摘要
针对生物催化不对称还原反应中必需辅酶再生及反应效率问题,以抗过敏药物的重要中间体化合物光学活性4-甲氧基-1-苯乙醇为目标产物,构建了羰基还原酶基因SCRII-A220D和葡萄糖脱氢酶基因gdh共表达的重组菌株E.coli BL21/p ET-SCRII-A220D-SD-AS-gdh。该双酶共表达耦联系统中,羰基还原酶与葡萄糖脱氢酶酶活水平相当,有利于双酶耦联催化反应的进行。通过考察反应体系和环境条件对该共表达耦联系统催化反应的影响,发现底物质量浓度、细胞质量浓度,及反应体系温度和p H值等条件对最终产物合成效率的影响规律。在较优条件下,产物(R)-4-甲氧基-1-苯乙醇的光学纯度达到98%,产率达到82%。
Concerning the issues of required coenzyme regeneration and reaction efficiency of biocatalytic asymmetric reduction, in this study,the recombinant Escherichia coli BL21/pET-SCRII- A220D-SD-AS-gdh from the co-expressing of the carbonyl reductase （SCRII-A220D） and the glucose dehydrogenase （GDH） was constructed to catalyze asymmetric synthesis of optically active （R）-4-methoxyl-l-phenylethanol. In this coupled recombinant system,SCRII-A220D and GDH exhibited the activities in the similar level,which help two coupled enzymes to function synergistically. By investigating the effects of bio-mediated reaction conditions,the involvedconditions including substrate concentrations,cell concentrations,reaction temperatures and initial pH values were optimized for enhancement of the yield of desired product. Consequently,the optically pure （R）-4-methoxyl-1-phenylethanol was obtained with optical purity of 98% e.e. and the yield up to 82%. In summary,the coupled recombinant system of co-expressing of SCRII-A220D and GDH would be promising in synthesis of chiral aryl alcohols with high efficiency.
作者
李鸣
聂尧
张荣珍
穆晓清
徐岩
LI Min NIE Yao ZHANG Rongzhen MU Xiaoqing XU Yan(School of Biotechnology,Key Laboratory of Industrial Biotechnology of Ministry of Education,Jiangnan University, Wuxi 214122 ,China State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China)
出处
《食品与生物技术学报》
CAS
CSCD
北大核心
2016年第6期569-576,共8页
Journal of Food Science and Biotechnology
基金
国家自然科学基金项目(21376107,21336009)
国家973计划项目(2011CB710800)
国家863计划项目(2011AA02A209,2011AA02A210)
教育部高等学校学科创新引智计划(111计划,111-2-06)
国家高端外国专家项目(GDW20133200113)
江苏高校优势学科建设工程资助项目
关键词
羰基还原酶
葡萄糖脱氢酶
共表达
不对称还原
芳基手性醇
carbonyl reductase,glucose dehydrogenase,co-expression,asymmetric reduction,chiral aryl alcohols
