Mihoko; Hori, Ryuji; Imagi, Jun; Miyazaki, Yosuke; Nagai, Toshiharu; Saito, Katsuyoshi; Sano, Takashi; Sasaki, Akiko; Sato, Chiemi; Tsukahara, Yuki; Yamashita, Atsushi; Watanabe, Shimpei; Watanabe, Yomi

    2016-01-01

    The positional distributions of fatty acids (FAs) in milk fat containing short- and medium-chain FAs were analyzed by sn-1(3)-selective transesterification of triacylglycerols (TAGs) with ethanol using immobilized Candida antarctica lipase B (CALB), in a collaborative study conducted by 10 laboratories. The mean C4:0, C6:0, and C8:0 FA contents, when analyzed as propyl esters (PEs) using gas chromatography (GC) with a DB-23 capillary column, were found to be 3.0, 2.0, and, 1.3 area%, respectively. Their reproducibility standard deviations were 0.33, 0.18, and 0.19, respectively. The mean C4:0, C6:0, and C8:0 contents at the sn-2 position were 0.3, 0.4, and 1.0 area%, respectively. Their reproducibility standard deviations were 0.17, 0.11, and 0.19, respectively. The reproducibility standard deviations of C4:0, C6:0, and C8:0 FAs at the sn-2 position were either the same as or smaller than those for milk fat, although the FA contents at the sn-2 position were smaller than those in the milk fat. Therefore, it was concluded that the CALB method for estimating the regiospecific distribution is applicable to TAGs containing short- and medium-chain FAs. When estimating the short-chain (SC) FA contents in fats and oils by GC, it is better to analyze SCFAs as PEs or butyl esters, and not as methyl esters, in order to prevent loss of SCFAs during the experimental procedure because of their volatility and water solubility. This study also revealed that the stationary phase of the GC capillary column affected the flame ionization detector (FID) response of SCFAs. The theoretical FID correction factor (MWFA / active carbon number / atomic weight of carbon) fitted well with the actual FID responses of C4:0-C12:0 FAs when they were analyzed as PEs using a DB-23 column; however, this was not the case when the GC analysis was performed using wax-type columns. PMID:26972465

  12. Cellulase immobilized on modified nylon for saccharification of cellulose

    SciTech Connect

    Jain, P.; Wilkins, E.S.

    1987-01-01

    The present study deals with the immobilization of cellulase on nylon and nylon incorporated with glass. The immobilized and free enzymes were compared in terms of their yields, using untreated sawdust (yellow pine wood) and carboxymethylcellulose (CMC) as substrates, at a standard pH and temperature. Also, the sawdust was pretreated with 1% alkaline hydrogen peroxide and the yield compared with the untreated sawdust hydrolysis to determine the importance of the substrate pretreatment.

  13. [Investigation of micro-aqueous covalent immobilization of horseradish peroxidase by "conformation memory"].

    PubMed

    Cai, Yixuan; Chen, Junhua; Yao, Dongsheng; Liu, Daling

    2009-12-01

    We has studied the feasibility of preventing protein from denature during covalent immobilization by "conformation memory", which was achieved by freeze-drying under enzyme active conformation and cross-linked with carrier under micro-aqueous media (MAM). Horseradish peroxidase (HRP) and chitosan beads have been used as the model enzyme and carrier. The MAM consisted of 99% dioxane and 1% water. We compared the immobilized HRP under MAM with that under traditional aqueous solvent, found that the optimum temperature of both was raised to 60 degrees C, and the optimum pH was 6.5. However, the MAM-immobilized HRP had shown less activity loss during usage and six times higher activity than that immobilized under aqueous solvent. After 30 min incubation at 70 degrees C, the MAM-immobilized HRP remained 75.42% activity while the aqueous-media-immobilized enzyme only 15.4%. The MAM-immobilized HRP has shown a better operation stability with 77.69% residue activity after 5 times of repeat operation while the aqueous-media-immobilized enzyme only 16.67%. In addition, the MAM-immobilized HRP had also shown more advantages when used in phenol removal. We constructed enzyme electrodes (CS-HRP-SWCNTs/Au) to further display the different properties of the two immobilized HRP. MAM-immobilized HRP-electrode has shown two times stronger response signal to H2O2 than that immobilized under aqueous media, which indicated a better enzyme activity of MAM-immobilized HRP. Our research demonstrated that the conformation memory, to some extent, did contribute to preventing protein from denaturing when use HRP as a model, and it is feasible to immobilize enzyme by covalent cross-linking method under micro-aqueous media.

  14. Status of plutonium ceramic immobilization processes and immobilization forms

    SciTech Connect

    Ebbinghaus, B.B.; Van Konynenburg, R.A.; Vance, E.R.; Jostsons, A.

    1996-05-01

    Immobilization in a ceramic followed by permanent emplacement in a repository or borehole is one of the alternatives currently being considered by the Fissile Materials Disposition Program for the ultimate disposal of excess weapons-grade plutonium. To make Pu recovery more difficult, radioactive cesium may also be incorporated into the immobilization form. Valuable data are already available for ceramics form R&D efforts to immobilize high-level and mixed wastes. Ceramics have a high capacity for actinides, cesium, and some neutron absorbers. A unique characteristic of ceramics is the existence of mineral analogues found in nature that have demonstrated actinide immobilization over geologic time periods. The ceramic form currently being considered for plutonium disposition is a synthetic rock (SYNROC) material composed primarily of zirconolite (CaZrTi{sub 2}O{sub 7}), the desired actinide host phase, with lesser amounts of hollandite (BaAl{sub 2}Ti{sub 6}O{sub 16}) and rutile (TiO{sub 2}). Alternative actinide host phases are also being considered. These include pyrochlore (Gd{sub 2}Ti{sub 2}O{sub 7}), zircon (ZrSiO{sub 4}), and monazite (CePO{sub 4}), to name a few of the most promising. R&D activities to address important technical issues are discussed. Primarily these include moderate scale hot press fabrications with plutonium, direct loading of PuO{sub 2} powder, cold press and sinter fabrication methods, and immobilization form formulation issues.

  15. Continuous fermentation: improvement of cell immobilization by zeta potential measurement

    SciTech Connect

    Michaux, M.; Paquot, M.; Baijot, B.; Thonart, P.

    1982-01-01

    The influence of various agents such as inulin, proteins, and cationic polymers on the zeta potential of yeast cells has been studied. With various agents, it appears feasible to give a positive charge to the cell and a negative one to the carrier. Under these conditions, the yield of immobilization (mg cells/g support) is significantly improved. This is the case with various agents such as proteins and cationic polymers. For example, the yeild of cell immobilization (Saccharomyces) on sawdust varied from 94.2 to 145.8 mg/g of support with addition of gelatin (0.05%). The results demonstrate the influence of zeta potential on cell immobilization by adsorption. These immobilized cells are advantageously used in continuous production of ethanol. 3 figures, 8 tables.

  16. Immobilization of enzyme on dimethyl-aminated nylon gels

    SciTech Connect

    Miyama, H.; Kawate, M.; Nosaka, Y.

    1985-10-01

    Miyama and co-workers have reported that dimethyl-aminated Nylon is a good material for the immobilization of enzyme after being quaternized. Also, they found a good correlation between the apparent Michaelis constant of the immobilized enzyme and the diffusion constant of the substrate in the matrix, and that between activity of the immobilized enzyme and interfacial free energy of the matrix. In their experiments, Nylon films of low degree of dimethylamination were used. In the present study, dimethylaminated Nylon of high degree of dimethylamination was quaternized by cationic oligomers having two functional groups. Thus, hydrogels were obtained, where the degree of cross-linking and the density of cation could be varied by changing the feeding amount and the length of cross-linking oligomers. These gels were then tested for the immobilization of urease and glucose oxidase. 9 references.

  17. Rational surface silane modification for immobilizing glucose oxidase.

    PubMed

    Tian, Feibao; Guo, Yi; Lin, Feifei; Zhang, Yumei; Yuan, Qipeng; Liang, Hao

    2016-06-01

    Glucose oxidase (GOx) has many significant applications in biosensor and biocatalysis. In this study, we firstly quantitatively analyzed the binding efficiency of (3-aminopropyl) trimethoxysilane (APTES) modified onto the surface of GOx. It was found that the contents of the grafted silane did not significantly influence the relative activities and tertiary structures of all surface modified GOxs. Immobilization ratio and relative activity of all instances of APTES modified GOx increased, compared with those of native enzyme. However, good stability of immobilized GOx at extreme pH and high temperature could only be obtained when modified protein with low binding silane content. At pH 2.0, the immobilized GOx with low binding content showed a more than 600% activity, compared to the free enzyme. Therefore, rational surface modification would be beneficial to improving the activity and stability of immobilized enzyme as well as increasing loading amount.

  18. Acid phosphatase and protease activities in immobilized rat skeletal muscles

    NASA Technical Reports Server (NTRS)

    Witzmann, F. A.; Troup, J. P.; Fitts, R. H.

    1982-01-01

    The effect of hind-limb immobilization on selected Iysosomal enzyme activities was studied in rat hing-limb muscles composed primarily of type 1. 2A, or 2B fibers. Following immobilization, acid protease and acid phosphatase both exhibited signifcant increases in their activity per unit weight in all three fiber types. Acid phosphatase activity increased at day 14 of immobilization in the three muscles and returned to control levels by day 21. Acid protease activity also changed biphasically, displaying a higher and earlier rise than acid phosphatase. The pattern of change in acid protease, but not acid phosphatase, closely parallels observed muscle wasting. The present data therefore demonstrate enhanced proteolytic capacity of all three fiber types early during muscular atrophy. In addition, the data suggest a dependence of basal hydrolytic and proteolytic activities and their adaptive response to immobilization on muscle fiber composition.

  19. Preparation of polyphosphazene hydrogels for enzyme immobilization.

    PubMed

    Qian, Yue-Cheng; Chen, Peng-Cheng; He, Gui-Jin; Huang, Xiao-Jun; Xu, Zhi-Kang

    2014-01-01

    We report on the synthesis and application of a new hydrogel based on a methacrylate substituted polyphosphazene. Through ring-opening polymerization and nucleophilic substitution, poly[bis(methacrylate)phosphazene] (PBMAP) was successfully synthesized from hexachlorocyclotriphosphazene. By adding PBMAP to methacrylic acid solution and then treating with UV light, we could obtain a cross-linked polyphosphazene network, which showed an ultra-high absorbency for distilled water. Lipase from Candida rugosa was used as the model lipase for entrapment immobilization in the hydrogel. The influence of methacrylic acid concentration on immobilization efficiency was studied. Results showed that enzyme loading reached a maximum of 24.02 mg/g with an activity retention of 67.25% when the methacrylic acid concentration was 20% (w/w). PMID:25006790

  20. [Recovery of platinum with immobilized Citrobacter freudii XP05 biomass].

    PubMed

    Hu, Hong-Bo; Liu, Yue-Ying; Fu, Jin-Kun; Xue, Ru; Gu, Ping-Ying

    2003-07-01

    The objective of this work was to develop a valuable adsorbent for recovery of platinum by studying the properties of Pt4+ -adsorption with immobilized Citrobacter freudii XP05 biomass. Five methods for immobilization of Citrobacter freudii XP05 biomass were compared. The method with gelatin-alginate sodium as entrapment matrix was considered to be the optimal. Spherical and uniform beads were produced and the SEM micrograph indicated that the cell of strain XP08 were uniformly dispersed within the matrix. The adsorption of Pt4+ by immobilized XP05 biomass was affected with adsorptive time, pH value of the solution, immobilized biomass concentration, Pt4+ initial concentration The adsorption was a rapid process. The optimal pH value for Pt4+ adsorption was 1.5, and its adsorptive capacity increased linearly with increasing Pt4+ initial concentrations in the range of 50 - 250 mg/L. The experimental data could be fitted to Langmuir and Freundlich models of adsorption isotherm. The adsorptive capacity reached 35.2 mg/g under the conditions of 250 Pt4+ mg/L, 2.0 g/L immobilized biomass, pH 1.5 and 30 degrees C for 60 min. 98.7% of Pt4+ adsorbed on immobilized biomass could be desorbed with 0.5 mol HC1/L. The characteristics of dynamic adsorption and desorption of immobilized XP05 biomass in packed-bed reactor were investigated. The saturation uptake was 24.66 mg Pt4+ /g under the conditions of flow rate 1.2 mL/min, pH 1.5, 50 mg Pt4+/L and 1.85 g biomass(dry weight) . Adsorptive efficiency of Pt4 + by the immobilized XP05 biomass was above 78% for 4 cycles of adsorption and desorption. The recovery of platinum from waste platinum catalyst was studied. The adsorptive capacity was 20.94 mg Pt4+/g immobilized biomass under the conditions of 4.0 g/L immobilized XP05 biomass, 117.76 mg Pt4+/L and pH 1.5 for 60 min. The immobilized XP05 biomass is potentially applicable to the recovery of platinum from waste and wastewater containing platinum. PMID:15969064

  1. [Immobilization of introduced bacteria and degradation of pyrene and benzo(alpha) pyrene in soil by immobilized bacteria].

    PubMed

    Wang, Xin; Li, Peijun; Song, Shouzhi; Zhong, Yong; Zhang, Hui; Verkhozina, E V

    2006-11-01

    In this study, introduced bacteria were applied in the bioremediation of pyrene and benzo (alpha) pyrene in organic pollutants-contaminated soils, aimed to test whether it was feasible to introduce bacteria to environmental engineering. Three introduced bacteria were immobilized separately or together to degrade the pyrene and benzo (alpha) pyrene in soil, taking dissociated bacteria as the control, and comparing with three indigenous bacteria. The results showed that immobilized introduced bacteria, either single or mixed, had higher degradation efficiency than dissociated bacteria. Compared with indigenous bacteria, some introduced bacteria had predominance to some degree. The introduced bacteria-mixture had better degradation efficiency after being immobilized. The degradation rate of pyrene and benzo(alpha) pyrene after treated with immobilized bacteria-( B61-B67)-mixture for 96 hours was 43.49% and 38.55%, respectively.

  2. Immobilization of enzyme on a polymer surface

    NASA Astrophysics Data System (ADS)

    Shen, Lei; Cheng, Kenneth Chun Kuen; Schroeder, McKenna; Yang, Pei; Marsh, E. Neil G.; Lahann, Joerg; Chen, Zhan

    2016-06-01

    We successfully immobilized enzymes onto polymer surfaces via covalent bonds between cysteine groups of the enzyme and dibromomaleimide functionalities present at the polymer surface. In this work, we used nitroreductase (NfsB) as a model enzyme molecule. The polymers were prepared by chemical vapor deposition (CVD) polymerization, resulting in surfaces with dibromomaleimide groups. NfsB variants were engineered so that each NfsB molecule only has one cysteine group on the enzyme surface. Two different NfsB constructs were studied, with cysteines at the positions of H360 and V424, respectively. A combination of sum frequency generation (SFG) vibrational and attenuated total reflectance Fourier transformed infrared (ATR-FTIR) spectroscopies were used to deduce the orientation of the immobilized enzymes on the surface. It was found that the orientation of the immobilized enzymes is controlled by the position of the cysteine residue in the protein. The NfsB H360C construct exhibited a similar orientational behavior on the polymer surface as compared to that on the self-assembled monolayer surface, but the NsfB V424C construct showed markedly different orientations on the two surfaces.

  3. Effect of limb immobilization on skeletal muscle

    NASA Technical Reports Server (NTRS)

    Booth, F. W.

    1982-01-01

    Current knowledge and questions remaining concerning the effects of limb immobilization on skeletal muscle is reviewed. The most dramatic of these effects is muscle atrophy, which has been noted in cases of muscles fixed at or below their resting length. Immobilization is also accompanied by a substantial decrease in motoneuronal discharges, which results in the conversion of slow-twitch muscle to muscle with fast-twitch characteristics. Sarcolemma effects include no change or a decrease in resting membrane potential, the appearance of extrajunctional acetylcholine receptors, and no change in acetylcholinesterase activity. Evidence of changes in motoneuron after hyperpolarization characteristics suggests that the muscle inactivity is responsible for neuronal changes, rather than vice versa. The rate of protein loss from atrophying muscles is determined solely by the first-order rate constant for degradation. Various other biochemical and functional changes have been noted, including decreased insulin responsiveness and protein synthesis. The model of limb immobilization may also be useful for related studies of muscle adaptation.

  4. Gold and silver nanoparticles for biomolecule immobilization and enzymatic catalysis

    NASA Astrophysics Data System (ADS)

    Petkova, Galina A.; Záruba, Кamil; Žvátora, Pavel; Král, Vladimír

    2012-06-01

    In this work, a simple method for alcohol synthesis with high enantiomeric purity was proposed. For this, colloidal gold and silver surface modifications with 3-mercaptopropanoic acid and cysteamine were used to generate carboxyl and amine functionalized gold and silver nanoparticles of 15 and 45 nm, respectively. Alcohol dehydrogenase from Thermoanaerobium brockii (TbADH) and its cofactor (NADPH) were physical and covalent (through direct adsorption and using cross-linker) immobilized on nanoparticles' surface. In contrast to the physical and covalent immobilizations that led to a loss of 90% of the initial enzyme activity and 98% immobilization, the use of a cross-linker in immobilization process promoted a loss to 30% of the initial enzyme activity and >92% immobilization. The yield of NADPH immobilization was about 80%. The best results in terms of activity were obtained with Ag-citr nanoparticle functionalized with carboxyl groups (Ag-COOH), Au-COOH(CTAB), and Au-citr functionalized with amine groups and stabilized with CTAB (Au-NH2(CTAB)) nanoparticles treated with 0.7% and 1.0% glutaraldehyde. Enzyme conformation upon immobilization was studied using fluorescence and circular dichroism spectroscopies. Shift in ellipticity at 222 nm with about 4 to 7 nm and significant decreasing in fluorescence emission for all bioconjugates were observed by binding of TbADH to silver/gold nanoparticles. Emission redshifting of 5 nm only for Ag-COOH-TbADH bioconjugate demonstrated change in the microenvironment of TbADH. Enzyme immobilization on glutaraldehyde-treated Au-NH2(CTAB) nanoparticles promotes an additional stabilization preserving about 50% of enzyme activity after 15 days storage. Nanoparticles attached-TbADH-NADPH systems were used for enantioselective ( ee > 99%) synthesis of ( S)-7-hydroxy-2-tetralol.

  5. Immobilized glycosylated Fmoc-amino acid for SPR: comparative studies of lectin-binding to linear or biantennary diLacNAc structures.

    PubMed

    Nakamura, Kosuke; Sakagami, Hiromi; Asanuma-Date, Kimie; Nagasawa, Nao; Nakahara, Yoshiaki; Akiyama, Hiroshi; Ogawa, Haruko

    2013-12-15

    A method to immobilize glycan-linked amino acids with protected α-amino groups, which are key intermediates to produce the desired neoglycoprotein, to a Biacore sensor chip was developed and its utility for interaction analyses was demonstrated. Two types of diN-acetyllactosamine (diLacNAc)-containing glycans, a core 2 hexasaccharide involving linear diLacNAc that is O-linked to N-(9-fluorenyl)methoxycarbonyl (Fmoc)-Thr and a biantennary diLacNAc that is N-linked to Fmoc-Asn, were used as ligands. For immobilization, the free carboxyl groups of the amino acid residues were activated with EDC/NHS, then reacted with the ethylenediamine-derivatized carboxymethyldextran sensor chip to obtain the desired ligand concentrations. Interactions of the ligands with five plant lectins were analyzed by surface plasmon resonance, and the bindings were compared. The resonance unit of each lectin was corrected by subtracting that of the reference cell on which the Fmoc-Thr-core 1 or Fmoc-Asn was immobilized as a ligand. The carbohydrate specificities of interactions were verified by preincubating lectins with their respective inhibitory sugar before injection. By steady state analysis, the Lycopersicon esculentum lectin showed a 27-fold higher affinity to linear diLacNAc than to biantennary diLacNAc, while Datura stramonium and Solanum tuberosum lectins both showed low Ka,apps of 10(6)M(-1) for these two ligands. In contrast, Ricinus communis agglutinin-120 showed a 3.2-fold higher Ka,app to biantennary LacNAc than to linear diLacNAc. A lectin purified from Pleurocybella porrigens mushroom interacted at the high affinity of 10(8)M(-1) with both linear and biantennary diLacNAcs, which identified it as a unique probe. This method provides a useful and sensitive system to analyze interactions by simulating the glycans on the cell surface. PMID:24211369

  6. [Growth inhibition effect of immobilized pectinase on Microcystis aeruginosa].

    PubMed

    Shen, Qing-Qing; Peng, Qian; Lai, Yong-Hong; Ji, Kai-Yan; Han, Xiu-Lin

    2012-12-01

    To confirm the growth inhibition effect of immobilized pectinase on algae, co-cultivation method was used to investigate the effect of immobilized pectinase on the growth of Microcystis aeruginosa. After co-cultivation, the damage status of the algae was observed through electron microscope, and the effect of immobilized pectase on the physiological and biochemical characteristics of the algae was also measured. The results showed that the algae and immobilized pectase co-cultivated solution etiolated distinctly on the third day and there was a significantly positive correlation between the extent of etiolation and the dosage as well as the treating time of the immobilized pectinase. Under electron microscope, plasmolysis was found in the slightly damaged cells, and the cell surface of these cells was rough, uneven and irregular; the severely damaged cells were collapsed or disintegrated completely. The algal yield and the chlorophyll a content decreased significantly with the increase of the treating time. The measurement of the malondiadehyde (MDA) value showed that the antioxidation system of the treated algal cells was destroyed, and their membrane lipid was severely peroxidated. The study indicated that the immobilized pectinase could efficiently inhibit the growth of M. aeruginosa, and the inhibitory rate reached up to 96%. PMID:23379158

  7. Nucleosome immobilization strategies for single-pair FRET microscopy.

    PubMed

    Koopmans, Wiepke J A; Schmidt, Thomas; van Noort, John

    2008-10-01

    All genomic transactions in eukaryotes take place in the context of the nucleosome, the basic unit of chromatin, which is responsible for DNA compaction. Overcoming the steric hindrance that nucleosomes present for DNA-processing enzymes requires significant conformational changes. The dynamics of these have been hard to resolve. Single-pair Fluorescence Resonance Energy Transfer (spFRET) microscopy is a powerful technique for observing conformational dynamics of the nucleosome. Nucleosome immobilization allows the extension of observation times to a limit set only by photobleaching, and thus opens the possibility of studying processes occurring on timescales ranging from milliseconds to minutes. It is crucial however, that immobilization itself does not introduce artifacts in the dynamics. Here we report on various nucleosome immobilization strategies, such as single-point attachment to polyethylene glycol (PEG) or surfaces coated with bovine serum albumin (BSA), and confinement in porous agarose or polyacrylamide gels. We compare the immobilization specificity and structural integrity of immobilized nucleosomes. A crosslinked star polyethylene glycol coating performs best with respect to tethering specificity and nucleosome integrity, and enables us to reproduce for the first time bulk nucleosome unwrapping kinetics in single nucleosomes without immobilization artifacts.

  8. Covalent immobilization of a flavoprotein monooxygenase via its flavin cofactor.

    PubMed

    Krzek, Marzena; van Beek, Hugo L; Permentier, Hjalmar P; Bischoff, Rainer; Fraaije, Marco W

    2016-01-01

    A generic approach for flavoenzyme immobilization was developed in which the flavin cofactor is used for anchoring enzymes onto the carrier. It exploits the tight binding of flavin cofactors to their target apo proteins. The method was tested for phenylacetone monooxygenase (PAMO) which is a well-studied and industrially interesting biocatalyst. Also a fusion protein was tested: PAMO fused to phosphite dehydrogenase (PTDH-PAMO). The employed flavin cofactor derivative, N6-(6-carboxyhexyl)-FAD succinimidylester (FAD*), was covalently anchored to agarose beads and served for apo enzyme immobilization by their reconstitution into holo enzymes. The thus immobilized enzymes retained their activity and remained active after several rounds of catalysis. For both tested enzymes, the generated agarose beads contained 3 U per g of dry resin. Notably, FAD-immobilized PAMO was found to be more thermostable (40% activity after 1 h at 60 °C) when compared to PAMO in solution (no activity detected after 1 h at 60 °C). The FAD-decorated agarose material could be easily recycled allowing multiple rounds of immobilization. This method allows an efficient and selective immobilization of flavoproteins via the FAD flavin cofactor onto a recyclable carrier.

  9. Immobilization of pectin depolymerising polygalacturonase using different polymers.

    PubMed

    Ur Rehman, Haneef; Aman, Afsheen; Nawaz, Muhammad Asif; Karim, Asad; Ghani, Maria; Baloch, Abdul Hameed; Ul Qader, Shah Ali

    2016-01-01

    Polygalacturonase catalyses the hydrolysis of pectin substances and widely has been used in food and textile industries. In current study, different polymers such as calcium alginate beads, polyacrylamide gel and agar-agar matrix were screened for the immobilization of polygalacturonase through entrapment technique. Polyacrylamide gel was found to be most promising one and gave maximum (89%) immobilization yield as compared to agar-agar (80%) and calcium alginate beads (46%). The polymers increased the reaction time of polygalacturonase and polymers entrapped polygalacturonases showed maximum pectinolytic activity after 10 min of reaction as compared to free polygalacturonase which performed maximum activity after 5.0 min of reaction time. The temperature of polygalacturonase for maximum enzymatic activity was increased from 45°C to 50°C and 55°C when it was immobilized within agar-agar and calcium alginate beads, respectively. The optimum pH (pH 10) of polygalacturonase was remained same when it was immobilized within polyacrylamide gel and calcium alginate beads, but changed from pH 10 to pH 9.0 after entrapment within agar-agar. Thermal stability of polygalacturonase was improved after immobilization and immobilized polygalacturonases showed higher tolerance against different temperatures as compared to free enzyme. Polymers entrapped polygalacturonases showed good reusability and retained more than 80% of their initial activity during 2nd cycles.

  10. Determination of the Enantiomerization Barrier of the Residual Enantiomers of C3 -Symmetric Tris[3-(1-Methyl-2-Alkyl)Indolyl]Phosphane Oxides: Case Study of a Multitasking HPLC Investigation Based on an Immobilized Polysaccharide Stationary Phase.

    PubMed

    Rizzo, Simona; Menta, Sergio; Benincori, Tiziana; Ferretti, Rosella; Pierini, Marco; Cirilli, Roberto; Sannicolò, Francesco

    2015-12-01

    The residual enantiomers of three tris-(3-indolyl)-phosphane oxides bearing different alkyl groups (methyl, ethyl or i-propyl) in position 2 of the indole rings constituting the blades were separated on the immobilized type Chiralpak IC column in polar organic and reversed-phase modes. The good enantioselectivity and versatility of the IC CSP allowed easy isolation of the enantiomerically highly enriched samples suitable for configurational stability studies. The enantiomerization barriers of residual phosphane oxides were evaluated both by off-column techniques (CD signal and enantiomeric purity decay kinetics) and by dynamic enantioselective high-performance liquid chromatography (HPLC). PMID:26402152

  11. Determination of the Enantiomerization Barrier of the Residual Enantiomers of C3 -Symmetric Tris[3-(1-Methyl-2-Alkyl)Indolyl]Phosphane Oxides: Case Study of a Multitasking HPLC Investigation Based on an Immobilized Polysaccharide Stationary Phase.

    PubMed

    Rizzo, Simona; Menta, Sergio; Benincori, Tiziana; Ferretti, Rosella; Pierini, Marco; Cirilli, Roberto; Sannicolò, Francesco

    2015-12-01

    The residual enantiomers of three tris-(3-indolyl)-phosphane oxides bearing different alkyl groups (methyl, ethyl or i-propyl) in position 2 of the indole rings constituting the blades were separated on the immobilized type Chiralpak IC column in polar organic and reversed-phase modes. The good enantioselectivity and versatility of the IC CSP allowed easy isolation of the enantiomerically highly enriched samples suitable for configurational stability studies. The enantiomerization barriers of residual phosphane oxides were evaluated both by off-column techniques (CD signal and enantiomeric purity decay kinetics) and by dynamic enantioselective high-performance liquid chromatography (HPLC).

  12. Selected transuranic waste immobilization systems

    SciTech Connect

    Timmerman, C.L.; Treat, R.L.; Ross, W.A.

    1981-12-01

    Waste contaminated with transuranic (TRU) elements may require treatment prior to final disposal. Pacific Northwest Laboratory has conducted research and development to identify and characterize the wastes; to evaluate the possible immobilization requirements and treatment alternatives; and to develop immobilization process technologies. This paper describes systems that are anticipated to be capable of immobilizing a selected TRU waste stream consisting of a blend of process sludge and incinerator ash. The selected waste streams are based on the waste compositions generated at the Rocky Flats Plant, Golden, Colorado. The specific process and waste forms are: in-can glass melting, borosilicate glass monolith; joule-heated glass melting, borosilicate/aluminosilicate glass monolith; glass marble, borosilicate/aluminosilicate glass marble; basalt glass-ceramic, basalt glass-ceramic monolith; cast cement, cast cement monolith; pressed cement, pressed cement pellet; and cold-pressed sintered ceramic, pressed ceramic pellet.

  13. Hydrolysis of triacetin catalyzed by immobilized lipases: effect of the immobilization protocol and experimental conditions on diacetin yield.

    PubMed

    Hernandez, Karel; Garcia-Verdugo, Eduardo; Porcar, Raul; Fernandez-Lafuente, Roberto

    2011-05-01

    The effect of the immobilization protocol and some experimental conditions (pH value and presence of acetonitrile) on the regioselective hydrolysis of triacetin to diacetin catalyzed by lipases has been studied. Lipase B from Candida antarctica (CALB) and lipase from Rhizomucor miehei (RML) were immobilized on Sepabeads (commercial available macroporous acrylic supports) activated with glutaraldehyde (covalent immobilization) or octadecyl groups (adsorption via interfacial activation). All the biocatalysts accumulated diacetin. Covalently immobilized RML was more active towards rac-methyl mandelate than the adsorbed RML. However, this covalent RML preparation presented the lowest activity towards triacetin. For this reason, this preparation was discarded as biocatalyst for this reaction. At pH 7, acyl migration occurred giving a mixture of 1,2 and 1,3 diacetin, but at pH 5.5, only 1,2 diacetin was produced. Yields were improved at acidic pH values and in the presence of 20% acetonitrile (to over 95%). RML immobilized on octadecyl Sepabeads was proposed as optimal preparation, mainly due to its higher specific activity. Each enzyme preparation presented very different properties. Moreover, changes in the reaction conditions affected the various immobilized enzymes in a different way.

  14. Biodiesel production from Jatropha oil catalyzed by immobilized Burkholderia cepacia lipase on modified attapulgite.

    PubMed

    You, Qinghong; Yin, Xiulian; Zhao, Yuping; Zhang, Yan

    2013-11-01

    Lipase from Burkholderia cepacia was immobilized on modified attapulgite by cross-linking reaction for biodiesel production with jatropha oil as feedstock. Effects of various factors on biodiesel production were studied by single-factor experiment. Results indicated that the best conditions for biodiesel preparation were: 10 g jatropha oil, 2.4 g methanol (molar ratio of oil to methanol is 1:6.6) being added at 3h intervals, 7 wt% water, 10 wt% immobilized lipase, temperature 35°C, and time 24h. Under these conditions, the maximum biodiesel yield reached 94%. The immobilized lipase retained 95% of its relative activity during the ten repeated batch reactions. The half-life time of the immobilized lipase is 731 h. Kinetics was studied and the Vmax of the immobilized lipases were 6.823 mmol L(-1). This immobilized lipase catalyzed process has potential industrial use for biodiesel production to replace chemical-catalyzed method.

  15. Biodiesel production from Jatropha oil catalyzed by immobilized Burkholderia cepacia lipase on modified attapulgite.

    PubMed

    You, Qinghong; Yin, Xiulian; Zhao, Yuping; Zhang, Yan

    2013-11-01

    Lipase from Burkholderia cepacia was immobilized on modified attapulgite by cross-linking reaction for biodiesel production with jatropha oil as feedstock. Effects of various factors on biodiesel production were studied by single-factor experiment. Results indicated that the best conditions for biodiesel preparation were: 10 g jatropha oil, 2.4 g methanol (molar ratio of oil to methanol is 1:6.6) being added at 3h intervals, 7 wt% water, 10 wt% immobilized lipase, temperature 35°C, and time 24h. Under these conditions, the maximum biodiesel yield reached 94%. The immobilized lipase retained 95% of its relative activity during the ten repeated batch reactions. The half-life time of the immobilized lipase is 731 h. Kinetics was studied and the Vmax of the immobilized lipases were 6.823 mmol L(-1). This immobilized lipase catalyzed process has potential industrial use for biodiesel production to replace chemical-catalyzed method. PMID:24055964

  16. Tissue stiffness induced by prolonged immobilization of the rat knee joint and relevance of AGEs (pentosidine).

    PubMed

    Lee, Sachiko; Sakurai, Takashi; Ohsako, Masafumi; Saura, Ryuichi; Hatta, Hideo; Atomi, Yoriko

    2010-12-01

    Joints, connective tissues consisting of extracellular matrix (ECM) with few blood vessels, transfer tension to the skeleton in response to environmental demand. Therefore, joint immobilization decreases active and passive mechanical stress, resulting in increased joint stiffness and tissue degeneration; however, the cause of joint stiffness is obscure. Using a rat knee immobilization model, we examined the relationship between range of motion (ROM) and cell numbers and ECM cross-links by accumulation of advanced glycation end products, pentosidine, in the posterior joint capsule of immobilized joints during 16 weeks of immobilization. The left knee joint was immobilized by internal fixation and compared with the non-immobilized right leg. As early as 2 weeks of immobilization, joint ROM and torque significantly decreased and in parallel, disordered alignment of collagen fiber bundles significantly increased, compared with non-immobilized joints. Those changes continued until 16 weeks of immobilization. Significant increases in pentosidine-positive areas after 8 weeks and significantly decreased cell numbers after 16 weeks of immobilization were also observed compared to the contralateral side. A significant negative correlation between tissue stiffness measured by restriction of ROM and accumulation of pentosidine was observed. This study is the first to show that immobilization of knee joints induces articular contracture associated with sequential changes of ECM alignment, influencing ROM and later pentosidine accumulation and decreased cell numbers during the 16-week immobilization period. Pentosidine appears to be an indicator toward a chronic tissue stiffness leading to decreased cell number rather than a cause of ROM restriction induced by joint immobilization.

  17. Bioreduction and immobilization of uranium in situ: a case study at a USA Department of Energy radioactive waste site, Oak Ridge, Tennessee

    SciTech Connect

    Wu, Weimin; Carley, Jack M; Watson, David B; Gu, Baohua; Brooks, Scott C; Kelly, Shelly D; Kemner, Kenneth M; Van Nostrand, Joy; Wu, Liyou; Zhou, Jizhong; Luo, Jian; Cardenas, Erick; Fields, Matthew Wayne; Marsh, Terence; Tiedje, James; Green, Stefan; Kostka, Joel; Kitanidis, Peter K.; Jardine, Philip; Criddle, Craig

    2011-01-01

    Bioremediation of uranium contaminated groundwater was tested by delivery of ethanol as an electron donor source to stimulate indigenous microbial bioactivity for reduction and immobilization of uranium in situ, followed by tests of stability of uranium sequestration in the bioreduced area via delivery of dissolved oxygen or nitrate at the US Department of energy's Integrated Field Research Challenge site located at Oak Ridge, Tennessee, USA. After long term treatment that spanned years, uranium in groundwater was reduced from 40-60 mg {center_dot} L{sup -1} to <0.03 mg {center_dot} L{sup -1}, below the USA EPA standard for drinking water. The bioreduced uranium was stable under anaerobic or anoxic conditions, but addition of DO and nitrate to the bioreduced zone caused U remobilization. The change in the microbial community and functional microorganisms related to uranium reduction and oxidation were characterized. The delivery of ethanol as electron donor stimulated the activities of indigenous microorganisms for reduction of U(VI) to U(IV). Results indicated that the immobilized U could be partially remobilized by D0 and nitrate via microbial activity. An anoxic environmental condition without nitrate is essential to maintain the stability of bioreduced uranium.

  18. Immobilized Metal Affinity Electrophoresis: A Novel Method of Capturing Phosphoproteins by Electrophoresis

    PubMed Central

    Lee, Bao-Shiang; Lasanthi, G.D.; Jayathilaka, P.; Huang, Jin-Sheng; Gupta, Shalini

    2008-01-01

    An immobilized metal affinity electrophoresis (IMAEP) method is described here. In this method, metal ions are immobilized in a native polyacrylamide gel to capture phosphoproteins. The capture of phosphoproteins by IMAEP is demonstrated with immobilized metals like iron, aluminum, manganese, or titanium. In the case studies, phosphoproteins α-casein, β-casein, and phosvitin are successfully extracted from a protein mixture by IMAEP. PMID:19137092

  19. Role of immobilization of irradiated rats in the protective effect of bone marrow shielding

    NASA Technical Reports Server (NTRS)

    Gronskaya, N. F.; Strelin, G. S.

    1982-01-01

    Rats were exposed to X-radiation to study the influence of immobilization and shielding of part of bone marrow during exposure on survival. It is concluded that (1) the beneficial effect of the stress factor (created by the immobilization of rats during exposure) can aggregate with the effect of bone marrow shielding and, under certain conditions, imitate the latter; and (2) the probability of the protective effect of immobilization should be taken into account when assessing the influence of bone marrow shielding.

  20. Effects of immobilization on articular cartilage: Autohistoradiographic findings with S35

    NASA Technical Reports Server (NTRS)

    Digiovanni, C.; Desantis, E.

    1980-01-01

    The effect of immobilization on the articular cartilage of rabbits was studied by light microscope. The knee joint of each rabbit was immobilized in a plaster in a position midway between flexion and extension for a 10 to 120 days period. Degenerative changes in the articular cartilage of increasing severity were observed. The fixation of the labeled SO4 by cartilage cells was decreased in advanced immobilization.

  1. Radiation immobilization of catalase and its application

    NASA Astrophysics Data System (ADS)

    Guanghui, Wang; Hongfei, Ha; Xia, Wang; Jilan, Wu

    Catalase was immobilized by chemical method on porous polyacrylamide particles which produced through radiation polymerization of acrylamide monomer at low temperature (-78°C). Activity of immobilized catalase was enhanced distinctly by joining a chemical "arm" to the support. The method of recovery of catalase activity on immobilized polymer was found by soaking it in certain buffer. The treatment of H 2O 2 both in aqueous solution and alcoholic solution by using the immobilized catalase was performed.

  2. Enzyme immobilization and biocatalysis of polysiloxanes

    NASA Astrophysics Data System (ADS)

    Poojari, Yadagiri

    Lipases have been proven to be versatile and efficient biocatalysts which can be used in a broad variety of esterification, transesterification, and ester hydrolysis reactions. Due to the high chemo-, regio-, and stereo-selectivity and the mild conditions of lipase-catalyzed reactions, the vast potential of these biocatalysts for use in industrial applications has been increasingly recognized. Polysiloxanes (silicones) are well known for their unique physico-chemical properties and can be prepared in the form of fluids, elastomers, gels and resins for a wide variety of applications. However, the enzymatic synthesis of silicone polyesters and copolymers is largely unexplored. In the present investigations, an immobilized Candida antarctica lipase B (CALB) on macroporous acrylic resin beads (Novozym-435 RTM) has been successfully employed as a catalyst to synthesize silicone polyesters and copolymers under mild reaction conditions. The silicone aliphatic polyesters and the poly(dimethylsiloxane)--poly(ethylene glycol) (PDMS-PEG) copolymers were synthesized in the bulk (without using a solvent), while the silicone aromatic polyesters, the silicone aromatic polyamides and the poly(epsilon-caprolactone)--poly(dimethylsiloxane)--poly(epsilon-caprolactone) (PCL-PDMS-PCL) triblock copolymers were synthesized in toluene. The synthesized silicone polyesters and copolymers were characterized by Gel Permeation Chromatography (GPC), Fourier Transform Infrared Spectroscopy (FTIR), Thermogravimetric Analysis (TGA), Differential Scanning Calorimetry (DSC) and Wide Angle X-ray Diffraction (WAXD). This dissertation also describes a methodology for physical immobilization of the enzyme pepsin from Porcine stomach mucosa in silicone elastomers utilizing condensation-cure room temperature vulcanization (RTV) of silanol-terminated poly(dimethylsiloxane) (PDMS). The activity and the stability of free pepsin and pepsin immobilized in silicone elastomers were studied with respect to p

  3. Mineral induction by immobilized phosphoproteins

    NASA Technical Reports Server (NTRS)

    Saito, T.; Arsenault, A. L.; Yamauchi, M.; Kuboki, Y.; Crenshaw, M. A.

