反-Prelog规则羰基还原酶产生菌Debaryomyces castellii ZJB-12032产酶条件优化研究  

Optimization of fermentation conditions for Debaryomyces castellii ZJB-12032 displaying anti-Prelog diastereoselective carbonyl reductase activity

作　　者：姚震[1,2] 罗希[1,2] 王亚军[1,2] 郑裕国[1,2] 

机构地区：[1]浙江工业大学生物工程研究所,浙江杭州310014 [2]生物转化与生物净化教育部工程研究中心,浙江杭州310014

出　　处：《发酵科技通讯》2014年第2期1-5,共5页Bulletin of Fermentation Science and Technology

基　　金：国家重点基础研究发展计划(2011CB710800);浙江省重点科技创新团队项目(2009R50043-06);浙江工业大学教学方法改革专项项目(JGZ1110)

摘　　要：6-氰基-(3R,5R)-二羟基己酸叔丁酯是合成阿托伐他汀的关键手性前体。实验室保藏菌株Debaryomyces castellii ZJB-12032能够不对称还原6-氰基-(5R)-羟基-3-羰基己酸叔丁酯β羰基,生成6-氰基-(3R,5R)-二羟基己酸叔丁酯,该生物还原反应符合反-prelog规则。通过优化D.castellii ZJB-12032发酵产酶条件,包括碳源、氮源、磷源、接种量、接种时间、发酵时间,获得其最佳培养基配方:麦芽糖50 g/L、NaNO315 g/L、NaH2PO4·2H2O 0.5 g/L、Na2HPO4·12H2O 0.5 g/L,最适发酵条件:接种量10%,种龄8 h,培养温度30℃,发酵时间24h。在上述最佳发酵条件下,发酵液体积酶活达到10.2 U/L,比酶活1.20 U/g dcw,产物d.e.值99.6%,D.castellii ZJB-12032表现出高差向选择性。Abstract：t-Butyl 6-cyano-（3R, 5R）-dihydroxyhexanoate is a vital chiral intermediate of ator-vastatin. Debaryomyces castellii ZJB-12032 isolated form soil sample and stored in our lab, can reduce t-butyl （R）-6-cyano-5-hydroxy-3- carbonylhexanoate to t-butyl 6-cyano-（3R, 5R）-di- hydroxyhexanoate, following the anti-Prelog＇s rule. Optimization of D. castellii ZJB-12032 culti- vation conditions was：onducted in this work. Results indicated that the optimal fermentationmedium was composed of maltose 50 g/L, NAN03 15g/L, NaH2P04·2H20 0.5 g/L, Na2HP04· 12H20 0.5 g/L; under the ideal euhivation conditions, 10 % inoculation size, inoeula age of 8 h, volume enzyme activity peaked at 10.2 U/L, with specific activity of 1.20 U/g dew and t-butyl 6-eyano- （3R, 5R）-dihydroxyhexanoate d.e. value over 99.5%, after grown at for 24 h, demonstrating Debaryomyces castellii ZJB-12032 has excellent diastereoselectivity towards （R）-6-cyano-5-hy- droxy-3 -earbonylhexanoate.

关 键 词：6-氰基-(3R 5R)-二羟基己酸叔丁酯 羰基还原酶 反-Prelog规则 

分 类 号：TS227[轻工技术与工程—粮食、油脂及植物蛋白工程]