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Alanine dehydrogenase catalyzed the conversion of 3-fluoropyruvate into 3-fluoro-L-alanine in the presence of NADH and ammonia. The optimum pH of the reaction was 7.8. The K(m) values of the enzyme for 3-fluoropyruvate, polyethylene glycol-bound NADH, and ammonia were 2.94, 0.56, and 105mM, respectively. 3-Fluoro-L-ala...
The effects of temperature on the hydrolysis of lactose by immobilized beta-galactosidase were studied in a continuous flow capillary bed reactor. Temperature affects the rates of enzymatic reactions in two ways. Higher temperatures increase the rate of the hydrolysis reaction, but also increase the rate of thermal dea...
Selected Digitalis lanata cell lines cultivated in 1-L shake flasks or 20-L airlift bioreactors converted beta-methyldigitoxin into beta-methyldigoxin with almost no side reactions. This biotransformation process was optimized with regard to substrate supply and culture medium composition, and was then scaled up to a v...
A structured kinetic model is developed to describe the dynamics of hybridoma growth and the production of monoclonal antibodies and metabolic waste products in suspension culture. The crucial details of known metabolic processes in hybridoma cells are incorporated by dividing the cell mass into four intracellular meta...
Theories of protein partitioning in two-phase polymer systems which account for the effects of different aspects of system composition-such as the choice of materials, protein size, polymer molecular weight, polymer concentration, salt concentration, and affinity ligands-are reviewed. Although the present models provid...
The antiviral activity of a surface-bonded quaternary ammonium chloride (QAC) was examined in this study. The mechanism of inactivation was elucidated by a combination of infectivity assay, radioactive labeling assay, and sedimentation analysis. Although the virions are still infectious when attached onto the chemicall...
Model experiments were performed with starved yeast (Saccharomyces cerevisiae) cultures in a batch reactor in order to develop a better understanding of NAD(P)H and flavin culture fluorescence. Fluorescence was monitored during aerobic-anaerobic-aerobic transitions and ethanol and glucose substrate addition experiments...
Measurements of mass transfer in a highly viscous pseudoplastic broth, which is typical to Xanthomonas campestris fermentations, are difficult to obtain by conventional methods and little data is available. A novel research method that uses bioluminescence for mass transfer studies has been developed. A plasmid carryin...
Near-homogeneous forms of glucoamylases I and II, previously purified from an industrial Aspergillus niger preparation, were used to hydrolyze a number of di- and trisaccharides linked by alpha-D-glucosidic bonds. Maximum rates and Michaelis constants were obtained at various temperatures and pH values with glucoamylas...
Biological processes may be used to convert gas phase substrates, such as H(2)S, CH(4), CO, H(2), and CO(2), to useful products. Utilization of these substrates is often a mass transfer limited process, first requiring absorption across the gas-liquid interface and diffusion through the culture medium to the cell surfa...
The aim of this article is to develop the on-line measurement system of enzyme reaction rates, using an advanced pH controller, which is called a repetitive PF(Programmed Controller/Feedback Compensator) System. In the esterolysis reaction of the N-acetyltyrosine ethyl ester by alpha-chymotrypsin, the enzyme reaction r...
A pH-dependent "series-type" enzyme deactivation model using rapid protonation and deprotonation equilibria and the relatively slower inactivation rates is presented. From the enzyme activity-time trajectories at different pH the models presented permit the evaluation of some of the protonation and inactivation rate co...
The efficiency of two different agitation systems (airlift and paddlewheel) in the biomass photoproduction of a nitrogen-fixing filamentous blue-green alga was evaluated outdoors, and the elemental and molecular composition of the cells grown with each system was analyzed. With the paddlewheel system, the productivity ...
In view of the biochemical reaction catalyzed by enzyme powder suspended in a water-insoluble organic solvent, an equation was derived to estimate the amount of water bound to the enzyme powder. With this equation, an apparent adsorption isotherm between free water (water freely dissolved in benzene) and bound water (w...