    1997-01-01

    Dentin phosphoproteins are thought to have a primary role in the deposition of mineral on the collagen of dentin. In this study we determined the type of binding between collagen and phosphoproteins necessary for mineral formation onto collagen fibrils and whether the phosphate esters are required. Bovine dentin phosphophoryn or phosvitin from egg yolk were immobilized on reconstituted skin type I collagen fibrils by adsorption or by covalent cross-linking. In some samples the ester phosphate was removed from the covalently cross-linked phosphoproteins by treatment with acid phosphatase. All samples were incubated at 37 degrees C in metastable solutions that do not spontaneously precipitate. Reconstituted collagen fibrils alone did not induce mineral formation. The phosphoproteins adsorbed to the collagen fibrils desorbed when the mineralization medium was added, and mineral was not induced. The mineral induced by the cross-linked phosphoproteins was apatite, and the crystals were confined to the surface of the collagen fibrils. With decreasing medium saturation the time required for mineral induction increased. The interfacial tensions calculated for apatite formation by either phosphoprotein cross-linked to collagen were about the same as that for phosphatidic acid liposomes and hydroxyapatite. This similarity in values indicates that the nucleation potential of these highly phosphorylated surfaces is about the same. It is concluded that phosphoproteins must be irreversibly bound to collagen fibrils for the mineralization of the collagen network in solutions that do not spontaneously precipitate. The phosphate esters of phosphoproteins are required for mineral induction, and the carboxylate groups are not sufficient.

  4. Growth and by-product profiles of Kluyveromyces marxianus cells immobilized in foamed alginate.

    PubMed

    Wilkowska, Agnieszka; Kregiel, Dorota; Guneser, Onur; Karagul Yuceer, Yonca

    2015-01-01

    The aim of this research was to study how the yeast cell immobilization technique influences the growth and fermentation profiles of Kluyveromyces marxianus cultivated on apple/chokeberry and apple/cranberry pomaces. Encapsulation of the cells was performed by droplet formation from a foamed alginate solution. The growth and metabolic profiles were evaluated for both free and immobilized cells. Culture media with fruit waste produced good growth of free as well as immobilized yeast cells. The fermentation profiles of K. marxianus were different with each waste material. The most varied aroma profiles were noted for immobilized yeast cultivated on apple/chokeberry pomace. PMID:25277269

  5. Degradation of mix hydrocarbons by immobilized cells of mix culture using a trickle fluidized bed reactor

    SciTech Connect

    Chapatwala, K.D.

    1993-01-01

    The microorganisms, capable of degrading mix hydrocarbons were isolated from the soil samples collected from the hydrocarbon contaminated sites. The mix cultures were immobilized in calcium alginate solution in the form of beads. A trickle fluidized bed air-uplift-type reactor designed to study the degradation of mix hydrocarbons was filled with 0.85% normal saline containing the immobilized cells of mix culture. The immobilized beads were aerated with CO[sub 2]-free air at 200 ml/min. The degradation of different concentrations of hydrocarbons in the presence/absence of commercially available fertilizers by the immobilized cells of mix culture is now in progress.

  6. A High-Resolution Magic Angle Spinning NMR Study of the Enantiodiscrimination of 3,4-Methylenedioxymethamphetamine (MDMA) by an Immobilized Polysaccharide-Based Chiral Phase

    PubMed Central

    Paixão, Márcio W.; Lourenço, Tiago C.

    2016-01-01

    This paper reports the investigation of the chiral interaction between 3,4-methylenedioxy-methamphetamine (MDMA) enantiomers and an immobilized polysaccharide-based chiral phase. For that, suspended-state high-resolution magic angle spinning nuclear magnetic resonance spectroscopy (1H HR-MAS NMR) was used. 1H HR-MAS longitudinal relaxation time and Saturation Transfer Difference (STD NMR) titration experiments were carried out yielding information at the molecular level of the transient diastereoisomeric complexes of MDMA enantiomers and the chiral stationary phase. The interaction of the enantiomers takes place through the aromatic moiety of MDMA and the aromatic group of the chiral selector by π-π stacking for both enantiomers; however, a stronger interaction was observed for the (R)-enantiomer, which is the second one to elute at the chromatographic conditions. PMID:27668862

  7. Metal immobilization and soil amendment efficiency at a contaminated sediment landfill site: a field study focusing on plants, springtails, and bacteria.

    PubMed

    Bert, Valérie; Lors, Christine; Ponge, Jean-François; Caron, Lucie; Biaz, Asmaa; Dazy, Marc; Masfaraud, Jean-François

    2012-10-01

    Metal immobilization may contribute to the environmental management strategy of dredged sediment landfill sites contaminated by metals. In a field experiment, amendment effects and efficiency were investigated, focusing on plants, springtails and bacteria colonisation, metal extractability and sediment ecotoxicity. Conversely to hydroxylapatite (HA, 3% DW), the addition of Thomas Basic Slag (TBS, 5% DW) to a 5-yr deposited sediment contaminated with Zn, Cd, Cu, Pb and As resulted in a decrease in the 0.01 M Ca(NO(3))(2)-extractable concentrations of Cd and Zn. Shoot Cd and Zn concentration in Calamagrostis epigejos, the dominant plant species, also decreased in the presence of TBS. The addition of TBS and HA reduced sediment ecotoxicity and improved the growth of the total bacterial population. Hydroxylapatite improved plant species richness and diversity and decreased antioxidant enzymes in C. Epigejos and Urtica dïoica. Collembolan communities did not differ in abundance and diversity between the different treatments. PMID:22647548

  8. Evaluation of an organo-layered double hydroxide and two organic residues as amendments to immobilize metalaxyl enantiomers in soils: A comparative study.

    PubMed

    López-Cabeza, Rocío; Cornejo, Juan; Celis, Rafael

    2016-10-01

    Many pollutants released into the environment as a result of human activities are chiral. Pollution control strategies generally consider chiral compounds as if they were achiral and rarely consider enantiomers separately. We compared the performance of three different materials, an organically-modified anionic clay (HT-ELA) and two organic agro-food residues (ALP and ALPc), as amendments to immobilize the chiral fungicide metalaxyl in two soils with different textures, addressing the effects of the amendments on the sorption, persistence, and leaching of each of the two enantiomers of metalaxyl (R-metalaxyl and S-metalaxyl) separately. The effects of the amendments were both soil- and amendment-dependent, as well as enantiomer-selective. The organo-clay (HT-ELA) was much more efficient in increasing the sorption capacity of the soils for the two enantiomers of metalaxyl than the agro-food residues (ALP and ALPc), even when applied at a reduced application rate. The enhanced sorption in HT-ELA-amended soils reduced the bioavailability of metalaxyl enantiomers and their leaching in the soils, mitigating the particularly high leaching potential of the more persistent S enantiomer. The immobilizing capacity of the agro-food residues was more variable, mainly because their addition did not greatly ameliorate the sorption capacity of the soils and had variable effects on the enantiomers degradation rates. HT-ELA showed potential to reduce the bioavailability and mobility of metalaxyl enantiomers in soil and to mitigate the contamination problems particularly associated with the higher leaching potential of the more persistent enantiomer.

  9. Evaluation of an organo-layered double hydroxide and two organic residues as amendments to immobilize metalaxyl enantiomers in soils: A comparative study.

    PubMed

    López-Cabeza, Rocío; Cornejo, Juan; Celis, Rafael

    2016-10-01

    Many pollutants released into the environment as a result of human activities are chiral. Pollution control strategies generally consider chiral compounds as if they were achiral and rarely consider enantiomers separately. We compared the performance of three different materials, an organically-modified anionic clay (HT-ELA) and two organic agro-food residues (ALP and ALPc), as amendments to immobilize the chiral fungicide metalaxyl in two soils with different textures, addressing the effects of the amendments on the sorption, persistence, and leaching of each of the two enantiomers of metalaxyl (R-metalaxyl and S-metalaxyl) separately. The effects of the amendments were both soil- and amendment-dependent, as well as enantiomer-selective. The organo-clay (HT-ELA) was much more efficient in increasing the sorption capacity of the soils for the two enantiomers of metalaxyl than the agro-food residues (ALP and ALPc), even when applied at a reduced application rate. The enhanced sorption in HT-ELA-amended soils reduced the bioavailability of metalaxyl enantiomers and their leaching in the soils, mitigating the particularly high leaching potential of the more persistent S enantiomer. The immobilizing capacity of the agro-food residues was more variable, mainly because their addition did not greatly ameliorate the sorption capacity of the soils and had variable effects on the enantiomers degradation rates. HT-ELA showed potential to reduce the bioavailability and mobility of metalaxyl enantiomers in soil and to mitigate the contamination problems particularly associated with the higher leaching potential of the more persistent enantiomer. PMID:27341374

  10. Algal polysaccharides as matrices for the immobilization of urease in lipid ultrathin films studied with tensiometry and vibrational spectroscopy: Physical-chemical properties and implications in the enzyme activity.

    PubMed

    de Brito, Audrey Kalinouski; Nordi, Cristina S F; Caseli, Luciano

    2015-11-01

    Currently, many biological substances extracted from algae have received special attention because of their intrinsic characteristics, which can be applied to different areas of biotechnology. Therefore, in the current study, exopolysaccharides (EPS) from the microalgae Cryptomonas tetrapirenoidosa were employed as an aqueous subphase of a monolayer formed by the lipid dioctadecyldimethylammonium bromide (DODAB). The primary objective of this approach was to evaluate whether EPS could serve as a matrix for the immobilization of the enzyme urease to produce biosensors for urea. After DODAB was spread on the EPS solutions, urease was injected into the aqueous subphase, and the surface was submitted to compression using lateral barriers. The monolayers were subsequently characterized by surface pressure-area isotherms and polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS). The results indicated that EPS enhanced the adsorption of the enzyme on the lipid monolayer. The mixed films were later transferred to solid supports using the Langmuir-Blodgett (LB) technique and were characterized by transfer ratio, PM-IRRAS, quartz crystal microbalance, and atomic force microscopy. The immobilization of the enzyme on solid supports was fundamental for providing an ideal geometrical accommodation of urease because the interaction of EPS with urease in solution causes a decrease of the relative activity of urease. Therefore, these LB films are promising for the fabrication of future urea biosensors, the architecture of which can be manipulated and enhanced at the molecular level.

  11. Recent developments and applications of immobilized laccase.

    PubMed

    Fernández-Fernández, María; Sanromán, M Ángeles; Moldes, Diego

    2013-12-01

    Laccase is a promising biocatalyst with many possible applications, including bioremediation, chemical synthesis, biobleaching of paper pulp, biosensing, textile finishing and wine stabilization. The immobilization of enzymes offers several improvements for enzyme applications because the storage and operational stabilities are frequently enhanced. Moreover, the reusability of immobilized enzymes represents a great advantage compared with free enzymes. In this work, we discuss the different methodologies of enzyme immobilization that have been reported for laccases, such as adsorption, entrapment, encapsulation, covalent binding and self-immobilization. The applications of laccase immobilized by the aforementioned methodologies are presented, paying special attention to recent approaches regarding environmental applications and electrobiochemistry.

  12. Immobilization of Heparin: Approaches and Applications

    PubMed Central

    Murugesan, Saravanababu; Xie, Jin; Linhardt, Robert J.

    2014-01-01

    Heparin, an anticoagulant, has been used in many forms to treat various diseases. These forms include soluble heparin and heparin immobilized to supporting matrices by physical adsorption, by covalent chemical methods and by photochemical attachment. These immobilization methods often require the use of spacers or linkers. This review examines and compares various techniques that have been used for the immobilization of heparin as well as applications of these immobilized heparins. In the applications reviewed, immobilized heparin is compared with soluble heparin for efficient and versatile use in each of the various applications. PMID:18289079

  13. Immobilization of diastase α-amylase on nano zinc oxide.

    PubMed

    Antony, Navya; Balachandran, S; Mohanan, P V

    2016-11-15

    Diastase α-amylase extracted from malt, catalyses break down of starch into maltose. It is commonly used in food and fermentation industry. In the present study nano zinc oxide is used as support for this starch hydrolyzing enzyme. IR study revealed that the enzyme got adsorbed via electrostatic interaction with the functional groups on the support. The immobilized enzyme possessed a better heat-resistance than free enzyme. The kinetic parameters were determined using Lineweaver-Burk plot. The immobilized enzyme showed higher Km 2.08mg/ml than the free enzyme whose Km is 0.45±.05mg/ml. The Vmax of immobilized enzyme was about 2.92±.02mg/ml/min and that of free enzyme was 7.14±.02mg/ml/min, showing decrease in activity after immobilization. The immobilized enzyme showed 70% activity after 30days of storage while free enzyme lost its activity within 7days. About 80% of enzyme retained activity after 4 cycles of reuse.

  14. Effects of acoustic wave resonance oscillation on immobilized enzyme

    NASA Astrophysics Data System (ADS)

    Nishiyama, Hiroshi; Watanabe, Tomoya; Inoue, Yasunobu

    2014-03-01

    In aiming at developing a new method to artificially activate enzyme catalysts immobilized on surface, the effects of resonance oscillation of bulk acoustic waves were studied. Glucose oxidase (GOD) was immobilized by a covalent coupling method on a ferroelectric lead zirconate titanate (PZT) device that was able to generate thickness-extensional resonance oscillation (TERO). Glucose oxidation by the GOD enzyme was studied in a microreactor. The generation of TERO immediately increased the catalytic activity of immobilized GOD by a factor of 2-3. With turn-off of TERO, no significant activity decrease occurred, and 80-90% of the enhanced activity was maintained while the reaction proceeded. The almost complete reversion of the activity to the original low level before TERO generation was observed when the immobilized GOD was exposed to a glucose substrate-free solution. These results indicated that the presence of glucose substrate was essential for TERO-induced GOD activation and preservation of the increased activity level. The influences of reaction temperature, glucose concentration, pH, and rf electric power on the TERO activation showed that TERO strengthened the interactions of the immobilized enzyme with glucose substrate and hence promoted the formation of an activation complex.

  15. Biodiesel production with immobilized lipase: A review.

    PubMed

    Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

    2010-01-01

    Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored. PMID:20580809

  16. Biodiesel production with immobilized lipase: A review.

    PubMed

    Tan, Tianwei; Lu, Jike; Nie, Kaili; Deng, Li; Wang, Fang

    2010-01-01

    Fatty acid alkyl esters, also called biodiesel, are environmentally friendly and show great potential as an alternative liquid fuel. Biodiesel is produced by transesterification of oils or fats with chemical catalysts or lipase. Immobilized lipase as the biocatalyst draws high attention because that process is "greener". This article reviews the current status of biodiesel production with immobilized lipase, including various lipases, immobilization methods, various feedstocks, lipase inactivation caused by short chain alcohols and large scale industrialization. Adsorption is still the most widely employed method for lipase immobilization. There are two kinds of lipase used most frequently especially for large scale industrialization. One is Candida antartica lipase immobilized on acrylic resin, and the other is Candida sp. 99-125 lipase immobilized on inexpensive textile membranes. However, to further reduce the cost of biodiesel production, new immobilization techniques with higher activity and stability still need to be explored.

  17. Comparative evaluation of pumice stone as an alternative immobilization material for 1,3-propanediol production from waste glycerol by immobilized Klebsiella pneumoniae.

    PubMed

    Gonen, Cagdas; Gungormusler, Mine; Azbar, Nuri

    2012-12-01

    In this study, pumice stone (PS), which is a vastly available material in Turkey, was evaluated as an alternative immobilization material in comparison to other commercially available immobilization materials such as glass beads and polyurethane foam. All immobilized bioreactors resulted in much better 1,3-propanediol production from waste glycerol in comparison to the suspended cell culture bioreactor. It was also demonstrated that the locally available PS material is as good as the commercially available immobilization material. The maximum volumetric productivity (8.5 g L(-1) h(-1)) was obtained by the PS material, which is 220 % higher than the suspended cell system. Furthermore, the immobilized bioreactor system was much more robust against cell washout even at very low hydraulic retention time values.

  18. Immobilization of Yarrowia lipolytica for aroma production from castor oil.

    PubMed

    Braga, Adelaide; Belo, Isabel

    2013-04-01

    The main aim of this study was to compare different materials for Y. lipolytica immobilization that could be used in the production of γ-decalactone (a peach-like aroma) in order to prevent the toxic effect both of the substrate and the aroma upon the cells. Therefore, cells adsorption onto pieces of methyl polymethacrylate and of DupUM(®) was studied and further used in the biotransformation of castor oil into γ-decalactone. The highest aroma concentration was obtained with immobilized cells in DupUM(®), where reconsumption of the aroma by the cells was prevented, contrarily to what happens with free cells. This is a very promising result for γ-decalactone production, with potential to be used at an industrial level since the use of immobilized cells system will facilitate the conversion of a batch process into a continuous mode keeping high cell density and allowing easier recovery of metabolic products.

  19. Immobilization of glucosyltransferase from Erwinia sp. using two different techniques.

    PubMed

    Contesini, Fabiano Jares; Ibarguren, Carolina; Grosso, Carlos Raimundo Ferreira; Carvalho, Patrícia de Oliveira; Sato, Hélia Harumi

    2012-04-15

    Two different techniques of glucosyltransferase immobilization were studied for the conversion of sucrose into isomaltulose. The optimum conditions for immobilization of Erwinia sp. glucosyltransferase onto Celite 545, determined using response surface methodology, was pH 4.0 and 170 U of glucosyltransferase/g of Celite 545. Using this conditions more than 60% conversion of sucrose into isomaltulose can be obtained. The immobilization of glucosyltransferase was also studied by its entrapment in microcapsules of low-methoxyl pectin and fat (butter and oleic acid). The non-lyophilized microcapsules of pectin, containing the enzyme and fat, showed higher glucosyltransferase activity, compared with lyophilized microcapsules containing enzyme plus fat, and also lyophilized microcapsules containing enzyme without fat addition. The non-lyophilized microcapsules of pectin containing the glucosyltransferase and fat, converted 30% of sucrose into isomaltulose in the first batch. However the conversion decreased to 5% at the 10th batch, indicating inactivation of the enzyme.

  20. Immobilization of iodine in concrete

    DOEpatents

    Clark, Walter E.; Thompson, Clarence T.

    1977-04-12

    A method for immobilizing fission product radioactive iodine recovered from irradiated nuclear fuel comprises combining material comprising water, Portland cement and about 3-20 wt. % iodine as Ba(IO.sub.3).sub.2 to provide a fluid mixture and allowing the fluid mixture to harden, said Ba(IO.sub.3).sub.2 comprising said radioactive iodine. An article for solid waste disposal comprises concrete prepared by this method. BACKGROUND OF THE INVENTION This invention was made in the course of, or under a contract with the Energy Research and Development Administration. It relates in general to reactor waste solidification and more specifically to the immobilization of fission product radioactive iodine recovered from irradiated nuclear fuel for underground storage.

  1. Plutonium Immobilization Project -- Can loading

    SciTech Connect

    Kriikku, E.

    2000-01-18

    The Savannah River Site (SRS) will immobilize excess plutonium in the proposed Plutonium Immobilization Project (PIP). The PIP scope includes unloading transportation containers, preparing the feed streams, converting the metal feed to an oxide, adding the ceramic precursors, pressing the pucks, inspecting pucks, and sintering pucks. The PIP scope also includes loading the pucks into metal cans, sealing the cans, inspecting the cans, loading the cans into magazines, loading magazines into Defense Waste Processing Facility (DWPF) canisters, and transporting the canisters to the DWPF. The DWPF fills the canister with a mixture of high level radioactive waste and glass for permanent storage. Due to the radiation, remote equipment must perform PIP operations in a contained environment.

  2. [Immobilization of heavy metal Pb2+ with geopolymer].

    PubMed

    Jin, Man-tong; Jin, Zan-fang; Huang, Cai-ju

    2011-05-01

    A series of geopolymers were synthesized by mixing metakaolinite, water glass, sodium hydroxide and water, and the lead ion solidification experiments were performed with the geopolymer. Then, the immobilization efficiency was characterized by monitoring the leaching concentration and compressive strength of solidified products. Additionally, the structure and properties of the solidified products were studied by X-ray diffraction (XRD), scan electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy. Furthermore, based on the analysis of immobilization efficiency, microstructure and mineral structure, the difference between geopolymer and cement on the performance of immobilizing heavy metals was discussed. The results of lead ion immobilization experiments showed that over 99.7% of heavy metal was captured by the geopolymer as the doping concentration of lead ion was less than 3%. Meanwhile, the compressive strength of the solidified product ranged from 40 MPa to 50 MPa. Furthermore, by using the same Pb2+ concentration, the geopolymer showed higher compressive strength and lower leaching concentration compared to the cement. Because lead ion participated in constitution of structure of geopolymer, or Pb2+ was adsorbed by the aluminium ions on the geopolymeric skeleton and held in geopolymer. However, cement mainly solidified lead ion by physical encapsulation and adsorption mechanism. Therefore, both from the compressive strength and leaching concentration and from the microstructure characterization as well as the mechanism of the geopolymerization reaction, the geopolymer has more advantages in immobilizing Pb2+ than the cement.

  3. In –Situ Spectroscopic Investigation of Immobilized Organometallic Catalysts

    SciTech Connect

    Davis, Robert, J.

    2007-11-14

    Immobilized organometallic catalysts, in principle, can give high rates and selectivities like homogeneous catalysts with the ease of separation enjoyed by heterogeneous catalysts. However, the science of immobilized organometallics has not been developed because the field lies at the interface between the homogeneous and heterogeneous catalysis communities. By assembling an interdisciplinary research team that can probe all aspects of immobilized organometallic catalyst design, the entire reacting system can be considered, where the transition metal complex, the complex-support interface and the properties of the support can all be considered simultaneously from both experimental and theoretical points of view. Researchers at Georgia Tech and the University of Virginia are studying the fundamental principles that can be used to understand and design future classes of immobilized organometallic catalysts. In the framework of the overall collaborative project with Georgia Tech, our work focused on (a) the X-ray absorption spectroscopy of an immobilized Pd-SCS-O complex (b) the mode of metal leaching from supported Pd catalysts during Heck catalysis and (c) the mode of deactivation of Jacobsen’s Co-salen catalysts during the hydrolytic kinetic resolution of terminal epoxides. Catalysts containing supported Pd pincer complexes, functionalized supports containing mercapto and amine groups, and oligomeric Co-salen catalysts were synthesized at Georgia Tech and sent to the University of Virginia. Incorporation of Pd onto several different kinds of supports (silica, mercapto-functionalized silica, zeolite Y) was performed at the University of Virginia.

  4. Immobilization as a route to surplus fissile materials disposition

    SciTech Connect

    Gray, L.W.; Kan, T.

    1995-04-27

    In the aftermath of the Cold War, the US and Russia have agreed to large reductions in nuclear weapons. To aid in the selection of long-term management options, DOE has undertaken a multifaceted study to select options for storage and disposition of plutonium (Pu) in keeping with the national policy that Pu must be subjected to the highest standards of safety, security, and accountability. One alternative being considered is immobilization. To arrive at a suitable immobilization form, the authors first reviewed published information on high-level waste (HLW) immobilization technologies in order to identify 72 possible Pu immobilization forms to be prescreened. Surviving forms were screened using multiattribute analysis to determine the most promising technologies. Promising immobilization families were further evaluated to identify chemical, engineering, environmental, safety, and health problems that remain to be solved prior to making technical decisions as to the viability of using the form for long-term disposition of plutonium. All data, analyses, and reports are being provided to the DOE Fissile Materials Disposition Project Office to support the Record of Decision that is anticipated in the fourth quarter of FY96.

  5. [Novel Immobilized Biocatalyst for Microbiological Synthesis of Pharmaceutical Steroids].

    PubMed

    Andryushina, V A; Karpova, N V; Druzhinina, A V; Stytsenko, T S; Podorozhko, E A; Ryabev, A N; Lozinskii, V I

    2015-01-01

    The steroid-transforming activity of free and immobilized cells of Pimelobacter simplex VKPM As-1632 entrapped in an operationally stable macroporous polyvinyl alcohol cryogel was studied. It was shown that the macroporous matrix of the carrier did not create any diffusional limitations for steroid access to the cells or the removal of the transformation products from them. The optimal conditions for the hydrocortisone 1,2-dehydration into prednisolone by free and immobilized cells were elucidated. The immobilized biocatalyst was obtained in a granulated form and used in 32 successive cycles of steroid transformation. The average cycle duration was 45 min, and the prednisolone yield of during the first 20 cycles was 98%. It was established that the immobilized cells of the actinobacteria P. simplex retained high steroid-transforming activity over all of the transformation cycles. The physicochemical and diffusion characteristics of the polyvinyl alcohol gels and its granules were determined, and their high stability during repeated cycles of steroid transformation was shown. The results indicated that P. simplex immobilized cells represent an effective catalyst suitable for multiple use. Biomass consumption decreased upon its use, and product isolation, as well as culture storage, was much easier. PMID:26596083

  6. Functionalized Polyacrylonitrile Nanofibrous Membranes for Covalent Immobilization of Glucose Oxidase.

    PubMed

    Manuel, James; Kim, Miso; Dharela, Rohini; Chauhan, Ghanshyam S; Fapyane, Deby; Lee, Soo-Jin; Chang, In Seop; Kang, Seo-Hee; Kim, Seon-Won; Ahn, Jou-Hyeon

    2015-01-01

    Nanofibrous membrane (NFM) with uniform morphology and large surface area was prepared from 10% solution of polyacrylonitrile (PAN) in N,N-dimethylformamide by electrospinning technique. NFM was chemically modified for use as a support for the immobilization of glucose oxidase. Chemical modification of NFM was carried out by two different methods. In the first method, the cyano groups of PAN were modified to amino groups by a two-step process, while in the second method the carboxylic groups were generated first and then further reacted with hexamethylene diamine to create a reactive spacer arm for the immobilization of enzyme. Scanning electron microscopy studies showed that the surface morphology of NFM was not changed by chemical modification and its mechanical strength was improved. The immobilized glucose oxidase (GOx) retained 54 and 60% of its original activity up to 25 cycles with the PAN NFMs modified by the first and the second method, respectively. The GOx-immobilized NFM from the second method showed promising performance with higher enzyme immobilization, activity retention, and favorable kinetic parameters. PMID:26301308

  7. Spine Immobilizer for Accident Victims

    NASA Technical Reports Server (NTRS)

    Vykukal, H. C.; Lampson, K.

    1983-01-01

    Proposed conformal bladder filled with tiny spheres called "microballoons," enables spine of accident victim to be rapidly immobilized and restrained and permit victim to be safely removed from accident scene in extremely short time after help arrives. Microballoons expand to form rigid mass when pressure within bladder is less than ambient. Bladder strapped to victim is also strapped to rescue chair. Void between bladder and chair is filled with cloth wedges.

  8. Industrial applications of immobilized cells

    SciTech Connect

    Linko, P.; Linko, Y.Y.

    1984-01-01

    Although the application of the natural attraction of many microorganisms to surfaces has been applied in vinegar production since the early 1980s, and has long been utilized in waste water purification, the development of microbial cell immobilization techniques for special applications dates back only to the early 1960s. The immobilization may involve whole cells, cell fragments, or lysed cells. Whole cells may retain their metabolic activity with their complex multienzyme systems and cofactor regeneration mechanisms intact, or they may be killed in the process with only a few desired enzymes remaining active in the final biocatalyst. Cells may also be coimmobilized with an enzyme to carry out special reactions. Although relatively few industrial scale applications exist today, some are of very large scale. Current applications vary from relatively small scale steroid conversions to amino acid production and high fructose syrup manufacture. A vast number of potential applications are already known, and one of the most interesting applications may be in continuous fermentation such as ethanol production by immobilized living microorganisms. 373 references.

  9. Photo induced surface heparin immobilization.

    PubMed

    Nakayama, Y; Matsuda, T

    1993-01-01

    This paper describes a novel method providing durable layering of heparin immobilized hydrogels on fabricated devices. The preparation method is based on photochemistry of a dithiocarbamate group that is dissociated into a highly reactive radical pair upon ultraviolet (UV) irradiation. By taking advantage of characteristics of the photo generated radicals, hydrogel formation and its fixation onto a substrate surface were attained. The immobilization of heparin onto poly(ethylene terephtalate) was demonstrated. First, a mixed aqueous solution containing a photoreactive water soluble poly(N,N-dimethylacrylamide-covinylbenzyl N,N-diethyldithiocarbamate) and heparin was coated on the substrate. Subsequent UV irradiation resulted in the simultaneous formation of a heparin immobilized hydrogel and its chemical fixation onto the substrate. No delamination was found after vigorous washing with water. Significant inhibition of platelet adhesion and markedly prolonged blood coagulation times were observed, which are apparently derived from the surface hydrogel, and from released and chemically fixed surface heparin. Thus, it is expected that the photochemical method developed here provides potent antithrombogenicity to artificial organs. PMID:8268639

  10. Microarray immobilization of biomolecules using a fast trans-cyclooctene (TCO)-tetrazine reaction.

    PubMed

    Wang, Ping; Na, Zhenkun; Fu, Jiaqi; Tan, Chelsea Y J; Zhang, Huatang; Yao, Shao Q; Sun, Hongyan

    2014-10-14

    By anchoring 1,2,4,5-tetrazine-containing biomolecules onto trans-cyclooctene (TCO)-functionalized slides, a site-specific microarray immobilization approach is described in this study. Compared with existing immobilization methods, our approach offers several distinctive features, including fast kinetics and high chemoselectivity.

  11. Venous Thrombosis Risk after Cast Immobilization of the Lower Extremity: Derivation and Validation of a Clinical Prediction Score, L-TRiP(cast), in Three Population-Based Case–Control Studies

    PubMed Central

    Nemeth, Banne; van Adrichem, Raymond A.; van Hylckama Vlieg, Astrid; Bucciarelli, Paolo; Martinelli, Ida; Baglin, Trevor; Rosendaal, Frits R.; le Cessie, Saskia; Cannegieter, Suzanne C.

    2015-01-01

    Background Guidelines and clinical practice vary considerably with respect to thrombosis prophylaxis during plaster cast immobilization of the lower extremity. Identifying patients at high risk for the development of venous thromboembolism (VTE) would provide a basis for considering individual thromboprophylaxis use and planning treatment studies. The aims of this study were (1) to investigate the predictive value of genetic and environmental risk factors, levels of coagulation factors, and other biomarkers for the occurrence of VTE after cast immobilization of the lower extremity and (2) to develop a clinical prediction tool for the prediction of VTE in plaster cast patients. Methods and Findings We used data from a large population-based case–control study (MEGA study, 4,446 cases with VTE, 6,118 controls without) designed to identify risk factors for a first VTE. Cases were recruited from six anticoagulation clinics in the Netherlands between 1999 and 2004; controls were their partners or individuals identified via random digit dialing. Identification of predictor variables to be included in the model was based on reported associations in the literature or on a relative risk (odds ratio) > 1.2 and p ≤ 0.25 in the univariate analysis of all participants. Using multivariate logistic regression, a full prediction model was created. In addition to the full model (all variables), a restricted model (minimum number of predictors with a maximum predictive value) and a clinical model (environmental risk factors only, no blood draw or assays required) were created. To determine the discriminatory power in patients with cast immobilization (n = 230), the area under the curve (AUC) was calculated by means of a receiver operating characteristic. Validation was performed in two other case–control studies of the etiology of VTE: (1) the THE-VTE study, a two-center, population-based case–control study (conducted in Leiden, the Netherlands, and Cambridge, United Kingdom

  12. Use of immobilized enzymes in automated clinical analysis: determination of uric acid and glucose using immobilized enzymes in column form.

    PubMed

    Endo, J; Tabata, M; Okada, S; Murachi, T

    1979-07-16

    We studied the use of immobilized enzymes, covalently bound to alkylaminosilane derivative of porous glass, to automated clinical analysis on uric acid and glucose in blood, serum and urine. A microcolumn with an immobilized enzyme was prepared and used in an AutoAnalyzer I continuous flow system. Uricase (EC 1.7.3.3) from Candida utilis and glucose oxidase (EC 1.1.3.4) from Aspergillus niger were immobilized for the determination of uric acid and glucose, respectively. Hydrogen peroxide produced by these oxidases was colorimetrically determined using horse-radish peroxidase (EC 1.11.1.7) and a hydrogen acceptor in solution. Sensitivity and wash charactertistics of a column with immobilized enzyme, 1.5 mm of inner diameter and up to 40 mm in length, were satisfactory at an assay speed of 50 samples per hour. The results correlated well with those obtained by other well established methods utilizing the AutoAnalyzer system. The immobilized enzymes were sufficiently stable for at least two months of 2000 tests when used repeatedly. Clinical trials proved that this method is capable of replacing the soluble enzyme method, giving reliable and reproducible results at lower cost.

  13. Immobilized enzymes in blood plasma exchangers via radiation grafting

    NASA Astrophysics Data System (ADS)

    Gombotz, Wayne; Hoffman, Allan; Schmer, Gottfried; Uenoyama, Satoshi

    The enzyme asparaginase was immobilized onto a porous hollow polypropylene (PP) fiber blood plasma exchange device for the treatment of acute lymphocytic leukemia. The devices were first radiation grafted with polymethacrylic acid (poly(MAAc)). This introduces carboxyl groups onto the surface of the fibers. Several variables were studied in the grafting reaction including the effects of solvent type and monomer concentration. The carboxyl groups were activated with N-hydroxy succinimide (NHS) using carbodiimide chemistry. Asparaginase was then covalently immobilized on the activated surfaces. Quantitative relationships were found relating the percent graft to the amount of immobilized enzyme which was active. The enzyme reactor was tested both in vitro and in vivo using a sheep as an animal model.

  14. Purification, immobilization, and characterization of nattokinase on PHB nanoparticles.

    PubMed

    Deepak, Venkataraman; Pandian, Suresh babu Ram Kumar; Kalishwaralal, Kalimuthu; Gurunathan, Sangiliyandi

    2009-12-01

    In this study, nattokinase was purified from Bacillus subtilis using ion exchange chromatography and immobilized upon polyhydroxybutyrate (PHB) nanoparticles. A novel strain isolated from industrial dairy waste was found to synthesize polyhydroxyalkanoates (PHA) and the strain was identified as Brevibacterium casei SRKP2. PHA granules were extracted from 48 h culture and the FT-IR analysis characterized them as PHB, a natural biopolymer from B. casei. Nanoprecipitation by solvent displacement technique was used to synthesize PHB nanoparticles. PHB nanoparticles were characterized using transmission electron microscopy and particle size ranged from 100-125 nm. Immobilization of nattokinase upon PHB nanoparticles resulted in a 20% increase in the enzyme activity. Immobilization also contributed to the enhanced stability of the enzyme. Moreover, the activity was completely retained on storage at 4 degrees C for 25 days. The method has proven to be highly simple and can be implemented to other enzymes also.

  15. Production of biodiesel using immobilized lipase--a critical review.

    PubMed

    Jegannathan, Kenthorai Raman; Abang, Sariah; Poncelet, Denis; Chan, Eng Seng; Ravindra, Pogaku

    2008-01-01

    Increase in volume of biodiesel production in the world scenario proves that biodiesel is accepted as an alternative to conventional fuel. Production of biodiesel using alkaline catalyst has been commercially implemented due to its high conversion and low production time. For the product and process development of biodiesel, enzymatic transesterification has been suggested to produce a high purity product with an economic, environment friendly process at mild reaction conditions. The enzyme cost being the main hurdle can be overcome by immobilization. Immobilized enzyme, which has been successfully used in various fields over the soluble counterpart, could be employed in biodiesel production with the aim of reducing the production cost by reusing the enzyme. This review attempts to provide an updated compilation of the studies reported on biodiesel production by using lipase immobilized through various techniques and the parameters, which affect their functionality.

  16. Straightforward protein immobilization on Sylgard 184 PDMS microarray surface.

    PubMed

    Heyries, Kevin A; Marquette, Christophe A; Blum, Loïc J

    2007-04-10

    In this work, a straightforward technique for protein immobilization on Sylgard 184 is described. The method consists of a direct transfer of dried protein/salt solutions to the PDMS interface during the polymer curing. Such non-conventional treatment of proteins was found to have no major negative consequence on their integrity. The mechanisms of this direct immobilization were investigated using a lysine modified dextran molecule as a model. Clear experimental results suggested that both chemical bounding and molding effect were implicated. As a proof of concept study, three different proteins were immobilized on a single microarray (Arachis hypogaea lectin, rabbit IgG, and human IgG) and used as antigens for capture of chemiluminescent immunoassays. The proteins were shown to be easily recognized by their specific antibodies, giving antibody detection limits in the fmol range.

  17. Utilization of immobilized urease for waste water treatment

    NASA Technical Reports Server (NTRS)

    Husted, R. R.

    1974-01-01

    The feasibility of using immobilized urease for urea removal from waste water for space system applications is considered, specifically the elimination of the urea toxicity problem in a 30-day Orbiting Frog Otolith (OFO) flight experiment. Because urease catalyzes the hydrolysis of urea to ammonia and carbon dioxide, control of their concentrations within nontoxic limits was also determined. The results of this study led to the use of free urease in lieu of the immobilized urease for controlling urea concentrations. An ion exchange resin was used which reduced the NH3 level by 94% while reducing the sodium ion concentration only 10%.

  18. Analysis of alternatives for immobilized low activity waste disposal

    SciTech Connect

    Burbank, D.A.

    1997-10-28

    This report presents a study of alternative disposal system architectures and implementation strategies to provide onsite near-surface disposal capacity to receive the immobilized low-activity waste produced by the private vendors. The analysis shows that a flexible unit strategy that provides a suite of design solutions tailored to the characteristics of the immobilized low-activity waste will provide a disposal system that best meets the program goals of reducing the environmental, health, and safety impacts; meeting the schedule milestones; and minimizing the life-cycle cost of the program.

  19. Maltodextrin hydrolysis in a fluidized-bed immobilized enzyme reactor

    SciTech Connect

    Vallat, I.; Monsan, P.; Riba, J.P.

    1986-02-01

    The present work deals with maltodextrin hydrolysis by glucoamylase immobilized onto corn stover in a fluidized bed reactor. An industrial enzyme preparation was convalently grafted onto corn stover, yielding an activity of up to 372 U/g and 1700 U/g for support particle sizes of 0.8 and 0.2 mm, respectively. A detailed kinetic study, using a differntial reactor, allowed the characterization of the influence of mass transfer resistance on the reaction catalyzed by immobilized glucoamylase. A simple and general mathematical model was then developed to describe the experimental conversion data and found to be vaild.

  20. Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres

    PubMed Central

    Ibrahim, Abdelnasser Salah Shebl; Al-Salamah, Ali A.; El-Toni, Ahmed M.; Almaary, Khalid S.; El-Tayeb, Mohamed A.; Elbadawi, Yahya B.; Antranikian, Garabed

    2016-01-01

    The stability and reusability of soluble enzymes are of major concerns, which limit their industrial applications. Herein, alkaline protease from Bacillus sp. NPST-AK15 was immobilized onto hollow core-mesoporous shell silica (HCMSS) nanospheres. Subsequently, the properties of immobilized proteases were evaluated. Non-, ethane- and amino-functionalized HCMSS nanospheres were synthesized and characterized. NPST-AK15 was immobilized onto the synthesized nano-supports by physical and covalent immobilization approaches. However, protease immobilization by covalent attachment onto the activated HCMSS–NH2 nanospheres showed highest immobilization yield (75.6%) and loading capacity (88.1 μg protein/mg carrier) and was applied in the further studies. In comparison to free enzyme, the covalently immobilized protease exhibited a slight shift in the optimal pH from 10.5 to 11.0, respectively. The optimum temperature for catalytic activity of both free and immobilized enzyme was seen at 60 °C. However, while the free enzyme was completely inactivated when treated at 60 °C for 1 h the immobilized enzyme still retained 63.6% of its initial activity. The immobilized protease showed higher Vmax, kcat and kcat/Km, than soluble enzyme by 1.6-, 1.6- and 2.4-fold, respectively. In addition, the immobilized protease affinity to the substrate increased by about 1.5-fold. Furthermore, the enzyme stability in various organic solvents was significantly enhanced upon immobilization. Interestingly, the immobilized enzyme exhibited much higher stability in several commercial detergents including OMO, Tide, Ariel, Bonux and Xra by up to 5.2-fold. Finally, the immobilized protease maintained significant catalytic efficiency for twelve consecutive reaction cycles. These results suggest the effectiveness of the developed nanobiocatalyst as a candidate for detergent formulation and peptide synthesis in non-aqueous media. PMID:26840303

  1. Enhancement of Alkaline Protease Activity and Stability via Covalent Immobilization onto Hollow Core-Mesoporous Shell Silica Nanospheres.

