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Using benzyl glycidyl ether as substrate, this paper screened 31 strains with epoxide hydrolases from several kinds of soil collected from many places. 11 strains showed enantioselectivity toward epoxide. A fungi with best enantioselectivity was chosed as an object of study and identified as Talaromyces flavus based on morphology and ITS sequence analysis. It’s named Talaromyces flavus ZJUTZQ159.Based on single factor experiments, the fermentation conditions for Talaromyces flavus ZJUTZQ159 were investigated, including carbon sources, nitrogen sources, metal ions, temperature, time for fermentation, the initial pH of medium and inducement, which influenced the biomass, enzyme activity, e.e.s value and E value. The best results were obtained under the following conditions: pH 4.0, temperature 30℃and 66h. And the fungi was found as constitute enzyme, unwanted any inducement. Simultaneously, the additive amounts of carbon sources, nitrogen sources and metal ions were get through orthogonal test. The optimum fermentation medium: soluble starch 20g/L, soybean flour 15g/L, NaNO3 1g/L、K2HPO4·3H2O 1.0g/L、KCl 0.5g/L、MgSO4·7H2O 1.476g/L. Under this condition, Talaromyces flavus ZJUTZQ159 could give the activity of enzyme 18.66U/gDCM. The conditions for kinetic resolution of racemic benzyl glycidyl ether by Talaromyces flavus ZJUTZQ159 were studied. It showed the best resolution conditions as follows: racemic BGE （15mmol/L） in DMSO （1/10, v/v） was hydrolyzed in 2mL KPB （0.1M, pH8.0） using wet mycelium of Talaromyces flavus ZJUTZQ159 （1.0g） by shaking the mixture （200 rpm） at 25℃for 4h. And it gave the best e.e.s value 96.4% and E value 7.16 under the best resolution condition. The research on substrate specificity of this enzyme indicated that this fungi showed low enantioselectivity toward benzyl glycidyl ether with methyl on the o-, m-, or p- position.
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