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Initial Activity (initial + activity)
Selected AbstractsPRODUCTION AND BIOCHEMICAL CHARACTERIZATION OF SCLEROTINIA SCLEROTIORUM ,-AMYLASE ScAmy1: ASSAY IN STARCH LIQUEFACTION TREATMENTSJOURNAL OF FOOD BIOCHEMISTRY, Issue 5 2008IMEN BEN ABDELMALEK KHEDHER ABSTRACT Among the lytic enzymes secreted by the phytopathogen fungus Sclerotinia sclerotiorum, a starch-degrading activity has been isolated and characterized. Two extracellular ,-amylases were produced in culture medium in presence of oats flour as carbons sources. An endoamylase named ScAmy1 was purified to homogeneity by ammonium sulfate precipitation, phosphocellulose and cation exchange high performance liquid chromatographies. Molecular mass of purified ScAmy1 was estimated as 54 kDa. Amylase exhibits maximal activity at pH 5 to 6 and at temperature 60C. ScAmy1 was stable in a pH range of (5,11) and at 50C. Initial activity was still conserved 40%, after heating at 60C during 30 min. In addition, Ca2+activate and stabilize the enzyme. Starch end products were determined as low molecular oligoglucanes, the liquefying power of ScAmy1 was also tested with the Amylograph Brabender, results suggest a suitable application of ScAmy1 in several industrial process. PRACTICAL APPLICATIONS ,-Amylase ScAmy1 was highly produced from Sclerotinia sclerotiorum on oats flour , a cheaper by-agro-substrate product. The enzyme was purified and biochemical characterized. ScAmy1 was applied in starch liquefaction treatments assay. The enzyme allows a decrease in peak viscosity after gelatinization and therefore has an important liquefying power. ScAmy1 has a nearly liquefaction effect on flour compared to the commercial enzyme Novamyl, from Novozymes, donated by Novo Nordisk Co. (Denmark). Enzyme end products were analyzed and identified as oligoglucanes and dextrins. Those are widely applied in food, paper, textile and pharmacological industries. Oligosaccharides are useful as prebiotics as dietary fiber or slowly digestible starch derivatives, and they can be used in form of supplement to certain foodstuffs. [source] Incorporation of a (Cyclopentadienyl)molybdenum Oxo Complex in MCM-41 and Its Use as a Catalyst for Olefin EpoxidationEUROPEAN JOURNAL OF INORGANIC CHEMISTRY, Issue 24 2004Marta Abrantes Abstract The tricarbonyl complex [(,5 -C5H4 -COOMe)Mo(CO)3Cl] was prepared from the reaction of sodium (methoxycarbonyl)cyclopentadienide, (C5H4 -CO2Me)Na, with (Bu4N)[Mo(CO)5I]. Heating the ester with 3-(triethoxysilyl)propylamine gave the amide derivative {[,5 -C5H4 -CONH-C3H6Si(OEt)3]Mo(CO)3Cl}. The functionalised tricarbonyl complex was immobilised in the ordered mesoporous silica MCM-41 with a loading of 13 wt.-% Mo (1.4 mmol·g,1) by carrying out a grafting reaction in dichloromethane. Powder X-ray diffraction and nitrogen adsorption,desorption analysis indicated that the structural integrity of the support was preserved during the grafting and that the channels remained accessible, despite significant reductions in surface area, pore volume and pore size. The success of the coupling reaction was confirmed by 29Si and 13C (CP) MAS NMR spectroscopy. A supported dioxo complex of the type [(,5 -C5H4R)MoO2Cl] was subsequently prepared by oxidative decarbonylation of the tethered tricarbonyl complex using tert -butyl hydroperoxide (TBHP). The oxidised material is an active catalyst for the liquid phase epoxidation of cyclooctene with TBHP as the oxygen source. Similar catalytic results were obtained using the tethered tricarbonyl complex directly as a pre-catalyst since fast oxidative decarbonylation occurs under the reaction conditions used. For both systems, the desired epoxide was the only product and the initial activities were about 13 mol·molMo,1·h,1. The solid catalysts were recycled several times. Some activity was lost between the first and second runs but thereafter tended to stabilise. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source] Functionalized Multi-Wall Carbon Nanotubes for Lipase Immobilization,ADVANCED ENGINEERING MATERIALS, Issue 5 2010I. V. Pavlidis Abstract We examine the immobilization of lipase B from Candida antarctica on functionalized multi-wall carbon nanotubes (MWCNTs) through physical adsorption. MWCNTs functionalized with carboxyl-, amine- and ester- terminal groups on their surface are used as immobilization carriers. Dispersion of the nanotubes and the immobilization procedure take place in aqueous and low-water media. High enzyme loadings are attained, up to 25% of the weight of the carbon nanotubes. These novel biomaterials are characterized though FT-IR and Raman spectroscopy. The MWCNT,lipase bioconjugates exhibit high catalytic activity and increased storage and operational stability. The biomaterials retain more than 55% of their initial activity after 6 months at 4,°C, while they retain approximately 25% of their initial activity after 30 d of incubation in hexane at 60,°C. The catalytic behaviour of the immobilized enzyme depends on the terminal group of the carbon nanotubes, the concentration of the enzyme and the immobilization method employed. [source] Modelling ecological half-lives for radiocaesium in Norwegian brown trout populationsJOURNAL OF APPLIED ECOLOGY, Issue 1 2000Dag O. Hessen Summary 1.,Models of ecological half-life may be valuable and cost-effective predictive tools for authorities setting restrictions on human consumption of freshwater fish after environmental releases of radioactivity. This work aimed to validate such a model for radioactive caesium (134Cs and 137Cs) in brown trout Salmo trutta populations. Data were drawn from lakes with a wide variability in abiotic and biotic factors and initial caesium load. 2.,In Norway, the highest fallout (more than 150 kBq m,2 of 137Cs) from the Chernobyl accident occurred in Oppland county, in south central Norway. Radioactivity was measured in more than 1800 samples of brown trout in nearly 100 localities in this region during 1986,95. 3.,The back-calculated maximum initial radioactivity on 1 January 1987 showed a strong regional variability (range 443,13 370; average 3855 Bq kg,1). Large variation in initial radioactivity was also recorded on a local scale (within 50 km). 4.,The ecological half-life model for caesium in brown trout populations for 1987,94/95 gave a close fit to real data from all localities with sufficient time series. Predicted half-lives ranged from 1·2 to 4·2 years (average 2·5) but 95% confidence limits were narrow (2·7 and 2·3 years). 5.,The overall variability in radioactivity levels over time was almost entirely related to the initial load and, with few exceptions, 88% of the changes in radioactivity was explained by the simple regression model. Modest variability in ecological half-life was not correlated with initial activity, and no clear effects of water quality or season could be detected. For most lakes, levels of radioactivity in brown trout appeared to be predictable, with high accuracy after a fallout event, without extensive information on population ecology and water quality. However, more detailed work may be required to assess patterns within individual lakes. [source] Influence of carbohydrates on stability of papain in aqueous tetrahydrofuran mixtureJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 1 2009András Szabó Abstract BACKGROUND: The use of enzymes in organic solvents has extended the scale of their practical applications. Papain has been widely used in chemical syntheses because of its broad substrate specificity. The aim of the present study was to improve the stability of papain in aqueous tetrahydrofuran (THF) by using different saccharides. The effects of these carbohydrates on the structure of papain were followed by means of circular dichroism (CD) and fluorescence spectroscopic measurements. RESULTS: In contrast with most organic solvents, 60% (i.e. 600 mL L,1) THF practically inactivated the enzyme within 30 min. Sugars protected papain from THF-induced inactivation in the sequence D -ribose > D -fructose > D -glucose > D -saccharose > D -raffinose. Ribose at 1.6 mol L,1 proved the