    PubMed

    Ibrahim, Abdelnasser Salah Shebl; Al-Salamah, Ali A; El-Toni, Ahmed M; Almaary, Khalid S; El-Tayeb, Mohamed A; Elbadawi, Yahya B; Antranikian, Garabed

    2016-01-29

    The stability and reusability of soluble enzymes are of major concerns, which limit their industrial applications. Herein, alkaline protease from Bacillus sp. NPST-AK15 was immobilized onto hollow core-mesoporous shell silica (HCMSS) nanospheres. Subsequently, the properties of immobilized proteases were evaluated. Non-, ethane- and amino-functionalized HCMSS nanospheres were synthesized and characterized. NPST-AK15 was immobilized onto the synthesized nano-supports by physical and covalent immobilization approaches. However, protease immobilization by covalent attachment onto the activated HCMSS-NH₂ nanospheres showed highest immobilization yield (75.6%) and loading capacity (88.1 μg protein/mg carrier) and was applied in the further studies. In comparison to free enzyme, the covalently immobilized protease exhibited a slight shift in the optimal pH from 10.5 to 11.0, respectively. The optimum temperature for catalytic activity of both free and immobilized enzyme was seen at 60 °C. However, while the free enzyme was completely inactivated when treated at 60 °C for 1 h the immobilized enzyme still retained 63.6% of its initial activity. The immobilized protease showed higher V(max), k(cat) and k(cat)/K(m), than soluble enzyme by 1.6-, 1.6- and 2.4-fold, respectively. In addition, the immobilized protease affinity to the substrate increased by about 1.5-fold. Furthermore, the enzyme stability in various organic solvents was significantly enhanced upon immobilization. Interestingly, the immobilized enzyme exhibited much higher stability in several commercial detergents including OMO, Tide, Ariel, Bonux and Xra by up to 5.2-fold. Finally, the immobilized protease maintained significant catalytic efficiency for twelve consecutive reaction cycles. These results suggest the effectiveness of the developed nanobiocatalyst as a candidate for detergent formulation and peptide synthesis in non-aqueous media.

  2. Drop drying on surfaces determines chemical reactivity - the specific case of immobilization of oligonucleotides on microarrays

    PubMed Central

    2013-01-01

    Background Drop drying is a key factor in a wide range of technical applications, including spotted microarrays. The applied nL liquid volume provides specific reaction conditions for the immobilization of probe molecules to a chemically modified surface. Results We investigated the influence of nL and μL liquid drop volumes on the process of probe immobilization and compare the results obtained to the situation in liquid solution. In our data, we observe a strong relationship between drop drying effects on immobilization and surface chemistry. In this work, we present results on the immobilization of dye labeled 20mer oligonucleotides with and without an activating 5′-aminoheptyl linker onto a 2D epoxysilane and a 3D NHS activated hydrogel surface. Conclusions Our experiments identified two basic processes determining immobilization. First, the rate of drop drying that depends on the drop volume and the ambient relative humidity. Oligonucleotides in a dried spot react unspecifically with the surface and long reaction times are needed. 3D hydrogel surfaces allow for immobilization in a liquid environment under diffusive conditions. Here, oligonucleotide immobilization is much faster and a specific reaction with the reactive linker group is observed. Second, the effect of increasing probe concentration as a result of drop drying. On a 3D hydrogel, the increasing concentration of probe molecules in nL spotting volumes accelerates immobilization dramatically. In case of μL volumes, immobilization depends on whether the drop is allowed to dry completely. At non-drying conditions, very limited immobilization is observed due to the low oligonucleotide concentration used in microarray spotting solutions. The results of our study provide a general guideline for microarray assay development. They allow for the initial definition and further optimization of reaction conditions for the immobilization of oligonucleotides and other probe molecule classes to different

  3. Immobilization of the urease on eggshell membrane and its application in biosensor.

    PubMed

    D'Souza, S F; Kumar, Jitendra; Jha, Sandeep Kumar; Kubal, B S

    2013-03-01

    Eggshell membrane is a natural material, essentially made up of protein fibers having flexibility in the aqueous solution and possessing gas and water permeability. It is used as a biomembrane for immobilization of urease for the development of a potentiometric urea biosensor. Eggshell membrane was treated with polyethyleneimine (PEI) to impart polycation characteristics. Urease was immobilized on the PEI treated eggshell membrane through adsorption. SEM study was carried out to observe the changes in surface morphology after immobilization. FTIR study of membrane was carried out to observe the changes in IR spectra after immobilization of enzyme. Immobilized membrane was associated with ammonium ion selective electrode. Biosensor exhibited sigmoidal responses for the urea concentration range from 0.5 to 10mM. The response time of the biosensor was 120 s. A single membrane was reused for 270 reactions without loss of activity. The urease-eggshell membranes were stable for 2 months when stored in buffer even at room temperature.

  4. pH-dependent immobilization of urease on glutathione-capped gold nanoparticles.

    PubMed

    Garg, Seema; De, Arnab; Mozumdar, Subho

    2015-05-01

    Urease is a nickel-dependent metalloenzyme that catalyzes the hydrolysis of urea to form ammonia and carbon dioxide. Although the enzyme serves a significant role in several detoxification and analytical processes, its usability is restricted due to high cost, availability in small amounts, instability, and a limited possibility of economic recovery from a reaction mixture. Hence, there is a need to develop an efficient, simple, and reliable immobilization strategy for the enzyme. In this study, the carboxyl terminated surface of glutathione-capped gold nanoparticles have been utilized as a solid support for the covalent attachment of urease. The immobilization has been carried out at different pH conditions so as to elucidate its effect on the immobilization efficiency and enzyme bioactivity. The binding of the enzyme has been quantitatively and qualitatively analyzed through techniques like ultraviolet-visible spectroscopy, intrinsic steady state fluorescence, and circular dichorism. The bioactivity of the immobilized enzyme was investigated with respect to the native enzyme under different thermal conditions. Recyclability and shelf life studies of the immobilized enzyme have also been carried out. Results reveal that the immobilization is most effective at pH of 7.4 followed by that in an acidic medium and is least in alkaline environment. The immobilized enzyme also exhibits enhance activity in comparison to the native form at physiological temperature. The immobilized urease (on gold glutathione nanoconjugates surface) can be effectively employed for biosensor fabrication, immunoassays and as an in vivo diagnostic tool in the future.

  5. Immobilization and controlled release of β-galactosidase from chitosan-grafted hydrogels.

    PubMed

    Facin, Bruno R; Moret, Bruna; Baretta, Dilmar; Belfiore, Laurence A; Paulino, Alexandre T

    2015-07-15

    Chitosan-grafted hydrogels were employed for immobilization and controlled released of β-galactosidase. These hydrogels containing immobilized enzymes were employed to simulate the production of lactose-free food and controlled release of β-galactosidase into lactose-intolerant individuals. The degree of swelling, efficiency of immobilization (i.e., fractional uptake of enzyme), and controlled release were studied as a function of pH and temperature. The degrees of swelling decreased in acidic media: 49.4 g absorbed water per g hydrogel at pH 7.0, and 8.4 g absorbed water per g hydrogel at pH 3.5. The immobilization efficiency was 19%, indicating that chitosan-grafted hydrogels are promising matrices for enzyme adsorption and immobilization. Cyclic experiments reveal that chitosan-grafted hydrogels containing immobilized enzymes can be reused several times without introducing additional enzyme prior to each cycle. There is no significant decrease in the activity of the immobilized enzyme during reutilization studies. All results were conducted in triplicate by considering t-tests at a 95% significance level. Analysis of β-galactosidase activity and controlled release reveals that chitosan-grafted hydrogels containing immobilized enzymes are useful for the production of lactose-free food and controlled enzyme release with high performance. PMID:25722137

  6. Immobilization and controlled release of β-galactosidase from chitosan-grafted hydrogels.

    PubMed

    Facin, Bruno R; Moret, Bruna; Baretta, Dilmar; Belfiore, Laurence A; Paulino, Alexandre T

    2015-07-15

    Chitosan-grafted hydrogels were employed for immobilization and controlled released of β-galactosidase. These hydrogels containing immobilized enzymes were employed to simulate the production of lactose-free food and controlled release of β-galactosidase into lactose-intolerant individuals. The degree of swelling, efficiency of immobilization (i.e., fractional uptake of enzyme), and controlled release were studied as a function of pH and temperature. The degrees of swelling decreased in acidic media: 49.4 g absorbed water per g hydrogel at pH 7.0, and 8.4 g absorbed water per g hydrogel at pH 3.5. The immobilization efficiency was 19%, indicating that chitosan-grafted hydrogels are promising matrices for enzyme adsorption and immobilization. Cyclic experiments reveal that chitosan-grafted hydrogels containing immobilized enzymes can be reused several times without introducing additional enzyme prior to each cycle. There is no significant decrease in the activity of the immobilized enzyme during reutilization studies. All results were conducted in triplicate by considering t-tests at a 95% significance level. Analysis of β-galactosidase activity and controlled release reveals that chitosan-grafted hydrogels containing immobilized enzymes are useful for the production of lactose-free food and controlled enzyme release with high performance.

  7. Immobilization of α-amylase onto a calix[4]arene derivative: Evaluation of its enzymatic activity.

    PubMed

    Veesar, Irshad Ali; Solangi, Imam Bakhsh; Memon, Shahabuddin

    2015-06-01

    In order to enhance the cost-effectiveness practicability of enzymes in many industries such as pharmaceutical, food, medical and some other technological processes, there is great need to immobilize them onto a solid supports. In this study, a new and efficient immobilization of α-amylase from Saccharomyces cerevisiae has been developed by using the surface functionalization of calix[4]arene as support. A glutaraldehyde-containing amino group functionalized calix[4]arene was used to immobilize α-amylase covalently. In this procedure, imide bonds are formed between amino groups on the protein and aldehyde groups on the calix[4]arene surface. The surface modified support was characterized using Fourier transform infrared spectroscopy (FT-IR), scanning electron microscopy (SEM). The effect of various preparation conditions on the immobilized α-amylase process such as immobilization time, enzyme concentration, temperature and pH were investigated. The influence of pH and temperature on the activity of free and immobilized α-amylase was also studied using starch as substrate. The optimum reaction temperature and pH value for the enzymatic conversion catalyzed by the immobilized α-amylase were 25°C and 7, respectively. Compared to the free enzyme, the immobilized α-amylase retained 85% of its original activity and exhibited significant thermal stability than the free one and excellent durability.

  8. Optimization, isotherm, kinetic and thermodynamic studies of Pb(II) ions adsorption onto N-maleated chitosan-immobilized TiO₂ nanoparticles from aqueous media.

    PubMed

    Shaker, Medhat A; Yakout, Amr A

    2016-02-01

    Chitosan, CS was chemically engineered by maleic anhydride via simple protocol to produce N-maleated chitosan, MCS which immobilized on anatase TiO2 to synthesize novel eco-friendly nanosorbent (51±3.8 nm), MCS@TiO2 for cost-effective and efficient removal of Pb(II) ions from aqueous media. The chemical structure, surface properties and morphology of MCS@TiO2 were recognized by FTIR, (1)H NMR, XRD, TEM, DLS and zeta-potential techniques. The relations between %removal of Pb(II) and different analytical parameters such as solution acidity (pH), MCS@TiO2 dosage, time of contact and initial Pb(II) concentration were optimized using response surface methodology (RSM) and Box-Behnken design (BBD) statistical procedures. The fitting of the experimental data to four different isotherm models at optimized conditions was carried out by various statistical treatments including the correlation coefficient (r), coefficient of determination (r(2)) and non-linear Chi-square (χ(2)) test analyses which all confirm the suitability of Langmuir model to explain the adsorption isotherm data. Also, statistics predicted that the pseudo-second-order model is the optimum kinetic model among four applied kinetic models to closely describe the rate equation of the adsorption process. Thermodynamics viewed the adsorption as endothermic and feasible physical process. EDTA could release the sorbed Pb(II) ions from MCS@TiO2 with a recovery above 92% after three sorption-desorption cycles. The novel synthesized nanosorbent is evidenced to be an excellent solid phase extractor for Pb(II) ions from wastewaters.

  9. Optimization, isotherm, kinetic and thermodynamic studies of Pb(II) ions adsorption onto N-maleated chitosan-immobilized TiO2 nanoparticles from aqueous media

    NASA Astrophysics Data System (ADS)

    Shaker, Medhat A.; Yakout, Amr A.

    2016-02-01

    Chitosan, CS was chemically engineered by maleic anhydride via simple protocol to produce N-maleated chitosan, MCS which immobilized on anatase TiO2 to synthesize novel eco-friendly nanosorbent (51 ± 3.8 nm), MCS@TiO2 for cost-effective and efficient removal of Pb(II) ions from aqueous media. The chemical structure, surface properties and morphology of MCS@TiO2 were recognized by FTIR, 1H NMR, XRD, TEM, DLS and zeta-potential techniques. The relations between %removal of Pb(II) and different analytical parameters such as solution acidity (pH), MCS@TiO2 dosage, time of contact and initial Pb(II) concentration were optimized using response surface methodology (RSM) and Box-Behnken design (BBD) statistical procedures. The fitting of the experimental data to four different isotherm models at optimized conditions was carried out by various statistical treatments including the correlation coefficient (r), coefficient of determination (r2) and non-linear Chi-square (χ2) test analyses which all confirm the suitability of Langmuir model to explain the adsorption isotherm data. Also, statistics predicted that the pseudo-second-order model is the optimum kinetic model among four applied kinetic models to closely describe the rate equation of the adsorption process. Thermodynamics viewed the adsorption as endothermic and feasible physical process. EDTA could release the sorbed Pb(II) ions from MCS@TiO2 with a recovery above 92% after three sorption-desorption cycles. The novel synthesized nanosorbent is evidenced to be an excellent solid phase extractor for Pb(II) ions from wastewaters.

  10. Optimization, isotherm, kinetic and thermodynamic studies of Pb(II) ions adsorption onto N-maleated chitosan-immobilized TiO₂ nanoparticles from aqueous media.

    PubMed

    Shaker, Medhat A; Yakout, Amr A

    2016-02-01

    Chitosan, CS was chemically engineered by maleic anhydride via simple protocol to produce N-maleated chitosan, MCS which immobilized on anatase TiO2 to synthesize novel eco-friendly nanosorbent (51±3.8 nm), MCS@TiO2 for cost-effective and efficient removal of Pb(II) ions from aqueous media. The chemical structure, surface properties and morphology of MCS@TiO2 were recognized by FTIR, (1)H NMR, XRD, TEM, DLS and zeta-potential techniques. The relations between %removal of Pb(II) and different analytical parameters such as solution acidity (pH), MCS@TiO2 dosage, time of contact and initial Pb(II) concentration were optimized using response surface methodology (RSM) and Box-Behnken design (BBD) statistical procedures. The fitting of the experimental data to four different isotherm models at optimized conditions was carried out by various statistical treatments including the correlation coefficient (r), coefficient of determination (r(2)) and non-linear Chi-square (χ(2)) test analyses which all confirm the suitability of Langmuir model to explain the adsorption isotherm data. Also, statistics predicted that the pseudo-second-order model is the optimum kinetic model among four applied kinetic models to closely describe the rate equation of the adsorption process. Thermodynamics viewed the adsorption as endothermic and feasible physical process. EDTA could release the sorbed Pb(II) ions from MCS@TiO2 with a recovery above 92% after three sorption-desorption cycles. The novel synthesized nanosorbent is evidenced to be an excellent solid phase extractor for Pb(II) ions from wastewaters. PMID:26520475

  11. Synthesis and heavy metal immobilization behaviors of slag based geopolymer.

    PubMed

    Yunsheng, Zhang; Wei, Sun; Qianli, Chen; Lin, Chen

    2007-05-01

    In this paper, two aspects of studies are carried out: (1) synthesis of geopolymer by using slag and metakaolin; (2) immobilization behaviors of slag based geopolymer in a presence of Pb and Cu ions. As for the synthesis of slag based geopolymer, four different slag content (10%, 30%, 50%, 70%) and three types of curing regimes (standard curing, steam curing and autoclave curing) are investigated to obtain the optimum synthesis condition based on the compressive and flexural strength. The testing results showed that geopolymer mortar containing 50% slag that is synthesized at steam curing (80 degrees C for 8h), exhibits higher mechanical strengths. The compressive and flexural strengths of slag based geopolymer mortar are 75.2 MPa and 10.1 MPa, respectively. Additionally, Infrared (IR), X-ray diffraction (XRD) and scanning electron microscopy (SEM) techniques are used to characterize the microstructure of the slag based geopolymer paste. IR spectra show that the absorptive band at 1086 cm(-1) shifts to lower wave number around 1007 cm(-1), and some six-coordinated Als transforms into four-coordination during the synthesis of slag based geopolymer paste. The resulting slag based geopolymeric products are X-ray amorphous materials. SEM observation shows that it is possible to have geopolymeric gel and calcium silicate hydrate (C-S-H) gel forming simultaneously within slag based geopolymer paste. As for immobilization of heavy metals, the leaching tests are employed to investigate the immobilization behaviors of the slag based geopolymer mortar synthesized under the above optimum condition. The leaching tests show that slag based geopolymer mortar can effectively immobilize Cu and Pb heavy metal ions, and the immobilization efficiency reach 98.5% greater when heavy metals are incorporated in the slag geopolymeric matrix in the range of 0.1-0.3%. The Pb exhibits better immobilization efficiency than the Cu in the case of large dosages of heavy metals.

  12. Immobilization of enzymes: a literature survey.

    PubMed

    Brena, Beatriz; González-Pombo, Paula; Batista-Viera, Francisco

    2013-01-01

    The term immobilized enzymes refers to "enzymes physically confined or localized in a certain defined region of space with retention of their catalytic activities, and which can be used repeatedly and continuously." Immobilized enzymes are currently the subject of considerable interest because of their advantages over soluble enzymes. In addition to their use in industrial processes, the immobilization techniques are the basis for making a number of biotechnology products with application in diagnostics, bioaffinity chromatography, and biosensors. At the beginning, only immobilized single enzymes were used, after 1970s more complex systems including two-enzyme reactions with cofactor regeneration and living cells were developed. The enzymes can be attached to the support by interactions ranging from reversible physical adsorption and ionic linkages to stable covalent bonds. Although the choice of the most appropriate immobilization technique depends on the nature of the enzyme and the carrier, in the last years the immobilization technology has increasingly become a matter of rational design. As a consequence of enzyme immobilization, some properties such as catalytic activity or thermal stability become altered. These effects have been demonstrated and exploited. The concept of stabilization has been an important driving force for immobilizing enzymes. Moreover, true stabilization at the molecular level has been demonstrated, e.g., proteins immobilized through multipoint covalent binding. PMID:23934795

  13. Immobilized lipid-bilayer materials

    DOEpatents

    Sasaki, Darryl Y.; Loy, Douglas A.; Yamanaka, Stacey A.

    2000-01-01

    A method for preparing encapsulated lipid-bilayer materials in a silica matrix comprising preparing a silica sol, mixing a lipid-bilayer material in the silica sol and allowing the mixture to gel to form the encapsulated lipid-bilayer material. The mild processing conditions allow quantitative entrapment of pre-formed lipid-bilayer materials without modification to the material's spectral characteristics. The method allows for the immobilization of lipid membranes to surfaces. The encapsulated lipid-bilayer materials perform as sensitive optical sensors for the detection of analytes such as heavy metal ions and can be used as drug delivery systems and as separation devices.

  14. Preparation of magnetite-fullerene nanocomposite with enzyme immobilization.

    PubMed

    Kalska-Szostko, B; Rogowska, M

    2012-09-01

    This study presents modification of magnetite nanoparticles and fullerene for biocompatibility. It show also specific fabrication of magnetite-carbon nanocomposite with immobilized biomolecule. The composites were created by joining individual components step-by-step manner (fullerene to magnetite and glucose oxidase or glucose oxidase to magnetite and fullerene). The resulting nanocomposites were characterized by infrared spectroscopy (IR) and transmission electron microscopy (TEM).

  15. Tunable control of antibody immobilization using electric field

    PubMed Central

    Emaminejad, Sam; Javanmard, Mehdi; Gupta, Chaitanya; Chang, Shuai; Davis, Ronald W.; Howe, Roger T.

    2015-01-01

    The controlled immobilization of proteins on solid-state surfaces can play an important role in enhancing the sensitivity of both affinity-based biosensors and probe-free sensing platforms. Typical methods of controlling the orientation of probe proteins on a sensor surface involve surface chemistry-based techniques. Here, we present a method of tunably controlling the immobilization of proteins on a solid-state surface using electric field. We study the ability to orient molecules by immobilizing IgG molecules in microchannels while applying lateral fields. We use atomic force microscopy to both qualitatively and quantitatively study the orientation of antibodies on glass surfaces. We apply this ability for controlled orientation to enhance the performance of affinity-based assays. As a proof of concept, we use fluorescence detection to indirectly verify the modulation of the orientation of proteins bound to the surface. We studied the interaction of fluorescently tagged anti-IgG with surface immobilized IgG controlled by electric field. Our study demonstrates that the use of electric field can result in more than 100% enhancement in signal-to-noise ratio compared with normal physical adsorption. PMID:25650429

  16. Natural impacted freshwaters: in situ use of alginate immobilized algae to the assessment of algal response.

    PubMed

    Corrêa, A X R; Tamanaha, M S; Horita, C O; Radetski, M R; Corrêa, R; Radetski, C M

    2009-05-01

    The objective of this study was to investigate the feasibility of an in situ phytotoxicity test using alginate-immobilized algae for 60 days, in the assessment of water quality in an impacted small peri-urban stream. After laboratory optimization of algae immobilization/de-immobilization processes, the performance of immobilized/de-immobilized algae was compared to the performance of free algae in terms of specific algal growth and sensitivity. This was done by comparing 72 h EC50 values obtained with zinc and the pesticides clomazone and carbofuran. The results showed a similar performance, which allow us to conclude that immobilization for 60 days do not cause any significant alteration in algae physiology. In the field, immobilized algae were exposed at different times (2, 4 and 7 days) to water samples in both disturbed and undisturbed sites. Both laboratory and field experiments indicated that alginate-immobilized algae for 60 days were sufficiently sensitive for use in the in situ assessment of water quality.

  17. Enzymatic decolorization of anthraquinone and diazo dyes using horseradish peroxidase enzyme immobilized onto various polysulfone supports.

    PubMed

    Celebi, Mithat; Kaya, Mehmet Arif; Altikatoglu, Melda; Yildirim, Huseyin

    2013-10-01

    In this study, covalent immobilization of the horseradish peroxidase (HRP) onto various polysulfone supports was investigated. For this purpose, different polysulfones were methacrylated with methacryloyl chloride, and then, nonwoven fabric samples were coated by using solutions of these methacrylated polysulfones. Finally, support materials were immersed into aquatic solution of HRP enzyme for covalent immobilization. Structural analysis of enzyme immobilization onto various polysulfones was confirmed with Fourier transform infrared spectroscopy, atomic force microscopy, and proton nuclear magnetic resonance spectroscopy. Decolorization of textile diazo (Acid Black 1) and anthraquinone (Reactive Blue 19) dyes was investigated by UV-visible spectrophotometer. Covalently immobilized enzyme has been used seven times in freshly prepared dye solutions through 63 days. Dye decolorization performance of the immobilized systems was observed that still remained high (70%) after reusing three times. Enzyme activities of immobilized systems were determined and compared to free enzyme activity at different conditions (pH, temperature, thermal stability, storage stability). Enzyme activities of immobilized systems and free enzyme were also investigated at the different temperatures and effects of temperature and thermal resistance for different incubation time at 50 °C. In addition, storage activity of free and immobilized enzymes was determined at 4 °C at different incubation days.

  18. Immobilization of Pseudomonas sp. DG17 onto sodium alginate–attapulgite–calcium carbonate

    PubMed Central

    Wang, Hong Qi; Hua, Fei; Zhao, Yi Cun; Li, Yi; Wang, Xuan

    2014-01-01

    A strain of Pseudomonas sp. DG17, capable of degrading crude oil, was immobilized in sodium alginate–attapulgite–calcium carbonate for biodegradation of crude oil contaminated soil. In this work, proportion of independent variables, the laboratory immobilization parameters, the micromorphology and internal structure of the immobilized granule, as well as the crude oil biodegradation by sodium alginate–attapulgite–calcium carbonate immobilized cells and sodium alginate–attapulgite immobilized cells were studied to build the optimal immobilization carrier and granule-forming method. The results showed that the optimal concentrations of sodium alginate–attapulgite–calcium carbonate and calcium chloride were 2.5%–3.5%, 0.5%–1%, 3%–7% and 2%–4%, respectively. Meanwhile, the optimal bath temperature, embedding cell amount, reaction time and multiplication time were 50–60 °C, 2%, 18 h and 48 h, respectively. Moreover, biodegradation was enhanced by immobilized cells with a total petroleum hydrocarbon removal ranging from 33.56% ± 3.84% to 56.82% ± 3.26% after 20 days. The SEM results indicated that adding calcium carbonate was helpful to form internal honeycomb-like pores in the immobilized granules. PMID:26019567

  19. Magnetic catechol-chitosan with bioinspired adhesive surface: preparation and immobilization of ω-transaminase.

    PubMed

    Ni, Kefeng; Zhou, Xu; Zhao, Li; Wang, Hualei; Ren, Yuhong; Wei, Dongzhi

    2012-01-01

    The magnetic chitosan nanocomposites have been studied intensively and been used practically in various biomedical and biological applications including enzyme immobilization. However, the loading capacity and the remained activity of immobilized enzyme based on existing approaches are not satisfied. Simpler and more effective immobilization strategies are needed. Here we report a simple catechol modified protocol for preparing a novel catechol-chitosan (CCS)-iron oxide nanoparticles (IONPs) composites carrying adhesive moieties with strong surface affinity. The ω-transaminase (ω-TA) was immobilized onto this magnetic composite via nucleophilic reactions between catechol and ω-TA. Under optimal conditions, 87.5% of the available ω-TA was immobilized on the composite, yielding an enzyme loading capacity as high as 681.7 mg/g. Furthermore, the valuation of enzyme activity showed that ω-TA immobilized on CCS-IONPs displayed enhanced pH and thermal stability compared to free enzyme. Importantly, the immobilized ω-TA retained more than 50% of its initial activity after 15 repeated reaction cycles using magnetic separation and 61.5% of its initial activity after storage at 4°C in phosphate buffered saline (PBS) for 15 days. The results suggested that such adhesive magnetic composites may provide an improved platform technology for bio-macromolecules immobilized.

  20. Magnetic catechol-chitosan with bioinspired adhesive surface: preparation and immobilization of ω-transaminase.

    PubMed

    Ni, Kefeng; Zhou, Xu; Zhao, Li; Wang, Hualei; Ren, Yuhong; Wei, Dongzhi

    2012-01-01

    The magnetic chitosan nanocomposites have been studied intensively and been used practically in various biomedical and biological applications including enzyme immobilization. However, the loading capacity and the remained activity of immobilized enzyme based on existing approaches are not satisfied. Simpler and more effective immobilization strategies are needed. Here we report a simple catechol modified protocol for preparing a novel catechol-chitosan (CCS)-iron oxide nanoparticles (IONPs) composites carrying adhesive moieties with strong surface affinity. The ω-transaminase (ω-TA) was immobilized onto this magnetic composite via nucleophilic reactions between catechol and ω-TA. Under optimal conditions, 87.5% of the available ω-TA was immobilized on the composite, yielding an enzyme loading capacity as high as 681.7 mg/g. Furthermore, the valuation of enzyme activity showed that ω-TA immobilized on CCS-IONPs displayed enhanced pH and thermal stability compared to free enzyme. Importantly, the immobilized ω-TA retained more than 50% of its initial activity after 15 repeated reaction cycles using magnetic separation and 61.5% of its initial activity after storage at 4°C in phosphate buffered saline (PBS) for 15 days. The results suggested that such adhesive magnetic composites may provide an improved platform technology for bio-macromolecules immobilized. PMID:22815930

  1. Cagelike mesoporous silica encapsulated with microcapsules for immobilized laccase and 2, 4-DCP degradation.

    PubMed

    Yang, Junya; Huang, Yan; Yang, Yuxiang; Yuan, Hongming; Liu, Xiangnong

    2015-12-01

    In this study, cage-like mesoporous silica was used as the carrier to immobilize laccase by a physical approach, followed by encapsulating with chitosan/alginate microcapsule membranes to form microcapsules of immobilized laccase based on layer-by-layer technology. The relationship between laccase activity recovery/leakage rate and the coating thickness was simultaneously investigated. Because the microcapsule layers have a substantial network of pores, they act as semipermeable membranes, while the laccase immobilized inside the microcapsules acts as a processing plant for degradation of 2,4-dichlorophenol. The microcapsules of immobilized laccase were able to degrade 2,4-dichlorophenol within a wide range of 2,4-dichlorophenol concentration, temperature and pH, with mean degradation rate around 62%. Under the optimal conditions, the thermal stability and reusability of immobilized laccase were shown to be improved significantly, as the removal rate and degradation rate remained over 40.2% and 33.8% respectively after 6cycles of operation. Using mass spectrometry (MS) and nuclear magnetic resonance (NMR), diisobutyl phthalate and dibutyl phthalate were identified as the products of 2,4-dichlorophenol degradation by the microcapsules of immobilized laccase and laccase immobilized by a physical approach, respectively, further demonstrating the degradation mechanism of 2,4-dichlorophenol by microcapsule-immobilized laccase.

  2. Influence of acetylcholinesterase immobilization on the photoluminescence properties of mesoporous silicon surface

    NASA Astrophysics Data System (ADS)

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2014-07-01

    Acetylcholinesterase immobilized p-type porous silicon surface was prepared by covalent attachment. The immobilization procedure was based on support surface chemical oxidation, silanization, surface activation with cyanuric chloride and finally covalent attachment of free enzyme on the cyanuric chloride activated porous silicon surface. Different pore diameter of porous silicon samples were prepared by electrochemical etching in HF based electrolyte solution and appropriate sample was selected suitable for enzyme immobilization with maximum trapping ability. The surface modification was studied through field emission scanning electron microscope, EDS, FT-IR analysis, and photoluminescence measurement by utilizing the fluctuation in the photoluminescence of virgin and enzyme immobilized porous silicon surface. Porous silicon showed strong photoluminescence with maximum emission at 643 nm and immobilization of acetylcholinesterase on porous silicon surface cause considerable increment on the photoluminescence of porous silicon material while acetylcholinesterase free counterpart did not exhibit any fluorescence in the range of 635-670 nm. The activities of the free and immobilized enzymes were evaluated by spectrophotometric method by using neostigmine methylsulfate as standard enzyme inhibitor. The immobilized enzyme exhibited considerable response toward neostigmine methylsulfate in a dose dependent manner comparable with that of its free counterpart alongside enhanced stability, easy separation from the reaction media and significant saving of enzyme. It was believed that immobilized enzyme can be exploited in organic and biomolecule synthesis possessing technical and economical prestige over free enzyme and prominence of easy separation from the reaction mixture.

  3. Degradation of raffinose oligosaccharides in soymilk by immobilized alpha-galactosidase of Aspergillus oryzae.

    PubMed

    Kotiguda, Girigowda; Kapnoor, Shankar S; Kulkarni, Dhananjay; Mulimani, Veerappa H

    2007-09-01

    Alpha-galactosidase was immobilized in a mixture of k-carrageenan and locust bean gum. The properties of the free and immobilized enzyme were then determined. The optimum pH for both the soluble and immobilized enzyme was 4.8. The optimum temperature for the soluble enzymes was 50 degrees C, whereas that for the immobilized enzyme was 55 degrees C. The immobilized enzyme was used in batch, repeated batch, and continuous modes to degrade the raffinose-family sugars present in soymilk. Two hours of incubation with the free and immobilized alpha-galactosidases resulted in an 80% and 68% reduction in the raffinose oligosaccharides in the soymilk, respectively. In the repeated batch, a 73% reduction was obtained in the fourth cycle. A fluidized bed reactor was also designed to treat soymilk continuously and the performance of the immobilized alpha-galactosidase tested at different flow rates, resulting in a 90% reduction of raffinose-family oligosaccharides in the soymilk at a flow rate 40 ml/h. Therefore, the present study demonstrated that immobilized alpha-galactosidase in a continuous mode is efficient for reducing the oligosaccharides present in soymilk, which may be of considerable interest for industrial application.

  4. Improved performance of α-amylase immobilized on poly(glycidyl methacrylate-co-ethylenedimethacrylate) beads.

    PubMed

    He, Tian; Tian, Yong-Le; Qi, Liang; Zhang, Jing; Zhang, Zhi-Qi

    2014-04-01

    α-Amylase was successfully immobilized onto poly(glycidyl methacrylate-co-ethylenedimethacrylate) (PGMA/EDMA) beads with high immobilization efficiency of 87.8%. PGMA/EDMA beads with a relatively uniform diameter of 2-3 μm were prepared by single-step swelling polymerization. After amination with ethanediamine and activation with glutaraldehyde, PGMA/EDMA beads showed commendable α-amylase immobilization capacity of 35.1 mg g(-1) carrier. Compared with free form, immobilized α-amylase had increasement of 12.94 mg mL(-1) for Km and 0.124 mmol mL(-1) min(-1) for Vmax, improved acid resistance (the optimal pH from 7 to 5), presented better thermal stability by retaining 61% activity than 40% at 90 °C, and displayed high operational reusability by retaining 58% of its initial activity after nine uses. Moreover, less than 10% of the free enzyme and more than 80% of the immobilized enzyme retained activity after 180 min pre-incubation at 50 °C. The easy modification, high immobilization efficiency and good properties of immobilized α-amylase in the present study indicate that PGMA/EDMA beads are suitable for α-amylase immobilization. The enhancement of acid resistance and thermo stability is doubtless of benefit for the industrial use of α-amylase.

  5. Acetylcholinesterase immobilization and characterization, and comparison of the activity of the porous silicon-immobilized enzyme with its free counterpart

    PubMed Central

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2016-01-01

    A successful prescription is presented for acetylcholinesterase physically adsorbed on to a mesoporous silicon surface, with a promising hydrolytic response towards acetylthiocholine iodide. The catalytic behaviour of the immobilized enzyme was assessed by spectrophotometric bioassay using neostigmine methyl sulfate as a standard acetycholinesterase inhibitor. The surface modification was studied through field emission SEM, Fourier transform IR spectroscopy, energy-dispersive X-ray spectroscopy, cathode luminescence and X-ray photoelectron spectroscopy analysis, photoluminescence measurement and spectrophotometric bioassay. The porous silicon-immobilized enzyme not only yielded greater enzyme stability, but also significantly improved the native photoluminescence at room temperature of the bare porous silicon architecture. The results indicated the promising catalytic behaviour of immobilized enzyme compared with that of its free counterpart, with a greater stability, and that it aided reusability and easy separation from the reaction mixture. The porous silicon-immobilized enzyme was found to retain 50% of its activity, promising thermal stability up to 90°C, reusability for up to three cycles, pH stability over a broad pH of 4–9 and a shelf-life of 44 days, with an optimal hydrolytic response towards acetylthiocholine iodide at variable drug concentrations. On the basis of these findings, it was believed that the porous silicon-immobilized enzyme could be exploited as a reusable biocatalyst and for screening of acetylcholinesterase inhibitors from crude plant extracts and synthesized organic compounds. Moreover, the immobilized enzyme could offer a great deal as a viable biocatalyst in bioprocessing for the chemical and pharmaceutical industries, and bioremediation to enhance productivity and robustness. PMID:26839417

  6. Acetylcholinesterase immobilization and characterization, and comparison of the activity of the porous silicon-immobilized enzyme with its free counterpart.

    PubMed

    Saleem, Muhammad; Rafiq, Muhammad; Seo, Sung-Yum; Lee, Ki Hwan

    2016-01-01

    A successful prescription is presented for acetylcholinesterase physically adsorbed on to a mesoporous silicon surface, with a promising hydrolytic response towards acetylthiocholine iodide. The catalytic behaviour of the immobilized enzyme was assessed by spectrophotometric bioassay using neostigmine methyl sulfate as a standard acetycholinesterase inhibitor. The surface modification was studied through field emission SEM, Fourier transform IR spectroscopy, energy-dispersive X-ray spectroscopy, cathode luminescence and X-ray photoelectron spectroscopy analysis, photoluminescence measurement and spectrophotometric bioassay. The porous silicon-immobilized enzyme not only yielded greater enzyme stability, but also significantly improved the native photoluminescence at room temperature of the bare porous silicon architecture. The results indicated the promising catalytic behaviour of immobilized enzyme compared with that of its free counterpart, with a greater stability, and that it aided reusability and easy separation from the reaction mixture. The porous silicon-immobilized enzyme was found to retain 50% of its activity, promising thermal stability up to 90°C, reusability for up to three cycles, pH stability over a broad pH of 4-9 and a shelf-life of 44 days, with an optimal hydrolytic response towards acetylthiocholine iodide at variable drug concentrations. On the basis of these findings, it was believed that the porous silicon-immobilized enzyme could be exploited as a reusable biocatalyst and for screening of acetylcholinesterase inhibitors from crude plant extracts and synthesized organic compounds. Moreover, the immobilized enzyme could offer a great deal as a viable biocatalyst in bioprocessing for the chemical and pharmaceutical industries, and bioremediation to enhance productivity and robustness. PMID:26839417

  7. Viability, ultrastructure and cytokinin metabolism of free and immobilized tobacco chloroplasts.

    PubMed

    Polanská, Lenka; Vicánková, Anna; Dobrev, Petre I; Cková, Ivana Macháv; Vanková, Radomíra

    2004-10-01

    Cytokinins play a decisive role in regulation of plastid development and differentiation, but their metabolism in plastids is not known. Metabolic studies using intact chloroplasts are prevented by their instability once they are isolated from leaf cells. Chloroplasts of Nicotiana tabacum L. cv. Petit Havana SR1 were therefore immobilized into low-viscosity alginate. Their intactness was assessed by a glyceraldehyde-3-phosphate dehydrogenase assay which indicated that free chloroplasts totally disintegrated within 7 h, while more than 50% of immobilized chloroplasts remained intact after 24 h. The immobilization had no marked impact on ultrastructure and postponed final destruction. The metabolite profile was similar in free and immobilized chloroplasts after 4 h incubation with tritiated zeatin. Nevertheless, the yield of conversion products decreased twice in immobilized chloroplasts, which was probably the outcome of mass transfer limitations and/or the sorption to polysaccharide matrix. PMID:15604795

  8. [Immobilization of E. coli cells in polyacrylamide-based microporous cryogels].

    PubMed

    Lusta, K A; Starostina, N G; Gorkina, N B; Fikhte, B A; Lozinskiĭ, V I

    1988-01-01

    E. coli cells were immobilized in polyacrylamide cryogel by three ways: (1) introduction of cells in the reaction mixture followed by cryopolymerization; (2) the filling of the cryogel pores followed by cell fixation with diluted glutaric dialdehyde (GDA), and (3) the filling of the macropores of the polymeric matrix with modified surface. The ultrastructure of the gels and immobilized cells as well as distribution of attachment of the cells immobilized by different techniques were studied. The first type of immobilization was characterized by the highest quantity of the biomass in the gel (by protein) and by a sharp decrease of the cell viability. The second failed to retain the cells in the pores, and the GDA treatment significantly decreased the viability index. The latter technique was the mildest and completely maintained the viability of the population. However, the biomass content was lower as compared to the first type of immobilization, but could be considerably increased by the GDA treatment.

  9. Polystyrene Attached Pt(IV)–Azomethine, Synthesis and Immobilization of Glucose Oxidase Enzyme

    PubMed Central

    Sarı, Nurşen; Antepli, Esin; Nartop, Dilek; Yetim, Nurdan Kurnaz

    2012-01-01

    Modified polystyrene with Pt(IV)–azomethine (APS–Sch–Pt) was synthesized by means of condensation and demonstrated to be a promising enzyme support by studying the enzymatic properties of glucose oxidase enzyme (GOx) immobilized on it. The characteristics of the immobilized glucose oxidase (APS–Sch–Pt–GOx) enzyme showed two optimum pH values that were pH = 4.0 and pH = 7. The insertion of stable Pt(IV)–azomethine spacers between the polystyrene backbone and the immobilized GOx, (APS–Sch–Pt–GOx), increases the enzymes’ activity and improves their affinity towards the substrate even at pH = 4. The influence of temperature, reusability and storage capacity on the free and immobilized glucose oxidase enzyme was investigated. The storage stability of the immobilized glucose oxidase was shown to be eleven months in dry conditions at +4 °C. PMID:23109888

  10. Effect of consecutive cooling and immobilization on catecholamine metabolism in rat tissues

    NASA Technical Reports Server (NTRS)

    Matlina, E. S.; Waysman, S. M.; Zaydner, I. G.; Kogan, B. M.; Nozdracheva, L. V.

    1979-01-01

    The combined effect of two stressor stimuli--cooling and immobilization--acting successively on the sympathetic-adrenaline system was studied experimentally in rats that were cooled for 8 hours at 7 C on the first day and immobilized for 6 hours on the next day. The biochemical and histochemical methods used and the experimental technique involved are described in detail. The following conclusions were formulated: (1) the successive action of cooling and immobilization results in a stronger decrease in the adrenaline and noradrenaline content in the adrenal gland than that which could be due to a simple summation of the cooling and immobilization effects; (2) successive cooling and immobilization are followed by activation of catecholamine synthesis in the adrenal gland; and (3) 1-DOPA administration (45 mg/kg 3 times in 2 days) intraabdominally activated catecholamine synthesis in the adrenal glands in both the control and test animals.

