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1.
Preparation and characterization of urease immobilized onto porous chitosan beads for urea hydrolysis 总被引:2,自引:0,他引:2
Urease was covalently immobilized onto porous chitosan beads via primary amine groups connected to the backbone via a six-carbon linear alkyl spacer. The optimum conditions for enzyme immobilization are activating the beads with 1%(w/w) glutaraldehyde, reacting the activated beads in pH 7 buffer with the enzyme, using an enzyme to bead weight ratio of 25, and without lyophilization. Chitosan-bound urease was found to fully retain its specific activity. Properties of the immobilized urease were characterized under batch and flow conditions. Increased optimum reaction temperature, enhanced thermal stability and storage stability, and excellent reusability were found after enzyme immobilization. Continuous hydrolysis of urea solution was studied in a column packed with the enzyme-containing beads for its possible application in regenerating dialysate solution during hemodialysis. 相似文献
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The stability of native and immobilized urease isolated from Staphylococcus saprophyticus was studied at 4 degrees and 25 degrees C. The activity yield was 20% and 1.4% on the enzyme immobilization in albumin gel and latex membrane, respectively. Inactivation of native microbial urease proceeded 10 times slower in the solution containing 1 mM EDTA and 30 mM sodium sulfite. This solution contributed to a great extent to stabilization of immobilized urease both during storage in the phosphate buffer solution and in case of lyophilization. 相似文献
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The results of experiments in a fixed-bed reactor and a CSTR containing urease immobilized on a nonporous support and conducted in the absence of diffusional limitations are reported. Kinetic parameters were established by separate batch experiments. The key observation was that the product ammonia attacked the free form of the enzyme and thereby illustrates the importance of mechanism in determining deactivation kinetics. 相似文献
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Preparation and characterization of immobilized lipase on magnetic hydrophobic microspheres 总被引:21,自引:0,他引:21
A novel magnetic poly(vinyl acetate (VAc)–divinyl benzene (DVB)) material (8–34 μm) was synthesized by copolymerization of vinyl acetate and divinyl benzene using oleic acid-stabilized magnetic colloids as magnetic cores. The magnetic colloids and the copolymer microspheres were characterized with transmission and scanning electron microscopes, respectively. Magnetization of the microspheres could be described by the Langevin function. All the observations indicated that the microspheres were superparamagnetic. Magnetic sedimentation of the microspheres was achieved within 3 min, over 300 times faster than the gravitational sedimentation. Candida cylindracea lipase (CCL) was immobilized to the porous carrier at up to 6750 IU/g carrier, remarkably higher than the previous studies. The pH and temperature dependencies of the immobilized CCL were investigated and the optimum temperature and pH for the immobilized CCL were determined. Activity amelioration of the immobilized CCL for the hydrolysis of olive oil was observed, indicating an interfacial activation of the enzyme after immobilization. Moreover, the immobilized CCL showed enhanced thermal stability and good durability in the repeated use after recovered by magnetic separations. 相似文献
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Glucoamylase (1,4-α-d-glucan glucohydrolase, EC 3.2.1.3) has been covalently immobilized on a polyacrylamide-type support containing carboxylic groups activated by water-soluble carbodiimide. The activity was 5.5– 6.0 units g?1solid. The optimum pH for catalytic activity was pH 3.8. The apparent optimum temperature was found at 60°C. With soluble starch as substrate the Km value was 14 mg ml?1. The pH for maximum stability was pH 4.0–4.5. In the presence of 8 m urea the immobilized glucoamylase retained most of its catalytic activity but it was more susceptible to guanidinium hydrochloride than the soluble enzyme. The practical applicability of immobilized glucoamylase was tested in batch process and continuous operation. 相似文献