most effective stabiliser: in 60% THF in the presence of ribose, papain preserved about 55% of its initial activity after 2 h. Fluorescence and near-UV CD spectroscopic measurements revealed local changes in the papain conformation. With decrease in the free amino group content of the enzyme, protein-carbohydrate interactions (Schiff base formation) were detected. CONCLUSION: These results demonstrate that the catalytic activity and stability of papain may be increased in aqueous THF by using different carbohydrates, when a more compact structure of the enzyme is formed. Copyright © 2008 Society of Chemical Industry [source] Simultaneous purification and immobilization of bitter gourd (Momordica charantia) peroxidases on bioaffinity supportJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 2 2005Suhail Akhtar Abstract This paper demonstrates the construction of an inexpensive bioaffinity adsorbent by simply incubating Sephadex G 50 matrix with jack bean meal extract at room temperature. Sephadex G 50 adsorbed 17 mg Con A (concanavalin A) per g of the matrix. Con A-adsorbed Sephadex was employed for the immobilization of glycoenzymes directly from ammonium sulfate-fractionated proteins of bitter gourd. The obtained bioaffinity support was very efficient for high yield immobilization of peroxidases from bitter gourd and it bound nearly 425 enzyme units per g of the matrix. Bitter gourd peroxidase immobilized on lectin,Sephadex support showed a very high effectiveness factor, ,,,' of 1.25. Immobilized BGP preparation was quite stable against the denaturation induced by pH, heat, urea, Triton X 100, Tween 20, SDS, Surf Excel and water-miscible organic solvents: dimethyl sulfoxide and dimethyl formamide. Low concentration of detergents like SDS, Tween 20, and Triton X 100 enhanced the activity of soluble and immobilized bitter gourd peroxidase. Peroxidase bound to the bioaffinity support exhibited very high resistance to proteolysis caused by the trypsin treatment. Con A,Sephadex-bound bitter gourd peroxidase retained 85% of its initial activity after treatment with 2.5 mg trypsin per cm3 of incubation mixture for 1 h at 37 °C while the soluble enzyme lost nearly 40% of the initial activity under similar incubation conditions. Immobilized bitter gourd peroxidase preparation appeared to be more rigid to proteolysis mediated by trypsin compared with soluble bitter gourd peroxidase. Copyright © 2004 Society of Chemical Industry [source] COVALENT IMMOBILIZATION OF INVERTASE ON CHEMICALLY ACTIVATED POLY (STYRENE-2-HYDROXYETHYL METHACRYLATE) MICROBEADSJOURNAL OF FOOD BIOCHEMISTRY, Issue 3 2008HAYDAR ALTINOK ABSTRACT A carrier for invertase enzyme was synthesized from styrene (S) and 2- hydroxyethyl methacrylate (HEMA) in the form of microbeads. These poly (styrene-2-hydroxyethyl methacrylate), P(S-HEMA) microbeads were activated by epichlorohydrin (ECH) treatment for covalent immobilization. The free and immobilized invertase were assayed in the hydrolysis of sucrose to glucose, and the obtained results were compared. The optimum pH was 4.5 for free and 5.5 for immobilized invertase. The optimum temperature of invertase shifted from 45C to 55C upon immobilization. For free and immobilized enzymes, kinetic parameters were calculated as 4.1 × 10,3 mol L,1and 9.2 × 10,3 mol L,1for Km, and 6.6 × 10,2 mol L,1 min,1and 4.1 × 10,1 mol L,1 min,1for Vmax, respectively. After 1 month of storage at 4C, free enzyme retained 36% of its initial activity, while for the ECH-activated P(S-HEMA) immobilized enzyme, P(S-HEMA)-E, this value was observed as 67%. In repeated batch use, i.e., 20 times in 3 days, 78% retention of the initial activity was observed for P(S-HEMA)-E system. PRACTICAL APPLICATIONS Immobilization of enzymes are very important for many industrial applications, e.g., food, medicine, pharmacology, etc. Invertase converts sucrose to glucose and fructose, which have wide applications in food industry especially as sweeteners. Glucose,fructose mixture has much lower crystallinity compared to sucrose and therefore used in the production