  11. Pineapple stem bromelain immobilized on different supports: catalytic properties in model wine.

    PubMed

    Ilaria, Benucci; Marco, Esti; Katia, Liburdi; Maria Vittoria, Garzillo Anna

    2012-01-01

    Bromelain from pineapple stem has been covalently immobilized on different supports to select the more efficient biocatalyst that should be applied toward unstable proteins in real white wine. In this preliminary study, catalytic properties of different immobilized bromelain forms were compared under wine-like conditions, against a synthetic substrate (Bz-Phe-Val-Arg-pNA).Covalent immobilization affected protease kinetic properties, even if all immobilized forms presented both a better substrate affinity and higher half-life (with the exception of a few procedures) with respect to the free enzyme. Stem bromelain was successfully immobilized on chitosan beads without glutaraldehyde thus yielding a food-safe and promising biocatalyst for unstable real wine future application. PMID:23011793

  12. Immobilization of a Plant Lipase from Pachira aquatica in Alginate and Alginate/PVA Beads

    PubMed Central

    Bonine, Bárbara M.; Polizelli, Patricia Peres; Bonilla-Rodriguez, Gustavo O.

    2014-01-01

    This study reports the immobilization of a new lipase isolated from oleaginous seeds of Pachira aquatica, using beads of calcium alginate (Alg) and poly(vinyl alcohol) (PVA). We evaluated the morphology, number of cycles of reuse, optimum temperature, and temperature stability of both immobilization methods compared to the free enzyme. The immobilized enzymes were more stable than the free enzyme, keeping 60% of the original activity after 4 h at 50°C. The immobilized lipase was reused several times, with activity decreasing to approximately 50% after 5 cycles. Both the free and immobilized enzymes were found to be optimally active between 30 and 40°C. PMID:24818012

  13. Pineapple stem bromelain immobilized on different supports: catalytic properties in model wine.

    PubMed

    Ilaria, Benucci; Marco, Esti; Katia, Liburdi; Maria Vittoria, Garzillo Anna

    2012-01-01

    Bromelain from pineapple stem has been covalently immobilized on different supports to select the more efficient biocatalyst that should be applied toward unstable proteins in real white wine. In this preliminary study, catalytic properties of different immobilized bromelain forms were compared under wine-like conditions, against a synthetic substrate (Bz-Phe-Val-Arg-pNA).Covalent immobilization affected protease kinetic properties, even if all immobilized forms presented both a better substrate affinity and higher half-life (with the exception of a few procedures) with respect to the free enzyme. Stem bromelain was successfully immobilized on chitosan beads without glutaraldehyde thus yielding a food-safe and promising biocatalyst for unstable real wine future application.

  14. Cellulose aerogel regenerated from ionic liquid solution for immobilized metal affinity adsorption.

    PubMed

    Oshima, Tatsuya; Sakamoto, Toshihiko; Ohe, Kaoru; Baba, Yoshinari

    2014-03-15

    Surface morphology of cellulosic adsorbents is expected to influence the adsorption behavior of biomacromolecules. In the present study, cellulose aerogel regenerated from ionic liquid solution was prepared for use as a polymer support for protein adsorption. Iminodiacetic acid groups were introduced to the aerogel for immobilized metal affinity adsorption of proteins. A Cu(II)-immobilized iminodiacetic acid cellulose aerogel (Cu(II)-IDA-CA), which has a large specific surface area, showed a higher adsorption capacity than Cu(II)-immobilized iminodiacetic acid bacterial cellulose (Cu(II)-IDA-BC) and Cu(II)-immobilized iminodiacetic acid plant cellulose (Cu(II)-IDA-PC). In contrast, the Cu(II)-immobilized cellulosic adsorbents showed similar adsorption capacities for smaller amino acid and peptides. The results show that cellulose aerogels are useful as polymer supports with high protein adsorption capacities.

  15. Stabilization of α-amylase by using anionic surfactant during the immobilization process

    NASA Astrophysics Data System (ADS)

    El-Batal, A. I.; Atia, K. S.; Eid, M.

    2005-10-01

    This work describes the entrapment of α-amylase into butylacrylate-acrylic acid copolymer (BuA/AAc) using γ irradiation. The effect of an anionic surfactant (AOT), the reuse efficiency, and kinetic behavior of immobilized α-amylase were studied. Covering of α-amylase with bis-(2-ethylhexyl)sulfosuccinate sodium salt (AOT) made the enzyme more stable than the uncovered form. The hydrolytic activity of the pre-coated immobilized α-amylase was increased below the critical micelle concentration (cmc) (10 mmol/L). The results showed an increase in the relative activity with increase in the degree of hydration. The pre-coated immobilized α-amylase showed a higher k/K and lower activation energy compared to the free and uncoated-immobilized preparation, respectively. The results suggest that the immobilization of α-amylase is a potentially useful approach for commercial starch hydrolysis in two-phase systems.

  16. Facile synthesis of amino-silane modified superparamagnetic Fe3O4 nanoparticles and application for lipase immobilization.

    PubMed

    Cui, Yanjun; Li, Yanfeng; Yang, Yong; Liu, Xiao; Lei, Lin; Zhou, Lincheng; Pan, Fei

    2010-10-01

    In present study, a facile prepared nano-sized magnetic support was successfully synthesized. Then this support was applied for lipase immobilization using glutaraldehyde as a coupling agent. Experimental data showed that the immobilized lipase exhibited good thermal stability and reusability. The lipase loading amount and activity recovery were found to be 43.6 mg/g support and 58.2%. Kinetic studies suggested it an acceptable degree of specificity retention for an immobilization process. PMID:20638425

  17. Cascade catalysis in membranes with enzyme immobilization for multi-enzymatic conversion of CO2 to methanol.

    PubMed

    Luo, Jianquan; Meyer, Anne S; Mateiu, R V; Pinelo, Manuel

    2015-05-25

    Facile co-immobilization of enzymes is highly desirable for bioconversion methods involving multi-enzymatic cascade reactions. Here we show for the first time that three enzymes can be immobilized in flat-sheet polymeric membranes simultaneously or separately by simple pressure-driven filtration (i.e. by directing membrane fouling formation), without any addition of organic solvent. Such co-immobilization and sequential immobilization systems were examined for the production of methanol from CO2 with formate dehydrogenase (FDH), formaldehyde dehydrogenase (FaldDH) and alcohol dehydrogenase (ADH). Enzyme activity was fully retained by this non-covalent immobilization strategy. The two immobilization systems had similar catalytic efficiencies because the second reaction (formic acid→formaldehyde) catalyzed by FaldDH was found to be the cascade bottleneck (a threshold substrate concentration was required). Moreover, the trade-off between the mitigation of product inhibition and low substrate concentration for the adjacent enzymes probably made the co-immobilization meaningless. Thus, sequential immobilization could be used for multi-enzymatic cascade reactions, as it allowed the operational conditions for each single step to be optimized, not only during the enzyme immobilization but also during the reaction process, and the pressure-driven mass transfer (flow-through mode) could overcome the diffusion resistance between enzymes. This study not only offers a green and facile immobilization method for multi-enzymatic cascade systems, but also reveals the reaction bottleneck and provides possible solutions for the bioconversion of CO2 to methanol.

  18. Effect of low-level helium-neon laser therapy on histological and ultrastructural features of immobilized rabbit articular cartilage.

    PubMed

    Bayat, Mohammad; Ansari, Enayatallah; Gholami, Narges; Bayat, Aghdas

    2007-05-25

    The present study investigates whether low-level helium-neon laser therapy can increase histological parameters of immobilized articular cartilage in rabbits or not. Twenty five rabbits were divided into three groups: the experiment group, which received low-level helium-neon laser therapy with 13J/cm(2) three times a week after immobilization of their right knees; the control group which did not receive laser therapy after immobilization of their knees; and the normal group which received neither immobilization nor laser therapy. Histological and electron microscopic examinations were performed at 4 and 7 weeks after immobilization. Depth of the chondrocyte filopodia in four-week immobilized experiment group, and depth of articular cartilage in seven-week immobilized experiment group were significantly higher than those of relevant control groups (exact Fisher test, p=0.001; student's t-test, p=0.031, respectively). The surfaces of articular cartilages of the experiment group were relatively smooth, while those of the control group were unsmooth. It is therefore concluded that low-level helium-neon laser therapy had significantly increased the depth of the chondrocyte filopodia in four-week immobilized femoral articular cartilage and the depth of articular cartilage in seven-week immobilized knee in comparison with control immobilized articular cartilage.

  19. Functional plasticity of sensorimotor representations following short-term immobilization of the dominant versus non-dominant hands.

    PubMed

    Meugnot, Aurore; Toussaint, Lucette

    2015-02-01

    The main purpose of the present study was to investigate the functional plasticity of sensorimotor representations for dominant versus non-dominant hands following short-term upper-limb sensorimotor deprivation. All participants were right-handed. A splint was placed either on the right hand or on the left hand of the participants during a brief period of 48 h and was used for the input/output signal restrictions. The participants were divided into 3 groups: right hand immobilization, left hand immobilization and control (without immobilization). The immobilized participants performed the hand laterality task before (pre-test) and immediately after (post-test) splint removal. The pre-/post-test procedure was similar for the control group. The main results showed a significant response time improvement when judging the laterality of hand stimuli in the control group. In contrast, the results showed a weaker response time improvement for the left-hand immobilization group and no significant improvement for the right-hand immobilization group. Overall, these results revealed that immobilization-induced effects were lower for the non-dominant hand and also suggested that 48 h of upper-limb immobilization led to an inter-limb transfer phenomenon regardless of the immobilized hand. The immobilization-induced effects were highlighted by the slowdown of the sensorimotor processes related to manual actions, probably due to an alteration in a general cognitive representation of hand movements.

  20. Covalent attachment of cholesterol oxidase and horseradish peroxidase on perlite through silanization: activity, stability and co-immobilization.

    PubMed

    Torabi, Seyed-Fakhreddin; Khajeh, Khosro; Ghasempur, Salehe; Ghaemi, Nasser; Siadat, Seyed-Omid Ranaei

    2007-08-31

    In the present work, co-immobilization of cholesterol oxidase (COD) and horseradish peroxidase (POD) on perlite surface was attempted. The surface of perlite were activated by 3-aminopropyltriethoxysilane and covalently bonded with COD and POD via glutaraldehyde. Enzymes activities have been assayed by spectrophotometric technique. The stabilities of immobilized COD and POD to pH were higher than those of soluble enzymes and immobilization shifted optimum pH of enzymes to the lower pH. Heat inactivation studies showed improved thermostability of the immobilized COD for more than two times, but immobilized POD was less thermostable than soluble POD. Also activity recovery of immobilized COD was about 50% since for immobilized POD was 11%. The K(m) of immobilized enzymes was found slightly lower than that of soluble enzymes. Immobilized COD showed inhibition in its activity at high cholesterol concentration which was not reported for soluble COD before. Co-immobilized enzymes retained 65% of its initial activity after 20 consecutive reactor batch cycles.

  1. Technetium Immobilization Forms Literature Survey

    SciTech Connect

    Westsik, Joseph H.; Cantrell, Kirk J.; Serne, R. Jeffrey; Qafoku, Nikolla

    2014-05-01

    Of the many radionuclides and contaminants in the tank wastes stored at the Hanford site, technetium-99 (99Tc) is one of the most challenging to effectively immobilize in a waste form for ultimate disposal. Within the Hanford Tank Waste Treatment and Immobilization Plant (WTP), the Tc will partition between both the high-level waste (HLW) and low-activity waste (LAW) fractions of the tank waste. The HLW fraction will be converted to a glass waste form in the HLW vitrification facility and the LAW fraction will be converted to another glass waste form in the LAW vitrification facility. In both vitrification facilities, the Tc is incorporated into the glass waste form but a significant fraction of the Tc volatilizes at the high glass-melting temperatures and is captured in the off-gas treatment systems at both facilities. The aqueous off-gas condensate solution containing the volatilized Tc is recycled and is added to the LAW glass melter feed. This recycle process is effective in increasing the loading of Tc in the LAW glass but it also disproportionally increases the sulfur and halides in the LAW melter feed which increases both the amount of LAW glass and either the duration of the LAW vitrification mission or the required supplemental LAW treatment capacity.

  2. Leukocyte responses to immobilized patterns of CXCL8.

    PubMed

    Girrbach, Maria; Rink, Ina; Ladnorg, Tatjana; Azucena, Carlos; Heißler, Stefan; Haraszti, Tamás; Schepers, Ute; Schmitz, Katja

    2016-06-01

    The attachment of neutrophils to the endothelial surface and their migration towards the site of inflammation following chemokine gradients play an essential role in the innate immune response. Chemokines adhere to glycosaminoglycans on the endothelial surface to be detected by leukocytes and trigger their movement along surface- bound gradients in a process called haptotaxis. In assays to systematically study the response of leukocytes to surface-bound compounds both the spatial arrangement of the compound as well as the mode of immobilization need to be controlled. In this study microcontact printing was employed to create patterns of hydrophobic or functionalized thiols on gold-coated glass slides and CXCL8 was immobilized on the thiol coated areas using three different strategies. Human neutrophils adhered to the CXCL8-coated lines but not to the PEG-coated background. We could show that more cells adhered to CXCL8 adsorbed to hydrophobic octadecanethiol than on CXCL8 covalently bound to amino undecanethiol or CXCL8 specifically bound to immobilized heparin on aminothiol. Likewise general cell activity such as lamellipodia formation and random migration were most pronounced for CXCL8 adsorbed on a hydrophobic surface which may be attributed to the larger amounts of protein immobilized on this type of surface. PMID:26970827

  3. Surface and Interface Control on Photochemically Initiated Immobilization

    SciTech Connect

    Liu, Li; Engelhard, Mark H.; Yan, Mingdi

    2006-11-01

    Surface and interface properties are important in controlling the yield and efficiency of the photochemically initiated immobilization. Using a silane-functionalized perfluorophenylazide (PFPA-silane) as the photoactive crosslinker, the immobilization of polymers was studied by adjusting the density of the surface azido groups. Dilution of the photolinker resulted in a gradual decrease in the density of surface azido groups as well as the thickness of the immobilized film. When a non-photoactive silane was added to PFPA-silane, the film thickness decreased more rapidly, indicating that the additive competed with PFPA-silane and effectively reduced the density of the surface azido groups. The effect of surface topography was studied by adding a non-photoactive silane with either a shorter (n-propyltrimethoxysilane (PTMS)) or a longer spacer (n-octadecyltrimethoxysilane (ODTMS)). In most cases the long chain ODTMS shielded the surface azido groups resulting in more rapid decrease in film thickness as compared to PTMS treated under the same conditions. As the density of the surface azido groups decreased, the immobilized polymer changed from smooth films to patched structures, and eventually single polymer molecules.

  4. Helicopter immobilization of elk in southcentral Washington

    SciTech Connect

    McCorquodale, S.M.; Eberhardt, L.E. ); Petron, S.E. )

    1988-01-01

    Free-ranging elk are commonly immobilized for research or management by rifle-fired darts shot from a helicopter. Compounds used for this purpose have included succinylcholine chloride (succinylcholine), etorphine hydrochloride (etorphine), and xylazine hydrochloride (xylazine). To assess the efficacy of various immobilizing drugs used in helicopter applications, we darted 38 elk from a helicopter on the Arid Lands Ecology Reserve, Washington from 1983 to 1987. We used either succinylcholine, etorphine hydrochloride, or xylazine hydrochloride a primary immobilants. Unsuccessful immobilizations were most common in elk darted with succinylcholine. Yohimbine was used to reverse xylazine immobilizations. The use of xylazine and yohimbine provides an efficient, cost-effective alternative to etorphine, diprenorphine immobilization and reversal in elk while increasing handler safety. Etorphine appeared to be the best immobilant when extended pain-producing procedures (such as surgical telemetry implantation) are planned because it induced the longest and deepest anesthesia. When the potential to lose contact with darted animals exist, we believe succinylcholine may be the preferred immobilant because of rapid, spontaneous recovery.

  5. Plutonium Immobilization Can Loading Equipment Review

    SciTech Connect

    Kriikku, E.; Ward, C.; Stokes, M.; Randall, B.; Steed, J.; Jones, R.; Hamilton, L.

    1998-05-01

    This report lists the operations required to complete the Can Loading steps on the Pu Immobilization Plant Flow Sheets and evaluates the equipment options to complete each operation. This report recommends the most appropriate equipment to support Plutonium Immobilization Can Loading operations.

  6. Protein immobilization strategies for protein biochips.

    PubMed

    Rusmini, Federica; Zhong, Zhiyuan; Feijen, Jan

    2007-06-01

    In the past few years, protein biochips have emerged as promising proteomic and diagnostic tools for obtaining information about protein functions and interactions. Important technological innovations have been made. However, considerable development is still required, especially regarding protein immobilization, in order to fully realize the potential of protein biochips. In fact, protein immobilization is the key to the success of microarray technology. Proteins need to be immobilized onto surfaces with high density in order to allow the usage of small amount of sample solution. Nonspecific protein adsorption needs to be avoided or at least minimized in order to improve detection performances. Moreover, full retention of protein conformation and activity is a challenging task to be accomplished. Although a large number of review papers on protein biochips have been published in recent years, few have focused on protein immobilization technology. In this review, current protein immobilization strategies, including physical, covalent, and bioaffinity immobilization for the fabrication of protein biochips, are described. Particular consideration has been given to oriented immobilization, also referred to as site-specific immobilization, which is believed will improve homogeneous surface covering and accessibility of the active site.

  7. Immobilized Lactase in the Biochemistry Laboratory

    NASA Astrophysics Data System (ADS)

    Allison, Matthew J.; Bering, C. Larry

    1998-10-01

    Immobilized enzymes have many practical applications. They may be used in clinical, industrial, and biotechnological laboratories and in many clinical diagnostic kits. For educational purposes, use of immobilized enzymes can easily be taught at the undergraduate or even secondary level. We have developed an immobilized enzyme experiment that combines many practical techniques used in the biochemistry laboratory and fits within a three-hour time frame. In this experiment, lactase from over-the-counter tablets for patients with lactose intolerance is immobilized in polyacrylamide, which is then milled into small beads and placed into a chromatography column. A lactose solution is added to the column and the eluant is assayed using the glucose oxidase assay, available as a kit. We have determined the optimal conditions to give the greatest turnover of lactose while allowing the immobilized enzymes to be active for long periods at room temperature.

  8. Immobilization of whole cells using polymeric coatings

    SciTech Connect

    Lawton, C.W.; Klei, H.E.; Sunstrom, D.V.; Voronka, P.J.; Scott, C.D.

    1986-01-01

    A cell immobilization procedure was developed using latex coatings on solid particles. The method's widespread applicability has been demonstrated by successfully immobilizing Saccharomyces cerevisiae (ethanol production), Bacillus subtilis (tryptophan production). Penicillium chrysogenum (penicillin G production), and Escherichia coli (aspartic acid production). In contrast to other immobilization methods, this procedure produces a pellicular particle that is porous, allowing rapid substrate and gas transfer, has a hard core to avoid compression in large beds, and is dense to allow use in fluidized beds. The immobilization procedure was optimized with S. cerevisiae. Kinetic constants obtained were used to calculate effectiveness factors to show that there was minimal intraparticle diffusion resistance. Reactors utilizing the optimized particles were run for 300 hours to evaluate immobilized particle half-life which was 250 hours.

  9. Immobilized fluid membranes for gas separation

    SciTech Connect

    Liu, Wei; Canfield, Nathan L; Zhang, Jian; Li, Xiaohong Shari; Zhang, Jiguang

    2014-03-18

    Provided herein are immobilized liquid membranes for gas separation, methods of preparing such membranes and uses thereof. In one example, the immobilized membrane includes a porous metallic host matrix and an immobilized liquid fluid (such as a silicone oil) that is immobilized within one or more pores included within the porous metallic host matrix. The immobilized liquid membrane is capable of selective permeation of one type of molecule (such as oxygen) over another type of molecule (such as water). In some examples, the selective membrane is incorporated into a device to supply oxygen from ambient air to the device for electrochemical reactions, and at the same time, to block water penetration and electrolyte loss from the device.

  10. The effects of joint immobilization on articular cartilage of the knee in previously exercised rats

    PubMed Central

    Maldonado, Diogo Correa; da Silva, Marcelo Cavenaghi Pereira; Neto, Semaan El-Razi; Souza, Mônica Rodrigues; Souza, Romeu Rodrigues

    2013-01-01

    Studies have determined the effects of joint immobilization on the articular cartilage of sedentary animals, but we are not aware of any studies reporting the effects of joint immobilization in previously trained animals. The objective of the present study was to determine whether exercise could prevent degeneration of the articular cartilage that accompanies joint immobilization. We used light microscopy to study the thickness, cell density, nuclear size, and collagen density of articular cartilage of the femoral condyle of Wistar rats subjected to aerobic physical activity on an adapted treadmill five times per week. Four groups of Wistar rats were used: a control group (C), an immobilized group (I), an exercised group (E), and an exercised and then immobilized group (EI). The right knee joints from rats in groups I and EI were immobilized at 90 °C of flexion using a plastic cast for 8 weeks. Cartilage thickness decreased significantly in group I (mean, 120.14 ± 15.6 μm, P < 0.05), but not in group EI (mean, 174 ± 2.25), and increased significantly in group E (mean, 289.49 ± 9.15) compared with group C (mean, 239.20 ± 6.25). The same results were obtained for cell density, nuclear size, and collagen density (in all cases, P < 0.05). We concluded that exercise can prevent degenerative changes in femoral articular cartilage caused by immobilization of the knee joint. PMID:23480127

  11. Cellobiose hydrolysis using Pichia etchellsii cells immobilized in calcium alginate

    SciTech Connect

    Jain, D.; Ghose, T.K.

    1984-04-01

    Cellulose degradation rates can be increased by the hydrolysis of cellobiose using immobilized beta-glucosidase. Production of beta-glucosidase in four yeasts was studied and a maximum activity of 1.22 IU/mg cells was obtained in cells of Pichia etchellsii grown on 3% cellobiose. The immobilization of beta-glucosidase containing cells on various solid supports was studied and entrapment in calcium alginate gel beads was found to be the best method. After ten sequential batch uses of the preparation, 96.5% of the initial activity was retained. The pH and temperature optima for free and immobilized cells were pH 6.5 (0.05M Maleate buffer) and 50/sup 0/C however, the enzyme has a better thermal stability at 45/sup 0/C. Beads stored at 4/sup 0/C for six months retain 80% of their activity. Kinetic studies performed on free and immobilized cells show that glucose is a noncompetitive product inhibitor. The immobilized preparation was limited by pore diffusion but exhibited no film-diffusion resistance during packed bed reactor operation. Good plug flow characteristics were observed. A model for reaction with pore diffusion for a noncompetitive type of inhibited system was developed and applied to this system. The reation rate with diffusional limitations was determined by using the model and effectiveness factors were calculated for different particle sizes. The modified rate expression using the effectiveness factor represented batch and packed bed reactor operation satisfactorily. The productivity in the packed bed column fell rapidly with an increase in conversion rate indicating that the operating conditions of the column would have to balance high conversion rates with acceptable productivity. The half-life in the column was affected by temperature, increasing to over seventeen days at 40/sup 0/C and decreasing to less than two days at 50/sup 0/C.

  12. Arthrobacter sp. lipase immobilization for improvement in stability and enantioselectivity.

    PubMed

    Chaubey, Asha; Parshad, Rajinder; Koul, Surrinder; Taneja, Subhash C; Qazi, Ghulam N

    2006-12-01

    Arthrobacter sp. lipase (ABL, MTCC no. 5125) is being recognized as an efficient enzyme for the resolution of drugs and their intermediates. The immobilization of ABL on various matrices for its enantioselectivity, stability, and reusability has been studied. Immobilization by covalent bonding on sepharose and silica afforded a maximum of 380 and 40 IU/g activity, respectively, whereas sol-gel entrapment provided a maximum of 150 IU/g activity in dry powder. The immobilized enzyme displayed excellent stability in the pH range of 4-10 and even at higher temperature, i.e., 50-60 degrees C, compared to free enzyme, which is unstable under extreme conditions. The resolution of racemic auxiliaries like 1-phenyl ethanol and an intermediate of antidepressant drug fluoxetine, i.e., ethyl 3-hydroxy-3-phenylpropanoate alkyl acylates, provided exclusively R-(+) products ( approximately 99% ee, E=646 and 473), compared to cell free extract/whole cells which gave a product with approximately 96% ee (E=106 and 150). The repeated use (ten times) of covalently immobilized and entrapped ABL resulted in no loss in activity, thus demonstrating its prospects for commercial applications. PMID:16896604

  13. On-chip immobilization of planarians for in vivo imaging.

    PubMed

    Dexter, Joseph P; Tamme, Mary B; Lind, Christine H; Collins, Eva-Maria S

    2014-09-17

    Planarians are an important model organism for regeneration and stem cell research. A complete understanding of stem cell and regeneration dynamics in these animals requires time-lapse imaging in vivo, which has been difficult to achieve due to a lack of tissue-specific markers and the strong negative phototaxis of planarians. We have developed the Planarian Immobilization Chip (PIC) for rapid, stable immobilization of planarians for in vivo imaging without injury or biochemical alteration. The chip is easy and inexpensive to fabricate, and worms can be mounted for and removed after imaging within minutes. We show that the PIC enables significantly higher-stability immobilization than can be achieved with standard techniques, allowing for imaging of planarians at sub-cellular resolution in vivo using brightfield and fluorescence microscopy. We validate the performance of the PIC by performing time-lapse imaging of planarian wound closure and sequential imaging over days of head regeneration. We further show that the device can be used to immobilize Hydra, another photophobic regenerative model organism. The simple fabrication, low cost, ease of use, and enhanced specimen stability of the PIC should enable its broad application to in vivo studies of stem cell and regeneration dynamics in planarians and Hydra.

  14. LPS interactions with immobilized and soluble antimicrobial peptides.

    PubMed

    Gustafsson, Anna; Olin, Anders I; Ljunggren, Lennart

    2010-04-19

    A promising approach in sepsis therapy is the use of peptides truncated from serum- and membrane-proteins with binding domains for LPS: antimicrobial peptides (AMPs). AMPs can be useful in combination with conventional antibiotics to increase killing and neutralize LPS. Although many AMPs show a high specificity towards bacterial membranes, they can also exhibit toxicity, i.e. non-specific membrane lysis, of mammalian cells such as erythrocytes and therefore, unsuitable as systemic drugs. A way to overcome this problem may be an extracorporeal therapy with immobilized peptides. This study will compare neutralization of LPS using different AMPs in solution and when immobilized on to solid phases. The peptides ability to neutralize LPS-induced cytokine release in whole blood will also be tested. The peptides are truncated derivates from the known AMPs LL-37, SC4, BPI, S3 Delta and CEME. Two different methods were used to immobilize peptides, biomolecular interaction analysis, and Pierce SulfoLink Coupling Gel. To investigate LPS binding in solution the LAL test was used. After whole blood incubation with LPS and AMPs ELISA was used to measure TNFalpha, IL-1 beta and IL-6 production. The results suggest that immobilization of antimicrobial peptides does not inhibit their capacity to neutralize LPS, although there are differences between the peptides tested. Thus, peptides derived from LL-37 and CEME were more efficient both in LPS binding and neutralizing LPS-induced cytokine production. PMID:20233038

  15. Cell immobilization in kappa-carrageenan for ethanol production.

    PubMed

    Luong, J H

    1985-12-01

    A combination of extended Monod kinetics and the diffusional equation was used for evaluating the effectiveness factor of entrapped immobilized cells. Based on the kinetics of Zymomonas mobilis reported in the literature, the numerical results have revealed that the problem of mass transfer diffusional restrictions can be neglected by using small beads (1 mm in diameter) with a corresponding cell loading up to 276 g/L gel. On the basis of the numerical results obtained, the application of immobilized cells for continuous ethanol production was investigated. The kappa-carrageenan method was utilized to entrap Z. mobilis CP4, a potential ethanol producer. A two stage fermentation process has also been developed for ethanol production by the Z. mobilis carrageenan-bound cells. About 90 g/L ethanol was produced by immobilized cells at a total residence time of 1.56 h. The ethanol yield was estimated to be 93% of theoretical. The results obtained in this study also indicated that the control of optimum pH in an immobilized cell column is necessary to enhance the rate of ethanol production. PMID:18553626

  16. Recent advances in covalent, site-specific protein immobilization

    PubMed Central

    Meldal, Morten; Schoffelen, Sanne

    2016-01-01

    The properties of biosensors, biomedical implants, and other materials based on immobilized proteins greatly depend on the method employed to couple the protein molecules to their solid support. Covalent, site-specific immobilization strategies are robust and can provide the level of control that is desired in this kind of application. Recent advances include the use of enzymes, such as sortase A, to couple proteins in a site-specific manner to materials such as microbeads, glass, and hydrogels. Also, self-labeling tags such as the SNAP-tag can be employed. Last but not least, chemical approaches based on bioorthogonal reactions, like the azide–alkyne cycloaddition, have proven to be powerful tools. The lack of comparative studies and quantitative analysis of these immobilization methods hampers the selection process of the optimal strategy for a given application. However, besides immobilization efficiency, the freedom in selecting the site of conjugation and the size of the conjugation tag and the researcher’s expertise regarding molecular biology and/or chemical techniques will be determining factors in this regard. PMID:27785356

  17. Bioflavour production from orange peel hydrolysate using immobilized Saccharomyces cerevisiae.

    PubMed

    Lalou, Sofia; Mantzouridou, Fani; Paraskevopoulou, Adamantini; Bugarski, Branko; Levic, Steva; Nedovic, Victor

    2013-11-01

    The rising trend of bioflavour synthesis by microorganisms is hindered by the high manufacturing costs, partially attributed to the cost of the starting material. To overcome this limitation, in the present study, dilute-acid hydrolysate of orange peel was employed as a low-cost, rich in fermentable sugars substrate for the production of flavour-active compounds by Saccharomyces cerevisiae. With this purpose, the use of immobilized cell technology to protect cells against the various inhibitory compounds present in the hydrolysate was evaluated with regard to yeast viability, carbon and nitrogen consumption and cell ability to produce flavour active compounds. For cell immobilization the encapsulation in Ca alginate beads was used. The results were compared with those obtained using free-cell system. Based on the data obtained immobilized cells showed better growth performance and increased ability for de novo synthesis of volatile esters of "fruity" aroma (phenylethyl acetate, ethyl hexanoate, octanoate, decanoate and dodecanoate) than those of free cells. The potential for in situ production of new formulations containing flavour-active compounds derive from yeast cells and also from essential oil of orange peel (limonene, α-terpineol) was demonstrated by the fact that bioflavour mixture was found to accumulate within the beads. Furthermore, the ability of the immobilized yeast to perform efficiently repeated batch fermentations of orange peel hydrolysate for bioflavour production was successfully maintained after six consecutive cycles of a total period of 240 h. PMID:23995224

  18. Ceramic Hosts for Fission Products Immobilization

    SciTech Connect

    Peter C Kong

    2010-07-01

    metal ions, Mg and Ca, in the ceramic host phases. The immobilization of rear earth (lanthanide series) fission products in these ceramic host phases will also be studied this year. Cerium oxide is chosen to represent the rear earth fission product for substitution studies in spinel, perovskite and zirconolite ceramic hosts. Cerium has +3 and +4 oxidation states and it can replace some of the trivalent or tetravalent host ions to produce the substitution ceramics such as MgAl2-xCexO4, CaTi1-xCexO3, CaZr1-xCexTi2O7 and CaZrTi2-xCexO7. X-ray diffraction analysis will be used to compare the crystalline structures of the pure ceramic hosts and the substitution phases. SEM-EDX analysis will be used to study the Ce distribution in the ceramic host phases. The range of cerium doping is planned to reach the full substitution of the trivalent or tetravalent ions, Al, Ti and Zr, in the ceramic host phases.

  19. [Activity of aldehyde scavenger enzymes in the heart of rats of different age during immobilized stress].

    PubMed

    Grabovetskaia, E R; Davydov, V V

    2009-01-01

    This study was made to determine the activity of aldehyde scavenger enzymes in the heart's postmitochondrial fraction of rats of different age during immobilization stress. Our study demonstrated, that immobilization of 1.5-, 2- and 12-month rats was accompanied by inhibiting activity of aldehyde dehydrogenase and aldehyde reductase. At the same time we observed an increase in glutathione transferase activity in immobilized 1.5-month-old rats and that in reductase activity in 24-month-old rats. The revealed changes can lead to a decrease in the rate of endogenous aldehyde utilization in the heart during stress at puberty.

  20. Plutonium Immobilization Can Loading Concepts

    SciTech Connect

    Kriikku, E.; Ward, C.; Stokes, M.; Randall, B.; Steed, J.; Jones, R.; Hamilton, L.; Rogers, L.; Fiscus, J.; Dyches, G.

    1998-05-01

    The Plutonium Immobilization Facility will encapsulate plutonium in ceramic pucks and seal the pucks inside welded cans. Remote equipment will place these cans in magazines and the magazines in a Defense Waste Processing Facility (DWPF) canister. The DWPF will fill the canister with glass for permanent storage. This report discusses five can loading conceptual designs and the lists the advantages and disadvantages for each concept. This report identifies loading pucks into cans and backfilling cans with helium as the top priority can loading development areas. The can loading welder and cutter are very similar to the existing Savannah River Site (SRS) FB-Line bagless transfer welder and cutter and thus they are a low priority development item.

  1. Plutonium Immobilization Project Baseline Formulation

    SciTech Connect

    Ebbinghaus, B.

    1999-02-01

    A key milestone for the Immobilization Project (AOP Milestone 3.2a) in Fiscal Year 1998 (FY98) is the definition of the baseline composition or formulation for the plutonium ceramic form. The baseline formulation for the plutonium ceramic product must be finalized before the repository- and plant-related process specifications can be determined. The baseline formulation that is currently specified is given in Table 1.1. In addition to the baseline formulation specification, this report provides specifications for two alternative formulations, related compositional specifications (e.g., precursor compositions and mixing recipes), and other preliminary form and process specifications that are linked to the baseline formulation. The preliminary specifications, when finalized, are not expected to vary tremendously from the preliminary values given.

  2. Application of granular activated carbon/MnFe₂O₄ composite immobilized on C. glutamicum MTCC 2745 to remove As(III) and As(V): Kinetic, mechanistic and thermodynamic studies.

    PubMed

    Podder, M S; Majumder, C B

    2016-01-15

    The main objective of the present study was to investigate the efficiency of Corynebacterium glutamicum MTCC 2745 immobilized on granular activated carbon/MnFe2O4 (GAC/MnFe2O4) composite to treat high concentration of arsenic bearing wastewater. Non-linear regression analysis was done for determining the best-fit kinetic model on the basis of three correlation coefficients and three error functions and also for predicting the parameters involved in kinetic models. The results showed that Fractal-like mixed 1,2 order model for As(III) and Brouser-Weron-Sototlongo as well as Fractal-like pseudo second order models for As(V) were proficient to provide realistic description of biosorption/bioaccumulation kinetic. Applicability of mechanistic models in the current study exhibited that the rate governing step in biosorption/bioaccumulation of both As(III) and As(V) was film diffusion rather than intraparticle diffusion. The evaluated thermodynamic parameters ΔG(0), ΔH(0) and ΔS(0) revealed that biosorption/bioaccumulation of both As(III) and As(V) was feasible, spontaneous and exothermic under studied conditions.

  3. Application of granular activated carbon/MnFe2O4 composite immobilized on C. glutamicum MTCC 2745 to remove As(III) and As(V): Kinetic, mechanistic and thermodynamic studies

    NASA Astrophysics Data System (ADS)

    Podder, M. S.; Majumder, C. B.

    2016-01-01

    The main objective of the present study was to investigate the efficiency of Corynebacterium glutamicum MTCC 2745 immobilized on granular activated carbon/MnFe2O4 (GAC/MnFe2O4) composite to treat high concentration of arsenic bearing wastewater. Non-linear regression analysis was done for determining the best-fit kinetic model on the basis of three correlation coefficients and three error functions and also for predicting the parameters involved in kinetic models. The results showed that Fractal-like mixed 1,2 order model for As(III) and Brouser-Weron-Sototlongo as well as Fractal-like pseudo second order models for As(V) were proficient to provide realistic description of biosorption/bioaccumulation kinetic. Applicability of mechanistic models in the current study exhibited that the rate governing step in biosorption/bioaccumulation of both As(III) and As(V) was film diffusion rather than intraparticle diffusion. The evaluated thermodynamic parameters ΔG0, ΔH0 and ΔS0 revealed that biosorption/bioaccumulation of both As(III) and As(V) was feasible, spontaneous and exothermic under studied conditions.

  4. [Progress in co-immobilization of multiple enzymes].

    PubMed

    Wang, Jindan; Zhang, Guangya

    2015-04-01

    Enzyme immobilization is the core technology of biocatalysis. Over the past few decades, enzyme immobilization research mainly focused on single enzyme immobilization. In recent years, multi-enzyme immobilization attracts more and more attention as it could increase the local concentration of reaction and improve the reaction yield. In this review, a summary of the recent progress, together with our research, is presented. Special emphasis is placed on four methods in multi-enzymes co-immobilization, namely, the nonspecific covalent co-immobilization, the nonspecific non-covalent co-immobilization, the non-covalent encapsulation co-immobilized and the site specificity co-immobilized. Finally, some industrial uses of immobilized multi-enzymes were addressed and the application prospect of multi-enzyme immobilization was highlighted.

  5. Effective Immobilization of Agrobacterium sp. IFO 13140 Cells in Loofa Sponge for Curdlan Biosynthesis.

    PubMed

    Martinez, Camila Ortiz; Ruiz, Suelen Pereira; Nogueira, Marcela Tiemi; Bona, Evandro; Portilho, Márcia; Matioli, Graciette

    2015-05-04

    Curdlan production by Agrobacterium sp. IFO13140 immobilized on loofa sponge, alginate and loofa sponge with alginate was investigated. There was no statistically-significant difference in curdlan production when the microorganism was immobilized in different matrices. The loofa sponge was chosen because of its practical application and economy and because it provides a high stability through its continued use. The best conditions for immobilization on loofa sponge were 50 mg of cell, 200 rpm and 72 h of incubation, which provided a curdlan production 1.50-times higher than that obtained by free cells. The higher volumetric productivity was achieved by immobilized cells (0.09 g/L/h) at 150 rpm. The operating stability was evaluated, and until the fourth cycle, immobilized cells retained 87.40% of the production of the first cycle. The immobilized cells remained active after 300 days of storage at 4 °C. The results of this study demonstrate success in immobilizing cells for curdlan biosynthesis, making the process potentially suitable for industrial scale-up. Additional studies may show a possible contribution to the reduction of operating costs.

  6. Antimicrobial Properties of Lysosomal Enzymes Immobilized on NH₂Functionalized Silica-Encapsulated Magnetite Nanoparticles.

    PubMed

    Bang, Seung Hyuck; Sekhon, Simranjeet Singh; Cho, Sung-Jin; Kim, So Jeong; Le, Thai-Hoang; Kim, Pil; Ahn, Ji-Young; Kim, Yang-Hoon; Min, Jiho

    2016-01-01

    The immobilization efficiency, antimicrobial activity and recovery of lysosomal enzymes on NH2 functionalized magnetite nanoparticles have been studied under various conditions. The immobi- lization efficiency depends upon the ratio of the amount of enzyme and magnetite and it shows an increase with magnetite concentration which is due to the presence of amine group at the magnetite surface that leads to a strong attraction. The optimized reaction time to immobilize the lysosomal enzymes on magnetite was determined by using a rolling method. The immobilization efficiency increases with reaction time and reached a plateau after 5 minutes and then remained constant for 10 minutes. However, after 30 minutes the immobilization efficiency decreased to 85%, which is due to the weaker electrostatic interactions between magnetite and detached lysosomal enzymes. The recovery and stability of immobilized lysosomal enzymes has also been studied. The antimicrobial activity was almost 100% but it decreased upon reuse and no activity was observed after its reuse for seven times. The storage stability of lysosomal enzymes as an antimicrobial agent was about 88%, which decreased to 53% after one day and all activity of immobilized lysosomal enzymes was maintained after five days. Thus, the lysosomal enzymes immobilized on magnetite nanoparticles could potentially be used as antimicrobial agents to remove bacteria. PMID:27398573

  7. Antimicrobial Properties of Lysosomal Enzymes Immobilized on NH₂Functionalized Silica-Encapsulated Magnetite Nanoparticles.

    PubMed

    Bang, Seung Hyuck; Sekhon, Simranjeet Singh; Cho, Sung-Jin; Kim, So Jeong; Le, Thai-Hoang; Kim, Pil; Ahn, Ji-Young; Kim, Yang-Hoon; Min, Jiho

    2016-01-01

    The immobilization efficiency, antimicrobial activity and recovery of lysosomal enzymes on NH2 functionalized magnetite nanoparticles have been studied under various conditions. The immobi- lization efficiency depends upon the ratio of the amount of enzyme and magnetite and it shows an increase with magnetite concentration which is due to the presence of amine group at the magnetite surface that leads to a strong attraction. The optimized reaction time to immobilize the lysosomal enzymes on magnetite was determined by using a rolling method. The immobilization efficiency increases with reaction time and reached a plateau after 5 minutes and then remained constant for 10 minutes. However, after 30 minutes the immobilization efficiency decreased to 85%, which is due to the weaker electrostatic interactions between magnetite and detached lysosomal enzymes. The recovery and stability of immobilized lysosomal enzymes has also been studied. The antimicrobial activity was almost 100% but it decreased upon reuse and no activity was observed after its reuse for seven times. The storage stability of lysosomal enzymes as an antimicrobial agent was about 88%, which decreased to 53% after one day and all activity of immobilized lysosomal enzymes was maintained after five days. Thus, the lysosomal enzymes immobilized on magnetite nanoparticles could potentially be used as antimicrobial agents to remove bacteria.