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Polymethylglutamate (PMG), a synthetic polypeptide, was used as a new carrier to immobilize urease (EC 3.5.1.5) and uricase (EC 1.7.3.3) by the azide method. The enzymes could be immobilized onto PMG in various forms, such as film, fiber, coating on various beads, and a silicon tube. The retained activities of the immobilized enzymes were excellent (more than 95%), therefore it was possible to immobilized almost all activities of the enzymes added in the coupling mixtures. Heat stabilities of the resulting immobilized enzymes were markedly improved, while the optimal pH and Km values remained almost unchanged. The urease immobilized on the PMG-coated glass beads packed in a column, was found to retain its activity more than 80% of the initial value, even after the occasional use for a year. In view of the improved retained activities and stabilities of the immobilized enzymes, PMG may therefore be a very versatile matrix for the immobilized enzymes. 相似文献
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Conidia of Penicillium chrysogenum were immobilized in K-carrageenan beads and then incubated in a growth-supporting medium to yield a penicillin producing immobilized cell mass. These in situ grown immobilized cells were used for the semicontinuous (replacement cultures)and continuous (fluidized bioreactor culture) production of penicillin-G. When periodically replaced into a minimal production medium, immobilized cells exhibited a half-life for penicillin production which was ninefold greater than that exhibited by free cells. The half-life of penicillin production and the yield of penicillin from glucose in such a replacement culture were greatly affected by the frequency of replacement and by the production medium's pH and concentration of glucose, phosphate, and trace metal nutrients. A penicillin-producing continuous flow bioreactor (150 mL), employing immobilized cells, was operated for up to 16 days. The best specific penicillin productivity (1.2 mg/g cells/h)yield from glucose (7.0 mg/g glucose) and half-life of production (15 days) were obtained when the feed medium contained 10 g/L of glucose, the pH was maintained at 7.0, the relative dissolved oxygen concentration was ca. 40%; and the residence time was 20 h. 相似文献
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The behavior of an enzyme/membrane system containing urease is studied when an external electric field is applied. The device using a potential difference across the enzyme/membrane system is first described. Optimal operating conditions with respect to substrate concentration, ionic strength and pH are studied. Possible mechanisms of the change in membrane activity by electric field are discussed. 相似文献
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Preparation and properties of solid-supported urease 总被引:1,自引:0,他引:1
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Purified skeletal muscle myosin (EC 3.6.1.3) has been covalently bound to Sepharose 4B by the cyanogen bromide procedure. The resulting complex, Sepharose-Myosin, possesses adenosine triphosphatase activity and is relatively stable for long periods of time. Under optimal binding conditions, approximately 33% of the specific ATPase activity of the bound myosin is retained. Polyacrylamide gel electrophoresis of polypeptides released from denatured Sepharose-Myosin indicates that 85% of the myosin is attached to the agarose beads through the heavy chains and the remainder through the light chains, in agreement with predictions of binding and release based upon either the lysine contents or molecular weights of themyosin subunits. The adenosine triphosphatase of the immobilized myosin has been investigated under conditions of varying pH, ionic strength, and cation concentration. The ATPase profiles of immobilized myosin are quite similar to those for free myosin, however subtle differences are found. The Sepharose-Myosin ATPase is not as sensitive as myosin to alterations in salt concentration and the apparent KM is approximately two-fold higher than that of myosin. These differences are probably due to chemical modification in the region of the attachment site(s) to the agarose beads and hydration and diffusion limitations imposed by the polymeric agarose matrix. 相似文献
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L. D'Angiuro P. Cremonesi G. Mazzola B. Focher G. Vecchio 《Biotechnology and bioengineering》1980,22(11):2251-2272
A method of enzyme immobilization by graft copolymerization on polysaccharides is reported. Glycidylmethacrylate was used as a vinylating reagent and the reaction product with enzymes (HRP, GOD, Am, ChT) was copolymerized with different matrices (cellulose, Sepharose, Sephadex, Starch). Various factors affect the final activity of copolymers; these include the redox system, the type of support, and the quantity and type of vinyl monomer added. Using a fixed quantity of enzyme and support (3 mg enzyme, 100 mg support), the coupling efficiency varied from 2 to 50%. The most important characteristics in these immobilized systems were tested (stability in continuous washing, kinetic characteristics, storage, thermal, and lyophilization stability). Immobilized-enzyme graft copolymers have very similar kinetic behavior to that of the free enzyme. Diffusion is not seriously limited, as shown by kinetic parameters and energy activation values, and this indicates that the immobilization reaction does not alter the enzymatic activity. 相似文献
