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Disaccharides were microbaially transformed to their corresponding 3-keto-derivatives by resting cells of Agrobacterium tumefaciens NCPPB 396. The kinetics and yield of this highly specific oxidation depend on several factors. The oxygen concentration especially has a major influence on the production of 3-keto-derivatives and was investigated kinetically with respect to low stationary oxygen concentrations in solution. Experiments showed unconventional results that conflicted with normal Michaelis-Menten kinetics. A kinetic model was developed and the kinetic constants were calculated. The model and experimental data for sucrose, maltose, iso-maltulose (palatinose), and leucrose are in good agreement with each other. Initial reaction rates with different sugars using constant oxygen concentrations resulted in a Michaelis-Mentent type function. The complete kinetics, including the effect of disaccharide and oxygen concentrations, are presented. (c) 1995 John Wiley & Sons, Inc. 相似文献
43.
Mass balancing in kinetic resolution: calculating yield and enantiomeric excess using chiral balance
When kinetic resolution is applied for the production of enantiomerically pure compounds, process options may be used which involve more than one chiral substrate and one chiral product, such as sequential or parallel enzymatic kinetic resolutions or hydrolysis of diastereomers. Although the relation between the yields (y) of the chiral compounds is straightforward in these cases, the relation between their enantiomeric excess (ee) values is not. Combining mass balances into a so-called chiral balance (Sigma y . ee(R) = 0) provides the relation between enantiomeric excess values in a useful manner. This chiral balance easily shows which nonmeasured enantiomeric excess values and yields can be calculated from measured values. The chiral balance is only valid when configurations at chiral centers are conserved. (c) 1995 John Wiley & Sons, Inc. 相似文献
44.
Schlatmann JE Koolhaas CM Vinke JL Ten Hoopen HJ Heijnen JJ 《Biotechnology and bioengineering》1995,47(5):525-534
The role of glucose in ajmalicine production by Catharanthus roseus was investigated in the second stage of a two-stage batch process. Activities of tryptophan decar-boxylate (TDC) and anthranilate synthase (AS), two enzymes In the pathway leading to ajmalicine, were higher after induction with 40 g/L glucose than after induction with 60 or 80 g/L glucose. Experiments with different media containing mixtures of glucose and the nonpermeating osmotic agent xylose, and using an already induced culture as inoculum, revealed that a minimum amount of glucose is required to support ajmalicine production after enzyme induction. This requirement was not an osmotic effect. The relation between the glucose concentration and the specific ajmalicine production rate, q(p), was investigated in seven (fed-)batch cultures with constant glucose concentrations: 23, 29, 35, 53, 57, 75, and 98 g/L. In the cultures with a low glucose concentration (23, 29, and 35 g/L) the q(p) was 2.7-times higher than the cultures with 53 and 57 g/L, and almost six times higher than the cultures with a high glucose concentration (75 and 98 g/L). A glucose perturbation experiment (from 53 to 32 g/L) demonstrated that the ajmalicine production rate was adjusted without much delay. A kinetic equation is proposed for the relationship between the glucose concentration and q(p). Differences in enzyme induction and ajmalicine production at different glucose levels could not be explained by the intracellular concentrations of glucose, fructose, sucrose, or starch. (c) 1995 John Wiley & Sons Inc. 相似文献
45.
Derivation and testing of pair potentials for protein folding. When is the quasichemical approximation correct? 总被引:7,自引:6,他引:1
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J. Skolnick L. Jaroszewski A. Kolinski A. Godzik 《Protein science : a publication of the Protein Society》1997,6(3):676-688
Many existing derivations of knowledge-based statistical pair potentials invoke the quasichemical approximation to estimate the expected side-chain contact frequency if there were no amino acid pair-specific interactions. At first glance, the quasichemical approximation that treats the residues in a protein as being disconnected and expresses the side-chain contact probability as being proportional to the product of the mole fractions of the pair of residues would appear to be rather severe. To investigate the validity of this approximation, we introduce two new reference states in which no specific pair interactions between amino acids are allowed, but in which the connectivity of the protein chain is retained. The first estimates the expected number of side-chain contracts by treating the protein as a Gaussian random coil polymer. The second, more realistic reference state includes the effects of chain connectivity, secondary structure, and chain compactness by estimating the expected side-chain contrast probability by placing the sequence of interest in each member of a library of structures of comparable compactness to the native conformation. The side-chain contact maps are not allowed to readjust to the sequence of interest, i.e., the side chains cannot repack. This situation would hold rigorously if all amino acids were the same size. Both reference states effectively permit the factorization of the side-chain contact probability into sequence-dependent and structure-dependent terms. Then, because the sequence distribution of amino acids in proteins is random, the quasichemical approximation to each of these reference states is shown to be excellent. Thus, the range of validity of the quasichemical approximation is determined by the magnitude of the side-chain repacking term, which is, at present, unknown. Finally, the performance of these two sets of pair interaction potentials as well as side-chain contact fraction-based interaction scales is assessed by inverse folding tests both without and with allowing for gaps. 相似文献
46.
