具有差向选择性还原(R)-6-氰基-5-羟基-3-羰基己酸叔丁酯活性的微生物菌株筛选和鉴定  

Screening for microbe displaying diastereoselective ketoreductase activity towards t-butyl 6-cyano-(5R)-hydroxyl-3-oxohexanoate

作　　者：盛骏桢[1] 王亚军[1] 罗希[1] 郑裕国[1] 

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

出　　处：《生物加工过程》2013年第1期54-59,共6页Chinese Journal of Bioprocess Engineering

基　　金：国家重点基础研究发展计划(973计划)资助(2011CB710800)

摘　　要：从土样中分离得到一株具有差向选择性还原(R)-6-氰基-5-羟基-3-羰基己酸叔丁酯活性的微生物菌株ZJB-09225,经生理生化特征鉴定和18S rDNA测序后鉴定为卡里比克毕赤酵母(Pichia caribbic ZJB-09225)。研究结果发现,在最适发酵条件下培养32 h,生物量为8.8 g/L,体积酶活达7.2 U/L;P.caribbic ZJB-09225最适作用温度、最适作用pH值分别为35℃和7.5。在最适的催化条件下,P.caribbic ZJB-09225细胞催化50.0 g/L(R)-6-氰基-5-羟基-3-羰基己酸叔丁酯3 h后,产物6-氰基-(3R,5R)-二羟基己酸叔丁酯得率3.4%,产物d.e.值99.5%以上。A strain ZJB-09225 displaying diastereoselective ketoreductasc activity towards t-butyl 6-cyano- （5R）-hydroxyl-3-oxohexanoate was isolated and identified as Pichia caribbic by phenotypic, biochemical characteristics and alignment analysis of 18S rDNA sequence. Results revealed that after 32h culture under the optimized fermentation conditions, the volumetric activity and the biomass density of P. caribbic ZJB-09225 were 7.2 U/L and 8.8 g/L, respectively. P. caribbic ZJB-09225 was most active to t-butyl 6- cyano-（5R）-bydroxyl-3-oxohexanoate at 35 ℃ and pH 7.5. In addition,50. 0 g/L t-butyl 6-eyano-（5R）- hydroxyl-3-oxohexanoate was diastereoselectively reduced to t-butyl 6-cyano-（ 3R, 5R ）- dihydroxyhexanoate by P. caribbic ZJB-09225 for 3h, in a yield of 3.4% with d. e. value over 99.5%.

关 键 词：羰基还原酶 卡里比克毕赤酵母 阿托伐他汀 (R)-6-氰基-5-羟基-3-羰基己酸叔丁酯 

分 类 号：Q621.2[生物学—生物物理学] R329.26[医药卫生—人体解剖和组织胚胎学]