High Substrate Concentration (high + substrate_concentration)

Distribution by Scientific Domains

Selected Abstracts

Enantioselective Reduction of 4-Fluoroacetophenone at High Substrate Concentration using a Tailor-Made Recombinant Whole-Cell Catalyst

Harald Gröger
Abstract A practical and highly efficient biocatalytic synthesis of optically active (R)-4-fluorophenylethan-1-ol has been developed based on reduction of the corresponding 4-fluoroacetophenone in the presence of a tailor-made recombinant whole-cell biocatalyst, containing an alcohol dehydrogenase and a glucose dehydrogenase. The reaction proceeds in a pure aqueous solvent media at a substrate concentration of ca. 0.5,M, and gives the desired product with high conversion (>95,%), good yield (87,%) and with an excellent enantioselectivity of >99,% ee. In addition, activity tests further showed that also the analogous 2- and 3-fluoroacetophenones are promising substrates. [source]

Enzymatic Production of l -Menthol by a High Substrate Concentration Tolerable Esterase from Newly Isolated Bacillus subtilis ECU0554

Gao-Wei Zheng
Abstract Enzymatic preparation of l -menthol has been attracting much attention in the flavor and fragrance industry. A new ideal strain, Bacillus subtilis ECU0554, which exhibited high hydrolytic activity and excellent enantioselectivity towards l -menthyl ester, has been successfully isolated from soil samples through enrichment culture and identified as Bacillus subtilis by 16S rDNA gene sequencing. The esterase extracted from B. subtilis ECU0554 (BSE) showed the best catalytic properties (E>200) for dl -menthyl acetate among the five menthyl esters examined. Enantioselective hydrolysis of 100,mM dl -menthyl acetate at 30°C and pH,7.0, using crude BSE as biocatalyst and 10% ethanol (v/v) as cosolvent, resulted in 49.0% conversion (3,h) and 98.0% ee for the l -menthol produced, which were much better than those using commercial enzymes tested. Moreover, BSE exhibited strong tolerance against high substrate concentration (up to 500,mM), and the concentration of l -menthol produced could reach as high as 182,mM, and more importantly, the optical purity of l -menthol produced was kept above 97% ee, which were not found in previous reports. These results imply that BSE is a potentially promising biocatalyst for the large-scale enzymatic preparation of l -menthol. Using this excellent biocatalyst, the enzymatic production of l -menthol will become a mild, efficient, inexpensive and easy-to-use "green chemistry" methodology. [source]

Properties of wheat bran polyphenol oxidase

dem Soysal
Abstract Polyphenol oxidase (PPO) obtained from wheat bran catalyzed the oxidation of 4-methyl catechol. Phenolic compounds found naturally in crude extract played role as an endogeneous substrate and activity of crude extract needed correction. Activity versus enzyme concentration gave a linear plot at high substrate concentration whereas a nonlinear plot was obtained at low substrate concentration which proved the presence of endogeneous substrate. Adsorption on celite and extraction with polyvinylpyrrolidone (PVPP) caused the removal of phenols. Adsorption of PPO on celite yielded a 4-fold increase in specific activity whereas extraction with PVPP yielded a 2.5-fold increase in specific activity compared to the crude extract. The kinetics of PPO catalyzed oxidation obeyed Michaelis-Menten model; Km and Vmax values were found as 218 mM and 99 ,M/min, respectively. The enzyme was inhibited by ethyl alcohol, dithiothreitol (DTT) and isoproterenol and exhibited heat stability up to a temperature of 90°C. The optimum pH of the enzyme was found to be 5.0. [source]

Production of delta-endotoxin by Bacillus thuringiensis subsp kurstaki and overcoming of catabolite repression by using highly concentrated gruel and fish meal media in 2- and 20-dm3 fermenters

Nabil Zouari
Abstract Delta-endotoxin production by a strain of Bacillus thuringiensis subsp kurstaki exhibiting larvicidal toxicity towards lepidoptera was investigated in 2- and 20-dm3 fermenters, using gruel- and fish meal-based media. The results show clearly that in such complex media, aeration plays an important role in bioinsecticide production. Optimal aeration led to improvement of delta-endotoxin concentrations with decreases of final spore count and proteolytic activity. Moreover, in order to use high gruel concentrations, a fermenter configuration with an efficient aeration system should be used. In a 20-dm3 Biolafite fermenter, 59,g,dm,3 or 75,g,dm,3 gruel was used to produce bioinsecticides with a significant reduction of carbon catabolite repression of delta-endotoxin synthesis. This result is very interesting in order to produce high final delta-endotoxin concentrations in the culture broth. It was also concluded, by considering the key role of oxidative pathways in delta-endotoxin synthesis, that oxygen supply must be adequate for bioinsecticide production at high substrate concentrations. Moreover, the role of sodium chloride in improving delta-endotoxin production is dependent not only on protease synthesis and its effect on crystal stability, but also on the aeration level of the production medium. © 2002 Society of Chemical Industry [source]

Biokinetic models for representing the complete inhibition of microbial activity at high substrate concentrations

Gunaseelan Alagappan
Abstract This paper reintroduces the Wayman and Tseng model for representing substrate inhibition effects on specific growth rate by further documenting its potential predictive capabilities. It also introduces a modification to this model in which an Andrews inhibition function is used in place of the Monod noninhibitory substrate function. This modification better represents the relationship between specific growth rate and substrate concentration for those substrates that show Andrews type inhibition at lower substrate concentrations, rather than the Monod type noninhibitory behavior described in the model of Wayman and Tseng. Results from nonlinear, least squares regression analysis are used to evaluate the ability of these models to empirically represent experimental data (both new and from the literature). The statistical goodness of fit is evaluated by comparing the regression results against those obtained using other empirical models. Finally, possible mechanisms of toxicity responsible for the observed inhibition trends are used to further justify use of these empirical models. The dominant mechanism considered to be relevant for conceptually explaining complete inhibition at high concentrations of solvents is the deterioration of cell membrane integrity. Literature citations are used to support this argument. This work should lead to improvements in the mathematical modeling of contaminant fate and transport in the environment and in the simulation of microbial growth and organic compound biodegradation in engineered systems. © 2001 John Wiley & Sons, Inc. Biotechnol Bioeng 75: 393,405, 2001. [source]