W. A. Fenton A. L. Horwich 《Protein science : a publication of the Protein Society》1997,6(4):743-760
I. Architecture of GroEL and GroES and the reaction pathway A. Architecture of the chaperonins B. Reaction pathway of GroEL-GroES-mediated folding II. Polypeptide binding A. A parallel network of chaperones binding polypeptides in vivo B. Polypeptide binding in vitro 1. Role of hydrophobicity in recognition 2. Homologous proteins with differing recognition-differences in primary structure versus effects on folding pathway 3. Conformations recognized by GroEL a. Refolding studies b. Binding of metastable intermediates c. Conformations while stably bound at GroEL 4. Binding constants and rates of association 5. Conformational changes in the substrate protein associated with binding by GroEL a. Observations b. Kinetic versus thermodynamic action of GroEL in mediating unfolding c. Crossing the energy landscape in the presence of GroEL III. ATP binding and hydrolysis-driving the reaction cycle IV. GroEL-GroES-polypeptide ternary complexes-the folding-active cis complex A. Cis and trans ternary complexes B. Symmetric complexes C. The folding-active intermediate of a chaperonin reaction-cis ternary complex D. The role of the cis space in the folding reaction E. Folding governed by a "timer" mechanism F. Release of nonnative polypeptides during the GroEL-GroES reaction G. Release of both native and nonnative forms under physiologic conditions H. A role for ATP binding, as well as hydrolysis, in the folding cycle V. Concluding remarks. 相似文献
47.
沈平 《生物化学与生物物理进展》1997,24(5):477-478
利用BIA技术来观察DNA之间的任何相互反应.包括:DNA的延长、连接和退火等.无需任何标记并可测定相互作用的动态参数. 相似文献
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49.
应用非结构的逻辑增殖模型研究了两种酵母的单碳源和双碳源单细胞蛋白间歇培养的动力学,用改进的逻辑增殖模型研究了双碳源流加培养过程的动力学,从实验数据拟合了动力学模型参数,模型计算值与实验数据吻合良好。 相似文献
50.
IZUMI WASHITANI 《Plant, cell & environment》1984,7(9):655-659
Abstract The germination responses of a nondormant fraction of a seed population of Taraxacum officinale Weber at constant temperatures in the range 7–34°C were analysed through a time-course study. Maximal percentage germination (approximately 90%) was attained at temperatures 10–18°C, where simple linear relationships were observed between the temperature and the germination rates, i.e. the reciprocals of the time taken to germinate by subpopulations with 20–80% germination. There was a variation in the required ‘thermal times’ (θ) which characterized the linear relationships, the distribution of which could be approximated for the seed population by the following distribution function: where m is the median of the distribution, and A is a shape parameter characterizing the pattern of the distribution. Final percentage germination decreased with increasing temperature from 20 to 32°C, where the final percentage germination vs. temperature plotted on a normal probability scale yielded a straight line, indicating the normality of the distribution of the upper limit temperature in the seed population. The estimated mean and standard deviation were 27.25 ± 3.75°C. The rate of germination for the subpopulation with 20–80% germination also decreased with increases in the temperature from 22 to 30°C. If the relationships between the temperature within this range and the rate for the subpopulations with 20–80% germination were approximated by the regression lines, the negative ‘thermal time’ characterizing the yielded linear relationship would have a distribution which could be approximated by the same function with the required thermal time for the relationship of suboptimal range. The parameters m and A for the negative ‘thermal time’ were determined to be 2870 K h and 1.7 × 10-10 K-3 h-3. 相似文献