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101.
Purification of bacterial L-methionine gamma-lyase 总被引:1,自引:0,他引:1
A chromatographic procedure using sequential ion-exchange columns is described for separating choline, trimethylamine, trimethylamine oxide, and betaine extracted from marine fish tissues; added exogenous carnitine can also be separated by the system. Choline with its positive charge binds to the AG 50W-X8 (Na+, pH 9) column. The column is first eluted with 0.1 N NaOH to collect trimethylamine, trimethylamine oxide, and betaine; choline is then eluted with 0.5 N NaOH. The amines collected with 0.1 N NaOH are subsequently separated using an AG 50W-X8 (H+, pH 4) column eluted with a linear 0-1 M NaC1 gradient. 相似文献
102.
Effect of plant growth and addition of plant residues on the phosphatase activity in soil 总被引:1,自引:0,他引:1
R. C. Dalal 《Plant and Soil》1982,66(2):265-269
Summary The phosphatase activity of the soil amended with roots and tops of clover (Trifolium repens) plant material (0.1% by weight) remained essentially constant in the absence of growing plants but changed considerably in the presence of plants (Avena sativa) grown for 10 weeks. There was a significant relationship between the phosphatase activity and organic and inorganic P in the soil solution only in the presence of growing plants. The differences in phosphatase activity between roots and tops amended soil were attributed to total C as well as differences in the degree of availability of C added through plant materials. This may also apply to the carpet grass (Axonopus affinis) amended soil. 相似文献
103.
Summary Organic (e.g. chitin, green manure, cellulose) amendments to soil induced quantitative and qualitative changes in the rhizosphere microflora of antirrhinum plants infected withVerticillium dahliae Kleb. Whereas reduction in disease severity occurred with chitin and green manure amendments, an increase in disease severity was observed with the application of cellulose. The reduction of the disease severity with chitin and green manure may be correlated with the increased population of actinomycetes in the antirrhinum rhizosphere. 相似文献
104.
Karlheinz Altendorf Margot Lukas Brigitte Kohl Clemens R. Müuller Heinrich Sandermann 《Journal of cellular biochemistry》1977,6(2):229-238
Techniques for the solubilization and fractionation of integral membrane proteins have been developed in recent years. A small portion of membrane protein (about 2%, proteolipid fraction) will partition into chloroform or 1-butanol, and, in several cases, these proteins retain functional activity. A virtually complete solubilization can be achieved at neutral pH by use of aprotic solvents, like hexamethylphosphoric triamide or N-methylpyrrolidone. At relatively low concentrations (< 3 M) aprotic solvents inhibited β-D-galactoside transport by whole cells and the derivative membrane vesicles of Escherichia coli, but this inhibition could be largely reversed by a simple washing procedure. At higher concentrations of aprotic solvent (5–6 M), 50–80% of the total protein of lactose transport-positive membrane vesicles was solubilized. When these extracts were added to intact lactose transport-negative membrane vesicles, lactose transport was reconstituted, the required energy being provided by either respiration (e.g., addition of D-lactate) or by a K+ diffusion potential established with the aid of valinomycin. The dicyclohexylcarbodiimide (DCCD)-reactive subunit of the E. coli ATPase complex was found to partition into chloroform, and to be amenable to further purification in organic solvent. Ether precipitation and chromatography on DEAE-cellulose and hydroxypropyl-Sephadex G-50 yielded an homogeneous polypeptide of an apparent molecular weight of 9,000. The purified and unlabeled DCCD-reactive protein was incorporated into K+-loaded liposomes, and a membrane potential was generated by the addition of valinomycin. There are indications that the DCCD-reactive protein alone made the membrane specifically permeable for protons. 相似文献
105.
106.
Christian Larroche Catherine Creuly Jean-Bernard Gros 《Biocatalysis and Biotransformation》1992,5(3):163-173
The production of 2-heptanone from octanoic acid may be performed by free and entrapped spores of Penicillium roquefortii in a water-organic solvent two phase system.
An industrial, isoparafflnic solvent, i.e. Hydrosol IP 230 O.S., which may be considered as tetradecane, is well suited for the process. Activities nearly double those achieved with aqueous systems are observed using an initial fatty acid content in the organic layer close to 100 mM and a ratio of the volume of the organic phase to the total volume of the medium of 0.88. The presence of the solvent allows a better recovery of the metabolite by lowering its activity coefficient.
Fed-batch experiments performed in an aerated, stirred reactor show that the bioconversion may proceed in the two-phase system for at least 300 h. These conditions allow conversion of 750 mM (108 g · 1-1) fatty acid, and production of 600 mM (68.5 g · 1-1) 2-heptanone. 相似文献
An industrial, isoparafflnic solvent, i.e. Hydrosol IP 230 O.S., which may be considered as tetradecane, is well suited for the process. Activities nearly double those achieved with aqueous systems are observed using an initial fatty acid content in the organic layer close to 100 mM and a ratio of the volume of the organic phase to the total volume of the medium of 0.88. The presence of the solvent allows a better recovery of the metabolite by lowering its activity coefficient.
Fed-batch experiments performed in an aerated, stirred reactor show that the bioconversion may proceed in the two-phase system for at least 300 h. These conditions allow conversion of 750 mM (108 g · 1-1) fatty acid, and production of 600 mM (68.5 g · 1-1) 2-heptanone. 相似文献
107.