  8. Effect of sulfonylureas administered centrally on the blood glucose level in immobilization stress model.

    PubMed

    Sharma, Naveen; Sim, Yun-Beom; Park, Soo-Hyun; Lim, Su-Min; Kim, Sung-Su; Jung, Jun-Sub; Hong, Jae-Seung; Suh, Hong-Won

    2015-05-01

    Sulfonylureas are widely used as an antidiabetic drug. In the present study, the effects of sulfonylurea administered supraspinally on immobilization stress-induced blood glucose level were studied in ICR mice. Mice were once enforced into immobilization stress for 30 min and returned to the cage. The blood glucose level was measured 30, 60, and 120 min after immobilization stress initiation. We found that intracerebroventricular (i.c.v.) injection with 30 µg of glyburide, glipizide, glimepiride or tolazamide attenuated the increased blood glucose level induced by immobilization stress. Immobilization stress causes an elevation of the blood corticosterone and insulin levels. Sulfonylureas pretreated i.c.v. caused a further elevation of the blood corticosterone level when mice were forced into the stress. In addition, sulfonylureas pretreated i.c.v. alone caused an elevation of the plasma insulin level. Furthermore, immobilization stress-induced insulin level was reduced by i.c.v. pretreated sulfonylureas. Our results suggest that lowering effect of sulfonylureas administered supraspinally against immobilization stress-induced increase of the blood glucose level appears to be primarily mediated via elevation of the plasma insulin level. PMID:25954123

  9. Activity of Laccase Immobilized on TiO2-Montmorillonite Complexes

    PubMed Central

    Wang, Qingqing; Peng, Lin; Li, Guohui; Zhang, Ping; Li, Dawei; Huang, Fenglin; Wei, Qufu

    2013-01-01

    The TiO2-montmorillonite (TiO2-MMT) complex was prepared by blending TiO2 sol and MMT with certain ratio, and its properties as an enzyme immobilization support were investigated. The pristine MMT and TiO2-MMT calcined at 800 °C (TiO2-MMT800) were used for comparison to better understand the immobilization mechanism. The structures of the pristine MMT, TiO2-MMT, and TiO2-MMT800 were examined by HR-TEM, XRD and BET. SEM was employed to study different morphologies before and after laccase immobilization. Activity and kinetic parameters of the immobilized laccase were also determined. It was found that the TiO2 nanoparticles were successfully introduced into the MMT layer structure, and this intercalation enlarged the “d value” of two adjacent MMT layers and increased the surface area, while the calcination process led to a complete collapse of the MMT layers. SEM results showed that the clays were well coated with adsorbed enzymes. The study of laccase activity revealed that the optimum pH and temperature were pH = 3 and 60 °C, respectively. In addition, the storage stability for the immobilized laccase was satisfactory. The kinetic properties indicated that laccase immobilized on TiO2-MMT complexes had a good affinity to the substrate. It has been proved that TiO2-MMT complex is a good candidate for enzyme immobilization. PMID:23771020

  10. Preparation and characterization of a thermostable enzyme (Mn-SOD) immobilized on supermagnetic nanoparticles.

    PubMed

    Song, Chongfu; Sheng, Liangquan; Zhang, Xiaobo

    2012-10-01

    Superoxide dismutase (SOD) has been widely applied in medical treatments, cosmetic, food, agriculture, and chemical industries. In industry, the immobilization of enzymes can offer better stability, feasible continuous operations, easy separation and reusing, and significant decrease of the operation costs. However, little attention has focused on the immobilization of the SOD, as well as the immobilization of thermostable enzymes. In this study, the recombinant thermostable manganese superoxide dismutase (Mn-SOD) of Thermus thermophilus wl was purified and covalently immobilized onto supermagnetic 3-APTES-modified Fe(3)O(4)@SiO(2) nanoparticles using glutaraldehyde method to prepare the Mn-SOD bound magnetic nanoparticles. The Mn-SOD nanoparticles were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, X-ray diffraction, transmission electron microscopy, and vibrating sample magnetometer analysis. The results indicated that the diameter of Mn-SOD nanoparticles was 40 (± 5) nm, and its saturation magnetization value was 27.9 emu/g without remanence or coercivity. By comparison with the free Mn-SOD, it was found that the immobilized Mn-SOD on nanoparticles exhibited better resistance to temperature, pH, metal ions, enzyme inhibitors, and detergents. The results showed that the immobilized Mn-SOD on nanoparticles could be reused ten times without significant decrease of enzymatic activity. Therefore, our study presented a novel strategy for the immobilization of thermostable Mn-SOD and for the application of thermostable enzymes. PMID:22237672

  11. Thermostable α-amylase immobilization: Enhanced stability and performance for starch biocatalysis.

    PubMed

    Kumar, Gudi Satheesh; Rather, Gulam Mohmad; Gurramkonda, Chandrasekhar; Reddy, Bontha Rajasekhar

    2016-01-01

    The uses of thermostable starch hydrolytic biocatalysts are steadily increasing for the industrial application because of their obvious need for biocatalytic performance at elevated temperatures. The starch liquefaction and saccharification can be carried out simultaneously by the use of thermostable starch hydrolytic biocatalysts, thus minimizing the unit operations, time, and efforts. The cost factor hampers the industrialization of expensive soluble (free) enzymes for biocatalytic applications and the immobilization of enzymes offers promising alternative to the hurdle. The present investigation was aimed for immobilization of thermostable α-amylase using calcium alginate, and statistical optimization studies were carried out for enhanced biocatalytic performance. Initially, one-parameter at a time optimization studies were carried out for identification of significant factors influencing the immobilization. Furthermore, a statistical approach, response surface methodology, was applied for immobilization of α-amylase. The immobilized α-amylase in alginate microbeads showed enhanced stability to temperature and reusable property for up to seven cycles (with the retention of 50% initial activity). Finally, the kinetic behavior of free and immobilized enzyme showed the Km value of 1.2% and 2.6% (w/v) and Vmax of 1,020 and 1,030 U, respectively. Fifty percent reduction in affinity of the immobilized enzyme toward substrate was compensated by its longer stability. PMID:25604037

  12. Thermostable α-amylase immobilization: Enhanced stability and performance for starch biocatalysis.

    PubMed

    Kumar, Gudi Satheesh; Rather, Gulam Mohmad; Gurramkonda, Chandrasekhar; Reddy, Bontha Rajasekhar

    2016-01-01

    The uses of thermostable starch hydrolytic biocatalysts are steadily increasing for the industrial application because of their obvious need for biocatalytic performance at elevated temperatures. The starch liquefaction and saccharification can be carried out simultaneously by the use of thermostable starch hydrolytic biocatalysts, thus minimizing the unit operations, time, and efforts. The cost factor hampers the industrialization of expensive soluble (free) enzymes for biocatalytic applications and the immobilization of enzymes offers promising alternative to the hurdle. The present investigation was aimed for immobilization of thermostable α-amylase using calcium alginate, and statistical optimization studies were carried out for enhanced biocatalytic performance. Initially, one-parameter at a time optimization studies were carried out for identification of significant factors influencing the immobilization. Furthermore, a statistical approach, response surface methodology, was applied for immobilization of α-amylase. The immobilized α-amylase in alginate microbeads showed enhanced stability to temperature and reusable property for up to seven cycles (with the retention of 50% initial activity). Finally, the kinetic behavior of free and immobilized enzyme showed the Km value of 1.2% and 2.6% (w/v) and Vmax of 1,020 and 1,030 U, respectively. Fifty percent reduction in affinity of the immobilized enzyme toward substrate was compensated by its longer stability.

  13. Metabolic Responses of Bacterial Cells to Immobilization.

    PubMed

    Żur, Joanna; Wojcieszyńska, Danuta; Guzik, Urszula

    2016-01-01

    In recent years immobilized cells have commonly been used for various biotechnological applications, e.g., antibiotic production, soil bioremediation, biodegradation and biotransformation of xenobiotics in wastewater treatment plants. Although the literature data on the physiological changes and behaviour of cells in the immobilized state remain fragmentary, it is well documented that in natural settings microorganisms are mainly found in association with surfaces, which results in biofilm formation. Biofilms are characterized by genetic and physiological heterogeneity and the occurrence of altered microenvironments within the matrix. Microbial cells in communities display a variety of metabolic differences as compared to their free-living counterparts. Immobilization of bacteria can occur either as a natural phenomenon or as an artificial process. The majority of changes observed in immobilized cells result from protection provided by the supports. Knowledge about the main physiological responses occurring in immobilized cells may contribute to improving the efficiency of immobilization techniques. This paper reviews the main metabolic changes exhibited by immobilized bacterial cells, including growth rate, biodegradation capabilities, biocatalytic efficiency and plasmid stability. PMID:27455220

  14. Enzymatic removal of flatulence-inducing sugars in chickpea milk using free and polyvinyl alcohol immobilized alpha-galactosidase from Aspergillus oryzae.

    PubMed

    Patil, Aravind Goud G; Kote, Naganagoud V; Mulimani, Veerappa

    2009-01-01

    The treatment of chickpea milk was carried out in batch, repeated batch and continuous reaction by soluble and polyvinyl alcohol (PVA) immobilized Aspergillus oryzae alpha-galactosidase for the removal of raffinose family oligosaccharides (RFOs). In the batch mode of treatment 96 and 92% of RFOs hydrolysis was observed by soluble and immobilized enzyme, respectively. In repeated batch experiments, immobilized enzyme showed 70% RFOs hydrolysis up to sixth cycle. Polyvinyl alcohol immobilized alpha-galactosidase in fluidized bed reactor showed highest reduction of 94% at a flow rate of 30 ml/h. The results obtained from the present study are very interesting for industrial use of PVA-immobilized enzyme.

  15. Covalent immobilization of p-selectin enhances cell rolling.

    PubMed

    Hong, Seungpyo; Lee, Dooyoung; Zhang, Huanan; Zhang, Jennifer Q; Resvick, Jennifer N; Khademhosseini, Ali; King, Michael R; Langer, Robert; Karp, Jeffrey M

    2007-11-20

    Cell rolling is an important physiological and pathological process that is used to recruit specific cells in the bloodstream to a target tissue. This process may be exploited for biomedical applications to capture and separate specific cell types. One of the most commonly studied proteins that regulate cell rolling is P-selectin. By coating surfaces with this protein, biofunctional surfaces that induce cell rolling can be prepared. Although most immobilization methods have relied on physisorption, chemical immobilization has obvious advantages, including longer functional stability and better control over ligand density and orientation. Here we describe chemical methods to immobilize P-selectin covalently on glass substrates. The chemistry was categorized on the basis of the functional groups on modified glass substrates: amine, aldehyde, and epoxy. The prepared surfaces were first tested in a flow chamber by flowing microspheres functionalized with a cell surface carbohydrate (sialyl Lewis(x)) that binds to P-selectin. Adhesion bonds between P-selectin and sialyl Lewis(x) dissociate readily under shear forces, leading to cell rolling. P-selectin immobilized on the epoxy glass surfaces exhibited enhanced long-term stability of the function and better homogeneity as compared to that for surfaces prepared by other methods and physisorbed controls. The microsphere rolling results were confirmed in vitro with isolated human neutrophils. This work is essential for the future development of devices for isolating specific cell types based on cell rolling, which may be useful for hematologic cancers and certain metastatic cancer cells that are responsive to immobilized selectins.

  16. Protein hydrolysis by immobilized and stabilized trypsin.

    PubMed

    Marques, Daniela; Pessela, Benavides C; Betancor, Lorena; Monti, Rubens; Carrascosa, Alfonso V; Rocha-Martin, Javier; Guisán, Jose M; Fernandez-Lorente, Gloria

    2011-01-01

    The preparation of novel immobilized and stabilized derivatives of trypsin is reported here. The new derivatives preserved 80% of the initial catalytic activity toward synthetic substrates [benzoyl-arginine p-nitroanilide (BAPNA)] and were 50,000-fold more thermally stable than the diluted soluble enzyme in the absence of autolysis. Trypsin was immobilized on highly activated glyoxyl-Sepharose following a two-step immobilization strategy: (a) first, a multipoint covalent immobilization at pH 8.5 that only involves low pK(a) amino groups (e.g., those derived from the activation of trypsin from trypsinogen) is performed and (b) next, an additional alkaline incubation at pH 10 is performed to favor an intense, additional multipoint immobilization between the high concentration of proximate aldehyde groups on the support surface and the high pK(a) amino groups at the enzyme surface region that participated in the first immobilization step. Interestingly, the new, highly stable trypsin derivatives were also much more active in the proteolysis of high molecular weight proteins when compared with a nonstabilized derivative prepared on CNBr-activated Sepharose. In fact, all the proteins contained a cheese whey extract had been completely proteolyzed after 6 h at pH 9 and 50°C, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Under these experimental conditions, the immobilized biocatalysts preserve more than 90% of their initial activity after 20 days. Analysis of the three-dimensional (3D) structure of the best immobilized trypsin derivative showed a surface region containing two amino terminal groups and five lysine (Lys) residues that may be responsible for this novel and interesting immobilization and stabilization. Moreover, this region is relatively far from the active site of the enzyme, which could explain the good results obtained for the hydrolysis of high-molecular weight proteins.

  17. Tendon and ligament adaptation to exercise, immobilization, and remobilization.

    PubMed

    Wren, T A; Beaupré, G S; Carter, D R

    2000-01-01

    This study provides a theoretical and computational basis for understanding and predicting how tendons and ligaments adapt to exercise, immobilization, and remobilization. In a previous study, we introduced a model that described the growth and development of tendons and ligaments. In this study, we use the same model to predict changes in the cross-sectional area, modulus, and strength of tendons and ligaments due to increased or decreased loading. The model predictions are consistent with the results of experimental exercise and immobilization studies performed by other investigators. These results suggest that the same fundamental principles guide both development and adaptation. A basic understanding of these principles can contribute both to prevention of tendon and ligament injuries and to more effective rehabilitation when injury does occur.

  18. Plutonium immobilization feed batching system concept report

    SciTech Connect

    Erickson, S.

    2000-07-19

    The Plutonium Immobilization Facility will encapsulate plutonium in ceramic pucks and seal the pucks inside welded cans. Remote equipment will place these cans in magazines and the magazines in a Defense Waste Processing Facility (DWPF) canister. The DWPF will fill the canister with high level waste glass for permanent storage. Feed batching is one of the first process steps involved with first stage plutonium immobilization. It will blend plutonium oxide powder before it is combined with other materials to make pucks. This report discusses the Plutonium Immobilization feed batching process preliminary concept, batch splitting concepts, and includes a process block diagram, concept descriptions, a preliminary equipment list, and feed batching development areas.

  19. Effects and mechanisms of hemopoiesis-stimulating activity of immobilized oligonucleotides under conditions of cytostatic myelosuppression.

    PubMed

    Dygai, A M; Goldberg, V E; Artamonov, A V; Bekarev, A A; Vereschagin, E I; Madonov, P G; Skurikhin, E G; Pershina, O V; Andreeva, T V; Khmelevskaya, E S; Ermakova, N N

    2012-02-01

    Hemopoiesis-stimulating activity of immobilized oligonucleotide preparation was studied on the model of cytostatic myelosuppression induced by injection of cyclophosphamide and 5-fluorouracil. Immobilized oligonucleotides stimulated regeneration of erythro- and granulocytopoiesis in the bone marrow under conditions of cytostatic treatment. The counts of neutrophilic granulocytes and platelets in the peripheral blood increased. The stimulatory effect of the drug was more manifest in animals with active behavior. The mechanism of immobilized oligonucleotide effect was based on stimulation of functional activity of erythroid and granulocytic macrophage precursors.

  20. Experimental joint immobilization in guinea pigs. Effects on the knee joint

    NASA Technical Reports Server (NTRS)

    Marcondesdesouza, J. P.; Machado, F. F.; Sesso, A.; Valeri, V.

    1980-01-01

    In young and adult guinea pigs, the aftermath experimentally induced by the immobilization of the knee joint in hyperextended forced position was studied. Joint immobilization which varied from one to nine weeks was attained by plaster. Eighty knee joints were examined macro and microscopically. Findings included: (1) muscular hypotrophy and joint stiffness in all animals, directly proportional to the length of immobilization; (2) haemoarthrosis in the first week; (3) intra-articular fibrous tissue proliferation ending up with fibrous ankylosis; (4) hyaline articular cartilage erosions; (5) various degrees of destructive menisci changes. A tentative explanation of the fibrous tissue proliferation and of the cartilage changes is offered.

  1. Sulfur-mediated palladium catalyst immobilized on a GaAs surface

    SciTech Connect

    Shimoda, M.; Konishi, T.; Nishiwaki, N.; Yamashita, Y.; Yoshikawa, H.

    2012-06-15

    We present a hard x-ray photoelectron spectroscopy study on the preparation process of palladium catalyst immobilized on an S-terminated GaAs(100) surface. It is revealed that Pd(II) species are reduced on the GaAs surface and yield Pd nanoparticles during the process of Pd immobilization and the subsequent heat treatment. A comparison with the results on GaAs without S-termination suggests that the reduction of Pd is promoted by hydroxy groups during the Pd immobilization and by S during the heat treatment.

  2. Development of a microfluidic “click chip” incorporating an immobilized Cu(I) catalyst

    PubMed Central

    Li, Hairong; Whittenberg, Joseph J.; Zhou, Haiying; Ranganathan, David; Desai, Amit V.; Koziol, Jan; Zeng, Dexing; Kenis, Paul J. A.; Reichert, David E.

    2015-01-01

    We have developed a microfluidic “click chip” incorporating an immobilized Cu(I) catalyst for click reactions. The microfluidic device was fabricated from polydimethylsiloxane (PDMS) bonded to glass and featured ~14,400 posts on the surface to improve catalyst immobilization. This design increased the immobilization efficiency and reduces the reagents’ diffusion time to active catalyst site. The device also incorporates five reservoirs to increase the reaction volume with minimal hydrodynamic pressure drop across the device. A novel water-soluble tris-(benzyltriazolylmethyl)amine (TBTA) derivative capable of stabilizing Cu(I), ligand 2, was synthesized and successfully immobilized on the chip surface. The catalyst immobilized chip surface was characterized by X-ray photoelectron spectroscopy (XPS). The immobilization efficiency was evaluated via radiotracer methods: the immobilized Cu(I) was measured as 1136±272 nmol and the surface immobilized Cu(I) density was 81±20 nmol cm−2. The active Cu(I)-ligand 2 could be regenerated up to five times without losing any catalyst efficiency. The “click” reaction of Flu568-azide and propargylamine was studied on chip for proof-of-principle. The on-chip reaction yields were ca. 82% with a 50 min reaction time or ca. 55% with a 15 min period at 37 °C, which was higher than those obtained in the conventional reaction. The on-chip “click” reaction involving a biomolecule, cyclo(RGDfK) peptide was also studied and demonstrated a conversion yield of ca. 98%. These encouraging results show promise on the application of the Cu(I) catalyst immobilized “click chip” for the development of biomolecule based imaging agents. PMID:25598970

  3. Production of xanthan gum by free and immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii.

    PubMed

    Niknezhad, Seyyed Vahid; Asadollahi, Mohammad Ali; Zamani, Akram; Biria, Davoud

    2016-01-01

    Production of xanthan gum using immobilized cells of Xanthomonas campestris and Xanthomonas pelargonii grown on glucose or hydrolyzed starch as carbon sources was investigated. Calcium alginate (CA) and calcium alginate-polyvinyl alcohol-boric acid (CA-PVA) beads were used for the immobilization of cells. Xanthan titers of 8.2 and 9.2g/L were obtained for X. campestris cells immobilized in CA-PVA beads using glucose and hydrolyzed starch, respectively, whereas those for X. pelargonii were 8 and 7.9 g/L, respectively. Immobilized cells in CA-PVA beads were successfully employed in three consecutive cycles for xanthan production without any noticeable degradation of the beads whereas the CA beads were broken after the first cycle. The results of this study suggested that immobilized cells are advantageous over the free cells for xanthan production. Also it was shown that the cells immobilized in CA-PVA beads are more efficient than cells immobilized in CA beads for xanthan production. PMID:26526173

  4. Immobilization of microbial cell and yeast cell and its application to biomass conversion using radiation techniques

    NASA Astrophysics Data System (ADS)

    Kaetsu, Isao; Kumakura, Minoru; Fujimura, Takashi; Kasai, Noboru; Tamada, Masao

    The recent results of immobilization of cellulase-producing cells and ethanol-fermentation yeast by radiation were reported. The enzyme of cellulase produced by immobilized cells was used for saccharification of lignocellulosic wastes and immobilized yeast cells were used for fermentation reaction from glucose to ethanol. The wastes such as chaff and bagasse were treated by γ-ray or electron-beam irradiation in the presence of alkali and subsequent mechanical crushing, to form a fine powder less than 50 μm in diameter. On the other hand, Trichoderma reesei as a cellulase-producing microbial cell was immobilized on a fibrous carrier having a specific porous structure and cultured to produce cellulase. The enzymatic saccharification of the pretreated waste was carried out using the produced cellulase. The enhanced fermentation process to produce ethanol from glucose with the immobilized yeast by radiation was also studied. The ethanol productivity of immobilized growing yeast cells thus obtained was thirteen times that of free yeast cells in a 1:1 volume of liquid medium to immobilized yeast cells.

  5. Immobilization of tyrosinase on modified diatom biosilica: enzymatic removal of phenolic compounds from aqueous solution.

    PubMed

    Bayramoglu, Gulay; Akbulut, Aydin; Arica, M Yakup

    2013-01-15

    Acid and plasma treated diatom-biosilica particles, were modified with 3-aminopropyl triethoxysilane (APTES), and activated with glutaraldehyde. Then, tyrosinase was immobilized onto the pre-activated biosilica by covalent bonding. The biosilica properties were determined using SEM, and FTIR. The enzyme system has been characterized as a function of pH, temperature and substrate concentration. Optimum pH of the free and immobilized enzyme was found to be pH 7.0. Optimum temperatures of the free and immobilized enzymes were determined as 35 and 45 °C respectively. The biodegradation of phenolic compounds (i.e., phenol, para-cresol and phenyl acetate) has been studied by means of immobilized tyrosinase in a batch system. The immobilized tyrosinase retained about 74% of its original activity after 10 times repeated use in the batch system. Moreover, the storage stability of the tyrosinase-biosilica system resulted excellent, since they maintained more than 67% of the initial activity after eighth week storage. Highly porous structure of biosilica can provide large surface area for immobilization of high quantity enzyme. The porous structure of the biosilica can decrease diffusion limitation both substrate phenols and their products. Finally, the immobilized tyrosinase was used in a batch system for degradation of three different phenols.

  6. Stereospecificity of mushroom tyrosinase immobilized on a chiral and a nonchiral support.

    PubMed

    Marín-Zamora, María Elisa; Rojas-Melgarejo, Francisco; García-Canovas, Francisco; García-Ruiz, Pedro Antonio

    2007-05-30

    Mushroom tyrosinase was immobilized from an extract onto glass beads covered with the cross-linked totally cinnamoylated derivates of d-sorbitol (sorbitol cinnamate) and glycerine (glycerine cinnamate). The enzyme was immobilized onto the support by direct adsorption, and the quantity of immobilized tyrosinase was higher for sorbitol cinnamate, the support with the higher number of esterified hydroxyls per unit of monosacharide, than for glycerine cinnamate. The results obtained from the stereospecificity study of the monophenolase and diphenolase activity of immobilized mushroom tyrosinase are reported. The enantiomers L-tyrosine, DL-tyrosine, D-tyrosine, L-dopa, DL-dopa, D-dopa, L-alpha-methyldopa, DL-alpha-methyldopa, L-isoprenaline, DL-isoprenaline, L-adrenaline, DL-adrenaline, L-noradrenaline, and D-noradrenaline were assayed with tyrosinase immobilized on a chiral support (sorbitol cinnamate), whereas L-tyrosine, DL-tyrosine, D-tyrosine, L-dopa, DL-dopa, D-dopa, L-alpha-methyldopa, and DL-alpha-methyldopa were assayed with tyrosinase immobilized on a nonchiral support (glycerine cinnamate). The same Vmax(app) values for each series of enantiomers were obtained. However, the Km(app) values were different, the l isomers showing lower values than the dl isomers, whereas the highest Km(app) value was obtained with d isomers. No difference was observed in the stereospecificity of tyrosinase immobilized on a chiral (sorbitol cinnamate) or nonchiral (glycerine cinnamate) support. PMID:17488091

  7. Arsenic immobilization in the contaminated soil using poorly crystalline Fe-oxyhydroxy sulfate.

    PubMed

    Yang, Zhihui; Liu, Lin; Chai, Liyuan; Liao, Yingping; Yao, Wenbin; Xiao, Ruiyang

    2015-08-01

    A low crystalline Fe-oxyhydroxy sulfate (FeOS) was used to immobilize arsenic (As) in soils in this study. The effects of FeOS amount, treatment time and soil moisture on As immobilization were investigated. The results showed that water-soluble and NaHCO3-extractable As were immobilized by 53.4-99.8 and 13.8-73.3% respectively, with 1-10% of FeOS addition. The highest immobilization of water-soluble (98.5%) and NaHCO3-extractable arsenic (47.2%) was achieved under condition of 4% of FeOS and 80% of soil moisture. Further, more amounts of FeOS addition resulted in less time requirement for As immobilization. Sequential chemical extraction experiment revealed that easily mobile arsenic phase was transferred to less mobile phase. The FeOS-bonded As may play a significant role in arsenic immobilization. Under leaching with simulated acid rain at 60 times pore volumes, accumulation amount of As release from untreated soil and soil amended with FeOS were 98.4 and 1.2 mg, respectively, which correspond to 7.69 and 0.09% of total As amounts in soil. The result showed that the low crystalline FeOS can be used as a suitable additive for arsenic immobilization in soils.

  8. Degradation of h-acid by free and immobilized cells of Alcaligenes latus

    PubMed Central

    Usha, M.S.; Sanjay, M.K.; Gaddad, S.M.; Shivannavar, C.T.

    2010-01-01

    Alcaligenes latus, isolated from industrial effluent, was able to grow in mineral salts medium with 50 ppm (0.15 mM) of H-acid as a sole source of carbon. Immobilization of Alcaligenes latus in Ca-alginate and polyurethane foam resulted in cells embedded in the matrices. When free cells and immobilized cells were used for biodegradation studies at concentration ranging from 100 ppm (0.3 mM) to 500 ppm (1.15 mM) degradation rate was enhanced with immobilized cells. Cells immobilized in polyurethane foam showed 100% degradation up to 350 ppm (1.05 mM) and 57% degradation at 500 ppm (1.5 mM). Degradation rate of Ca-alginate immobilized cells was less as compared to that of polyurethane foam immobilized cells. With Ca-alginate immobilized cells 100% degradation was recorded up to 200 ppm (0.6 mM) of H-acid and only 33% degradation was recorded at 500 ppm (1.5 mM) of H-acid. Spectral analysis of the products after H-acid utilization showed that the spent medium did not contain any aromatic compounds indicating H-acid degradation by A. latus. PMID:24031573

  9. Assessment of the ecological security of immobilized enzyme remediation process with biological indicators of soil health.

    PubMed

    Zhang, Ying; Dong, Xiaonan; Jiang, Zhao; Cao, Bo; Ge, Shijie; Hu, Miao

    2013-08-01

    This study used the enzymes extracted from an atrazine-degrading strain, Arthrobacter sp. DNS10, which had been immobilized by sodium alginate to rehabilitate atrazine-polluted soil. Meanwhile, a range of biological indices were selected to assess the ecological health of contaminated soils and the ecological security of this bioremediation method. The results showed that there was no atrazine detected in soil samples after 28 days in EN+AT (the soil containing atrazine and immobilized enzyme) treatment. However, the residual atrazine concentration of the sample in AT (the soil containing atrazine only) treatment was about 5.02 ± 0.93 mg kg(-1). These results suggest that the immobilized enzyme exhibits an excellent ability in atrazine degradation. Furthermore, the immobilized enzyme could relieve soil microbial biomass carbon and soil microbial respiration intensity to 772.33 ± 34.93 mg C kg(-1) and 5.01 ± 0.17 mg CO(2) g(-1) soil h(-1), respectively. The results of the polymerase chain reaction-degeneration gradient gel electrophoresis experiment indicated that the immobilized enzyme also could make the Shannon-Wiener index and evenness index of the soil sample increase from 1.02 and 0.74 to 1.51 and 0.84, respectively. These results indicated that the immobilized enzymes not only could relieve the impact from atrazine on the soil, but also revealed that the immobilized enzymes did no significant harm on the soil ecological health.

  10. Enhanced starch hydrolysis using α-amylase immobilized on cellulose ultrafiltration affinity membrane.

    PubMed

    Konovalova, Viktoriia; Guzikevich, Kateryna; Burban, Anatoliy; Kujawski, Wojciech; Jarzynka, Karolina; Kujawa, Joanna

    2016-11-01

    In order to prepare ultrafiltration membranes possessing biocatalytic properties, α-amylase has been immobilized on cellulose membranes. Enzyme immobilization was based on a covalent bonding between chitosan and a surface of cellulose membrane, followed by an attachment of Cibacron Blue F3G-A dye as affinity ligand. Various factors affecting the immobilization process, such as enzyme concentration, pH of modifying solution, zeta-potential of membrane surface, and stability of immobilized enzyme were studied. The applicability of immobilized α-amylase has been investigated in ultrafiltration processes. The immobilization of α-amylase on membrane surface allows to increase the value of mass transfer coefficient and to decrease the concentration polarization effect during ultrafiltration of starch solutions. The enzyme layer on the membrane surface prevents a rapid increase of starch concentration due to the amylase hydrolysis of starch in the boundary layer. The presented affinity immobilization technique allows also for the regeneration of membranes from inactivated enzyme. PMID:27516322

  11. Stereospecificity of mushroom tyrosinase immobilized on a chiral and a nonchiral support.

    PubMed

    Marín-Zamora, María Elisa; Rojas-Melgarejo, Francisco; García-Canovas, Francisco; García-Ruiz, Pedro Antonio

    2007-05-30

    Mushroom tyrosinase was immobilized from an extract onto glass beads covered with the cross-linked totally cinnamoylated derivates of d-sorbitol (sorbitol cinnamate) and glycerine (glycerine cinnamate). The enzyme was immobilized onto the support by direct adsorption, and the quantity of immobilized tyrosinase was higher for sorbitol cinnamate, the support with the higher number of esterified hydroxyls per unit of monosacharide, than for glycerine cinnamate. The results obtained from the stereospecificity study of the monophenolase and diphenolase activity of immobilized mushroom tyrosinase are reported. The enantiomers L-tyrosine, DL-tyrosine, D-tyrosine, L-dopa, DL-dopa, D-dopa, L-alpha-methyldopa, DL-alpha-methyldopa, L-isoprenaline, DL-isoprenaline, L-adrenaline, DL-adrenaline, L-noradrenaline, and D-noradrenaline were assayed with tyrosinase immobilized on a chiral support (sorbitol cinnamate), whereas L-tyrosine, DL-tyrosine, D-tyrosine, L-dopa, DL-dopa, D-dopa, L-alpha-methyldopa, and DL-alpha-methyldopa were assayed with tyrosinase immobilized on a nonchiral support (glycerine cinnamate). The same Vmax(app) values for each series of enantiomers were obtained. However, the Km(app) values were different, the l isomers showing lower values than the dl isomers, whereas the highest Km(app) value was obtained with d isomers. No difference was observed in the stereospecificity of tyrosinase immobilized on a chiral (sorbitol cinnamate) or nonchiral (glycerine cinnamate) support.

  12. Arsenic immobilization in the contaminated soil using poorly crystalline Fe-oxyhydroxy sulfate.

    PubMed

    Yang, Zhihui; Liu, Lin; Chai, Liyuan; Liao, Yingping; Yao, Wenbin; Xiao, Ruiyang

    2015-08-01

    A low crystalline Fe-oxyhydroxy sulfate (FeOS) was used to immobilize arsenic (As) in soils in this study. The effects of FeOS amount, treatment time and soil moisture on As immobilization were investigated. The results showed that water-soluble and NaHCO3-extractable As were immobilized by 53.4-99.8 and 13.8-73.3% respectively, with 1-10% of FeOS addition. The highest immobilization of water-soluble (98.5%) and NaHCO3-extractable arsenic (47.2%) was achieved under condition of 4% of FeOS and 80% of soil moisture. Further, more amounts of FeOS addition resulted in less time requirement for As immobilization. Sequential chemical extraction experiment revealed that easily mobile arsenic phase was transferred to less mobile phase. The FeOS-bonded As may play a significant role in arsenic immobilization. Under leaching with simulated acid rain at 60 times pore volumes, accumulation amount of As release from untreated soil and soil amended with FeOS were 98.4 and 1.2 mg, respectively, which correspond to 7.69 and 0.09% of total As amounts in soil. The result showed that the low crystalline FeOS can be used as a suitable additive for arsenic immobilization in soils. PMID:25911284

  13. Enzymatic removal of paracetamol from aqueous phase: horseradish peroxidase immobilized on nanofibrous membranes.

    PubMed

    Xu, Ran; Si, Yifang; Li, Fengting; Zhang, Bingru

    2015-03-01

    Paracetamol is a widely used as an analgesic and an antipyretic that can easily accumulate in aquatic environments. This study aimed to enhance paracetamol removal efficiency from water by combining the biocatalytic activity of horseradish peroxidase (HRP) with the adsorption of nanofibrous membrane. Poly(vinyl alcohol)/poly(acrylic acid)/SiO2 electrospinning nanofibrous membrane was prepared with fiber diameters of 200 to 300 nm. The membrane was made insoluble by the thermal cross-linking process. HRP, which was previously activated by 1,1'-carbonyldiimidazole, was covalently immobilized on the surface of nanofibers. Immobilized HRP retained 79.4 % of the activity of free HRP. The physical, chemical, and biochemical properties of the immobilized HRP and its application in paracetamol removal were comprehensively investigated. Immobilized HRP showed better storage capability and higher tolerance to the changes in pH and temperature than free HRP. Paracetamol removal rate by immobilized HRP (83.5 %) was similar to that of free HRP (84.4 %), but immobilized HRP showed excellent reusability. The results signify that enzyme immobilized on nanofibers has great application potential in water treatment.

  14. Immobilization of Bacillus amyloliquefaciens MBL27 cells for enhanced antimicrobial protein production using calcium alginate beads.

    PubMed

    Kumaravel, Vijayalakshmi; Gopal, Suseela Rajakumar

    2010-12-01

    Cell immobilization is one of the common techniques for increasing the overall cell concentration and productivity. Bacillus amyloliquefaciens MBL27 cells were immobilized in calcium alginate beads and it is a promising method for repeated AMP (antimicrobial protein) production. The present study aimed at determining the optimal conditions for immobilization of B. amyloliquefaciens MBL27 cells in calcium alginate beads and the operational stability for enhanced production of the AMP. AMP production with free and immobilized cells was also done. In batch fermentation, maximum AMP production (7300 AU (arbitrary units)/ml against Staphylococcus aureus) was obtained with immobilized cells in shake flasks under optimized parameters such as 3% (w/v) sodium alginate, 136 mM CaCl2 with 350 alginate beads/flask of 2.7-3.0 mm diameter. In repeated cultivation, the highest activity was obtained after the second cycle of use and approx. 94% production was noted up to the fifth cycle. The immobilized cells of B. amyloliquefaciens MBL27 in alginate beads are more efficient for the production of AMP and had good stability. The potential application of AMP as a wound healant and the need for development of economical methods for improved production make whole cell immobilization an excellent alternative method for enhanced AMP production.

  15. Site-specific immobilization of biotinylated proteins for protein microarray analysis.

    PubMed

    Lue, Rina Y P; Chen, Grace Y J; Zhu, Qing; Lesaicherre, Marie-Laure; Yao, Shao Q

    2004-01-01

    The postgenome era has led to a new frontier of proteomics that requires the development of protein microarray, which enables us to unravel the biological function of proteins in a massively parallel fashion. Several ways of immobilizing proteins onto surfaces have been reported, but many of these attachments are unspecific, resulting in the unfavorable orientation of the immobilized proteins. His6 tag has been used to site-specifically immobilize proteins onto nickel-coated slides, which presumably oriented proteins uniformly on the surface of the slide. However, the binding between Ni-NTA and His tag proteins is not strong, causing the immobilized proteins to dissociate from the slide even under simple wash conditions. The authors have developed a novel strategy of using an intein-mediated expression system to generate biotinylated proteins suitable for immobilization onto avidin-functionalized glass slides. Array-scan results not only show successful immobilization of proteins onto slides by antibody detection method but also full retention of biological activities of the immobilized proteins. The strong and specific interaction between biotin and avidin also permits the use of stringent incubation and washing conditions on the protein microchip, thus making it a highly robust method for array studies.

  16. Evaluation of free and immobilized Aspergillus niger NRC1ami pectinase applicable in industrial processes.

    PubMed

    Esawy, Mona A; Gamal, Amira A; Kamel, Zeinat; Ismail, Abdel-Mohsen S; Abdel-Fattah, Ahmed F

    2013-02-15

    The Aspergillus niger NRC1ami pectinase was evaluated according to its hydrolysis efficiency of dry untreated orange peels (UOP), HCl-treated orange peels and NaOH-treated orange peels (HOP and NOP). Pectinase was entrapped in polyvinyl alcohol (PVA) sponge and the optimum pH and temperature of the free and immobilized enzymes were shifted from 4, 40 °C to 6, 50 °C respectively. The study of pH stability of free and immobilized pectinase showed that the immobilization process protected the enzyme strongly from severe alkaline pHs. The immobilization process improved the enzyme thermal stability to great instant. The unique feature of the immobilization process is its ability to solve the orange juice haze problem completely. Immobilized enzyme was reused 12 times in orange juice clarification with 9% activity loss from the original activity. Maximum reaction rate (V(max)) and Michaelis-Menten constant (K(m)) of the partially purified form were significantly changed after immobilization.

  17. Novel functionalized fluorescent polymeric nanoparticles for immobilization of biomolecules

    NASA Astrophysics Data System (ADS)

    Jain, Swati; Chattopadhyay, Sruti; Jackeray, Richa; Abid, C. K. V. Zainul; Singh, Harpal

    2013-07-01

    Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable, monodisperse, spherical nano sized particles exhibiting high luminescence properties. Particles with 1% SLS (S1) showed good dispersion stability and fluorescence intensity and were chosen as ideal candidates for further immobilization studies. Steady state fluorescence studies showed 10 times higher fluorescence intensity of S1 nanoparticles than that of pyrene solution in solvent-toluene at the same concentration. Environmental factors such as pH, ionic strength and time were found to have no effect on fluorescence intensity of FPNPs. Surface β-di-ketone groups were utilized for the covalent immobilization of enzyme conjugated antibodies without any activation or pre-treatment of nanoparticles.Novel, size controlled fluorescent polymeric nanoparticles (FPNP) were synthesized having acetoacetoxy functionality on the surface for immobilization of biomolecules which can be utilized as biomarkers and labels in fluoroimmunoassays. Core-shell nanoparticles of poly(styrene, St-methyl methacrylate, MMA-acetoacetoxy ethyl methacrylate, AAEM), stabilized by various concentrations of surfactant, sodium lauryl sulphate (SLS), were obtained by facile miniemulsion co-polymerization encapsulated with pyrene molecules in their hydrophobic core. Analytical, spectroscopic and imaging characterization techniques revealed the formation of stable

  18. Simultaneous and sequential co-immobilization of glucose oxidase and catalase onto florisil.

    PubMed

    Ozyilmaz, Gul; Tukel, S Seyhan

    2007-06-01

    The co-immobilization of Aspergillus niger glucose oxidase (GOD) with bovine liver catalase (CAT) onto florisil (magnesium silicate-based porous carrier) was investigated to improve the catalytic efficiency of GOD against H2O2 inactivation. The effect of the amount of bound CAT on the GOD activity was also studied for 12 different initial combinations of GOD and CAT, using simultaneous and sequential coupling. The sequentially co-immobilized GOD-CAT showed a higher efficiency than the simultaneously co-immobilized GOD-CAT in terms of the GOD activity and economic costs. The highest activity was shown by the sequentially co-immobilized GOD-CAT when the initial amounts of GOD and CAT were 10 mg and 5 mg per gram of carrier. The optimum pH, buffer concentration, and temperature for GOD activity for the same co-immobilized GOD-CAT sample were then determined as pH 6.5, 50 mM, and 30 degrees C, respectively. When compared with the individually immobilized GOD, the catalytic activity of the co-immobilized GOD-CAT was 70% higher, plus the reusability was more than two-fold. The storage stability of the co-immobilized GOD-CAT was also found to be higher than that of the free form at both 5 degrees C and 25 degrees C. The increased GOD activity and reusability resulting from the co-immobilization process may have been due to CAT protecting GOD from inactivation by H2O2 and supplying additional O2 to the reaction system. PMID:18050914

  19. Pigeonpea (Cajanus cajan L.) urease immobilized on glutaraldehyde-activated chitosan beads and its analytical applications.

