厄斯考维菌产羰基还原酶的发酵条件优化

Fermentation optimization for enhanced production of carbonyl reductase by Oerskovia turbata

从土壤中筛到一株产羰基还原酶的菌种骚动厄斯考维菌(Oerskovia turbata)ZJPH1604,利用该微生物细胞可不对称催化3-氯苯乙酮合成(R)-1-(3-氯苯基)乙醇,对映体过量值>99.9%.通过单因素试验发现:酵母提取物、(NH_4)_2SO_4和KH_2PO_4为影响酶活的显著影响因子,继而采用响应面法对产酶条件进行了优化.结果表明:最优发酵培养基组成为葡萄糖15.0g/L,酵母提取物30.0g/L,(NH_4)_2SO_4 2.6g/L,KH_2PO_4 2.8g/L,NaCl 0.6g/L,MgSO_4 0.8g/L.在最适培养条件下,菌体羰基还原酶的酶活力达到32.0U/g,较优化前提高了166.7%.为高效合成(R)-1-(3-氯苯基)乙醇提供了新途径,丰富了生物不对称合成手性药物中间体的手段.

A bacterial strain Oerskovia turbata ZJPH1604 containing carbonyl reductase was screened out from soil,which can reduce 3-chloroacetophenone to（R）-1-（3-chlorophenyl）ethanol with enantiomeric excess（e.e.）reached more than 99.9%.By the single factor experiment,yeast extract,（NH4）2SO4 and KH2PO4 were found to be significant factors among six tested variables that influence the carbonyl reductase production.In order to improve the enzyme production,the medium components were further optimized by response surface methodology.The results showed that the enzyme activity was the highest when the fermentation medium was composed of glucose 15.0 g/L,yeast extract 30.0 g/L,（NH4）2SO4 2.6 g/L,KH2PO4 2.8 g/L,NaCl 0.6 g/L,MgSO4 0.8 g/L.Under the optimum conditions,the enzyme activity reached 32.0 U/g,with an increase of 166.7% compared to the original medium components.This study provides a new approach for the efficient synthesis of（R）-1-（3-chlorophenyl）ethanol,and enrich the methods for the synthesis of chiral intermediates of drugs.