As part of a process development campaign, studies have been conducted to determine the influence of induction temperature on the expression of two different malaria antigens, RN1 and RT2. Single-step temperature inductions, in which growth at 32.0 degrees C is followed by a shift in temperature to a desired setpoint, ...
Protein localization in Saccharomyces cerevisiae was studied with two plasmid systems used as a model: one containing the SUC2 structural gene fused with the MFalpha1 (alpha-factor) promoter and signal-sequence, the other containing the entire SUC2 gene. Special emphasis was placed on the effect of promoter/signal-sequ...
A method for the simultaneous extraction of oil and proteins from vegetable meals is presented. The method uses hydrocarbon reverse micelles, so that the oil is extracted directly into the hydrocarbon phase and the proteins are solubilized in the water pools of the reverse micelles. The surfactant used is bis (2-ethylh...
The enzymatic conversion of gaseous substrates represents a novel concept in bioprocessing. A critical parameter in such systems is the water activity, A(w) The present article reports the effect of A(w) on the catalytic performance of alcohol oxidase acting on ethanol vapors. Enzyme activity in the gas-phase reaction ...
Reactive polymers have been prepared by copolymeriz-ing N-isopropyl acrylamide (NIPAM) with N-acryloxy-succinimide (NASI) or glycidyl methacrylate (GMA). The amino groups of ligands could react with the residues of NASI or GMA and the polymers could be precipitated by temperature and/or salinity variation, since they c...
A novel process has been developed which uses reversed micelles to isolate denatured protein molecules from each other and allows them to refold individually. These reversed micelles are aqueous phase droplets stabilized by the surfactant AOT and suspended in isooctane. By adjusting conditions such that only one protei...
A novel process has been developed to improve the refolding yield of denatured proteins. It uses reversed micelles to isolate denatured protein molecules from each other and thus, upon refolding, reduces the intermolecular interactions which lead to aggregation. The feasibility of this process was first demonstrated wi...
The concept of pore size distribution is incorporated into the Clark model of enzyme immobilization in the present study. This refined model predicted that in the case of small harmonic pore radius with the same surface area and porosity of the support, more enzyme could be loaded in a support with nonuniform pores tha...
A new method for the immobilization of microbial cells has been developed. Whole cells of Escherichia coli with aspartase activity were immobilized by capture on the surface of cross-linked poly(N-benzyl-4-vinylpyridinium bromide) containing styrene (BVPS resin), an insoluble pyridinium-type resin. When a suspension of...
Targeted molecular therapies, mainly enzyme inhibitors and humanized antibodies, are being widely developed, especially in the area of lung cancer. Though often considered to be better tolerated than conventional cytotoxic chemotherapies, targeted molecular therapies induce specific toxicities that may have detrimental...
To study the effects of Hemp seeds protein (HSP) on antifatigue and immunnomodulation effect in mice.
To clone CPn0308 gene from Clamyida pneumonia and express its fusion protein, to make antibodies to fusion protein GST-CPn0308, and to further localize endogenous protein preliminarily using antibodies raised with CPn0308 fusion protein.
Although depression is a common finding in Parkinson's disease (PD), its neurobiological mechanism is still unknown. The purpose of this study was to determine whether there are specific spectral electroencephalographic (EEG) characteristics that distinguish depressed from non-depressed PD patients. The study was perfo...
One of the clinical features in the patients with Parkinson's disease (PD) is an apparent low incidence of colds. MxA protein is an interferon-induced protein which inhibits influenza A viral infection. MxA protein has been found in association with Lewy bodies in neurons in PD patients' brains. We performed a semiquan...
The aim of the authors is to examine the relationship between the cytokine levels that are thought to be involved in stroke etiopathogenesis (tumor necrosis factor [TNF]-alpha, interleukin [IL]-2, IL-6, IL-8, IL-11), soluble protein C receptor (sEPCR), and factor VIII (FVIII) levels. The study included 27 patients with...