of noncrystallizing jams and creams. They are also used as liquid sweeteners. Immobilization enables repeated use, provides significant reduction in the operation costs, facilitates easy separation and speeds up recovery of enzyme and extends the stability of enzyme by protecting the active material from deactivation. Industrial application of immobilized invertase may decrease the production cost of glucose,fructose mixture because it could be used repeatedly for long periods. Although invertase is not a very expensive enzyme, the technique can also be applied to expensive ones for biotechnological productions. [source] Is the failure to detect stimulus deviance during sleep due to a rapid fading of sensory memory or a degradation of stimulus encoding?JOURNAL OF SLEEP RESEARCH, Issue 2 2005MERAV SABRI Summary The mismatch negativity (MMN) is thought to reflect the outcome of a system responsible for the detection of change in an otherwise repetitive, homogenous acoustic environment. This process depends on the storage and maintenance of a sensory representation of the frequently presented stimulus to which the deviant stimulus is compared. Few studies have been able to record the MMN in non-rapid eye movement (NREM) sleep. This pattern of results might be explained by either a rapid fading of sensory memory or an inhibition of stimulus input prior to entry into the cortical MMN generator site. The present study used a very rapid rate of presentation in an attempt to capture mismatch-related negativity prior to the fading of sensory memory. Auditory event-related potentials were recorded from 12 subjects during a single sleep period. A 1000 Hz standard stimulus was presented every 150 ms. At random, on 6.6% of the trials, the standard was changed to either a large 2000 Hz or a small 1100 Hz deviant. In wakefulness, the large deviant elicited an extended negativity that was reduced in amplitude following the presentation of the small deviant. This negativity was also apparent during REM sleep following the presentation of the large deviant. These deviant-related negativities (DRNs) were probably a composite of N1 and MMN activity. During NREM sleep (stage 2 and slow-wave sleep), only the large deviant continued to elicit a DRN. However this DRN might be overlapped by the initial activity of a component that is unique to sleep, the N350. There was little evidence of the DRN or the MMN during sleep following the presentation of the small deviant. A rapid rate of presentation, therefore, does not preserve the MMN following small deviance within sleep. It is possible that inhibition of sensory input occurs before entry into the MMN generating system in the temporal cortex. [source] The Properties of Covalently Immobilized Trypsin on Soap-Free P(MMA-EA-AA) Latex ParticlesMACROMOLECULAR BIOSCIENCE, Issue 4 2005Kai Kang Abstract Summary: The covalent immobilization of trypsin onto poly[(methyl methacrylate)- co -(ethyl acrylate)- co -(acrylic acid)] latex particles, produced by a soap-free emulsion polymerization technique, was carried out using the carbodiimide method. The catalytic properties and kinetic parameters, as well as the stability of the immobilized enzyme were compared to those of the free enzyme. Results showed that the optimum temperature and pH for the immobilized trypsin in the hydrolysis of casein were 55,°C and 8.5, both of which were higher than that of the free form. It was found that Km (Michaelis constant) was 45.7 mg,·,ml,1 and Vmax (maximal reaction rate) was 793.0 ,g,·,min,1 for immobilized trypsin, compared to a Km of 30.0 mg,·,ml,1 and a Vmax of 5,467.5 ,g,·,min,1 for free trypsin. The immobilized trypsin exhibited much better thermal and chemical stabilities than its free counterpart and maintained over 63% of its initial activity after reusing ten times. TEM photograph of latex particles after trypsin immobilization. [source] A dimeric 5- enol -pyruvyl-shikimate-3-phosphate synthase from the cyanobacterium Spirulina platensisNEW PHYTOLOGIST, Issue 2 2001Giuseppe Forlani Summary ,,Isolation and biochemical