    PubMed

    Kayastha, A M; Srivastava, P K

    2001-01-01

    Urease from pigeonpea (Cajanus cajan L.) was covalently linked to crab shell chitosan beads using glutaraldehyde. The optimum immobilization (64% activity) was observed at 4 degrees C, with a protein concentration of 0.24 mg/bead and 3% glutaraldehyde. The immobilized enzyme stored in 0.05 M Tris-acetate buffer, pH 7.3, at 4 degrees C had a t(1/2) of 110 d. There was practically no leaching of enzyme (<3%) from the immobilized beads in 30 d. The immobilized urease was used 10 times at an interval of 24 h between each use with 80% residual activity at the end of the period. The chitosan-immobilized urease showed a significantly higher Michaelis constant (8.3 mM) compared to that of the soluble urease (3.0 mM). Its apparent optimum pH also shifted from 7.3 to 8.5. Immobilized urease showed an optimal temperature of 77 degrees C, compared with 47 degrees C for the soluble urease. Time-dependent kinetics of the thermal denaturation of immobilized urease was studied and found to be monophasic in nature compared to biphasic in nature for soluble enzyme. This immobilized urease was used to analyze blood urea of some of the clinical samples from the clinical pathology laboratories. The results compared favorably with those obtained by the various chemical/biochemical methods employed in the clinical pathology laboratories. A column packed with immobilized urease beads was also prepared in a syringe for the regular and continuous monitoring of serum urea concentrations.

  20. Immobilization of Rocky Flats Graphite Fines Residue

    SciTech Connect

    Rudisill, T.S.

    1999-04-06

    The development of the immobilization process for graphite fines has proceeded through a series of experimental programs. The experimental procedures and results from each series of experiments are discussed in this report.

  1. Plutonium Immobilization Bagless Transfer Can Size Evaluation

    SciTech Connect

    Kriikku, E.; Stokes, M.; Rogers, L.; Ward, C.

    1998-02-01

    This report identifies and documents the most appropriate bagless transfer can size to support Plutonium Immobilization Can Loading operations. Also, this report considers can diameter, can wall thickness, and can length.

  2. Immobilization Technologies in Probiotic Food Production

    PubMed Central

    Mitropoulou, Gregoria; Nedovic, Viktor; Goyal, Arun; Kourkoutas, Yiannis

    2013-01-01

    Various supports and immobilization/encapsulation techniques have been proposed and tested for application in functional food production. In the present review, the use of probiotic microorganisms for the production of novel foods is discussed, while the benefits and criteria of using probiotic cultures are analyzed. Subsequently, immobilization/encapsulation applications in the food industry aiming at the prolongation of cell viability are described together with an evaluation of their potential future impact, which is also highlighted and assessed. PMID:24288597

  3. Ceramification: A plutonium immobilization process

    SciTech Connect

    Rask, W.C.; Phillips, A.G.

    1996-05-01

    This paper describes a low temperature technique for stabilizing and immobilizing actinide compounds using a combination process/storage vessel of stainless steel, in which measured amounts of actinide nitrate solutions and actinide oxides (and/or residues) are systematically treated to yield a solid article. The chemical ceramic process is based on a coating technology that produces rare earth oxide coatings for defense applications involving plutonium. The final product of this application is a solid, coherent actinide oxide with process-generated encapsulation that has long-term environmental stability. Actinide compounds can be stabilized as pure materials for ease of re-use or as intimate mixtures with additives such as rare earth oxides to increase their degree of proliferation resistance. Starting materials for the process can include nitrate solutions, powders, aggregates, sludges, incinerator ashes, and others. Agents such as cerium oxide or zirconium oxide may be added as powders or precursors to enhance the properties of the resulting solid product. Additives may be included to produce a final product suitable for use in nuclear fuel pellet production. The process is simple and reduces the time and expense for stabilizing plutonium compounds. It requires a very low equipment expenditure and can be readily implemented into existing gloveboxes. The process is easily conducted with less associated risk than proposed alternative technologies.

  4. Immobilization of Fast Reactor First Cycle Raffinate

    SciTech Connect

    Langley, K. F.; Partridge, B. A.; Wise, M.

    2003-02-26

    This paper describes the results of work to bring forward the timing for the immobilization of first cycle raffinate from reprocessing fuel from the Dounreay Prototype Fast Reactor (PFR). First cycle raffinate is the liquor which contains > 99% of the fission products separated from spent fuel during reprocessing. Approximately 203 m3 of raffinate from the reprocessing of PFR fuel is held in four tanks at the UKAEA's site at Dounreay, Scotland. Two methods of immobilization of this high level waste (HLW) have been considered: vitrification and cementation. Vitrification is the standard industry practice for the immobilization of first cycle raffinate, and many papers have been presented on this technique elsewhere. However, cementation is potentially feasible for immobilizing first cycle raffinate because the heat output is an order of magnitude lower than typical HLW from commercial reprocessing operations such as that at the Sellafield site in Cumbria, England. In fact, it falls within the upper end of the UK definition of intermediate level waste (ILW). Although the decision on which immobilization technique will be employed has yet to be made, initial development work has been undertaken to identify a suitable cementation formulation using inactive simulant of the raffinate. An approach has been made to the waste disposal company Nirex to consider the disposability of the cemented product material. The paper concentrates on the process development work that is being undertaken on cementation to inform the decision making process for selection of the immobilization method.

  5. Surface cell immobilization within perfluoroalkoxy microchannels

    NASA Astrophysics Data System (ADS)

    Stojkovič, Gorazd; Krivec, Matic; Vesel, Alenka; Marinšek, Marjan; Žnidaršič-Plazl, Polona

    2014-11-01

    Perfluoroalkoxy (PFA) is one of the most promising materials for the fabrication of cheap, solvent resistant and reusable microfluidic chips, which have been recently recognized as effective tools for biocatalytic process development. The application of biocatalysts significantly depends on efficient immobilization of enzymes or cells within the reactor enabling long-term biocatalyst use. Functionalization of PFA microchannels by 3-aminopropyltriethoxysilane (ATPES) and glutaraldehyde was used for rapid preparation of microbioreactors with surface-immobilized cells. X-ray photoelectron spectroscopy and scanning electron microscopy were used to accurately monitor individual treatment steps and to select conditions for cell immobilization. The optimized protocol for Saccharomyces cerevisiae immobilization on PFA microchannel walls comprised ethanol surface pretreatment, 4 h contacting with 10% APTES aqueous solution, 10 min treatment with 1% glutaraldehyde and 20 min contacting with cells in deionized water. The same protocol enabled also immobilization of Escherichia coli, Pseudomonas putida and Bacillus subtilis cells on PFA surface in high densities. Furthermore, the developed procedure has been proved to be very efficient also for surface immobilization of tested cells on other materials that are used for microreactor fabrication, including glass, polystyrene, poly (methyl methacrylate), polycarbonate, and two olefin-based polymers, namely Zeonor® and Topas®.

  6. Herbaceous plants as filters: immobilization of particulates along urban street corridors.

    PubMed

    Weber, Frauke; Kowarik, Ingo; Säumel, Ina

    2014-03-01

    Among air pollutants, particulate matter (PM) is considered to be the most serious threat to human health. Plants provide ecosystem services in urban areas, including reducing levels of PM by providing a surface for deposition and immobilization. While previous studies have mostly addressed woody species, we focus on herbaceous roadside vegetation and assess the role of species traits such as leaf surface roughness or hairiness for the immobilization of PM. We found that PM deposition patterns on plant surfaces reflect site-specific traffic densities and that strong differences in particulate deposition are present among species. The amount of immobilized PM differed according to particle type and size and was related to specific plant species traits. Our study suggests that herbaceous vegetation immobilizes a significant amount of the air pollutants relevant to human health and that increasing biodiversity of roadside vegetation supports air filtration and thus healthier conditions along street corridors.

  7. Histological and compositional responses of bone to immobilization and other experimental conditions

    NASA Technical Reports Server (NTRS)

    Brown, R. J.; Niklowitz, W. J.

    1985-01-01

    Histological techniques were utilized for evaluating progressive changes in tibial compact bone in adult male monkeys during chronic studies of immobilization-associated osteopenia. The animals were restrained in a semirecumbent position which reduces normally occurring stresses in the lower extremities and results in bone mass loss. The longest immobilization studies were of seven months duration. Losses of haversian bone tended to occur predominatly in the proximal tibia and were characterized by increased activation with excessive depth of penetration of osteoclastic activity. There was no apparent regulation of the size and orientation of resorption cavities. Rapid bone loss seen during 10 weeks of immobilization appeared to be due to unrestrained osteoclastic activity without controls and regulation which are characteristic of adaptive systems. The general pattern of loss persisted throughout 7 months of immobilization. Clear cut evidence of a formation phase in haversian bone was seen only after two months of reambulation.

  8. Reversible immobilization of BSA on Cu-chelated PAMAM dendrimer modified iron oxide nanoparticles

    NASA Astrophysics Data System (ADS)

    Demir, M.; Şenel, M.; Baykal, A.

    2014-09-01

    In this study, polyamidoamine (PAMAM) dendrimer coated superparamagnetite nanoparticles were synthesized by growing of PAMAM on amino-silane coated iron oxide nanoparticles. The PAMAM modified superparamagnetite nanoparticles were used as reversible protein immobilization host materials. During the reversible immobilization studies the effect of different metal ions such as; Cu+2, Zn+2, Co+2, Ni+2 on immobilization efficiency of BSA were evaluated. The maximum BSA adsorption capacity of the PAMAM-MNP- Cu+2 beads was observed to be 52.84 mg/g (BSA/PAMAM-MNP) at pH 7.0. Various characteristics of immobilized BSA such as; effect of generation, effect of pH, BSA concentration, temperature, salt concentration and reusability of PAMAM-MNP were evaluated.

  9. Semi-permeable nanocapsules of konjac glucomannan-chitosan for enzyme immobilization.

    PubMed

    Wang, Rui; Xia, Bing; Li, Bang-Jing; Peng, Shu-Lin; Ding, Li-Sheng; Zhang, Sheng

    2008-11-19

    Carboxymethyl konjac glucomannan-chitosan (CKGM-CS) nanocapsules, spontaneously prepared under very mild conditions by electrostatic complexation, were used for immobilizing L-asparaginase. The matrix has semi-permeability to allow the substrate and product to pass through and to keep L-asparaginase in the matrix to prevent leaking. The cell-like hydrogel matrix was prepared in aqueous system without organic solvents and reagents. The process of the preparation does not denature the enzyme and the activity of the immobilized and native enzyme is very similar. The activity, stability, and characters of the enzyme-loaded nanocapsules were studied. The results indicated the immobilized enzyme has better stability and activity in contrast to the native enzyme. These studies may supply a new material for the immobilization of pH and temperature-sensitive enzyme. PMID:18725277

  10. Development of 170 MHz Electrodeless Quartz-Crystal Microbalance Immunosensor with Nonspecifically Immobilized Receptor Proteins

    NASA Astrophysics Data System (ADS)

    Hirotsugu Ogi,; Hironao Nagai,; Yuji Fukunishi,; Taiji Yanagida,; Masahiko Hirao,; Masayoshi Nishiyama,

    2010-07-01

    Staphylococcus aureus protein A (SPA) shows high nonspecific binding affinity on a naked quartz surface, and it can be used as the receptor protein for detecting immunoglobulin G (IgG), the most important immunoglobulin. The immunosensor ability, however, significantly depends on the immobilization procedure. In this work, the effect of the nonspecific immobilization procedure on the sensor sensitivity is studied using a home-built electrodeless quartz-crystal microbalance (QCM) biosensor. The pure-shear vibration of a 9.7-μm-thick AT-cut quartz plate is excited and detected in liquids by the line antenna located outside the flow channel. SPA molecules are immobilized on the quartz surfaces, and human IgG is injected to monitor the binding reaction between SPA and IgG. This study reveals that a long (nearly 24 h) immersion procedure is required for immobilizing SPA to achieve the tight biding with the quartz surfaces.

  11. Development of 170 MHz Electrodeless Quartz-Crystal Microbalance Immunosensor with Nonspecifically Immobilized Receptor Proteins

    NASA Astrophysics Data System (ADS)

    Ogi, Hirotsugu; Nagai, Hironao; Fukunishi, Yuji; Yanagida, Taiji; Hirao, Masahiko; Nishiyama, Masayoshi

    2010-07-01

    Staphylococcus aureus protein A (SPA) shows high nonspecific binding affinity on a naked quartz surface, and it can be used as the receptor protein for detecting immunoglobulin G (IgG), the most important immunoglobulin. The immunosensor ability, however, significantly depends on the immobilization procedure. In this work, the effect of the nonspecific immobilization procedure on the sensor sensitivity is studied using a home-built electrodeless quartz-crystal microbalance (QCM) biosensor. The pure-shear vibration of a 9.7-µm-thick AT-cut quartz plate is excited and detected in liquids by the line antenna located outside the flow channel. SPA molecules are immobilized on the quartz surfaces, and human IgG is injected to monitor the binding reaction between SPA and IgG. This study reveals that a long (nearly 24 h) immersion procedure is required for immobilizing SPA to achieve the tight biding with the quartz surfaces.

  12. Simulation and analysis of immobilized-cell fermentors

    SciTech Connect

    Heyman, E.C.; Nguyen, X.N.

    1982-06-01

    Immobilized cell bioreactors are important for developing high-productivity fermentation processes. A computer simulation program has been devised to aid in the design of three basic fermentor types including batch, tubular, and continuous stirred tank and the analysis of important process variables. Parametric studies on the sensitivity of fermentor residence time for the glucose to ethanol yeast fermentation system with changes kinetic constants, film/pore diffusion limitations, gel support characteristics and recycle of product flow are discussed.

  13. Immobilization of Proteins on a Glass Surface at High Density

    SciTech Connect

    Thomas, Marlon; Vullev, Valentine I.; Wan Jiandi

    2009-07-06

    We describe a rational molecular-level design of biocompatible surface coatings and immobilization of biological species onto them to produce biofunctional interfaces. Our method adapted a strategy for coating glass and other silica-type substrates with bioinert layers of polyethylene glycol (PEG). The introduction of {alpha}, {omega}-bifunctional polymers into the coatings allowed for covalent attachment of proteins to the PEGylated surfaces. Spectroscopic studies indicate that the surface-bound proteins had their biological activity preserved.

  14. [Immobilization and skeletal system of the human body].

    PubMed

    Kisała, Aleksander; Pluskiewicz, Wojciech

    2015-01-01

    Shaping the process of evolution musculoskeletal and nervous systems in animals has allowed these organisms steady increase mobility and mastery of new environments to life. Movement is the essence of life and health. But health is not a permanent condition. Its absence often results in limited mobility of the body. The aim of this study is to assess the impact of immobilization on the state of the skeletal system and the evaluation of the effectiveness of various measures to reduce this impact.

  15. Immobilization of uranium in contaminated soil by natural apatite addition

    SciTech Connect

    Mrdakovic Popic, Jelena; Stojanovic, Mirjana; Milosevic, Sinisa; Iles, Deana; Zildzovic, Snezana

    2007-07-01

    Available in abstract form only. Full text of publication follows: The goal of this study was to evaluate the effectiveness of Serbian natural mineral apatite as soil additive for reducing the migration of uranium from contaminated sediments. In laboratory study we investigated the sorption properties of domestic apatite upon different experimental conditions, such as pH, adsorbent mass, reaction period, concentration of P{sub 2}O{sub 5} in apatite, solid/liquid ratio. In second part of study, we did the quantification of uranium in soil samples, taken from uranium mine site 'Kalna', by sequential extraction method. The same procedure was, also, used for uranium determination in contaminated soil samples after apatite addition, in order to determine the changes in U distribution in soil fraction. The obtained results showed the significant level of immobilization (96.7%) upon certain conditions. Increase of %P{sub 2}O{sub 5} in apatite and process of mechano-chemical activation led to increase of immobilization capacity from 17.50% till 91.64%. The best results for uranium binding were obtained at pH 5.5 and reaction period 60 days (98.04%) The sequential extraction showed the presence of uranium (48.2%) in potentially available soil fractions, but with the apatite addition uranium content in these fractions decreased (30.64%), what is considering environmental aspect significant fact. In situ immobilization of radionuclide using inexpensive sequestering agents, such as apatite, is very adequate for big contaminated areas of soil with low level of contamination. This investigation study on natural apatite from deposit 'Lisina' Serbia was the first one of this type in our country. Key words: apatite, uranium, immobilization, soil, contamination. (authors)

  16. Immobilized high-level waste interim storage alternatives generation and analysis and decision report

    SciTech Connect

    CALMUS, R.B.

    1999-05-18

    This report presents a study of alternative system architectures to provide onsite interim storage for the immobilized high-level waste produced by the Tank Waste Remediation System (TWRS) privatization vendor. It examines the contract and program changes that have occurred and evaluates their impacts on the baseline immobilized high-level waste (IHLW) interim storage strategy. In addition, this report documents the recommended initial interim storage architecture and implementation path forward.

  17. [Effect of the Hydrogel Carrier Structure on the Activity of Immobilized Trypsin].

    PubMed

    Valuev, L I; Valuev, I L; Vanchugova, L V; Valueva, T A

    2015-01-01

    The dependence of the activity of trypsin immobilized in polyacrylamide hydrogel on the hydrogel swelling ratio, the size distribution of its pores, and the means of enzyme binding has been studied. It has been shown that the most favorable conditions for immobilized trypsin are provided upon its binding to hydrogel via trypsin macromonomer copolymerization with acrylamide and a linking agent in the presence of a modifier that limits polymer chain growth. PMID:26596090

  18. Immobilization of a Pleurotus ostreatus laccase mixture on perlite and its application to dye decolourisation.

    PubMed

    Pezzella, Cinzia; Russo, Maria Elena; Marzocchella, Antonio; Salatino, Piero; Sannia, Giovanni

    2014-01-01

    In the present study, a crude laccase preparation from Pleurotus ostreatus was successfully immobilized on perlite, a cheap porous silica material, and tested for Remazol Brilliant Blue R (RBBR) decolourisation in a fluidized bed recycle reactor. Results showed that RBBR decolourisation is mainly due to enzyme action despite the occurrence of dye adsorption-related enzyme inhibition. Fine tuning of immobilization conditions allowed balancing the immobilization yield and the resulting rate of decolourisation, with the adsorption capacity of the solid biocatalyst. In the continuous lab scale reactor, a maximum conversion degree of 56.1% was achieved at reactor space-time of 4.2 h. Stability and catalytic parameters of the immobilized laccases were also assessed in comparison with the soluble counterparts, revealing an increase in stability, despite a reduction of the catalytic performances. Both effects are most likely ascribable to the occurrence of multipoint attachment phenomena. PMID:24895564

  19. Developing an Efficient and General Strategy for Immobilization of Small Molecules onto Microarrays Using Isocyanate Chemistry.

    PubMed

    Zhu, Chenggang; Zhu, Xiangdong; Landry, James P; Cui, Zhaomeng; Li, Quanfu; Dang, Yongjun; Mi, Lan; Zheng, Fengyun; Fei, Yiyan

    2016-01-01

    Small-molecule microarray (SMM) is an effective platform for identifying lead compounds from large collections of small molecules in drug discovery, and efficient immobilization of molecular compounds is a pre-requisite for the success of such a platform. On an isocyanate functionalized surface, we studied the dependence of immobilization efficiency on chemical residues on molecular compounds, terminal residues on isocyanate functionalized surface, lengths of spacer molecules, and post-printing treatment conditions, and we identified a set of optimized conditions that enable us to immobilize small molecules with significantly improved efficiencies, particularly for those molecules with carboxylic acid residues that are known to have low isocyanate reactivity. We fabricated microarrays of 3375 bioactive compounds on isocyanate functionalized glass slides under these optimized conditions and confirmed that immobilization percentage is over 73%. PMID:26999137

  20. Production of gluconic acid by Aspergillus niger immobilized on polyurethane foam.

    PubMed

    Vassilev, N B; Vassileva, M C; Spassova, D I

    1993-06-01

    Production of gluconic acid by cells of Aspergillus niger immobilized on polyurethane foam was studied in repeated-batch shake-flask and bubble-column fermentations. For passive immobilization, various amounts of polyurethane foam and spore suspension were tested in order to obtain a suitable combination for optimal concentration of immobilized biomass. Immobilized cells were successfully reused with higher levels of product formation being maintained for longer period (65-70 h) than free cells. The highest gluconic acid concentration of about 143 g l-1 was reached on hydrol-based production medium with 0.3-cm3 foam cubes in the bubble column, where the effect of more suitable aeration and particle volume: medium volume ratio scheme was also investigated.

  1. Laccase immobilized on mesoporous SiO2 and its use for degradation of chlorophenol pesticides

    NASA Astrophysics Data System (ADS)

    Yang, Yuxiang; Xu, Yong; Yang, Yiwen; Yang, Huan; Yuan, Hongmin; Huang, Yan; Liu, Xiangnong

    2016-10-01

    In this paper, mesoporous silica with large specific surface area was used to immobilize laccase by the glutaraldehyde cross-linking method, and after screening and optimization experiments, the best enzyme immobilization process conditions were found (25°C, pH 5.4, 4% glutaraldehyde and 0.2 g/L laccase, treatment time 6 h). After that, the removal and degradation ratio of 2,4-dichlorophenol (abbreviated as DCP) under different conditions were also studied. After the degradation process was performed for 6 h at 30°C, pH 5.4, and DCP initial concentration of 50 mg/L in the presence of 0.1 g of immobilized laccase, the removal ratio and the degradation ratio were 42.28 and 15.93%, respectively. Compared with free laccase, the reusability of immobilized laccase is significantly improved.

  2. Immobilization of a Pleurotus ostreatus Laccase Mixture on Perlite and Its Application to Dye Decolourisation

    PubMed Central

    Salatino, Piero; Sannia, Giovanni

    2014-01-01

    In the present study, a crude laccase preparation from Pleurotus ostreatus was successfully immobilized on perlite, a cheap porous silica material, and tested for Remazol Brilliant Blue R (RBBR) decolourisation in a fluidized bed recycle reactor. Results showed that RBBR decolourisation is mainly due to enzyme action despite the occurrence of dye adsorption-related enzyme inhibition. Fine tuning of immobilization conditions allowed balancing the immobilization yield and the resulting rate of decolourisation, with the adsorption capacity of the solid biocatalyst. In the continuous lab scale reactor, a maximum conversion degree of 56.1% was achieved at reactor space-time of 4.2 h. Stability and catalytic parameters of the immobilized laccases were also assessed in comparison with the soluble counterparts, revealing an increase in stability, despite a reduction of the catalytic performances. Both effects are most likely ascribable to the occurrence of multipoint attachment phenomena. PMID:24895564

  3. Kinetic analysis of dihydroxyacetone production from crude glycerol by immobilized cells of Gluconobacter oxydans MTCC 904.

    PubMed

    Dikshit, Pritam Kumar; Moholkar, Vijayanand S

    2016-09-01

    The present study has investigated kinetic features of bioconversion of biodiesel-derived crude glycerol to dihydroxyacetone with immobilized Gluconobacter oxydans cells using modified Haldane substrate-inhibition model. The results have been compared against free cells and pure glycerol. Relative variations in the kinetic parameters KS, KI, Vmax, n and X reveal that immobilized G. oxydans cells (on PU foam substrate) with crude glycerol as substrate give higher order of inhibition (n) and lower maximum reaction velocities (Vmax). These results are essentially implications of substrate transport restrictions across immobilization matrix, which causes retention of substrate in the matrix and reduction in fractional available substrate (X) for the cells. This causes reduction in both KS (substrate concentration at Vmax/2) and KI (inhibition constant) as compared to free cells. For immobilized cells, substrate concentration (Smax) corresponding to Vmax is practically same for both pure and crude glycerol as substrate. PMID:27343447

  4. Decolorization of direct dyes by immobilized turnip peroxidase in batch and continuous processes.

    PubMed

    Matto, Mahreen; Husain, Qayyum

    2009-03-01

    An inexpensive immobilized turnip peroxidase has been employed for the decolorization of some direct dyes in batch and continuous reactors. Wood shaving was investigated as an inexpensive material for the preparation of bioaffinity support. Concanavalin A-wood shaving bound turnip peroxidase exhibited 67% of the original enzyme activity. Both soluble and immobilized turnip peroxidase could effectively remove more than 50% color from dyes in the presence of metals/salt and 0.6mM 1-hydroxybenzotriazole, after 1h of incubation. The columns containing immobilized peroxidase could decolorize 64% direct red 23% and 50% mixture of direct dyes at 4 and 3 months of operation, respectively. Total organic carbon analysis of treated dye or mixture of dyes revealed that these results were quite comparable to the loss of color from solutions. Thus, this study showed that the immobilized enzyme could be efficiently used for the removal of synthetic dyes from industrial effluents. PMID:18423852

  5. Developing an Efficient and General Strategy for Immobilization of Small Molecules onto Microarrays Using Isocyanate Chemistry

    PubMed Central

    Zhu, Chenggang; Zhu, Xiangdong; Landry, James P.; Cui, Zhaomeng; Li, Quanfu; Dang, Yongjun; Mi, Lan; Zheng, Fengyun; Fei, Yiyan

    2016-01-01

    Small-molecule microarray (SMM) is an effective platform for identifying lead compounds from large collections of small molecules in drug discovery, and efficient immobilization of molecular compounds is a pre-requisite for the success of such a platform. On an isocyanate functionalized surface, we studied the dependence of immobilization efficiency on chemical residues on molecular compounds, terminal residues on isocyanate functionalized surface, lengths of spacer molecules, and post-printing treatment conditions, and we identified a set of optimized conditions that enable us to immobilize small molecules with significantly improved efficiencies, particularly for those molecules with carboxylic acid residues that are known to have low isocyanate reactivity. We fabricated microarrays of 3375 bioactive compounds on isocyanate functionalized glass slides under these optimized conditions and confirmed that immobilization percentage is over 73%. PMID:26999137

  6. Developing an Efficient and General Strategy for Immobilization of Small Molecules onto Microarrays Using Isocyanate Chemistry.

    PubMed

    Zhu, Chenggang; Zhu, Xiangdong; Landry, James P; Cui, Zhaomeng; Li, Quanfu; Dang, Yongjun; Mi, Lan; Zheng, Fengyun; Fei, Yiyan

    2016-03-16

    Small-molecule microarray (SMM) is an effective platform for identifying lead compounds from large collections of small molecules in drug discovery, and efficient immobilization of molecular compounds is a pre-requisite for the success of such a platform. On an isocyanate functionalized surface, we studied the dependence of immobilization efficiency on chemical residues on molecular compounds, terminal residues on isocyanate functionalized surface, lengths of spacer molecules, and post-printing treatment conditions, and we identified a set of optimized conditions that enable us to immobilize small molecules with significantly improved efficiencies, particularly for those molecules with carboxylic acid residues that are known to have low isocyanate reactivity. We fabricated microarrays of 3375 bioactive compounds on isocyanate functionalized glass slides under these optimized conditions and confirmed that immobilization percentage is over 73%.

  7. Substitution of carbonate buffer by water for IgG immobilization in enzyme linked immunosorbent assay.

    PubMed

    Shrivastav, Tulsidas G; Basu, Anupam; Kariya, Kiran P

    2003-01-01

    The first step of enzyme linked immunosorbent assay (ELISA), namely, adsorption of antigen or antibody to the plastic microtiter well plate, was studied as a function of insolubility of IgG in water. Immobilization efficiency was assessed in terms of number of wells coated per milliliter of primary antiserum. We have compared different coating/immobilization protocols, i.e., direct and indirect immobilization of primary antibody to the plastic microtiter well plate using carbonate buffer and phosphate buffer with glutaraldehyde. We have observed efficient coating when the immobilization of primary antibody through an immunobridge technique was performed, where water was used as a coating medium. It gave a higher number of wells coated per milliliter of anti-serum (primary or secondary) than other compared coating protocols and it allowed the use of serum (non-immune) and anti-serum (primary and secondary antibody) dilutions, avoiding the need for gamma-globulin purification from normal and immunized serum. PMID:12778971

  8. Membranes for specific adsorption: immobilizing molecularly imprinted polymer microspheres using electrospun nanofibers.

    PubMed

    Büttiker, Roman; Ebert, Jürgen; Hinderling, Christian; Adlhart, Christian

    2011-01-01

    Molecularly imprinted polymer microspheres were immobilized within a polymer nanofiber membrane by electrospinning. Such membranes simplify the handling of functional microspheres and provide specific recognition capabilities for solid-phase extraction and filtration applications. In this study, microspheres were prepared by precipitation polymerization of methacrylic acid and divinylbenzene as a cross-linker with the target molecule (-)-cinchonidine and then, they were electrospun into a non-woven polyacrylonitrile nanofiber membrane. The composite membrane showed specific affinity for (-)-cinchonidine which was attributed to the functional microspheres as confirmed by Raman microscopy. The target molecule capturing capacity of the composite membrane was 5 mg/g or 25 mg/g immobilized functional microsphere. No difference in target affinity was observed between the immobilized microspheres and the free microspheres. These results reveal that electrospun composite membranes are a feasible approach to immobilizing functional microspheres.

  9. Membranes for specific adsorption: immobilizing molecularly imprinted polymer microspheres using electrospun nanofibers.

    PubMed

    Büttiker, Roman; Ebert, Jürgen; Hinderling, Christian; Adlhart, Christian

    2011-01-01

    Molecularly imprinted polymer microspheres were immobilized within a polymer nanofiber membrane by electrospinning. Such membranes simplify the handling of functional microspheres and provide specific recognition capabilities for solid-phase extraction and filtration applications. In this study, microspheres were prepared by precipitation polymerization of methacrylic acid and divinylbenzene as a cross-linker with the target molecule (-)-cinchonidine and then, they were electrospun into a non-woven polyacrylonitrile nanofiber membrane. The composite membrane showed specific affinity for (-)-cinchonidine which was attributed to the functional microspheres as confirmed by Raman microscopy. The target molecule capturing capacity of the composite membrane was 5 mg/g or 25 mg/g immobilized functional microsphere. No difference in target affinity was observed between the immobilized microspheres and the free microspheres. These results reveal that electrospun composite membranes are a feasible approach to immobilizing functional microspheres. PMID:21528654

  10. Effect of immobilization stress on the level of macroergic phosphates in the blood of rats

    NASA Technical Reports Server (NTRS)

    Pudov, V. I.; Sosenkov, V. A.

    1980-01-01

    The effect of immobilization for 1, 2, and 24 hours, and of daily (for 2 hours for 7 days) immobilization on the blood nucleotide (ATP, ADP, AMP) level was studied on 79 male rats. A progressive reduction of the ATP content was most pronounced in immobilization for 24 hours. This was accompanied by an increase of lymphopenia and eosinopenia. A fall of the relative weight of the thymus and a weight gain of the adrenal glands was observed along with a reduction of ascorbic acid concentration in both of them. In case of daily immobilization for 2 hours, the ATP and ADP content on the 1st and 2nd day was below the normal level, and then showed a gradual increase, with complete normalization in 6-7 days, except for inorganic phosphorus the level of which remained lower than normal.

  11. Linear alkyl benzene sulphonate (LAS) degradation by immobilized Pseudomonas aeruginosa under low intensity ultrasound.

    PubMed

    Lijun, Xiang; Bochu, Wang; Zhimin, Li; Chuanren, Duan; Qinghong, Wang; Liu, Liu

    2005-01-15

    We studied the LAS degradation of immobilized Pseudomonas aeruginosa with low-intensity ultrasonic and the influence of original LAS concentration, pH, rotary velocity and different conditions of low-intensity ultrasonic irradiation on the degradation of LAS. In our experiment, the degradation rate of LAS was the main index. We found that low-intensity ultrasonic irradiation could improve the metabolism of microorganism cells and promote the LAS biodegradation of immobilized cells. In the experiment, 50 mg/l LAS were used to simulate wastewater, and low-intensity ultrasonic was considered. We found the influence was obvious, and the optimal degradation rate was acquired when the conditions of ultrasonic were frequency 24 kHz, power 8 W, stimulation time 5 s, intermissive time 30 s, and total time 10 min. The LAS degradation rate of immobilized cells with ultrasonic were respectively 40% and 9.5% higher than that of the suspending cells and immobilized cells without irradiation.

  12. A radioimmunoassay method for detection of DNA based on chemical immobilization of anti-DNA antibody.

    PubMed

    Yoo, S K; Yoon, M; Park, U J; Han, H S; Kim, J H; Hwang, H J

    1999-09-30

    High selectivity provided by biomolecules such as antibodies and enzymes has been exploited during the last two decades for development of biosensors. Of particular importance are efficient immobilization methods for biomolecules in order to preserve their biological activities. In this study, we have evaluated immobilization strategies for an anti-DNA antibody on a self-assembled monolayer of omega-functionalized thiols. The antibody was immobilized via peptide bond formation between the primary amines in the antibody and the carboxyl groups on the self-assembled monolayer. The peptide bond coupling was achieved by activating COOH groups on the surface through N-Hydroxysuccimide (NHS)-ester formation, followed by acylation of NH2 group in the antibody. DNA binding activity of the immobilized antibody was examined by counting beta emission from 35S-labeled DNA. PMID:10551259

  13. Development of a new antibacterial biomaterial by tetracycline immobilization on calcium-alginate beads.

    PubMed

    Ozseker, Emine Erdogan; Akkaya, Alper

    2016-10-20

    In recent years, increasing risk of infection, caused by resistant microorganism to antibiotics, has become the limelight discovery of new and natural antibacterial materials. Heavy metals, such as silver, copper, mercury and titanium, have antibacterial activity. Products, which improved these metals, do not have stable antibacterial property. Therefore, use of these products is restricted. The aim of this study was to immobilize tetracycline to alginate and improve an antibacterial biomaterial. For this purpose, calcium-alginate beads were formed by dropping to calcium-chloride solution and tetracycline was immobilized to beads using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at optimum conditions. After immobilization, actualization of immobilization was investigated by analyzing ATR-FTIR spectrum and SEM images. Also, antibacterial property of obtained product was tested. Improved product demonstrated antibacterial property. It has potential for open wound, surgical drapes, bed and pillow sheath in hospitals and it may also be used for increasing human comfort in daily life. PMID:27474587

  14. Development of a new antibacterial biomaterial by tetracycline immobilization on calcium-alginate beads.

    PubMed

    Ozseker, Emine Erdogan; Akkaya, Alper

    2016-10-20

    In recent years, increasing risk of infection, caused by resistant microorganism to antibiotics, has become the limelight discovery of new and natural antibacterial materials. Heavy metals, such as silver, copper, mercury and titanium, have antibacterial activity. Products, which improved these metals, do not have stable antibacterial property. Therefore, use of these products is restricted. The aim of this study was to immobilize tetracycline to alginate and improve an antibacterial biomaterial. For this purpose, calcium-alginate beads were formed by dropping to calcium-chloride solution and tetracycline was immobilized to beads using 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide at optimum conditions. After immobilization, actualization of immobilization was investigated by analyzing ATR-FTIR spectrum and SEM images. Also, antibacterial property of obtained product was tested. Improved product demonstrated antibacterial property. It has potential for open wound, surgical drapes, bed and pillow sheath in hospitals and it may also be used for increasing human comfort in daily life.

  15. Immobilization of a Pleurotus ostreatus laccase mixture on perlite and its application to dye decolourisation.

    PubMed

    Pezzella, Cinzia; Russo, Maria Elena; Marzocchella, Antonio; Salatino, Piero; Sannia, Giovanni

    2014-01-01

    In the present study, a crude laccase preparation from Pleurotus ostreatus was successfully immobilized on perlite, a cheap porous silica material, and tested for Remazol Brilliant Blue R (RBBR) decolourisation in a fluidized bed recycle reactor. Results showed that RBBR decolourisation is mainly due to enzyme action despite the occurrence of dye adsorption-related enzyme inhibition. Fine tuning of immobilization conditions allowed balancing the immobilization yield and the resulting rate of decolourisation, with the adsorption capacity of the solid biocatalyst. In the continuous lab scale reactor, a maximum conversion degree of 56.1% was achieved at reactor space-time of 4.2 h. Stability and catalytic parameters of the immobilized laccases were also assessed in comparison with the soluble counterparts, revealing an increase in stability, despite a reduction of the catalytic performances. Both effects are most likely ascribable to the occurrence of multipoint attachment phenomena.

  16. Characterization of protein-immobilized polystyrene nanoparticles using impedance spectroscopy.

    PubMed

    Park, Soo-In; Lee, Sang-Yup

    2014-10-01

    A novel approach for characterization of non-conductive protein-immobilized nanoparticles using AC impedance spectroscopy combined with conductive atomic force microscopy was examined. As AC impedance spectroscopy can provide information on diverse electrical properties such as capacitance and inductance, it is applicable to the characterization of non-conductive substances. Several non-conductive protein-immobilized polystyrene nanoparticles were analyzed using AC impedance spectroscopy, and their impedance spectra were used as markers for nanoparticle identification. Analyses of impedance signals using an electrical circuit model established that the capacitance and inductance of each nanoparticle changed with the adsorbed protein and that impedance spectral differences were characteristic properties of the proteins. From this study, AC impedance spectroscopy was shown to be a useful tool for characterization of non-conductive nanoparticles and is expected to be applicable to the development of sensors for nanomaterials. PMID:25942903

  17. Active Acetylcholinesterase Immobilization on a Functionalized Silicon Surface.

    PubMed

    Khaldi, K; Sam, S; Gouget-Laemmel, A C; Henry de Villeneuve, C; Moraillon, A; Ozanam, F; Yang, J; Kermad, A; Ghellai, N; Gabouze, N

    2015-08-01

    In this work, we studied the attachment of active acetylcholinesterase (AChE) enzyme on a silicon substrate as a potential biomarker for the detection of organophosphorous (OP) pesticides. A multistep functionalization strategy was developed on a crystalline silicon surface: a carboxylic acid-terminated monolayer was grafted onto a hydrogen-terminated silicon surface by photochemical hydrosilylation, and then AChE was covalently attached through amide bonds using an activation EDC/NHS process. Each step of the modification was quantitatively characterized by ex-situ Fourier transform infrared spectroscopy in attenuated-total-reflection geometry (ATR-FTIR) and atomic force microscopy (AFM). The kinetics of enzyme immobilization was investigated using in situ real-time infrared spectroscopy. The enzymatic activity of immobilized acetylcholinesterase enzymes was determined with a colorimetric test. The surface concentration of active AChE was estimated to be Γ = 1.72 × 10(10) cm(-2).

  18. Immobilization of Rubia tinctorum L. Suspension Cultures and Biomass Production.

    PubMed

    Nartop, Pınar

    2016-01-01

    Plants are natural sources of valuable secondary metabolites used as pharmaceuticals, agrochemicals, flavors, fragrances, colors, biopesticides, and food additives. There is an increasing demand to obtain these metabolites through more productive plant tissue applications and cell culture methods due to the importance of secondary metabolites.Immobilization of plant cells is a method used in plant cell cultures to induce secondary metabolite production. In this method, plant cells are fixed in or on a supporting material or matrix such as agar, agarose, calcium alginate, glass, or polyurethane foam. In the present study, three natural lignocellulosic materials, loofah sponge, and the long fibers of sisal and jute, were used to immobilize suspended R. tinctorum cells. PMID:27108315

  19. Heparin immobilized chitosan--poly ethylene glycol interpenetrating network: antithrombogenicity.

    PubMed

    Beena, M S; Chandy, T; Sharma, C P

    1995-01-01

    This work deals with the synthesis and blood compatibility studies of Heparin immobilized chitosan--polyethyleneglycol (Chit-PEG) hydrogels for various biomedical applications. Chit-PEG interpenetrating net work (IPN) had been synthesised by crosslinking different ratios of chitosan with glutaraldehyde using schiffs base reaction mechanism and interpenetrating polyethyleneglycol (PEG) to form hydrogen bonding between the amino hydrogen in chitosan and polyether oxygen. An optimum gel combination was selected from the IPN of Chit-PEG and used for bonding heparin. This modified gel had dramatically improved its blood compatibility. The antithrombotic function of this gel and the release profile of heparin had been investigated using coagulation assays, and spectrophotometric quantitation. Recalcification times of plasma exposed to heparin immobilized Chit-PEG hydrogel were markedly increased as compared to heparin free gels. The anticoagulant function of this gel matrix may be due to partially released heparin and bonded heparin.

  20. Immobilization of Rubia tinctorum L. Suspension Cultures and Biomass Production.

    PubMed

    Nartop, Pınar

    2016-01-01

    Plants are natural sources of valuable secondary metabolites used as pharmaceuticals, agrochemicals, flavors, fragrances, colors, biopesticides, and food additives. There is an increasing demand to obtain these metabolites through more productive plant tissue applications and cell culture methods due to the importance of secondary metabolites.Immobilization of plant cells is a method used in plant cell cultures to induce secondary metabolite production. In this method, plant cells are fixed in or on a supporting material or matrix such as agar, agarose, calcium alginate, glass, or polyurethane foam. In the present study, three natural lignocellulosic materials, loofah sponge, and the long fibers of sisal and jute, were used to immobilize suspended R. tinctorum cells.