11beta-Hydroxysteroid dehydrogenase type 1 (11beta-HSD1) enzyme catalyzes interconversion of inactive cortisone to active cortisol. Its expression in adipose tissue has been associated with obesity and some of its metabolic disorders. Controversies regarding which fat depots [subcutaneous adipose tissue (SAT) or viscer...
Pirt's maintenance model has been widely accepted for the effects of growth rate and maintenance on growth yield. However, the interpretation of parameters in Pirt's model as biological constants is difficult for energy-sufficient culture growth. In this study, a mechanistic model for the growth energetics of energy-su...
Arthrobacter simplex cells, which convert the steroid hydrocortisone to prednisolone, have been entrapped in a thermally reversible hydrogel. Such hydrogels exhibit a lower critical solution temperature (LCST) wherein the gel shrinks and deswells when it is warmed through its LCST, and then reversibly expands and reswe...
Theoretical and experimental studies concerning the application of modern adaptive techniques for the control of continuous alcoholic fermentation of glucose by a yeast strain conducted in a multistage reactor are reported. The practical control objective was the regulation of the substrate concentration in the process...
Enhancement of the productivity of xylanase and beta-xy-losidase of Aspergillus ochraceus was investigated by multistep mutagenesis. The spores of the wild strain were subjected to UV and N-methyl-N-nitro-N-nitro-soguanidine (NTG). The hyperxylanolytic mutant (NG-13), which showed good clearing on the surface of the xy...
In this article we describe the use of bench-scale single-fiber dialyzers for the development and testing of an immobilized enzyme reactor for the treatment of leukemia. The treatment is based on the enzymatic removal of specific amino acids from the blood of leukemia patients. L-Lysine alpha-oxidase and catalase were ...
Lignin prepared by acid and enzyme hydrolysis of a softwood mixture adsorbs acetone, butanol, and other alcohols while showing only a slight uptake of glucose. Adsorption of butanol is independent of temperature in the range of 30-65 degrees C. The Polanyi theory fits adsorption for the linear alcohols methanol through...
Semicontinuous fermentations, in which a fraction of a culture is replaced with fresh media at regular intervals, have been previously used as a means of approximating continuous growth. In most cases deviations from continuous operation were erroneously estimated using Fencl's model, which is only valid when the speci...
The results of numerical simulations for a growing biological film are presented to justify the use of steady-state biofilm models for approximating the behavior of both unlimited and shear-limited biofilms. For an unlimited biofilm we show that although the total biofilm thickness may continue to increase over time, t...
This article describes the characteristics of silk fibroin membranes and glucose oxidase, immobilized in membranes as determined by a variety of physical methods, mainly the spin-label electron spin resonance (ESR) method. The properties of membranes insolubilized by different methods, i. e., immersion in 80% methanol ...
Water-insoluble proteases were prepared by immobilizing papain and chymotrypsin onto the surface of polyacrolein microspheres with and without oligoglycines as spacer. The activity of immobilized proteases was found to be still high toward small ester substrates, but very low toward casein, a high-molecular-weight subs...
A quantitative assay of beta-galactosidase activity in single cells of Saccharomyces cerevisiae has been developed using a fluorogenic substrate and flow cytometry [reported in Wittrup & Bailey, Cytometry, 9,394 (1988)]. The beta-galactosidase activity is expressed in yeast from the Escherichia coli lacZ gene under the...
Inactivation of alpha-chymotrypsin in aqueous solutions of alcohols and diols proceeds both reversibly and irreversibly. Reversible loss of the specific enzyme activity results from conformational changes (unfolding) of the enzyme detected by fluorescence spectroscopy. Multipoint covalent attachment to the matrix of po...
Experimental elucidation of the metabolic load placed on bacteria by the expression of foreign protein is presented. The host/vector system is Escherichia coli RR1/pBR329 (amp(r), cam(r), and let(r)). Plasmid content results, which indicate that the plasmid copy number monotonically increases with decreasing growth rat...
A mathematical model of metal affinity partitioning has been derived and used to describe protein partitioning in Cu (II)PEG/dextran systems. A working model has been extended to account for inhibition, which for metal affinity extraction is the inhibition of protein-metal binding by hydrogen ion. PEG/dextran partition...
