羰基还原酶产生菌Candida sorboxylosa的筛选与产酶条件优化  

Strain screening and culture condition optimization for production of carbonyl reductase by Candida sorboxylosa

作　　者：舒学香[1] 孙丽慧[1] 郑裕国[1] 

机构地区：[1]浙江工业大学生物工程研究所,浙江杭州310014

出　　处：《工业微生物》2013年第5期14-20,共7页Industrial Microbiology

基　　金：国家"973"计划资助项目(编号:2011CB710806)

摘　　要：从采自全国各地的200余份土壤样品中,筛选得到一株能够高效合成(S)-4-氯-3-羟基丁酸乙酯的菌株Candida sorboxylosa。对该菌株的产羰基还原酶条件进行的研究表明,最适发酵培养基配方:葡萄糖10 g/L,酵母提取粉50 g/L,KH_2PO_4 2.5 g/L,CuCl_2 0.02 g/L,NaCl 1 g/L。最适发酵培养条件:装液量30 mL/500 mL摇瓶,接种量1%,初始pH 5.0,该茵在此条件下发酵培养20 h,底物4-氯乙酰乙酸乙酯(COBE)羰基还原酶酶活达到202.42 U/L,与优化前相比,提高11.8倍。该茵在催化COBE的转化反应中,具有很好的立体选择性,产物光学纯度(e.e.%)始终大于99%。Candida sorboxylosa ZJB-12164 for the production of ethyl （S）-4-chloro-3-hydroxybutanoate with high enantioselectivity was isolated from soil samples. Then, the optimization of culture conditions for enzyme production by the strain ZJB-121CM was studied. The results showed that the optimum medium components were as follows： 10.0 g/L glucose, 50.0 g/L yeast extract powder, 2.5 g/L KH2PO4,0.02 g/L CuC12, 1.0 g/L NaCI. The optimal culture conditions were medium volume of 30 mL/500 mL, the inoculum size of 1%, and pH of 5.0. After optimization, the enzyme activity increased to 202.42 U/L （ about 11.8 fold）. In the reduction process of ethyl 4-chloroacetoacetate by using resting cell catalysis system, the optical purifies of （S）-CHBE were always higher than 99%.

关 键 词：（S）4-氯-3-羟基丁酸乙酯 CANDIDA sorboxylosa 羰基还原酶 4-氯乙酰乙酸乙酯 

分 类 号：TQ925[轻工技术与工程—发酵工程]