  1. In vitro refolding of porcine pepsin immobilized on agarose beads.

    PubMed

    Kurimoto, E; Harada, T; Akiyama, A; Sakai, T; Kato, K

    2001-08-01

    Since in vitro refolding of pepsin has long been attempted without success, it has been suspected that pepsin has no intrinsic refolding ability. In the present study, in order to eliminate unfavorable intermolecular interactions bringing about aggregation and autoproteolysis, we immobilized pepsin onto agarose beads. This technique enabled us to search extensively for appropriate refolding conditions without limitation of the refolding period. Renaturation of immobilized pepsin was observed exclusively at pH 3-5. This process was extremely slow and reached equilibrium after 300 h. Sixty percent of the proteolytic activity was recovered at pH 5. Addition of salts raised the recovery to 80% but had no significant effect on the refolding rate, suggesting that the salts mainly stabilize the native state of pepsin. This is the first report on the successful in vitro refolding of pepsin.

  2. Hydrogen production from biodiesel byproduct by immobilized Enterobacter aerogenes.

    PubMed

    Han, Jinmi; Lee, Dohoon; Cho, Jinku; Lee, Jeewon; Kim, Sangyong

    2012-01-01

    The recent rapid growth of the biodiesel industry has generated a significant amount of glycerol as a byproduct. As a result, the price of glycerol is currently relatively low, making it an attractive starting material for the production of chemicals with higher values. Crude glycerol can be directly converted through microbial fermentation into various chemicals such as hydrogen. In this study, we optimized immobilization of a facultative hydrogen producing microorganism, Enterobacter aerogenes, with the goal of developing biocatalysts that was appropriate for the continuous hydrogen production from glycerol. Several carriers were tested and agar was found to be the most effective. In addition, it was clearly shown that variables such as the carrier content and cell loading should be controlled for the immobilization of biocatalysts with high hydrogen productivity, stability, and reusability. After optimization of these variables, we were able to obtain reusable biocatalysts that could directly convert the byproduct stream from biodiesel processes into hydrogen in continuous processes.

  3. Synthesis, spectroscopic and catalytic studies of Cu(II), Co(II) and Ni(II) complexes immobilized on Schiff base modified chitosan

    NASA Astrophysics Data System (ADS)

    Antony, R.; Theodore David Manickam, S.; Saravanan, K.; Karuppasamy, K.; Balakumar, S.

    2013-10-01

    A new class of bidentate (N, O) Schiff base ligand (L) has been derived from the functional biopolymer (chitosan) and 1,2-diphenylethanedione in 1:1 M ratio. This ligand has been used to synthesise the new first row transition metal complexes of Cu(II), Co(II) and Ni(II). The structural properties of the ligand and the synthesized tetra-coordinated complexes have been investigated by elemental analysis, magnetic study, molar conductance measurement and spectroscopic methods viz. FT-IR, UV-Vis., 1H NMR, 13C NMR and XRD. The spectral evidences strongly suggested the square planar geometry to the complexes. The XRD studies proved that crystallinity of chitosan has been diminished after Schiff base formation and metal complexation of L. Thermal and surface properties of the complexes have been also discussed from the investigation of their TG-DTG curves and SEM images, respectively. In addition, the catalytic efficiency of these complexes has been studied in the cyclohexane oxidation reaction using H2O2 as oxidant at 70 °C.

  4. The study of the influence of temperature and initial glucose concentration on the fermentation process in the presence of Saccharomyces cerevisiae yeast strain immobilized on starch gels by reversed-flow gas chromatography.

    PubMed

    Lainioti, G Ch; Kapolos, J; Koliadima, A; Karaiskakis, G

    2012-01-01

    The technique of reversed-flow gas chromatography (RFGC) was employed for the determination of the alcoholic fermentation phases and of kinetic parameters for free and immobilized cell systems, at different initial glucose concentrations and temperature values. In addition to this, due to its considerable advantages over other techniques, RFGC was used for the characterization of a new biocatalyst, yeast cells immobilized on starch gel, and especially wheat starch gel. Immobilization of wine yeast Saccharomyces cerevisiae AXAZ-1 was accomplished on wheat and corn starch gels in order to prepare new biocatalysts with great interest for the fermentation industry. The RFGC led with great accuracy, resulting from a literature review, to the determination of reaction rate constants and activation energies at each phase of the fermentation processes. A maximum value of rate constants was observed at initial glucose concentration of 205 g/L, where a higher number of yeast cells was observed. The increase of glucose concentrations had a negative influence on the growth of AXAZ-1 cells and rate constants were decreased. The decrease of fermentation temperature caused a substantial reduction in the viability of immobilized cells as well as in rate constant values. Activation energies of corn starch gel presented lower values than those of wheat starch gel. However, the two supports showed higher catalytic efficiency than free cell systems, proving that starch gels may act as a promoter of the catalytic activity of the yeast cells involved in the fermentation process.

  5. Immobilization of lipase onto micron-size magnetic beads.

    PubMed

    Liu, Xianqiao; Guan, Yueping; Shen, Rui; Liu, Huizhou

    2005-08-01

    A novel and economical magnetic poly(methacrylate-divinylbenzene) microsphere (less than 8 microm in diameter) was synthesized by the modified suspension polymerization of methacrylate and cross-linker divinylbenzene in the presence of magnetic fluid. Then, surface aminolysis was employed to obtain a high content of surface amino groups (0.40-0.55 mmolg(-1) supports). The morphology and properties of these magnetic supports were characterized with scanning electron microscopy, transmission electron microscopy, Fourier transform infrared spectroscopy and a vibrating sample magnetometer. These magnetic supports exhibited superparamagnetism with a high specific saturation magnetization (sigma(s)) of 14.6 emicrog(-1). Candida cylindracea lipase was covalently immobilized on the amino-functionalized magnetic supports with the activity recovery up to 72.4% and enzyme loading of 34.0 mgg(-1) support, remarkably higher than the previous studies. The factors involved in the activity recovery and enzymatic properties of the immobilized lipase prepared were studied in comparison with free lipase, for which olive oil was chosen as the substrate. The results show that the immobilized lipase has good stability and reusability after recovery by magnetic separation within 20s. PMID:15998604

  6. Enhanced accumulation of starch and total carbohydrates in alginate-immobilized Chlorella spp. induced by Azospirillum brasilense: II. Heterotrophic conditions.

    PubMed

    Choix, Francisco J; de-Bashan, Luz E; Bashan, Yoav

    2012-10-10

    The effect of the bacterium Azospirillum brasilense jointly immobilized with Chlorella vulgaris or C. sorokiniana in alginate beads on total carbohydrates and starch was studied under dark and heterotrophic conditions for 144 h in synthetic growth medium supplemented with either d-glucose or Na-acetate as carbon sources. In all treatments, enhanced total carbohydrates and starch content per culture and per cell was obtained after 24h; only jointly immobilized C. vulgaris growing on d-glucose significantly increased total carbohydrates and starch content after 96 h. Enhanced accumulation of carbohydrate and starch under jointly immobilized conditions was variable with time of sampling and substrate used. Similar results occurred when the microalgae was immobilized alone. In both microalgae growing on either carbon sources, the bacterium promoted accumulation of carbohydrates and starch; when the microalgae were immobilized alone, they used the carbon sources for cell multiplication. In jointly immobilized conditions with Chlorella spp., affinity to carbon source and volumetric productivity and yield were higher than when Chlorella spp. were immobilized alone; however, the growth rate was higher in microalgae immobilized alone. This study demonstrates that under heterotrophic conditions, A. brasilense promotes the accumulation of carbohydrates in two strains Chlorella spp. under certain time-substrate combinations, producing mainly starch. As such, this bacterium is a biological factor that can change the composition of compounds in microalgae in dark, heterotrophic conditions.

  7. Influence of protein bulk properties on membrane surface coverage during immobilization.

    PubMed

    Militano, Francesca; Poerio, Teresa; Mazzei, Rosalinda; Piacentini, Emma; Gugliuzza, Annarosa; Giorno, Lidietta

    2016-07-01

    Biomolecules immobilization is a key factor for many biotechnological applications. For this purpose, the covalent immobilization of bovine serum albumin (BSA), lipase from Candida rugosa and protein G on differently functionalized regenerated cellulose membranes was investigated. Dynamic light scattering and electrophoresis measurements carried out on biomolecules in solution indicated the presence of monomers, dimers and trimers for both BSA and protein G, while large aggregates were observed for lipase. The immobilization rate and the surface coverage on functionalized regenerated cellulose membranes were studied as a function of biomolecule concentration. Results indicated that the saturation coverage of BSA and protein G was concentration independent (immobilized protein amount of 2.40±0.03mg/g and 2.65±0.07mg/g, respectively). Otherwise, a different immobilization kinetics trend was obtained for lipase, for which the immobilized amount increases as a function of time without reaching a saturation value. Atomic force microscopy (AFM) micrographs showed the formation of monolayers for both BSA and protein G on the membrane surface, while a multilayer structure is found for lipase, in agreement with the trends observed in the related immobilization kinetics. As a result, the morphology of the proteins layer on the membrane surface seems to be strictly dependent on the proteins behavior in solution. Besides, the surface coverage has been described for BSA and protein G by the pseudo second order models, the results indicating the surface reaction as the controlling step of immobilization kinetics. Finally, enzyme activity and binding capacity studies indicated the preservation of the biomolecule functional properties. PMID:27022871

  8. A Biomechanical Comparison of Intralaminar C7 Screw Constructs with and without Offset Connector Used for C6-7 Cervical Spine Immobilization : A Finite Element Study

    PubMed Central

    Qasim, Muhammad; Natarajan, Raghu N.; An, Howard S.

    2013-01-01

    Objective The offset connector can allow medial and lateral variability and facilitate intralaminar screw incorporation into the construct. The aim of this study was to compare the biomechanical characteristics of C7 intralaminar screw constructs with and without offset connector using a three dimensional finite element model of a C6-7 cervical spine segment. Methods Finite element models representing C7 intralaminar screw constructs with and without the offset connector were developed. Range of motion (ROM) and maximum von Mises stresses in the vertebra for the two techniques were compared under pure moments in flexion, extension, lateral bending and axial rotation. Results ROM for intralaminar screw construct with offset connector was less than the construct without the offset connector in the three principal directions. The maximum von Misses stress was observed in the C7 vertebra around the pedicle in both constructs. Maximum von Mises stress in the construct without offset connector was found to be 12-30% higher than the corresponding stresses in the construct with offset connector in the three principal directions. Conclusion This study demonstrated that the intralaminar screw fixation with offset connector is better than the construct without offset connector in terms of biomechanical stability. Construct with the offset connector reduces the ROM of C6-7 segment more significantly compared to the construct without the offset connector and causes lower stresses around the C7 pedicle-vertebral body complex. PMID:24003366

  9. [The combined action of drinking mineral water and low-intensity electromagnetic radiation under the immobilization stress conditions (an experimental study)].

    PubMed

    Korolev, Yu N; Bobrovnitsky, I P; Geniatulina, M S; Mikhailik, L V; Nikulina, L A; Bobkova, A S; Yakovlev, M Yu

    2015-01-01

    The present study carried out on white male rats in experiments with the use of biochemical, radioimmunological, and electron- microscopic methods. It was shown that the combined treatment with potable mineral water (MV) and low-intensity electromagnetic radiation (LIEMR) of ultrahigh frequency (power density less than 1 pW/cm2, the frequency about 1000 MHz) facilitated the activation of metabolic and intracellular regenerative processes in the liver and testes. One of the advantages of the combined application of MV and LIEMR over the single-factor treatment manifested itself as the weakening of stress reactions, the increase in the frequency of the plastic processes, and the more harmonious development of different forms of intracellular regeneration. The results of the study provide a deeper insight ino the mechanisms underlying the combined actions of drinking mineral water and low-intensity electromagnetic radiation; also, they justify the application of these factors for the protection of the reproductive system and the entire body from stress-induced disorders.

  10. TAT peptide immobilization on gold surfaces: a comparison study with a thiolated peptide and alkylthiols using AFM, XPS, and FT-IRRAS.

    PubMed

    Cho, Youngnam; Ivanisevic, Albena

    2005-04-01

    A TAT peptide was used to functionalize a gold surface by three different methods: adsorption from solution, microcontact printing, and dip-pen nanolithography (DPN). The composition and structure of the modified gold was characterized by atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), and Fourier transform -infrared reflection absorption spectroscopy (FT-IRRAS). We used two well-studied alkylthiols, mercaptohexadecanoic acid and 1-octadecanethiol, as a comparison in order to understand the structure of the TAT peptide monolayers prepared by the three methods. AFM studies allowed us to assess the homogeneity after each modification protocol. XPS was used to characterize the chemical composition of the gold surface after each functionalization procedure. The XPS results showed that surfaces modified with the TAT peptide by the three methods exhibit similar surface chemistry. Finally, FT-IRRAS experiments allowed us to conclude that the structure of the alkyl chains of the TAT peptides is fairly disordered and different after each procedure. Regardless of the type of surface functionalization method used, the monolayer of TAT peptide formed on the surface was of "liquidlike" nature.

  11. Cortical Reorganization after Hand Immobilization: The beta qEEG Spectral Coherence Evidences

    PubMed Central

    Fortuna, Marina; Teixeira, Silmar; Machado, Sérgio; Velasques, Bruna; Bittencourt, Juliana; Peressutti, Caroline; Budde, Henning; Cagy, Mauricio; Nardi, Antonio E.; Piedade, Roberto; Ribeiro, Pedro; Arias-Carrión, Oscar

    2013-01-01

    There is increasing evidence that hand immobilization is associated with various changes in the brain. Indeed, beta band coherence is strongly related to motor act and sensitive stimuli. In this study we investigate the electrophysiological and cortical changes that occur when subjects are submitted to hand immobilization. We hypothesized that beta coherence oscillations act as a mechanism underlying inter- and intra-hemispheric changes. As a methodology for our study fifteen healthy individuals between the ages of 20 and 30 years were subjected to a right index finger task before and after hand immobilization while their brain activity pattern was recorded using quantitative electroencephalography. This analysis revealed that hand immobilization caused changes in frontal, central and parietal areas of the brain. The main findings showed a lower beta-2 band in frontal regions and greater cortical activity in central and parietal areas. In summary, the coherence increased in the frontal, central and parietal cortex, due to hand immobilization and it adjusted the brains functioning, which had been disrupted by the procedure. Moreover, the brain adaptation upon hand immobilization of the subjects involved inter- and intra-hemispheric changes. PMID:24278213

  12. Bioinspired Immobilization of Glycerol Dehydrogenase by Metal Ion-Chelated Polyethyleneimines as Artificial Polypeptides

    PubMed Central

    Zhang, Yonghui; Ren, Hong; Wang, Yali; Chen, Kainan; Fang, Baishan; Wang, Shizhen

    2016-01-01

    In this study, a novel, simple and generally applicable strategy for multimeric oxidoreductase immobilization with multi-levels interactions was developed and involved activity and stability enhancements. Linear polyethyleneimines (PEIs) are flexible cationic polymers with molecular weights that span a wide range and are suitable biomimic polypeptides for biocompatible frameworks for enzyme immobilization. Metal ion-chelated linear PEIs were applied as a heterofunctional framework for glycerol dehydrogenase (GDH) immobilization by hydrogen bonds, electrostatic forces and coordination bonds interactions. Nanoparticles with diameters from 250–650 nm were prepared that exhibited a 1.4-fold enhancement catalytic efficiency. Importantly, the half-life of the immobilized GDH was enhanced by 5.6-folds in aqueous phase at 85 °C. A mechanistic illustration of the formation of multi-level interactions in the PEI-metal-GDH complex was proposed based on morphological and functional studies of the immobilized enzyme. This generally applicable strategy offers a potential technique for multimeric enzyme immobilization with the advantages of low cost, easy operation, high activity reservation and high stability. PMID:27053034

  13. “Fish-in-Net”, a Novel Method for Cell Immobilization of Zymomonas mobilis

    PubMed Central

    Niu, Xuedun; Wang, Zhi; Li, Yang; Zhao, Zijian; Liu, Jiayin; Jiang, Li; Xu, Haoran; Li, Zhengqiang

    2013-01-01

    Background Inorganic mesoporous materials exhibit good biocompatibility and hydrothermal stability for cell immobilization. However, it is difficult to encapsulate living cells under mild conditions, and new strategies for cell immobilization are needed. We designed a “fish-in-net” approach for encapsulation of enzymes in ordered mesoporous silica under mild conditions. The main objective of this study is to demonstrate the potential of this approach in immobilization of living cells. Methodology/Principal Findings Zymomonas mobilis cells were encapsulated in mesoporous silica-based materials under mild conditions by using a “fish-in-net” approach. During the encapsulation process, polyethyleneglycol was used as an additive to improve the immobilization efficiency. After encapsulation, the pore size, morphology and other features were characterized by various methods, including scanning electron microscopy, nitrogen adsorption-desorption analysis, transmission electron microscopy, fourier transform infrared spectroscopy, and elemental analysis. Furthermore, the capacity of ethanol production by immobilized Zymomonas mobilis and free Zymomonas mobilis was compared. Conclusions/Significance In this study, Zymomonas mobilis cells were successfully encapsulated in mesoporous silica-based materials under mild conditions by the “fish-in-net” approach. Encapsulated cells could perform normal metabolism and exhibited excellent reusability. The results presented here illustrate the enormous potential of the “fish-in-net” approach for immobilization of living cells. PMID:24236145

  14. A Biomechanical Comparison of Three Different Posterior Fixation Constructs Used for C6–C7 Cervical Spine Immobilization: A Finite Element Study

    PubMed Central

    HONG, Jae Taek; QASIM, Muhammad; ESPINOZA ORÍAS, Alejandro A.; NATARAJAN, Raghu N.; AN, Howard S.

    2014-01-01

    The intralaminar screw construct has been recently introduced in C6–C7 fixation. The aim of the study is to compare the stability afforded by three different C7 posterior fixation techniques using a three-dimensional finite element model of a C6–C7 cervical spine motion segment. Finite element models representing three different cervical anchor types (C7 intralaminar screw, C7 lateral mass screw, and C7 pedicle screw) were developed. Range of motion (ROM) and maximum von Mises stresses in the vertebra for the three screw techniques were compared under pure moments in flexion, extension, lateral bending, and axial rotation. ROM for pedicle screw construct was less than the lateral mass screw construct and intralaminar screw construct in the three principal directions. The maximum von Misses stress was observed in the C7 vertebra around the pedicle in all the three screw constructs. Maximum von Mises stress in pedicle screw construct was less than the lateral mass screw construct and intralaminar screw construct in all loading modes. This study demonstrated that the pedicle screw fixation is the strongest instrumentation method for C6–C7 fixation. Pedicle screw fixation resulted in least stresses around the C7 pedicle-vertebral body complex. However, if pedicle fixation is not favorable, the laminar screw can be a better option compared to the lateral mass screw because the stress around the pedicle-vertebral body complex and ROM predicted for laminar screw construct was smaller than those of lateral mass screw construct. PMID:24418790

  15. Immobilization of trypsin onto 1,4-diisothiocyanatobenzene-activated porous glass for microreactor-based peptide mapping by capillary electrophoresis: effect of calcium ions on the immobilization procedure.

    PubMed

    Dartiguenave, Catherine; Hamad, Hussein; Waldron, Karen C

    2010-03-24

    The immobilization conditions and kinetic behaviour of trypsin, covalently immobilized via the 1,4-diisothiocyanatobenzene (DITC) linker onto aminopropylated controlled pore glass (CPG) particles, have been evaluated to establish a rapid and efficient protocol for fabrication of an immobilized enzyme microreactor (IMER) for protein hydrolysis and subsequent peptide mapping. Addition of calcium ions to either the immobilization reaction solution or hydrolysis assay was studied for a synthetic substrate. Activity was slightly higher when immobilization was carried out in the presence of Ca(2+) whereas more enzyme could be immobilized in its absence. A protocol requiring less than 3 h was devised to obtain maximal enzymatic activity with the lowest ratio of soluble trypsin to DITC-CPG particles. The resulting immobilized enzyme was found to retain an acceptable percentage (ca. 35%) of its activity after immobilization. The particles were dry-packed into a capillary to make a microscale IMER. Repeatability, reusability and digestion efficiency of the microIMER were investigated for the substrate beta-casein using capillary electrophoretic-based peptide mapping. In initial tests, a single device showed reproducible peptide maps for 21 digestions lasting 2 h each, carried out over a period of 2 months. Complete digestion of beta-casein could be achieved in a few minutes (86 s residence time in the microIMER followed by a wash step).

  16. Screen-printed graphite macroelectrodes for the direct electron transfer of cytochrome c: a deeper study of the effect of pH on the conformational states, immobilization and peroxidase activity.

    PubMed

    Gómez-Mingot, Maria; Montiel, Vicente; Banks, Craig E; Iniesta, Jesús

    2014-03-21

    The direct electron transfer of cytochrome c has been studied at screen-printed graphite macroelectrodes without recourse to mediators or the need for any electrode pre-treatment as is commonly employed within the literature. A wide range of pH values from 2.0 to 11.0 have been explored upon the electrochemical response of cytochrome c and different voltammetric signatures have been observed. The direct electron transfer of the alkaline transition of cytochrome c was found impeded within alkaline media leading to either an irreversible redox process or even no voltammetric responses. In acidic aqueous media the electrochemical process is observed to undergo a mixed diffusion and adsorption controlled process rather than a purely diffusional process of the native conformation as observed at pH 7.0. Interestingly, at pH 3.5 a new conformational state is revealed in cooperation with the native conformation. The immobilization of the protein was satisfactorily obtained using a simple method by cycling the protein at specific solution pH values allowing amperometric responses to be obtained and gives rise to useful pseudo-peroxidase activity for sensing H2O2. Apparent Michaelis-Menten constant values (Km) were calculated via the Lineweaver-Burk method with deduced values of 25 ± 4, 98 ± 12 and 230 ± 30 mM, respectively for pH values of 2.0, 3.0 and 7.0. Such work is important for those utilising cytochrome c in bio-electrochemical and related applications.

  17. Immobilization of thermolysin to polyamide nonwoven materials.

    PubMed

    Moeschel, Klaus; Nouaimi, Meryem; Steinbrenner, Christa; Bisswanger, Hans

    2003-04-20

    In the last few years, an increasing number of biotechnological techniques have been applied to the restoration and conservation of works of art, paintings, old maps, and papers or books. Enzymes can solve problems that give restorers difficulties, although for many applications it is not possible to use soluble enzymes; therefore, it is necessary to look for suitable carriers for immobilization. Different methods for covalent immobilization of enzymes to polyamide nonwovens were tested, using thermolysin as an example. Two distinct strategies were pursued: (1). controlled, partial hydrolysis of the polymer and subsequent binding of the enzyme to the released amino and carboxy groups; and (2). attachment of reactive groups directly to the polyamide without disintegrating the polymeric structure (O-alkylation). Different spacers were used for covalent fixation of the enzyme in both cases. The enzyme was fixed to the released amino groups by glutaraldehyde, either with or without a spacer. Either way, active enzyme could be immobilized to the matrix. However, intense treatment caused severe damage to the stability of the nonwoven fabric, and reduced the mechanical strength. Conditions were investigated to conserve the nonwoven fabric structure while obtaining near-maximum immobilized enzyme activity. Immobilization of the enzyme to the released carboxy group after acid hydrolysis was performed using dicyclohexylcarbodiimide. In comparison to the enzyme bound via the amino group, the yield of immobilized enzyme activity was slightly lower when benzidine was taken as spacer and still lower with a 1,6-hexanediamine spacer. O-alkylation performed with dimethylsulfate caused severe damage to the nonwoven fabric structure. Considerably better results were obtained with triethyloxonium tetrafluoroborate. As the spacers 1,6-hexanediamine and adipic acid dihydrazide were used, activation for immobilizing thermolysin was performed with glutaraldehyde, adipimidate, and azide

  18. Studies with an immobilized metal affinity chromatography cassette system involving binuclear triazacyclononane-derived ligands: automation of batch adsorption measurements with tagged recombinant proteins.

    PubMed

    Petzold, Martin; Coghlan, Campbell J; Hearn, Milton T W

    2014-07-18

    This study describes the determination of the adsorption isotherms and binding kinetics of tagged recombinant proteins using a recently developed IMAC cassette system and employing automated robotic liquid handling procedures for IMAC resin screening. These results confirm that these new IMAC resins, generated from a variety of different metal-charged binuclear 1,4,7-triaza-cyclononane (tacn) ligands, interact with recombinant proteins containing a novel N-terminal metal binding tag, NT1A, with static binding capacities similar to those obtained with conventional hexa-His tagged proteins, but with significantly increased association constants. In addition, higher kinetic binding rates were observed with these new IMAC systems, an attribute that can be positively exploited to increase process productivity. The results from this investigation demonstrate that enhancements in binding capacities and affinities were achieved with these new IMAC resins and chosen NT1A tagged protein. Further, differences in the binding performances of the bis(tacn) xylenyl-bridged ligands were consistent with the distance between the metal binding centres of the two tacn moieties, the flexibility of the ligand and the potential contribution from the aromatic ring of the xylenyl group to undergo π/π stacking interactions with the tagged proteins.

  19. Catalysis of rice straw hydrolysis by the combination of immobilized cellulase from Aspergillus niger on β-cyclodextrin-Fe3O4 nanoparticles and ionic liquid.

    PubMed

    Huang, Po-Jung; Chang, Ken-Lin; Hsieh, Jung-Feng; Chen, Shui-Tein

    2015-01-01

    Cellulase from Aspergillus niger was immobilized onto β-cyclodextrin-conjugated magnetic particles by silanization and reductive amidation. The immobilized cellulase gained supermagnetism due to the magnetic nanoparticles. Ninety percent of cellulase was immobilized, but the activity of immobilized cellulase decreased by 10%. In this study, ionic liquid (1-butyl-3-methylimidazolium chloride) was introduced into the hydrolytic process because the original reaction was a solid-solid reaction. The activity of immobilized cellulase was improved from 54.87 to 59.11 U g immobilized cellulase(-1) at an ionic liquid concentration of 200 mM. Using immobilized cellulase and ionic liquid in the hydrolysis of rice straw, the initial reaction rate was increased from 1.629 to 2.739 g h(-1) L(-1). One of the advantages of immobilized cellulase is high reusability--it was usable for a total of 16 times in this study. Compared with free cellulase, magnetized cellulase can be recycled by magnetic field and the activity of immobilized cellulase was shown to remain at 85% of free cellulase without denaturation under a high concentration of glucose (15 g L(-1)). Therefore, immobilized cellulase can hydrolyze rice straw continuously compared with free cellulase. The amount of harvested glucose can be up to twentyfold higher than that from the hydrolysis by free cellulase. PMID:25874210

  20. Catalysis of rice straw hydrolysis by the combination of immobilized cellulase from Aspergillus niger on β-cyclodextrin-Fe3O4 nanoparticles and ionic liquid.

    PubMed

    Huang, Po-Jung; Chang, Ken-Lin; Hsieh, Jung-Feng; Chen, Shui-Tein

    2015-01-01

    Cellulase from Aspergillus niger was immobilized onto β-cyclodextrin-conjugated magnetic particles by silanization and reductive amidation. The immobilized cellulase gained supermagnetism due to the magnetic nanoparticles. Ninety percent of cellulase was immobilized, but the activity of immobilized cellulase decreased by 10%. In this study, ionic liquid (1-butyl-3-methylimidazolium chloride) was introduced into the hydrolytic process because the original reaction was a solid-solid reaction. The activity of immobilized cellulase was improved from 54.87 to 59.11 U g immobilized cellulase(-1) at an ionic liquid concentration of 200 mM. Using immobilized cellulase and ionic liquid in the hydrolysis of rice straw, the initial reaction rate was increased from 1.629 to 2.739 g h(-1) L(-1). One of the advantages of immobilized cellulase is high reusability--it was usable for a total of 16 times in this study. Compared with free cellulase, magnetized cellulase can be recycled by magnetic field and the activity of immobilized cellulase was shown to remain at 85% of free cellulase without denaturation under a high concentration of glucose (15 g L(-1)). Therefore, immobilized cellulase can hydrolyze rice straw continuously compared with free cellulase. The amount of harvested glucose can be up to twentyfold higher than that from the hydrolysis by free cellulase.

  1. Catalysis of Rice Straw Hydrolysis by the Combination of Immobilized Cellulase from Aspergillus niger on β-Cyclodextrin-Fe3O4 Nanoparticles and Ionic Liquid

    PubMed Central

    Huang, Po-Jung; Chang, Ken-Lin; Chen, Shui-Tein

    2015-01-01

    Cellulase from Aspergillus niger was immobilized onto β-cyclodextrin-conjugated magnetic particles by silanization and reductive amidation. The immobilized cellulase gained supermagnetism due to the magnetic nanoparticles. Ninety percent of cellulase was immobilized, but the activity of immobilized cellulase decreased by 10%. In this study, ionic liquid (1-butyl-3-methylimidazolium chloride) was introduced into the hydrolytic process because the original reaction was a solid-solid reaction. The activity of immobilized cellulase was improved from 54.87 to 59.11 U g immobilized cellulase−1 at an ionic liquid concentration of 200 mM. Using immobilized cellulase and ionic liquid in the hydrolysis of rice straw, the initial reaction rate was increased from 1.629 to 2.739 g h−1 L−1. One of the advantages of immobilized cellulase is high reusability—it was usable for a total of 16 times in this study. Compared with free cellulase, magnetized cellulase can be recycled by magnetic field and the activity of immobilized cellulase was shown to remain at 85% of free cellulase without denaturation under a high concentration of glucose (15 g L−1). Therefore, immobilized cellulase can hydrolyze rice straw continuously compared with free cellulase. The amount of harvested glucose can be up to twentyfold higher than that from the hydrolysis by free cellulase. PMID:25874210

  2. EFFECTS OF JOINT IMMOBILIZATION ON STANDING BALANCE

    PubMed Central

    de Freitas, Paulo B.; Freitas, Sandra M. S. F.; Duarte, Marcos; Latash, Mark L.; Zatsiorsky, Vladimir M.

    2009-01-01

    We investigated the effect of joint immobilization on the postural sway during quiet standing. We hypothesized that center of pressure (COP), rambling, and trembling trajectories could be affected by joint immobilization. Ten young adults stood on a force plate during 60 s without and with immobilized joints (only knees constrained, CK; knees and hips, CH; and knees, hips and trunk, CT), with their eyes opened (EO) or closed (EC). The root mean square deviation (RMS, the standard deviation from the mean) and mean speed of COP, rambling, and trembling trajectories in the anterior-posterior and medial-lateral directions were analyzed. Similar effects of vision were observed for both directions: larger amplitude for all variables was observed in the EC condition. In the anterior-posterior direction, postural sway increased only when the knees, hips and trunk were immobilized. For the medial-lateral direction, the RMS and the mean speed of the COP, rambling, and trembling displacements decreased after immobilization of knees and hips and knees, hips and trunk. These findings indicate that the inverted pendulum model is unable to completely explain the processes involved in the control of the quiet upright stance in the anterior-posterior and medial-lateral directions. PMID:19342114

  3. Accumulation of uranium by immobilized persimmon tannin

    SciTech Connect

    Sakaguchi, Takashi; Nakajima, Akira )

    1994-01-01

    We have discovered that the extracted juice of unripe astringent persimmon fruit, designated as kakishibu or shibuol, has an extremely high affinity for uranium. To develop efficient adsorbents for uranium, we tried to immobilize kakishibu (persimmon tannin) with various aldehydes and mineral acids. Persimmon tannin immobilized with glutaraldehyde can accumulate 1.71 g (14 mEq U) of uranium per gram of the adsorbent. The uranium accumulating capacity of this adsorbent is several times greater than that of commercially available chelating resins (2-3 mEq/g). Immobilized persimmon tannin has the most favorable features for uranium recovery; high selective adsorption ability, rapid adsorption rate, and applicability in both column and batch systems. The uranium retained on immobilized persimmon tannin can be quantitatively and easily eluted with a very dilute acid, and the adsorbent can thus be easily recycled in the adsorption-desorption process. Immobilized persimmon tannin also has a high affinity for thorium. 23 refs., 13 figs., 7 tabs.

  4. Reversible and oriented immobilization of ferrocene-modified proteins.

    PubMed

    Yang, Lanti; Gomez-Casado, Alberto; Young, Jacqui F; Nguyen, Hoang D; Cabanas-Danés, Jordi; Huskens, Jurriaan; Brunsveld, Luc; Jonkheijm, Pascal

    2012-11-21

    Adopting supramolecular chemistry for immobilization of proteins is an attractive strategy that entails reversibility and responsiveness to stimuli. The reversible and oriented immobilization and micropatterning of ferrocene-tagged yellow fluorescent proteins (Fc-YFPs) onto β-cyclodextrin (βCD) molecular printboards was characterized using surface plasmon resonance (SPR) spectroscopy and fluorescence microscopy in combination with electrochemistry. The proteins were assembled on the surface through the specific supramolecular host-guest interaction between βCD and ferrocene. Application of a dynamic covalent disulfide lock between two YFP proteins resulted in a switch from monovalent to divalent ferrocene interactions with the βCD surface, yielding a more stable protein immobilization. The SPR titration data for the protein immobilization were fitted to a 1:1 Langmuir-type model, yielding K(LM) = 2.5 × 10(5) M(-1) and K(i,s) = 1.2 × 10(3) M(-1), which compares favorably to the intrinsic binding constant presented in the literature for the monovalent interaction of ferrocene with βCD self-assembled monolayers. In addition, the SPR binding experiments were qualitatively simulated, confirming the binding of Fc-YFP in both divalent and monovalent fashion to the βCD monolayers. The Fc-YFPs could be patterned on βCD surfaces in uniform monolayers, as revealed using fluorescence microscopy and atomic force microscopy measurements. Both fluorescence microscopy imaging and SPR measurements were carried out with the in situ capability to perform cyclic voltammetry and chronoamperometry. These studies emphasize the repetitive desorption and adsorption of the ferrocene-tagged proteins from the βCD surface upon electrochemical oxidation and reduction, respectively.

  5. Protective Effects of Resveratrol against Chronic Immobilization Stress on Testis

    PubMed Central

    Bitgul, Gulsah; Tekmen, Isil; Keles, Didem; Oktay, Gulgun

    2013-01-01

    Objective. The aim of this study was to investigate protective effects of resveratrol, a strong antioxidant, against possible negative effects of chronic immobilization stress on testes of male rats histochemically, immunohistochemically, ultrastructurally, and biochemically. Material and Methods. Male Wistar rats were divided into 4 groups (n = 7). Group I, control group (C), was not exposed to stress. Group II, stress group (S), was exposed to chronic immobilization stress. In Group III, low dose resveratrol + stress group (LRS), rats were given 10 mg/kg/day resveratrol just before the stress application. In Group IV, high dose resveratrol + stress group (HRS), rats were given 20 mg/kg/day resveratrol just before the stress application. For chronic immobilization stress application animals were put in the plastic tubes (6 cm in diameter, 15 cm in length) during 32 days for 6 hours. All animals were sacrificed 18 hours after the last stress application. Results. Histochemical and ultrastructural investigations showed that in stress group there was germ cell deprivation in seminiferous tubules and increase of connective tissue on interstitial area. No significant changes were seen in low and high dose resveratrol groups. After immunohistochemical investigations, TUNEL (+) and Active Caspase-3 (+) cells were increased in seminiferous tubules of stress group compared with those control group, but they were decreased in low and high dose resveratrol groups. According to biochemically results, MDA, GSH, and testosterone levels in stress group showed no significant difference when compared with those of the other groups. Conclusion. The chronic immobilization stress increases oxidative stress and apoptosis and causes histological tissue damages; resveratrol can minimize the histological damage in testes significantly. PMID:24307953

  6. Airline Chair-rest Deconditioning: Induction of Immobilization Thromboemboli?

    NASA Technical Reports Server (NTRS)

    Greenleaf, J. E.; Rehrer, N. J.; Mohler, S. R.; Quach, D. T.; Evans, D. G.; Dalton, Bonnie P. (Technical Monitor)

    2002-01-01

    Air passenger miles will likely double by year 2020. The altered and restrictive environment in an airliner cabin can influence hematological homeostasis in passengers and crew. Flight-related deep various thromboemboli (DVT) have been associated with at least 577 deaths on 42 of 120 airlines from 1977 to 1984 (25 deaths/million departures), whereas many such cases go unreported. However, there are four major factors that could influence formation of possible flight-induced DVT: sleeping accomodations (via sitting immobilization), travelers' medical history (via tissue injury), cabin environmental factors (via lower partial pressure of oxygen and lower relative humidity), and the more encompassing chair-rest deconditioning (C-RD) syndrome. There is ample evidence that recent injury and surgery (especially in deconditioned hospitalized patients) facilitate thrombophlebitis and formation of DVT that may be exacerbated by the immobilization of prolonged air travel. In the healthy flying population immobilization factors associated with prolonged (> 5 hr) C-RID such as total body dehydration, hypovolemia and increased blood viscosity, and reduced various blood flow (pooling) in the legs may facilitate formation of DVT. However, data from at least four case-controlled epidemiological studies did not confirm a direct causative relationship between air travel and DART, but factors such as history of vascular thromboemboli, various insufficiency, chronic heart failure, obesity, immobile standing position, more than 3 pregnancies, infectious disease, long-distance travel, muscular trauma and violent physical effort were significantly more frequent in DVT patients than in controls. Thus, there is no clear, direct evidence yet that prolonged sitting in airliner seats, or prolonged experimental chair-rest- or bed- rest-deconditioning treatments cause deep various thromboemboli in healthy people.

  7. Cellobiose hydrolysis using Pichia etchellsii cells immobilized in calcium alginate

    SciTech Connect

    Jain, D.; Ghose, T.K.

    1984-01-01

    The rate of cellulose degradation, limited by inhibition by cellobiose, can be increased by hydrolysis of cellobiose to glucose using immobilized ..beta..-glucosidase. Production of ..beta..-glucosidase in four yeasts was studied and a maximum activity of 1.22 IU/mg cells was obtained in cells of Pichia etchellsii when grown on 3% cellobiose as the sole carbon source. Immobilization of ..beta..-glucosidase containing cells of Pichia etchellsii on various solid supports was conducted and immobilization by entrapment in calcium alginate gel beads was found to be the most simple and efficient method. The immobilized preparation was found to be limited by pore diffusion but exhibited no film-diffusion resistance during packed bed reactor operation. Good plug flow characteristics were observed in the packed bed column indicated by a low dispersion number of 0.1348. A model for reaction with pore diffusion for a noncompetitive type of inhibited system was developed and applied to the cellobiose hydrolysis system. The rate of reaction with diffusional limitations was determined by using the model and effectiveness factors were calculated for different particle sizes. An effectiveness factor of 0.49 was obtained for a particle diameter of 2.5 mm. The modified rate expression using the effectiveness factor represented batch and packed bed reactor operation satisfactorily. The productivity in the packed bed column was found to fall rapidly with increase in conversion rate indicating that the operating conditions of the column would have to be a compromise between high conversion rates and reasonable productivity. A half-life of over seven days was obtained at the operating temperature of 45/sup 0/C in continuous operation of the packed bed reactor. However, the half-life in the column was found to be greatly affected by temperature, increasing to over seve

  8. Immobilized rennin in TC/SG composite in cheese production.

    PubMed

    Barouni, Eleftheria; Petsi, Theano; Kolliopoulos, Dionysios; Vasileiou, Dimitrios; Panas, Panagiotis; Bekatorou, Argyro; Kanellaki, Maria; Koutinas, Athanasios A

    2016-06-01

    The object of the current study was to develop a new process for continuous Feta-type cheese production using a biocatalyst consisting of immobilized rennin on a tubular cellulose/starch gel (TC/SG) composite, which has been proven to be an appropriate carrier for enzyme immobilization. Different methodologies were used in order to prepare four biocatalysts. The most effective was selected for cheese production in a 1L continuous system, providing two economically useful results for the dairy industries: (i) increase of productivity by the continuous coagulation of milk, and (ii) saving of the rennin enzyme expenses of the batch coagulation of milk. The criteria used to choose the appropriate biocatalyst was based on the time of coagulation in successive batches, the concentration of immobilized rennin combined with the filter efficiency and its application in the continuous system. Physicochemical analyses of the cheeses at various stages of the ripening were performed. No significant differences compared to cheeses prepared with the traditional method were found. Aroma compounds were determined by SPME GC-MS.