Production of various extracellular enzymes (the beta-lactamases from Streptomyces albus G, Streptomyces cacaoi, Actinomadura R39, and the DD-carboxypeptidase from Streptomyces R61) by genetically engineered Streptomyces lividans TK24 in Lennox broth medium reached a maximum after 36 to 48 h. Subsequently, the enzyme a...
An enzyme sensor system has been developed to assess the freshness level in fish tissue. The system was designed to measure the K value, the concentration ratio of [Hx + HxR] and [Hx + HxR + IMP], where Hx, HxR, and IMP are hypoxanthine, inosine and inosine-5'-monophosphate, respectively. The [Hx + HxR] concentration i...
The stereoselective reduction of ethyl acetoacetate to (+)-(S)-ethyl 3-hydroxybutyrate catalyzed by Saccharomyces cerevisiae was optimized by means of chemometric methods. The quantitative effects of temperature, time of incubation, and concentrations of yeast and substrate on the optical purity and on the percent of r...
Amino groups of trypsin (EC 3.4.21.4) were reductively alkylated in solid phase to obtain a surface-active and biologically active enzyme in an o/w emulsion system. Trypsin adsorbed on a benzamidine-sepharose column was reductively alkylated with n-octanal in the presence of sodium borohydride, i.e., trypsin-C8. Activi...
Intracellular metabolite concentration and enzyme activity measurements were made to explain the new metabolic and growth phenomena seen in the micro-aerobic, continuous yeast cultures described in Part I. The results of these assays suggested mechanisms for the observed maximum in the specific ethanol productivity as ...
Mathematical models of the catabolic pathways, the utilization and waste of ATP, and the factors affecting yeast growth in a micro-aerobic chemostat are presented. The models incorporate the intracellular metabolite and enzyme activity assays performed in Part II to explain the unusual macroscopic chemostat behaviors r...
Hybridoma cells (S3H5/gamma2bA2) were cultivated in spinner flasks with 1% serum media and serum-free media. Monoclonal antibody productivity was maintained in 1% serum media. However, cells in serum-free media showed a decrease in antibody productivity, and it completely disappeared in IMDM-based low protein medium. T...
The production of IGF-1 (insulin-like growth factor 1), expressed in Escherichia coli as a secreted fusion protein with affinity for the Fc region of IgG, was monitored automatically during fermentations. A sampling device was used to automatically inject filtered culture medium from the fermentor onto a small affinity...
Various factors affecting the catalytic activity of pure lipase of Pseudomonas fluorescens in microaqueous benzene were investigated with respect to lactonization of 15-hydroxypentadecanoic acid. Without deposition of the enzyme or of the enzyme plus activity enhancer (additive) on celite powder, the pure enzyme was ve...
The protein releases, the particle size distribution and the viscosity of disrupted E. coli suspensions from Dyno Mill KDL, Manton Gaulin 15 M-8TA and Microfluidizer M-110 were determined. The effects of these parameters on separation of the cell debris from the protein solution by centrifugation and by filtration were...
In order to develop an efficient process for large-scale production of recombinant protein, various factors were studied which affect the productivity of Sf-9 (Spodoptera frugiperda) insect cells when using the baculovirus expression system. It was shown that upon infection with the Bac-BRV6L recombinant baculovirus, t...
Based on the well-known mechanism describing Michaelis-Menten kinetics, three rate expressions may be developed: the exact solution (Model 1), a rate equation resulting from the pseudo-steady-state assumption (Model 2), and Model 2 with the additional assumption that the amount of free substrate is approximately equal ...
A supported liquid membrane system was developed for the extraction of ethanol during semicontinuous fermentation of Saccharomyces bayanus. it consisted of a porous Teflon sheet as support, soaked with isotridecanol. This assembly permitted combining biocompatibility, permeation efficiency, and stability. The removal o...
