摩氏摩根菌产酯酶条件优化及在普瑞巴林手性中间体合成中的应用

为建立廉价、高效的普瑞巴林关键手性中间体生产工艺,经单因素优化和正交试验,确定了摩氏摩根菌CCTCC M 2011175最佳产酯酶培养基,组成(g/L)为葡萄糖15.0,牛肉膏7.0,Na2HPO41.0,Fe2(SO4)30.1,吐温-8010.0。优化后酯酶比酶活达到1 071.0 U/L,为出发培养基的2.5倍。以培养基优化后获得的摩氏摩根菌菌体为催化剂,立体选择性拆分外消旋2-羧乙基-3-氰基-5-甲基己酸乙酯水解,转化率达到45%,对映体过量值(e.e.)大于94%,为酶法生产普瑞巴林关键手性中间体奠定了良好基础。

Optimization of Morgarella morganii esterase production and its application in synthesis of chiral intermediate of Pregabalin

In order to develop an efficient and inexpensive biocatalytic process for （S）-2-carboxyethyl-3-cy- ano-5-methylhexanoic acid, key intermediate of Pregabalin, production of esterase by Morgarella morganii CCTCC M 2011175 was optimized by single factor method and orthogonal test. The optimal medium compo- sitions were as follows （ in g/L） ： glucose 15.0, beef extract 7. 0, Na2 HPO4 1.0, Fe2 （ SO4 ） 3 0. l, and Tween-80 10. 0. The esterase activity reached 1 071.0 U/L,which was 1.5 times higher than that before optimization. Kinetic resolutions of rac-2-earboxyethyl-3-eyano-5-methylhexanoic acid ethyl ester using whole cells of M. morganii CCTCC M 2011175 afforded the product in 45% conversion and 94% e.e. It provided a potentially promising bioprocess for the production of chiral intermediate of Pregabalin.