  9. Ex vivo model of an immobilized-enzyme reactor.

    PubMed Central

    Bernstein, H; Langer, R

    1988-01-01

    Immobilized-enzyme reactors are beginning to be studied for a variety of therapeutic applications. To facilitate the design of these devices for different clinical situations and a diverse patient population, mathematical models may be valuable. An immobilized-heparinase (EC 4.2.2.7) reactor was selected as a model system. The device removes heparin from blood that has been anticoagulated to prevent thrombus formation. Heparinase was immobilized to cross-linked agarose particles. A mathematical model was developed to describe the clearance of heparin by the reactor ex vivo and compared to experimental clearances measured in sheep. The model accounted for enzymatic degradation as well as the binding of heparin and its breakdown products to antithrombin. The device was modeled as a steady-state continuously stirred tank reactor. Molar conservation equations within the agarose particles accounted for simultaneous diffusion and chemical reaction. The model had no adjustable parameters and was able to predict the clearance of heparin within 5-25% for three different animals and 12 different perfusions. PMID:3186758

  10. Growth and immobilization of tripterygium wilfordii cultured cells.

    PubMed

    Pépin, M F; Chavarie, C; Archambault, J

    1991-12-20

    The plant Tripterygium wilfordii produces di- and triterpenes of interest for male contraception and treatment of arthritis and skin disorders. Cell line TRP4a obtained form this plant in 1981 was reported to produce these valuable compounds at yields ( approximately 0.04% of the biomass dry weight) higher than found in the plant (0.001%). In order to improve this production, studies were carried out to determine the feasibility of eliminating the troublesome component of coconut milk originally used to culture this cell line. A defined formulation suitable for growth ad maintenance has been developed. This medium consisted of Gamborg's PRL4 or B5 medium supplemented with 2 mg L(-1) 2,4-dichlorophenoxyacetic acid and 20 g L(-1) sucrose. Furthermore, monitoring of carbohydrate uptake revealed that T. wilfordii cells, contrary to many plant cell species, did not hydrolyze sucrose extra-cellularly before uptake. Replacement of this disaccharide by glucose or fructose increased specific growth rate from 0.15 to 0.25 day(-1). As tripdiolide is reported to be present in broth extract in significant amounts, plant cell immobilization technology offers a promising alternative to suspension cultures, especially in view to on line harvesting of the product. Surface immobilized T. wilfordii cell cultures were successfully carried out in 2-L bioreactors. Their biomass production and carbohydrate uptake were comparable to those observed for shake flask grown suspension cultures. Higher nitrate and ammonium uptake were found in immobilized cultures. PMID:18600729

  11. Potential Applications of Carbohydrases Immobilization in the Food Industry

    PubMed Central

    Contesini, Fabiano Jares; de Alencar Figueira, Joelise; Kawaguti, Haroldo Yukio; de Barros Fernandes, Pedro Carlos; de Oliveira Carvalho, Patrícia; Nascimento, Maria da Graça; Sato, Hélia Harumi

    2013-01-01

    Carbohydrases find a wide application in industrial processes and products, mainly in the food industry. With these enzymes, it is possible to obtain different types of sugar syrups (viz. glucose, fructose and inverted sugar syrups), prebiotics (viz. galactooligossacharides and fructooligossacharides) and isomaltulose, which is an interesting sweetener substitute for sucrose to improve the sensory properties of juices and wines and to reduce lactose in milk. The most important carbohydrases to accomplish these goals are of microbial origin and include amylases (α-amylases and glucoamylases), invertases, inulinases, galactosidases, glucosidases, fructosyltransferases, pectinases and glucosyltransferases. Yet, for all these processes to be cost-effective for industrial application, a very efficient, simple and cheap immobilization technique is required. Immobilization techniques can involve adsorption, entrapment or covalent bonding of the enzyme into an insoluble support, or carrier-free methods, usually based on the formation of cross-linked enzyme aggregates (CLEAs). They include a broad variety of supports, such as magnetic materials, gums, gels, synthetic polymers and ionic resins. All these techniques present advantages and disadvantages and several parameters must be considered. In this work, the most recent and important studies on the immobilization of carbohydrases with potential application in the food industry are reviewed. PMID:23344046

  12. Potential applications of carbohydrases immobilization in the food industry.

    PubMed

    Contesini, Fabiano Jares; de Alencar Figueira, Joelise; Kawaguti, Haroldo Yukio; de Barros Fernandes, Pedro Carlos; de Oliveira Carvalho, Patrícia; da Graça Nascimento, Maria; Sato, Hélia Harumi

    2013-01-01

    Carbohydrases find a wide application in industrial processes and products, mainly in the food industry. With these enzymes, it is possible to obtain different types of sugar syrups (viz. glucose, fructose and inverted sugar syrups), prebiotics (viz. galactooligossacharides and fructooligossacharides) and isomaltulose, which is an interesting sweetener substitute for sucrose to improve the sensory properties of juices and wines and to reduce lactose in milk. The most important carbohydrases to accomplish these goals are of microbial origin and include amylases (α-amylases and glucoamylases), invertases, inulinases, galactosidases, glucosidases, fructosyltransferases, pectinases and glucosyltransferases. Yet, for all these processes to be cost-effective for industrial application, a very efficient, simple and cheap immobilization technique is required. Immobilization techniques can involve adsorption, entrapment or covalent bonding of the enzyme into an insoluble support, or carrier-free methods, usually based on the formation of cross-linked enzyme aggregates (CLEAs). They include a broad variety of supports, such as magnetic materials, gums, gels, synthetic polymers and ionic resins. All these techniques present advantages and disadvantages and several parameters must be considered. In this work, the most recent and important studies on the immobilization of carbohydrases with potential application in the food industry are reviewed. PMID:23344046

  13. Immobilization of Lipid Substrates: Application on Phospholipase A2 Determination.

    PubMed

    Karkabounas, Athanassios; Georgiadou, Dimitra G; Argitis, Panagiotis; Psycharis, Vassilios; Nakos, George; Kosmas, Agni M; Lekka, Marilena E

    2015-12-01

    The purpose of the study was to assess a fluorimetric assay for the determination of total phospholipase A(2) (PLA(2)) activity in biological samples introducing the innovation of immobilized substrates on crosslinked polymeric membranes. The immobilized C(12)-NBD-PtdCho, a fluorescent analogue of phosphatidylcholine, exhibited excellent stability for 3 months at 4 °C and was not desorbed in the aqueous reaction mixture during analysis. The limit of detection was 0.5 pmol FA (0.2 pg) and the linear part of the response curve extended from 1 up to 190 nmol FA/h/mL sample. The intra- and inter-day relative standard deviations (%RSD), were ≤6 and ≤9 %, respectively. Statistical comparison with other fluorescent methods showed excellent correlation and agreement. Semiempirical calculations showed a fair amount of electrostatic interaction between the NBD-labeled substrate and the crosslinked polyvinyl alcohol with the styryl pyridinium residues (PVA-SbQ) material, from the plane of which, the sn-2 acyl chain of the phospholipid stands out and is accessible by PLA(2). Atomic Force Microscopy revealed morphological alterations of the immobilized substrate after the reaction with PLA(2). Mass spectrometry showed that only C(12)-NBD-FA, the PLA(2 )hydrolysis product, was detected in the reaction mixture, indicating that PLA(2) recognizes PVA-SbQ/C(12)-NBD-PtdCho as a surface to perform catalysis. PMID:26449236

  14. Tailoring immobilization of immunoglobulin by excimer laser for biosensor applications.

    PubMed

    Sima, Felix; Axente, Emanuel; Ristoscu, Carmen; Mihailescu, Ion N; Kononenko, Taras V; Nagovitsin, Ilya A; Chudinova, Galina; Konov, Vitaly I; Socol, Marcela; Enculescu, Ionut; Sima, Livia E; Petrescu, Stefana M

    2011-02-01

    The sheltered transfer and immobilization of rabbit anti-human antiserum immunoglobulin G (IgG) by matrix-assisted pulsed laser evaporation (MAPLE) are reported. The iced targets submitted to laser irradiation consisted of 0.2-2 mg/mL IgG blended or not with lipid (L-α-phosphatidylcholine dipalmitoyl) dissolved in distilled water-based saline buffer. Thin IgG coatings were obtained at room temperature onto glass, fused silica, or silicon substrates. Ten thousand subsequent laser pulses of 0.33, 0.5, or 0.67 J/cm(2) fluence were applied for the synthesis of each sample. Morphology and composition of the thin films were studied by optical, scanning, and atomic force microscopy and Fourier transformed infrared spectrometry. Optical labeling methods such as spectrofluorimetry and fluorescence microscopy were selected to verify the biosensor transduction principle because of their high sensitivity for detecting low amounts of antigen (IgG). Protein immobilization to the substrate surface was demonstrated for all obtained structures after immersion in the donkey anti-rabbit secondary antibody solution. The IgG transfer and immobilization onto substrates were improved by addition of lipid to MAPLE solutions.

  15. Protein synthesis rates in atrophied gastrocnemius muscles after limb immobilization

    NASA Technical Reports Server (NTRS)

    Tucker, K. R.; Seider, M. J.; Booth, F. W.

    1981-01-01

    Noting that protein synthesis declines in the gastrocnemius 6 hr after immobilization, the study sought to detect an increase of protein synthesis when the limb was freed, and to examine the effects of exercise on the rate of increase. Rats were used as subjects, with their hind legs in plaster of Paris in plantar flexion to eliminate strain on the gastrocnemius. Periods of immobilization were varied and samples of blood from the muscle were taken to track protein synthesis rates for different groups in immobilization and exercise regimens (running and weightlifting). Synthesis rates declined 3.6% during time in the cast, then increased 6.3%/day after the casts were removed. Both running and weightlifting were found to increase the fractional rate of protein formation in the gastrocnemius muscle when compared with contralateral muscles that were not exercised and were used as controls, suggesting that the mechanism controlling protein synthesis in skeletal muscles is rapidly responsive to changes in muscular contractile activity.

  16. Immobilization of lipid vesicles on polymer support via an amphiphilic peptidic anchor: application to a membrane enzyme.

    PubMed

    Percot, A; Zhu, X X; Lafleur, M

    2000-01-01

    To immobilize lipid vesicles on a polymer support, we have used a peptidic anchor with the following sequence: Ala-Ala-Leu-Leu-Leu-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Ala-Ala-A la-Ala-Ala-Ala-Ala-Ala-Ala-Trp-Lys-Lys-Lys-Lys-Lys-Lys. This amphiphilic peptide was previously designed in our group to interact spontaneously and strongly with vesicles without perturbing their permeability. At the end of the solid-phase peptide synthesis, the peptide was left on the polymer beads and this novel polymer-peptide system was used for vesicle immobilization. It was shown that this polymer-peptide system could immobilize as much as 200 micromol of lipids per gram of dry resin. The amount of immobilized vesicles was decreased by a reduction of the proportion of the negatively charged lipids in the vesicles, indicating the importance of electrostatic interactions in the immobilization of the vesicles. The integrity of the vesicles was mostly preserved after the immobilization. This new polymer-peptide system was used easily and successfully to immobilize a membrane-bound enzyme, gamma-glutamyl transpeptidase. The activity of the membrane-bound enzyme was studied by monitoring the release of p-nitroaniline. The activity of the enzyme was still retained, even after being re-used eight times, indicating the strong immobilization of the enzyme in its active form. The polymer-peptide support could be regenerated by washing with ethanol and reused.

  17. Reporter Immobilization Assay (REIA) for Bioconjugating Reactions.

    PubMed

    Schatte, Martin; Bocola, Marco; Roth, Teresa; Martinez, Ronny; Kopetzki, Erhard; Schwaneberg, Ulrich; Bönitz-Dulat, Mara

    2016-06-15

    Enzymes able to ligate biomolecules are emerging tools to generate site-specific bioconjugates. In this study we present a detection and screening method for bioconjugating enzymes which overcomes limitations of analytical methods such as HPLC or MS. These techniques are experimentally demanding and often limited in sensitivity and throughput compared to enzymatic assays. The principle of this Reporter Immobilization Assay (REIA) is the ligation of a reporter enzyme to a peptide carrying an affinity handle, which can be utilized for its isolation. The REIA system exhibits a high sensitivity with a linear range down to 1 μg/mL (55 nM), a variation coefficient of 6.5%, and can be performed cost-efficiently in 96-well microtiter plate format. The application of this assay allowed the characterization of a thiol transpeptidase sortase from S. aureus which is an important drug target and a biotechnological tool for ligation and modification of proteins. Thereby, yet-undetectable promiscuous activity of sortase could be detected, e.g., the acceptance of alanine as nucleophile. In addition, we were able to provide evidence that the REIA is suitable for high throughput screening of enzyme libraries using crude cellular extract with a throughput of 600 samples per hour. PMID:27182715

  18. Immobilized Bioluminescent Reagents in Flow Injection Analysis.

    NASA Astrophysics Data System (ADS)

    Nabi, Abdul

    Available from UMI in association with The British Library. Bioluminescent reactions exhibits two important characteristics from an analytical viewpoint; they are selective and highly sensitive. Furthermore, bioluminescent emissions are easily measured with a simple flow-through detector based on a photomultiplier tube and the rapid and reproducible mixing of sample and expensive reagent is best achieved by a flow injection manifold. The two most important bioluminescent systems are the enzyme (luciferase)/substrate (luciferin) combinations extracted from fireflies (Photinus pyralis) and marine bacteria (Virio harveyi) which requires ATP and NAD(P)H respectively as cofactors. Reactions that generate or consume these cofactors can also be coupled to the bioluminescent reaction to provide assays for a wide range of clinically important species. A flow injection manifold for the study of bioluminescent reactions is described, as are procedures for the extraction, purification and immobilization of firefly and bacterial luciferase and oxidoreductase. Results are presented for the determination of ATP using firefly system and the determination of other enzymes and substrates participating in ATP-converting reactions e.g. creatine kinase, ATP-sulphurylase, pyruvate kinase, creatine phosphate, pyrophosphate and phophoenolypyruvate. Similarly results are presented for the determination of NAD(P)H, FMN, FMNH_2 and several dehydrogenases which produce NAD(P)H and their substrates, e.g. alcohol, L-lactate, L-malate, L-glutamate, Glucose-6-phosphate and primary bile acid.

  19. Effect of passive stretching on the immobilized soleus muscle fiber morphology.

    PubMed

    Coutinho, E L; Gomes, A R S; França, C N; Oishi, J; Salvini, T F

    2004-12-01

    The aim of the present study was to determine the effect of stretching applied every 3 days to the soleus muscle immobilized in the shortened position on muscle fiber morphology. Eighteen 16-week-old Wistar rats were used and divided into three groups of 6 animals each: a) the left soleus muscle was immobilized in the shortened position for 3 weeks; b) during immobilization, the soleus was stretched for 40 min every 3 days; c) the non-immobilized soleus was only stretched. Left and right soleus muscles were examined. One portion of the soleus was frozen for histology and muscle fiber area evaluation, while the other portion was used to identify the number and length of serial sarcomeres. Immobilized muscles (group A) showed a significant decrease in weight (44 +/- 6%), length (19 +/- 7%), serial sarcomere number (23 +/- 15%), and fiber area (37 +/- 31%) compared to the contralateral muscles (P < 0.05, paired Student t-test). The immobilized and stretched soleus (group B) showed a similar reduction but milder muscle fiber atrophy compared to the only immobilized group (22 +/- 40 vs 37 +/- 31%, respectively; P < 0.001, ANOVA test). Muscles submitted only to stretching (group C) significantly increased the length (5 +/- 2%), serial sarcomere number (4 +/- 4%), and fiber area (16 +/- 44%) compared to the contralateral muscles (P < 0.05, paired Student t-test). In conclusion, stretching applied every 3 days to immobilized muscles did not prevent the muscle shortening, but reduced muscle atrophy. Stretching sessions induced hypertrophic effects in the control muscles. These results support the use of muscle stretching in sports and rehabilitation.

  20. From physical inactivity to immobilization: Dissecting the role of oxidative stress in skeletal muscle insulin resistance and atrophy.

    PubMed

    Pierre, Nicolas; Appriou, Zephyra; Gratas-Delamarche, Arlette; Derbré, Frédéric

    2016-09-01

    In the literature, the terms physical inactivity and immobilization are largely used as synonyms. The present review emphasizes the need to establish a clear distinction between these two situations. Physical inactivity is a behavior characterized by a lack of physical activity, whereas immobilization is a deprivation of movement for medical purpose. In agreement with these definitions, appropriate models exist to study either physical inactivity or immobilization, leading thereby to distinct conclusions. In this review, we examine the involvement of oxidative stress in skeletal muscle insulin resistance and atrophy induced by, respectively, physical inactivity and immobilization. A large body of evidence demonstrates that immobilization-induced atrophy depends on the chronic overproduction of reactive oxygen and nitrogen species (RONS). On the other hand, the involvement of RONS in physical inactivity-induced insulin resistance has not been investigated. This observation outlines the need to elucidate the mechanism by which physical inactivity promotes insulin resistance.

  1. Characterization of a new stationary phase based on microwave immobilized polybutadiene on titanium oxide-modified silica.

    PubMed

    Morais, Lais S R; Jardim, Isabel C S F

    2005-05-01

    Titanium oxide-modified silica was prepared by reaction of silica with titanium tetrabutoxide and then was used as support in the preparation of stationary phases with self-immobilized polybutadiene (PBD) and PBD immobilized through microwave radiation. Chromatographic performance of the stationary phases was evaluated in terms of the efficiency (plates/m), asymmetry (A(s)), retention factor (k) and resolution (R(s)) of two standard sample mixtures, one of then containing the basic compound N,N-dimethylaniline. A microwave irradiation of 30 min at 520 W gave the best efficiency (86,500 N m(-1)), greater than that of a 6-day self immobilized phase (69,500 N m(-1)). Self-immobilized stationary phases prepared with bare silica were also studied for comparison. These resulted in lower chromatographic performance, 43,800 N m(-1), when compared to the self-immobilized phase prepared with titanized silica.

  2. Wheat Bran Enhances the Cytotoxicity of Immobilized Alcaligenes aquatilis F8 against Microcystis aeruginosa.

    PubMed

    Sun, Pengfei; Lin, Hui; Wang, Guan; Zhang, Ximing; Zhang, Qichun; Zhao, Yuhua

    2015-01-01

    Algicidal bacteria offer a promising option for killing cyanobacteria. Therefore, a new Alcaligenes aquatilis strain F8 was isolated to control Microcystis aeruginosa in this study. The algicidal activity of strain F8 was dependent on the cell density of M. aeruginosa, and the maximal algicidal rate of the free bacterium reached 88.45% within 72 h. With a view to its application to the control of M. aeruginosa in the natural environment, strain F8 was immobilized in sodium alginate beads, but immobilization of the strain decreased its algicidal rate compared to that of the free bacterium. However, addition of wheat bran to the sodium alginate matrix used to immobilize strain F8 not only eliminated the adverse effects of immobilization on the bacteria but also resulted in an 8.83% higher algicidal rate of the immobilized than free bacteria. Exclusion and recovery methods were used to identify key ingredients of wheat bran and gain insight into the mechanism underlying the observed enhancement of algicidal activity. This analysis indicated that certain factors in wheat bran, including vitamins B1, B2, B9, and E were responsible for promoting bacterial growth and thereby improving the algicidal rate of immobilized strain F8. Our findings indicate that wheat bran is able to improve the algicidal efficiency of A. aquatilis strain F8 for killing M. aeruginosa and is a good source of not only carbon and nitrogen but also vitamins for bacteria. PMID:26295573

  3. Removal of arsenic(III) from aqueous solutions using fresh and immobilized plant biomass.

    PubMed

    Kamala, C T; Chu, K H; Chary, N S; Pandey, P K; Ramesh, S L; Sastry, A R K; Sekhar, K Chandra

    2005-08-01

    The ability of Garcinia cambogia, an indigenous plant found in many parts of India, to remove trivalent arsenic from solution was assessed. Batch experiments were carried out to characterize the As(III) removal capability of fresh and immobilized biomass of G. cambogia. It was found that the kinetic property and uptake capacity of fresh biomass were significantly enhanced by the immobilization procedure. The uptake of As(III) by fresh and immobilized biomass was not greatly affected by solution pH with optimal biosorption occurring at around pH 6--8. The presence of common ions such as Ca and Mg at concentrations up to 100mg/l had no effect on As(III) removal. However, the presence of Fe(III) at 100mg/l caused a noticeable drop in the extent of As(III) removal but the effect was minimal when Fe(III) was present at 10mg/l. The adsorption isotherms quantitatively predicted the extent of As(III) removal in groundwater samples collected from an arsenic-contaminated site in India. Immobilized biomass loaded with As(III) was amenable to efficient regeneration with NaOH solution. Column studies showed that immobilized biomass could be reused over five cycles of loading and elution. The excellent As(III) sequestering capability of fresh and immobilized G. cambogia biomass could lead to the development of a viable and cost-effective technology for arsenic removal in groundwater. PMID:15993920

  4. The removal of thermo-tolerant coliform bacteria by immobilized waste stabilization pond algae.

    PubMed

    Pearson, H W; Marcon, A E; Melo, H N

    2011-01-01

    This study investigated the potential of laboratory- scale columns of immobilized micro-algae to disinfect effluents using thermo-tolerant coliforms (TTC) as a model system. Cells of a Chlorella species isolated from a waste stabilization pond complex in Northeast Brazil were immobilized in calcium alginate, packed into glass columns and incubated in contact with TTC suspensions for up to 24 hours. Five to six log removals of TTC were achieved in 6 hours and 11 log removals in 12 hours contact time. The results were similar under artificial light and shaded sunlight. However little or no TTC removal occurred in the light in columns of alginate beads without immobilized algae present or when the immobilized algae were incubated in the dark suggesting that the presence of both algae and light were necessary for TTC decay. There was a positive correlation between K(b) values for TTC and increasing pH in the effluent from the immobilized algal columns within the range pH 7.2 and 8.9. The potential of immobilized algal technology for wastewater disinfection may warrant further investigation.

  5. Catalytic performance and molecular dynamic simulation of immobilized CC bond hydrolase based on carbon nanotube matrix.

    PubMed

    Zhou, Hao; Qu, Yuanyuan; Kong, Chunlei; Li, Duanxing; Shen, E; Ma, Qiao; Zhang, Xuwang; Wang, Jingwei; Zhou, Jiti

    2014-04-01

    Carbon nanotube (CNT) has been proved to be a kind of novel support for enzyme immobilization. In this study, we tried to find the relationship between conformation and catalytic performance of immobilized enzyme. Two CC bond hydrolases BphD and MfphA were immobilized on CNTs (SWCNT and MWCNT) via physical adsorption and covalent attachment. Among the conjugates, the immobilized BphD on chemically functionalized SWCNT (BphD-CSWCNT) retained the highest catalytic efficiency (kcat/Km value) compared to free BphD (92.9%). On the other hand, when MfphA bound to pristine SWCNT (MfphA-SWCNT), it was completely inactive. Time-resolved fluorescence spectrum indicated the formation of static ground complexes during the immobilization processes. Circular dichroism (CD) showed that the secondary structures of immobilized enzymes changed in varying degrees. In order to investigate the inhibition mechanism of MfphA by SWCNT, molecular dynamics simulation was employed to analyze the adsorption process, binding sites and time evolution of substrate tunnels. The results showed that the preferred binding sites (Trp201 and Met81) of MfphA for SWCNT blocked the main substrate access tunnel, thus making the enzyme inactive. The "tunnel-block" should be a novel possible inhibition mechanism for enzyme-nanotube conjugate.

  6. Immobilization of Microbes for Bioremediation of Crude Oil Polluted Environments: A Mini Review

    PubMed Central

    Bayat, Zeynab; Hassanshahian, Mehdi; Cappello, Simone

    2015-01-01

    Petroleum hydrocarbons are the most common environmental pollutants in the world and oil spills pose a great hazard to terrestrial and marine ecosystems. Oil pollution may arise either accidentally or operationally whenever oil is produced, transported, stored and processed or used at sea or on land. Oil spills are a major menace to the environment as they severely damage the surrounding ecosystems. To improve the survival and retention of the bioremediation agents in the contaminated sites, bacterial cells must be immobilized. Immobilized cells are widely tested for a variety of applications. There are many types of support and immobilization techniques that can be selected based on the sort of application. In this review article, we have discussed the potential of immobilized microbial cells to degrade petroleum hydrocarbons. In some studies, enhanced degradation with immobilized cells as compared to free living bacterial cells for the treatment of oil contaminated areas have been shown. It was demonstrated that immobilized cell to be effective and is better, faster, and can be occurred for a longer period PMID:26668662

  7. Wheat Bran Enhances the Cytotoxicity of Immobilized Alcaligenes aquatilis F8 against Microcystis aeruginosa

    PubMed Central

    Sun, Pengfei; Lin, Hui; Wang, Guan; Zhang, Ximing; Zhang, Qichun; Zhao, Yuhua

    2015-01-01

    Algicidal bacteria offer a promising option for killing cyanobacteria. Therefore, a new Alcaligenes aquatilis strain F8 was isolated to control Microcystis aeruginosa in this study. The algicidal activity of strain F8 was dependent on the cell density of M. aeruginosa, and the maximal algicidal rate of the free bacterium reached 88.45% within 72 h. With a view to its application to the control of M. aeruginosa in the natural environment, strain F8 was immobilized in sodium alginate beads, but immobilization of the strain decreased its algicidal rate compared to that of the free bacterium. However, addition of wheat bran to the sodium alginate matrix used to immobilize strain F8 not only eliminated the adverse effects of immobilization on the bacteria but also resulted in an 8.83% higher algicidal rate of the immobilized than free bacteria. Exclusion and recovery methods were used to identify key ingredients of wheat bran and gain insight into the mechanism underlying the observed enhancement of algicidal activity. This analysis indicated that certain factors in wheat bran, including vitamins B1, B2, B9, and E were responsible for promoting bacterial growth and thereby improving the algicidal rate of immobilized strain F8. Our findings indicate that wheat bran is able to improve the algicidal efficiency of A. aquatilis strain F8 for killing M. aeruginosa and is a good source of not only carbon and nitrogen but also vitamins for bacteria. PMID:26295573

  8. Immobilization of Microbes for Bioremediation of Crude Oil Polluted Environments: A Mini Review.

    PubMed

    Bayat, Zeynab; Hassanshahian, Mehdi; Cappello, Simone

    2015-01-01

    Petroleum hydrocarbons are the most common environmental pollutants in the world and oil spills pose a great hazard to terrestrial and marine ecosystems. Oil pollution may arise either accidentally or operationally whenever oil is produced, transported, stored and processed or used at sea or on land. Oil spills are a major menace to the environment as they severely damage the surrounding ecosystems. To improve the survival and retention of the bioremediation agents in the contaminated sites, bacterial cells must be immobilized. Immobilized cells are widely tested for a variety of applications. There are many types of support and immobilization techniques that can be selected based on the sort of application. In this review article, we have discussed the potential of immobilized microbial cells to degrade petroleum hydrocarbons. In some studies, enhanced degradation with immobilized cells as compared to free living bacterial cells for the treatment of oil contaminated areas have been shown. It was demonstrated that immobilized cell to be effective and is better, faster, and can be occurred for a longer period. PMID:26668662

  9. Investigating a galactomannan gel obtained from Cassia grandis seeds as immobilizing matrix for Cramoll lectin.

    PubMed

    Albuquerque, Priscilla B S; Silva, Caroline S; Soares, Paulo A G; Barros, Wilson; Correia, Maria T S; Coelho, Luana C B B; Teixeira, José A; Carneiro-da-Cunha, Maria G

    2016-05-01

    Characterization, with emphasis on the rheological properties, of Cassia grandis seeds galactomannan gel containing immobilized Cramoll 1-4 is presented. The gels, with and without immobilized Cramoll 1-4, were evaluated along time by rheometry, pH, color, microbial contamination and lectin hemagglutinating activity (HA). Rheological determinations confirmed the gels to be very stable up to 30 days with variations occurring after this period. Rheological data also showed that the gel/Cramoll 1-4 immobilizing matrix loses its elastic modulus substantially after 60 days. Both gels presented no microbial contamination as well as a pH close to neutral. Colorimetric parameters demonstrated the gels transparency with occasional yellowness. The opacity of the galactomannan gel did not change significantly along the study; the same did not occur for the gel with immobilized Cramoll 1-4 as a statistically significant reduction of its opacity was observed. In what concerns immobilized Cramoll 1-4 HA, up to 90% of its initial HA was maintained after 20 days, with a decrease to 60% after 60 days. These results combined with the thickening and stabilizing characteristics of the galactomannan gel make this gel a promising immobilizing matrix for Cramoll 1-4 that can be further exploited for clinical and cosmetic applications. PMID:26840177

  10. Trypsin immobilization on to polymer surface through grafted layer and its reaction with inhibitors.

    PubMed

    Kulik, E A; Kato, K; Ivanchenko, M I; Ikada, Y

    1993-08-01

    Trypsin was covalently immobilized and physically adsorbed on to the surface of poly(ethylene terephthalate) fibres using poly(acrylic acid) (PAAc) chains grafted on to the ozonized fibres. The covalent immobilization was accomplished through amide formation between amino groups of trypsin and carboxyl groups of grafted PAAc chains, with the use of water-soluble carbodiimide. A set of samples with surface concentrations of grafted polymer ranging from 0.03 to 2.5 micrograms/cm2 was used to study the effects of grafted layer on the enzymatic activity of immobilized trypsin and its inhibition by trypsin inhibitors of different molecular sizes. The amount of immobilized trypsin increased linearly with an increase in graft yield of fibres, but the activity of immobilized enzyme reached saturation at a certain graft yield, probably because of diffusion limitation for the transport of enzyme substrate molecules into the grafted PAAc layer. The reduction of inhibition with an increase in graft yield and in molecular weight of inhibitors was attributed to enhancement of steric hindrance and enzyme inactivation in the dense grafted layer. We also found that the adsorbed trypsin was inhibited more easily than the covalently immobilized at any concentration of the grafted PAAc and for any type of inhibitor used.

  11. Preparation of core-shell magnetic polydopamine/alginate biocomposite for Candida rugosa lipase immobilization.

    PubMed

    Hou, Chen; Qi, Zhigang; Zhu, Hao

    2015-04-01

    A flexible, biocompatible and bioadhesive enzyme immobilizing material, which was synthesized based on the covalent assembly of biomimetic polymer and oxidized polysaccharide on magnetic nanoparticles (NPs), has been developed in this feasibility study. In this work, the bio-inspired polymer, polydopamine (PDA), was used to modify the well-monodispersed Fe3O4 NPs (mPDA NPs) with a controllable thickness via a dip-coating process, then the alginate di-aldehyde (ADA) was covalently assembled on the mPDA NPs and employed as a naturally occurring linking agent for Candida rugosa lipase (CRL) immobilization. The resulting support material was characterized by means of the transmission electron microscope (TEM), Fourier transform infrared spectra (FT-IR), X-ray diffraction (XRD), thermogravimetry (TG) analyser, and vibrating sample magnetometer (VSM). It was verified that the prepared mPDA NPs possessed distinct core-shell structure with uniform size and high saturation magnetization. For further application, the mPDA NPs was utilized in CRL immobilizing procedures and demonstrated can facilitate improving the enzyme activities. The optimum amount of lipase was 200 mg g(-1) support, the optimal pH and temperature for the catalyse condition of the immobilized CRL was 7.0 and 40°C, respectively. Moreover, the immobilized CRL kept the high activity at 77% after 12 times of recycling for batch hydrolysis of olive oil emulsion. This magnetic bioadhesive composite with functionalized properties and adhesion strength presents a general strategy for the immobilization of macromolecules.

  12. Recovery of skeletal muscle after 3 mo of hindlimb immobilization in rats

    NASA Technical Reports Server (NTRS)

    Booth, F. W.; Seider, M. J.

    1979-01-01

    During immobilization, skeletal muscle undergoes decreases in size and strength with concomitant atrophic and degenerative changes in slow-twitch muscle fibers. Currently there are no objective data in slow-twitch muscle demonstrating recovery of biochemical or physiological indices following termination of immobilization. The purpose of this study was to determine whether the soleus, a slow-twitch muscle, could recover normal biochemical or physiological levels following termination of immobilization. Adenosine triphosphate, glycogen, and protein concentration (mg/g wet wt) all significantly decreased following 90 days of hindlimb immobilization, but these three values returned to control levels by the 60th recovery day. Similarly, soleus muscle wet weight and protein content (mg protein/muscle) returned to control levels by the 14th recovery day. In contrast, maximal isometric tension did not return to normal until the 120th day. These results indicate that following muscular atrophy, which was achieved through 90 days of hindlimb immobilization, several biochemical and physiological values in skeletal muscle are recovered at various times after the end of immobilization.

  13. [Effects of the size of magnetic particles of immobilized enzyme reactors on the digestion performance].

    PubMed

    Zhang, Jiao; Zhou, Lianqi; Tian, Fang; Zhang, Yangjun; Qian, Xiaohong

    2013-02-01

    We applied immobilized enzyme reactors prepared with different sizes of magnetic particles into protein and proteome digestion. In addition, the influences of different sizes of the magnetic particles were studied on the reunion, enzyme efficiency and leakage sites. The experimental results showed that in comparison with the submicron magnetic particles, the amount of trypsin immobilized on the magnetic nanoparticles was 3. 5 times more than that of the submicron magnetic particles. However, the enzymatic efficiency was at the same level when the same amount of trypsin was used, and the reunion phenomenon was obviously improved when the size of the magnetic nanoparticles increased. Taking the immobilized enzyme reactor of 20 nm magnetic nanoparticles as an example, the digestion performance was further examined. The experimental results showed that rapid digestion could be achieved within 1 mm when the mass ratio of the trypsin and bovine serum albumin was 1:1. The peptide number of 0 missed cleavage site and the sequence coverage changed little after the protein was digested for 10 mm. It was concluded that the digestion efficiency of the immobilized enzyme reactor was much better than that of the in-solution digestion. When the immobilized enzyme reactors and the free trypsin were used for digestion, little differences of the leakage sites were found. Therefore, the immobilized enzyme reactors prepared with different sizes of magnetic particles can be applied in proteomic research for quick and efficient digestion.

  14. Radiofrequency treatment enhances the catalytic function of an immobilized nanobiohybrid catalyst.

    PubMed

    San, Boi Hoa; Ha, Eun-Ju; Paik, Hyun-Jong; Kim, Kyeong Kyu

    2014-06-01

    Biocatalysis, the use of enzymes in chemical transformation, has undergone intensive development for a wide range of applications. As such, maximizing the functionality of enzymes for biocatalysis is a major priority to enable industrial use. To date, many innovative technologies have been developed to address the future demand of enzymes for these purposes, but maximizing the catalytic activity of enzymes remains a challenge. In this study, we demonstrated that the functionality of a nanobiocatalyst could be enhanced by combining immobilization and radiofrequency (RF) treatment. Aminopeptidase PepA-encapsulating 2 nm platinum nanoparticles (PepA-PtNPs) with the catalytic activities of hydrolysis and hydrogenation were employed as multifunctional nanobiocatalysts. Immobilizing the nanobiocatalysts in a hydrogel using metal chelation significantly enhanced their functionalities, including catalytic power, thermal-stability, pH tolerance, organic solvent tolerance, and reusability. Most importantly, RF treatment of the hydrogel-immobilized PepA-PtNPs increased their catalytic power by 2.5 fold greater than the immobilized PepA. Our findings indicate that the catalytic activities and functionalities of PepA-PtNPs are greatly enhanced by the combination of hydrogel-immobilization and RF treatment. Based on our findings, we propose that RF treatment of nanobiohybrid catalysts immobilized on the bulk hydrogel represents a new strategy for achieving efficient biocatalysis.

  15. Calcium alginate beads encapsulated PMMA-g-CS nano-particles for α-chymotrypsin immobilization.

    PubMed

    Abd El-Ghaffar, M A; Hashem, M S

    2013-02-15

    Chitosan grafted with polymethyl methacrylate (PMMA-g-CS) was prepared via a free-radicals polymerization technique as a carrier for enzyme immobilization. α-Chymotrypsin (CT), as an enzyme model in this study, was immobilized onto the prepared PMMA-g-CS via covalent bonding. Calcium alginate (CA) beads were developed for encapsulating PMMA-g-CS-CT to produce PMMA-g-CS-CT/CA composite beads. Morphology and size of PMMA-g-CS particles were investigated by TEM and found to be in the nanoscale. The structure and surface morphology of the beads before and after immobilization process were characterized by FT-IR and SEM, respectively. Both the bound CT content and relative activity of immobilized enzyme were measured. A higher retained activity (about 97.7%) obtained for the immobilized CT at pH 9 for 24 h. The results indicated that immobilized CT maintained excellent performance even after 25 reuses and retained 75% from its original activity after 60 days of storage at 25 °C.

  16. Improved nonporous magnetic supports for immobilized enzymes.

    PubMed

    Halling, P J; Dunnill, P

    1979-03-01

    Ni powders coated by deposition of TiO2 or controlled oxidation to NiO develop substantial resistance to corrosion. Chymotrypsin immobilized to these coated Ni supports shows very high stability of activity on storage. Chymotrypsin immobilized by adsorption and glutaraldehyde crosslinking was fairly rapidly eluted under operational conditions in the presence of substrate. If 3-aminopropyltriethoxysilane (APS) was used to produce a covalent linkage, desorption of enzyme still occurred because of relatively unstable bonding of the silane to the oxide surface. A more stable attachment was produced by joining together many silane links with a layer of polyglutaraldehyde. The mechanism of action of APS as a coupling agent under these conditions is discussed. gamma-Fe2O3, and particularly a Mn-Zn ferrite, are suitable magnetic support materials available with smaller particle sizes. Particles below 1 mum give the expected higher specific activities of immobilized enzymes.

  17. Immobilization of lipase from grey mullet.

    PubMed

    Aryee, Alberta N A; Simpson, Benjamin K

    2012-12-01

    Grey mullet (Mugil cephalus) lipase was isolated using para-aminobenzamidine agarose and immobilized on octyl Sepharose CL-4B (o-Sep). Immobilized grey mullet lipase (GMLi) had a 10 °C higher optimum temperature compared to the free enzyme and showed remarkable thermal stability. GMLi was most active within the pH range of 8.0-9.5 with an optimum at 8.5. Immobilization also enhanced the storage stability and reusability of the enzyme with minimal changes in efficiency during repeated batches. GMLi showed variable stabilities in various organic solvents. A signal in the amide I absorption region of the FTIR spectrum of GMLi was attributed to the protein layer on o-Sep. The surface morphology of o-Sep was visualized on a Zeiss stereomicroscope as globular-shaped beads.

  18. Immobilization of the Methanogenic bacterium methanosarcina barkeri

    SciTech Connect

    Scherer, P.; Kluge, M.; Klein, J.; Sahm, H.

    1981-05-01

    Whole cells of the methanogen Methanosarcina barkeri were immobilized in an alginate network which was crosslinked with Ca/sup 2+/ calcium ions. The rates of methanol conversion to methane of entrapped cells were found to be in the same range as the corresponding rates of free cells. Furthermore, immobilized cells were active for a longer period than free cells. The particle size of the spherical alginate beads and thus diffusion has no obvious influence on the turnover of methanol. The half-value period for methanol conversion activity determined in a buffer medium was approximately 4 days at 37/degree/C for entrapped cells. The high rates of methanol degradation indicated that the immobilization technique preserved the cellular functions of this methanogenic bacterium. 24 refs.

  19. Plutonium Immobilization Can Loading Conceptual Design

    SciTech Connect

    Kriikku, E.

    1999-05-13

    'The Plutonium Immobilization Facility will encapsulate plutonium in ceramic pucks and seal the pucks inside welded cans. Remote equipment will place these cans in magazines and the magazines in a Defense Waste Processing Facility (DWPF) canister. The DWPF will fill the canister with glass for permanent storage. This report discusses the Plutonium Immobilization can loading conceptual design and includes a process block diagram, process description, preliminary equipment specifications, and several can loading issues. This report identifies loading pucks into cans and backfilling cans with helium as the top priority can loading development areas.'

  20. Combined of ultrasound irradiation with high hydrostatic pressure (US/HHP) as a new method to improve immobilization of dextranase onto alginate gel.

    PubMed

    Bashari, Mohanad; Abbas, Shabbar; Xu, Xueming; Jin, Zhengyu

    2014-07-01

    In this research work, dextranase was immobilized onto calcium alginate beads by the combination of ultrasonic irradiation and high hydrostatic pressure (US/HHP) treatments. Effects of US/HHP treatments on loading efficiency and immobilization yield of dextranase enzyme onto calcium alginate beads were investigated. Furthermore, the activities of immobilized enzymes prepared with and without US/HHP treatments and that prepared with ultrasonic irradiation (US) and high hydrostatic pressure (HHP), as a function of pH, temperature, recyclability and enzyme kinetic parameters, were compared with that for free enzyme. The maximum loading efficiency and the immobilization yield were observed when the immobilized dextranase was prepared with US (40 W at 25 kHz for 15 min) combined with HHP (400 MPa for 15 min), under which the loading efficiency and the immobilization yield increased by 88.92% and 80.86%, respectively, compared to immobilized enzymes prepared without US/HHP treatment. On the other hand, immobilized enzyme prepared with US/HHP treatment showed Vmax, KM, catalytic and specificity constants values higher than that for the immobilized enzyme prepared with HHP treatment, indicated that, this new US/HHP method improved the catalytic kinetics activity of immobilized dextranase at all the reaction conditions studied. Compared to immobilized enzyme prepared either with US or HHP, the immobilized enzymes prepared with US/HHP method exhibited a higher: pH optimum, optimal reaction temperature, thermal stability and recyclability, and lower activation energy, which, illustrating the effectiveness of the US/HHP method. These results indicated that, the combination of US and HHP treatments could be an effective method for improving the immobilization of enzymes in polymers.