Recombinant cell growth and protein synthesis by a recombinant Escherichia coli under various inducing conditions are compared to the predictions of a mathematical model. The mathematical model used was a combination of two literature models: (1) an empirical kinetic model for recombinant growth and product formation a...
A technique for enzyme reuse and product recovery from enzymatic catalysis in microemulsions is demonstrated. The enzymatic reaction is performed in a homogeneous isotropic microemulsion; AOT (sodium bis-(2-ethyl- hexyl)sulfosuccinate)/isooctane/buffer or C(12)E(5)(penta ethylene glycol dodecyl ether)/heptane/buffer. B...
A bioenergetic model has been developed for the fermentation of glucose by Bacillus polymyxa. This model uses energy balances to determine which pathways are utilized by the substrate. The model can predict substrate consumption, biomass formation, and the product distribution for this fermentation. The products are ca...
The concept of a charged membrane bioreactor (CMBR) has been proposed for continuous reactions of enzymatic reduction dependent upon the nicotinamide coenzyme NADP(H). It was found that a composite membrane with a negative charge, NTR 7410, could retain NADP(H) selectively without any chemical modification. Several per...
A theoretical model was constructed in order to study charged membrane bioreactors (CMBRs). In this model, it was postulated that a native nicotinamide coenzyme NADP(H) can be partially retained by a charged membrane in continuous operation. A multienzyme system composed of NADPH-dependent aldose reductase (AR) and glu...
The notion that the regulated and flux-controlling enzyme in a metabolic network need not correspond suggests that the purpose of regulation may not be flux homeostasis under all physiological circumstances. Additionally, the fact that diversity in the function of intact metabolic networks exists suggests that in addit...
A stochastic model is proposed to simulate the growth of anchorage dependent cells on a flat surface. The model, based on representing the cell shapes on the surface as external irregular polygons with the nuclei distributed as a set of Poisson points (producing a modified Voronoi tessellation of 2 space) and incorpora...
A protein separation scheme combining affinity or ion exchange sorption with hollow fiber cross-flow filtration is described. Sorptive gel particles were loaded into the shell side of a hollow fiber membrane module. In the adsorption step, crude protein mixtures were passed through the lumen and permeating proteins pas...
A lipoprotein lipase (LPL) was made water insoluble by immobilizing onto the surface of polyacrolein (PAA) microspheres with and without oligoglycines as spacer. The activity of the immobilized LPL was found to remain high toward a small ester substrate, p-nitrophenyl laurate (pNPL). The relative activity of the immobi...
Trypsin either modified with polyethylene glycol or as a suspended powder was used to catalyze digestion of protein substrates in benzene in order to get insight into protein-protein interactions in water-immiscible organic media. Depending on whether suspended or soluble trypsin was used, catalysis was found to procee...
Lipase from Pseudomonas fluorescens biotype I was immobilized by adsorption of anion exchange resin using glutaraldehyde to enhance the adsorption. The activity yield of the immobilized lipase was very low (below 1%) when lipase activity was measured using emulsion substrate. The activity yield was 10-70% when lipase a...
The production of prourokinase (PUK) by a human kidney tumor cell line is studied in long term cultures. Cells are grown on microcarriers which are retained inside the reactor by sedimentation or with a spin filter. Two modes of operation are compared: feed harvest at an average medium exchange rate of 0.3 d(-1) and co...
In previous studies on the acidogenic phase of anaerobic fermentation of lactose, a pathway for the reaction and a rate equation have been proposed. The question then remained as to the effect of the protein in whole whey on the mechanism and on the overall organic substrate conversion. In this study, it was found that...
To express human HZF1 fusion protein in E. coli and to obtain an anti-HZF1 antibody.
To observe the effect of intra-articular injection of SB203580, a selective p38 mitogen-activated protein kinase inhibitor, on the expression of matrix metalloproteinase (MMP)-3, MMP-13 in a rat model of osteoarthritis (OA) and to explore the relationship between the MMP-3/MMP-13 expressions and the severity of OA.