characterization is reported here of 5- enol -pyruvyl-shikimate-3-phosphate (EPSP) synthase, the enzyme that catalyses the sixth step in the common prechorismate pathway of aromatic amino acid biosynthesis and the target of the widely used herbicide glyphosate, from the cyanobacterium Spirulina platensis. ,,Homogeneous enzyme preparations were obtained by ammonium sulphate fractionation, anion-exchange and substrate-elution chromatography, and chromatofocusing. Protein characterization was carried out by conventional kinetic analysis, PAGE and gel permeation. ,,A 2800-fold purification was achieved, with a recovery of 20% of initial activity. Unusually low apparent affinities for both substrates, phosphoenolpyruvate and shikimate-3-phosphate, did not correspond to decreased glyphosate sensitivity. During SDS-PAGE, the protein migrated as a single band corresponding to a molecular mass of c. 49 kDa. The behaviour of the protein upon gel permeation chromatography under nondenaturing conditions was, however, consistent with a mass of c. 91 kDa. ,,The native enzyme appears to be homodimeric, a remarkable feature that has not been previously reported for EPSP synthases from either cyanobacteria or higher plants. The presence of mono- and dimeric EPSP synthases could represent an important tool for cyanobacterial classification. [source] ,-Glucosidase and peroxidase stability in crude enzyme extracts from green beans of Vanilla planifolia AndrewsPHYTOCHEMICAL ANALYSIS, Issue 3 2001Mark J. W. Dignum Abstract The extraction method for ,-glucosidase from green vanilla beans has been studied. The effect of storage of green beans and protein extracts on ,-glucosidase and peroxidase activity was investigated: the best method, resulting in the highest enzyme activities, particularly for glucosidase, was through extraction of very fresh green beans in the presence of BisTris propane buffer at pH 8. The best method for storage of the extracts was at ,80°C after addition of 15% glycerol, when over 90% of initial activity was still present. Peroxidase activity did not change in frozen beans or in frozen extracts. Copyright © 2001 John Wiley & Sons, Ltd. [source] Fabrication of poly(ethylene glycol)-based hydrogels entrapping enzyme-immobilized silica nanoparticlesPOLYMERS FOR ADVANCED TECHNOLOGIES, Issue 7 2010Eunji Jang Abstract In this study, we immobilized enzymes by combining covalent surface immobilization and hydrogel entrapment. A model enzyme, glucose oxidase (GOX), was first covalently immobilized on the surface of silica nanoparticles (SNPs) via 3-aminopropyltriethoxysilane (APTES), and the resultant SNP-immobilized enzyme was physically entrapped within photopolymerized hydrogels prepared from two different molecular weights (MWs) (575 and 8000,Da) of poly(ethylene glycol)(PEG). The hydrogel entrapment resulted in a decrease in reaction rate and an increase in apparent Km of SNP-immobilized GOX, but these negative effects could be minimized by using hydrogel with a higher MW PEG, which provides higher water content and larger mesh size. The catalytic rate of the PEG 8000 hydrogel was about ten times faster than that of the PEG 575 hydrogel because of enhanced mass transfer. Long-term stability test demonstrated that SNP-immobilized GOX entrapped within hydrogel maintained more than 60% of its initial activity after a week, whereas non-entrapped SNP-immobilized GOX and entrapped GOX without SNP immobilization maintained less than 20% of their initial activity. Incorporation of SNPs into hydrogel enhanced the mechanical strength of the hydrogel six-fold relative to bare hydrogels. Finally, a hydrogel microarray entrapping SNP-immobilized GOX was fabricated using photolithography and successfully used for quantitative glucose detection. Copyright © 2009 John Wiley & Sons, Ltd. [source] Direct utilization of ethanol on ceria-based anodes for solid oxide fuel cellsASIA-PACIFIC JOURNAL OF CHEMICAL ENGINEERING, Issue 1 2009Massimiliano Cimenti Abstract The direct utilization of ethanol was investigated in CuCeO2, CuZr0.35Ce0.65O2 (ZDC) and Cu/RuZr0.35Ce0.65O2 anodes for solid oxide fuel cells (SOFC). The anodes were prepared by impregnation with nitrate precursors on a porous layer of yttria-stabilized zirconia (YSZ) obtained by tape casting, while (La0.8Sr0.2MnO3,,) LSM cathodes were screen-printed. The cells were tested in both hydrogen and ethanol. The outlet gas composition was monitored with a gas chromatograph, which showed that almost all the ethanol was decomposed, mainly to H2, CH4, CO, H2O and C2H4. The maximum power outputs obtained in ethanol were 0.075 and 0.400 W/cm2 on CuCeO2|YSZ|LSM and CuZDC|YSZ|LSM, respectively. All cells were more active in alcohol than in hydrogen with the peak performance occurring after approximately 4 h. That is, the power density initially increased, peaked and then decreased. This behavior was likely a consequence of carbon deposition that initially results in an improvement of the electronic conductivity in the anode but later results in the blocking of the active sites. Zirconia doping (in the ZDC anodes) resulted in better stability and, in addition, the initial activity of the ZDC anodes could be recovered after approximately 1 h of exposure to humidified hydrogen, whereas the initial activity of the ceria anodes could not be recovered. The addition of ruthenium (<0.5 wt%) further improved the stability by delaying the onset of carbon formation. Copyright © 2008 Curtin University of Technology and John Wiley & Sons, Ltd. [source] Plant cell calcium-rich environment enhances thermostability of recombinantly produced ,-amylase from the hyperthermophilic bacterium Thermotoga maritimeBIOTECHNOLOGY & BIOENGINEERING, Issue 5 2009Monica C. Santa-Maria Abstract In the industrial processing of starch for sugar syrup and ethanol production, a liquefaction step is involved where starch is initially solubilized at high temperature and partially hydrolyzed with a thermostable and thermoactive ,-amylase. Most amylases require calcium as a cofactor for their activity and stability, therefore calcium, along with the thermostable enzyme, are typically added to the starch mixture during enzymatic liquefaction, thereby increasing process costs. An attractive alternative would be to produce the enzyme directly in the tissue to be treated. In a proof of concept study, tobacco cell cultures were used as model system to test in planta production of a hyperthermophilic ,-amylase from Thermotoga maritima. While comparable biochemical properties to recombinant production in Escherichia coli were observed, thermostability of the plant-produced ,-amylase benefited significantly from high intrinsic calcium levels in the tobacco cells. The plant-made enzyme retained 85% of its initial activity after 3,h incubation at 100°C, whereas the E. coli -produced enzyme was completely inactivated after 30,min under the same conditions. The addition of Ca2+ or plant cell extracts from tobacco and sweetpotato to the E. coli -produced enzyme resulted in a similar stabilization, demonstrating the importance of a calcium-rich environment for thermostability, as well as the advantage of producing this enzyme directly in plant cells where calcium is readily available. Biotechnol. Bioeng. 2009; 104: 947,956. © 2009 Wiley Periodicals, Inc. [source] Stabilization of glucose oxidase in alginate microspheres with photoreactive diazoresin nanofilm coatingsBIOTECHNOLOGY & BIOENGINEERING, Issue 1 2005Rohit Srivastava Abstract The nanoassembly and photo-crosslinking of diazo-resin (DAR) coatings on small alginate microspheres for stable enzyme entrapment is described. Multilayer nanofilms of DAR with poly(styrene sulfonate) (PSS) were used in an effort to stabilize the encapsulation of glucose oxidase enzyme for biosensor applications. The activity and physical encapsulation of the trapped enzyme were measured over 24 weeks to compare the effectiveness of nanofilm coatings and crosslinking for stabilization. Uncoated spheres exhibited rapid loss of activity, retaining only 20% of initial activity after one week, and a dramatic reduction in effective activity over 