多酶偶联催化制备手性化合物(S)-环氧苯乙烷  

Multi-Enzyme Cascade Catalysis for the Preparation of Chiral Compound(S)-Styrene Oxide

作　　者：朱利娟[1] 胡蝶[2] 储亚琴 臧嘉[2] 余涛[1] 邬敏辰[3] ZHU Lijuan HU Die CHU Yaqin ZANG Jia YU Tao WU Minchen(School of Phannaceutical Science, Jiangnan University, Wuxi 214122, China School of Biotechnology Jiangnan University, Wuxi 214122, China Wuxi Medical School, Jiangnan University, Wuxi 214122, China)

机构地区：[1]江南大学药学院,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122 [3]江南大学无锡医学院,江苏无锡214122

出　　处：《食品与生物技术学报》2017年第5期473-478,共6页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(31271811);江苏省普通高校研究生科研创新计划项目(SJLX-0529)

摘　　要：在水/有机两相中将羰基还原酶SyS1、葡萄糖1-脱氢酶SyGDH和卤代醇脱卤酶CSyHheC偶联催化α-溴代苯乙酮以高效制备(S)-环氧苯乙烷(SO)。第1步反应催化α-溴代苯乙酮不对称还原为(S)-2-溴-1-苯基乙醇((S)-BPE)及NADPH的原位再生,其优化的反应条件:100 mmol/L磷酸钾缓冲液(p H 8.0)/正己烷(V∶V=4∶6)、40 mmol/Lα-溴代苯乙酮、80 mmol/LD-葡萄糖、0.2 mmol/L NADP+和70 mg/m L E.coli/Sygdh-Sys1湿菌体(共表达SyGDH和SyS1),终体积1.0 m L,35℃反应8 h。第2步反应催化(S)-BPE脱卤闭环为(S)-SO,其反应条件:在上述反应体系中加入磷酸钾缓冲液(p H 8.0)和0.3 U SyHheC,终体积2.0 m L,35℃反应30 min。通过连续2步酶催化反应,终产物(S)-SO的摩尔产率为99%、对映体过量值>99%。In an aqueous/organic two-phase,cascade catalysis was carried out for the efficient preparation of（S）-styrene oxide（SO） by a combination of carbonyl reductase（SyS1）,glucose 1-dehydrogenase（SyGDH） and halohydrin dehalogenase C（SyHheC）. The firststep was to asymmetrically reduceα-bromoacetophenone into（S）-2-bromo-1-phenylethanol（（S）-BPE） and to regenerate NADPH in situ. The optimized conditions were 100 mmol/L potassium phosphate buffer（pH 8.0）/n-hexane（v∶v=4∶6）,40 mmol/L α-bromoacetophenone,80 mmol/L D-glucose,0.2 mmol/L NADP＋ and 70 mg/mL wet E. coli/Sygdh-Sys1 cells（co-expressing both SyGDH and SyS1） in a volume of 1.0 mL at 35 ℃ for 8 h. The second step was dehalogenation ring-closure of（S）-BPE into（S）-SO by adding potassium phosphate buffer（pH 8.0） and 0.3 U SyHheC into the above reaction system, in a volume of 2.0 mL,at 35 ℃ for 30 min. Through two steps of enzyme catalysis,the final product（S）-SO was obtained with a molar yield of 99% and an enantiomeric excess value more than 99%.

关 键 词：多酶偶联 (S)-环氧苯乙烷 水/有机两相 不对称还原 脱卤闭环 

分 类 号：O621.3[理学—有机化学]