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In an effort to use a biocatalyst repeatedly with acceptable stability, amylosucrase from Neisseria polysaccharea (NpAS) was non-covalently immobilized on a pH-dependent autoprecipitating polymer, Eudragit L100. About 87% of the enzyme activity and 96% of the protein were recovered after the immobilization process. The immobilized NpAS showed significantly improved thermostability, whereas no difference was observed in the temperature and pH profiles of activity between the native and immobilized NpAS. To evaluate the reusability of the immobilized NpAS, repeated production of linear α-(1,4)-glucans was carried out at 35°C with 0.1M sucrose as substrate. Although only 71-74% of the protein was recovered after each reaction cycle, high stability index was observed from 0.93 to 0.96. Reusability of the immobilized NpAS was further confirmed by comparing its linear α-(1,4)-glucan products with that produced by native one. Almost identical production yield and molecular size distribution of the linear α-(1,4)-glucans were obtained between them. 相似文献
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《Process Biochemistry》2010,45(8):1422-1426
The starch hydrolyzing enzyme amyloglucosidase (AMG) from Rhizopus was immobilized onto the protonated salt (TS) and basic (TB) forms of chemically synthesized poly(o-toluidine) (POT) using adsorption and covalent binding. The polymers were activated with glutaraldehyde prior to covalent bonding. The immobilization efficiency was affected by the pH of the immobilization medium, contact time and amount of enzyme. After immobilization, the pH and temperature were changed to conditions under which the enzyme is most active. Immobilized AMG was more stable with respect to changes in pH and increases in temperature compared to free AMG. The immobilized enzyme retained high catalytic activity after multiple uses and showed enhanced stability with storage compared to free enzyme. 相似文献
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Li GY Huang KL Jiang YR Yang DL Ding P 《International journal of biological macromolecules》2008,42(5):405-412
The covalently immobilized of Saccharomyces cerevisiae alcohol dehydrogenase (SCAD) to magnetic Fe(3)O(4) nanoparticles via glutaraldehyde coupling reaction was studied. The magnetic Fe(3)O(4) nanoparticles were prepared by hydrothermal method using H(2)O(2) as an oxidizer. Functionalization of surface-modified magnetic particles was performed by the covalent binding of chitosan onto the surface. The amino functional group on the magnetic Fe(3)O(4)-chitosan particles surface and the amino group of the dehydrogenase were coupled with glutaraldehyde. The immobilization process did not affect the size and structure of magnetic nanoparticles. For the reduction of phenylglyoxylic acid by immobilized SCAD, the kinetic analysis data indicated that the immobilized SCAD retained 48.77% activity of its original activity. The activation energy within 20-40 degrees C, the maximum specific activity and the Michaelis constants for phenylglyoxylic acid were 7.79 KJ mol(-1), 279.33 nmol min(-1) and 37.77 mmol l(-1), respectively. Furthermore, the immobilized SCAD enhanced thermal stability and good durability in the repeated use after recovered by magnetic separations. 相似文献
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S. George M. Chellapandian B. Sivasankar P. V. Sundaram 《Bioprocess and biosystems engineering》1996,14(6):311-315
Gas holdup and liquid circulation velocity meassurements were made for a range of liquid viscosities in a 22 l external loop airlift column and 250 l pilot-scale concentric cylinder airlift bioreactor. The results showed that for a fixed superficial gas velocity, liquid circulation velocity decreased monotonically with increasing liquid viscosity. The gas holdup for a fixed gas flow rate showed an initial increase with liquid viscosity followed by a decrease when liquid viscosity increased beyond a critical value. These observations could not be described satisfactorily using the available models of gas holdup and liquid circulation.List of Symbols U
sg
m/s
Superficial gas velocity
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sl
m/s
Superficial liquid velocity in the riser
Greek Letters
Pas
Liquid viscosity
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g
Gas holdup in the riser 相似文献