24 weeks, whereas the uncrosslinked and crosslinked {DAR/PSS}-coated spheres retained more than 50% of their initial activity after 4 weeks, which remained stable even after 24 weeks for the two and three bilayer films. Nanofilms comprising more polyelectrolyte layers maintained higher overall activity compared to films of the same composition but fewer layers, and crosslinking the films increased retention of activity over uncrosslinked films after 24 weeks. These findings demonstrate that enzyme immobilization and stabilization can be achieved by using simple modifications to the layer-by-layer self-assembly technique. © 2005 Wiley Periodicals, Inc. [source] Enzyme immobilization on ultrafine cellulose fibers via poly(acrylic acid) electrolyte graftsBIOTECHNOLOGY & BIOENGINEERING, Issue 4 2005Hong Chen Abstract Ultrafine cellulose fiber (diameter 200,400 nm) surfaces were grafted with polyacrylic acid (PAA) via either ceric ion initiated polymerization or methacrylation of cellulose with methacrylate chloride (MACl) and subsequent free-radical polymerization of acrylic acid. PAA grafts by ceric ion initiated polymerization increased with increasing reaction time (2,24 h), monomer (0.3,2.4 M), and initiator (1,10 mM) concentrations, and spanned a broad range from 5.5,850%. PAA grafts on the methacrylated cellulose fibers also increased with increasing molar ratios of MACl to cellulosic hydroxyl groups (MACl/OH, 2,6.4) and monomer acrylic acid (AA) to initiator potassium persulfate (KPS) ratios ([AA]/[KPS], 1.5,6), and were in a much narrower range between 12.8% and 29.4%. The adsorption of lipase (at 1 mg/ml lipase and pH 7) and the activity of adsorbed lipase (pH 8.5, 30°C), in both cases decreased with increasing PAA grafts. The highest adsorption and activity of the lipase on the ceric ion initiated grafted fibers were 1.28 g/g PAA and 4.3 U/mg lipase, respectively, at the lowest grafting level of 5.5% PAA, whereas they were 0.33 g/g PAA and 7.1 U/mg lipase, respectively, at 12.8% PAA grafts on the methacrylated and grafted fibers. The properties of the grafted fibers and the absorption behavior and activity of lipase suggest that the PAA grafts are gel-like by ceric-initiated reaction and brush-like by methacrylation and polymerization. The adsorbed lipase on the ceric ion-initiated grafted surface possessed greatly improved organic solvent stability over the crude lipase. The adsorbed lipases exhibited 0.5 and 0.3 of the initial activity in the second and third assay cycles, respectively. © 2004 Wiley Periodicals, Inc. [source] Enhanced Glucose to Fructose Conversion in Acetone with Xylose Isomerase Stabilized by Crystallization and Cross-LinkingBIOTECHNOLOGY PROGRESS, Issue 5 2004Kati M. Vilonen The effects of acetone and ethanol on glucose to fructose conversion catalyzed by soluble and cross-linked crystalline (CLXIC) xylose isomerase were studied. Relative to pure buffer solvent, the fructose production rate was more than doubled in 50% acetone. The same kind of increase in the isomerization rate was not seen with ethanol. Increase both in acetone and in ethanol concentration in the reaction solvent enhanced the production of fructose. At 50 °C in pure buffer solvent the reaction mixture contained 49% fructose in equilibrium and in 90% acetone the fructose equilibrium content was 64%. Furthermore, CLXIC was relatively stable in the presence of high concentration of acetone: 70,80% of activity was left after incubation for 24 h at 50 °C in buffer solutions (pH 7.2) containing 10,90% acetone. In buffer containing 50% ethanol only 2% of the initial activity of CLXIC was retained after 24 h at 50 °C. Soluble xylose isomerase was considerably less stable than CLXIC in both acetone- and ethanol-containing solutions. These results show that the addition of acetone enhances the production of fructose from glucose by enhancing the reaction rate and shifting the equilibrium toward fructose. However, xylose isomerase must be in the form of cross-linked crystals for maximal activity and stability. [source] | ||||||||||||||||