To investigate the effects of 11, 12-epoxyeicosatrienoic acid (11, 12-EET) preconditioning and postconditioning on Ca(2+)-handling proteins in myocardial ischemia/reperfusion (IR) injury in rats and reveal the effects and mechanism of 11, 12-EET on cardioprotection. METHODS The IR injury model was built by stopping per...
Peroxisome proliferator-activated receptor-gamma (PPARgamma) exerts multiple functions in determination of cell fate, tissue metabolism, and host immunity. Two synthetic PPARgamma ligands (rosiglitazone and pioglitazone) were approved for the therapy of type-2 diabetes mellitus and are expected to serve as novel cures ...
The ion-dependent enzymes, activities of which are affected by ions, were entrapped in the inside aqueous phase of reverse-phase evaporation vesicles (REVs) to protect the enzyme activities from inhibitory cations. Their activities were still preserved in the presence of the inhibitory cations because the permeabilitie...
A mathematical model of steady state reactor behavior was developed, combining the tanks-in-series model with fermentation kinetic description. The model was tested with experimental data-from an immobilized cell reactor for propionic acid production operated under different residence times. In order to determine the e...
Lactate dehydrogenase (LDH) was recovered from a solution by affinity binding to an N(6)-(6-aminohexyl)-AMP-Sepharose gel. An enzyme thermistor unit was employed to continously measure the activity of the unbound LDH. The enzyme activity signal from the enzyme thermistor was used in a PID controller to regulate the add...
The use of the affinity interaction between Cibacron Blue F3G-A (CB) and NADH-dependent enzymes to selectively precipitate these enzymes has been examined. An attempt was made to form crosslinked precipitates of lactate dehydrogenase (LDH) using bis- and poly-CB conjugates. When precipitation was not observed, an exami...
Cells of Zymomonas mobilis were permeabilized with toluene in order to utilize the enzymes, glucose-fructose oxidoreductase and gluconolactonase, inside the intact cells. Permeabilized cells were immobilized in a gelatin membrane, and a whole cell enzyme electrode was constructed by fixing the membrane on pH electrode....
In order to produce a product with a high content of maltotetraose, dual-enzyme systems composed of immobilized maltotetraose-forming amylase (G(4)-forming amylase) and pullulanase were studied. The thermostability of individually immobilized enzymes was examined in continuous operation; studies revealed that the enzym...
In Part II, the process utility of a thermotolerant methylotrophic bacterium is evaluated with respect to its dynamic response when various substrate or nutrient pulses are imposed on it while growing under steady-state conditions in a chemostat. The pulses investigated were methanol pulses on methanol-, methanol/forma...
Microporous-membrane-based extractive product recovery in product-inhibited fermentations allows in situ recovery of inhibitory products in a nondispersive fashion. A tubular bioreactor with continuous strands of hydrophobic microporous hollow fibers having extracting solvent flowing in fiber lumen was utilized for yea...
Alcohol oxidase, an enzyme which exhibits relatively weak substrate specificity among short chain alcohols, forms the corresponding aldehyde and hydrogen peroxide as coproduct. The ability of alcohol oxidase from Pichia pastoris yeast to convert ethanol to acetaldehyde and hydrogen peroxide was examined in an oxygen pr...
Recombinant bacterial systems exhibit limited capacities for heterologous protein production. As seen with this and other systems, cloned-gene protein production reaches an upper limit despite further increases in gene dosage. A series of closely related plasmids which contain mutations affecting their copy number has ...
Escherichia coli cells were immobilized and grown in hollow-fiber reactors allowing simultaneous NMR spectroscopy and perfusion with nutrient medium. The extent to which the cells were starved due to inadequate mass transfer was predicted using a mathematical model of reaction and diffusion. Reactors were experimentall...
The heterogeneous nature of hollow fiber reactors for cell cultivation requires special considerations for proper design and operation. Downstream concentration of high-molecular-weight proteins has been measured in the shell side of ultrafiltration hollow fiber bioreactors. This distribution resulted from shell-side c...