Peter Miethe Ingeborg Jansen Uwe Niedermeyer Udo Kragl Regine Haftendorn Maria-Regina Kula Christian Wandrey Karl-Heinz Mohr Helmut-Walter Meyer 《Biocatalysis and Biotransformation》1992,7(1):61-73
The possibility of using the enzyme (R)-Oxynitrilase in a biphasic lyotropic liquid crystal/dibutylether system has been demonstrated. This reaction system is applicable for the continuous production of (R)-benzaldehydecyanohydrin in a fixed bed reactor. The optical purity was between 94 and 96% ee and independent of the flow rate. The space time yield was maximal (2650 g/(1*d)) at a flow rate of 1.6 ml/min. 相似文献
108.
Water Concentration and Activity Effects on Aminoacylase in Aqueous/Organic One-Liquid-Phase Systems
Aminoacylase has been employed as a model system to study its catalytic properties at low water concentrations/water activities with different water-miscible organic cosolvents. Cosolvents assayed were alcohols and polyols with pure logarithm of the partition coefficient (log P) values, on the standard water/octanol system, ranging between -5.2 and 0.24.
Experimental hydrolysis equilibrium constants (Kapp), at a constant water concentration, decreased with the fall in log P of the cosolvent, as well as with reduction of the water concentration/water activity, as would be expected. The enzyme hydrolytic and synthetic activities, measured at a constant water concentration/water activity value, followed a sigmoidal dependence on log P of the cosolvent employed when the water concentration or water activity values were lower than 50% (w/w) or 0.66, respectively. This became a hyperbolic relationship at higher water concentration/water activity values. A linear relationship between the logarithm of the limiting water activity necessary to maintain enzyme activity and log P was obtained. Both hydrolytic and synthetic activities were suppressed for water activities higher than 0.66 and cosolvents with log P lower than -1.6. 相似文献
Experimental hydrolysis equilibrium constants (Kapp), at a constant water concentration, decreased with the fall in log P of the cosolvent, as well as with reduction of the water concentration/water activity, as would be expected. The enzyme hydrolytic and synthetic activities, measured at a constant water concentration/water activity value, followed a sigmoidal dependence on log P of the cosolvent employed when the water concentration or water activity values were lower than 50% (w/w) or 0.66, respectively. This became a hyperbolic relationship at higher water concentration/water activity values. A linear relationship between the logarithm of the limiting water activity necessary to maintain enzyme activity and log P was obtained. Both hydrolytic and synthetic activities were suppressed for water activities higher than 0.66 and cosolvents with log P lower than -1.6. 相似文献
109.
The association between enzymatic and electrochemical reactions, enzymatic electrocatalysis, had proven to be a very powerful tooth in both analytical and synthetic fields. However, most of the combinations studied have involved enzymatic catalysis of irreversible or quasi-irreversible reaction. In the present work, we have investigated the possibility of applying enzymatic electrocatalysis to a case where the electrochemical reaction drives a thermodynamically unfavorable reversible reaction. Such thermodynamically unfavorable reactions include most of the oxidations catalyzed by dehydrogenases. Yeast alcohol dehydrogenase (E.C. 1.1.1.1) was chosen as a model enzyme because the oxidation of ethanol is thermodynamically very unfavorable and because its kinetics are well known. The electrochemical reaction was the oxidation of NADH which is particularly attractive as a method of cofactor regeneration. Both the electrochemical and enzymatic reactions occur in the same batch reactor in such a way that electrical energy is the only external driving force. Two cases were experimentally and theoretically developed with the enzyme either in solution or immobilized onto the electrode's surface. In both cases, the electrochemical reaction could drive the enzymatic reaction by NADH consumption in solution or directly in the enzyme's microenvironment. However even for a high efficiency of NADH consumption, the rate of enzymatic catalysis was limited by product (acetaldedehyde) inhibition. Extending this observation to the subject of organic synthesis catalyzed by dehydrogenases, we concluded that thermodynamically unfavorable reaction and can only be used in a process if efficient NAD regeneration and product elimination are simultaneously carried out within the reactor. 相似文献
110.
The stability of the serine proteases from Bacillus amyloliquefaciens (subtillisin BPN') and Bacillus licheniformis (subtilisin Carlsberg) was investigated in various anhydrous solvents at 45 degrees C. The half-life of subtilisin BPN' in dimethyl-formamide dramatically depends on the pH of the aqueous solutions from which the enzyme was lyophilized, increasing from 48 min to 20 h when the pH is raised from 6.0 to 7.9. Both subtilisins exhibited substantial inactivation during multihour incubations in tert-amyl alcohol and acetonitrile when enzymatic activities were also measured in these solvents; however, when the enzymes were assayed in water instead, hardly any loss of activity was detected. This surprising difference appears to stem from the partitioning of the bound water essential for catalytic activity from the enzymes into the solvents. When assayed in organic solvents, this time-dependent stripping of water results in decay of enzymatic activity; however, when assayed in water, where the dehydrated subtilisins can undergo rehydration thereby recovering catalytic activity, little inactivation is observed. In agreement with this hypothesis, the addition of small quantities of water tert-amyl alcohol stabilized the subtilisins in it even when enzymatic activity was measured in the nonaqueous solvent. Ester substrates (vinyl butyrate and trichloroethyl butyrate) greatly enhanced the stability of both subtilisins in organic solvents possibly because of the formation of the acyl-enzymes. 相似文献