The mode of n-hexadecane uptake by two organisms-Pseudomonas M 1 and Pseudomonas N 1-was studied. During the growth of Pseudomonas M 1 on n-hexadecane, no extracellular biosurfactant/bioemulsifier was produced, no significant n-hexadecane pseudosolubilization was observed, and the reduction of surface and interfacial t...
The ability of L-sorbose to stimulate cellulase production In shake flask culture of Trichoderma reesei was examined in mineral salts media (initial pH 5.0) containing either 1.0% D-xylose, 1.0% cellulose, and/or 0.1, 0.3, or 0.5% L-sorbose. When sorbose was the only carbon source, growth was limited, little substrate ...
Bacterial biofilm removal processes due to shear and catastrophic sloughing have been investigated in a turbulent flow system under conditions of carbon versus oxygen substrate limitations and varying aqueous phase calcium concentrations. Biofilm cellular and extracellular polymer carbon, total biofilm carbon and mass,...
Plasmid-encoded mercuric reduction involves transfer of Hg(2+) across the cellular envelope and reduction to Hg(0) by the cytoplasmic mercuric reductase using NADPH. A mathematical model was developed for the binding and transfer of Hg(2+) by transport proteins and the subsequent reduction of Hg(2+). The values of the ...
Escherichia coli HB101 harboring an expression plasmid that bears the calf prochymosin gene controlled by the tac promoter was cultivated under different conditions in order to find an optimal fermentation arrangement that would lead to maximal prochymosin yield. Our results indicate that it is advantageous to use lact...
Solvent selection tests were carried out for the Delta(1) dehydrogenation of 6-alpha-methylhydrocortisone-21-acetate by Arthrobacter simplex cells in the presence of organic solvents. Solubility limits were determined for substrate and product in dry and water-saturated solvents and solvent mixtures. Molecular toxicity...
A purified phosphotriesterase was successfully immobilized onto trityl agarose in a fixed bed reactor. A total of up to 9200 units of enzyme activity was immobilized onto 2.0 mL of trityl agarose (65 micromol trityl groups/mL agarose), where one unit is the amount of enzyme required to catalyze the hydrolysis of one mi...
Substrate transfer rates from organic to aqueous phases were measured in the presence and absence of biocatalyst in the reaction medium, using modified Lewis cells. These measurements, in combination with intrinsic aqueous phase biocatalytic reaction kinetics, were used to confirm that benzyl acetate hydrolysis by pig ...
A critical problem in the storage and delivery of pharmaceutical proteins is their aggregation induced by moisture. A model system has been elaborated and investigated to elucidate the mechanism of this phenomenon. When 10 mg of bovine serum albumin lyophilized from an aqueous solution of pH 7.3 are wetted with just 3 ...
Thermodynamic and kinetic calculations were performed in a model of mixed substrate metabolism. The model simulates the catabolic breakdown of a first substrate, glucose (S(1)), in the presence of a second substrate, formate (S(2)), which acts as an additional source of free energy. The principal results obtained with ...
An approach is presented for the stable covalent immobilization of proteins with a high retention of biological activity. First, chemical modification studies were used to establish enzyme structural and functional properties relevant to the covalent immobilization of an enzyme to agarose based supports. Heparinase was...
The insect cell (Sf9)-baculovirus (AcNPV) expression system was employed for the synthesis of beta-galactosidase, a model heterologous protein. In the recombinant virus studied, the lacZ gene is fused to a portion of the polyhedrin structural gene and is under the control of the polyhedrin promoter. The effect of the m...
When purified xylanases from Trichoderma harzianum E58 or from a clone of Bacillus circulans were incubated with various low-yield wood pulps, little of the original enzyme activity could be detected in the filtrate at the end of the reaction. Partial bleaching of the pulps prior to enzymatic treatment generally result...
The capability of binding enzymes adsorptively to unmodified and silanized silica and glass as well as modified polystyrene carriers was studied for alpha-amylase, beta-amylase, and alpha-chymotrypsin. In most cases a high percentage of protein was bound very firmly under considerable loss of activity. The leakage of p...