一株不对称水解N-癸酰草铵膦生产L-草铵膦菌株的筛选及催化性质研究  

Screening and catalytic property of a strain for asymmetrically hydrolyzing racemic N-decanoyl-glufosinate to produce L-glufosinate

作　　者：史泰龙 赵文卓 张朝晖[1] SHI Tailong;ZHAO Wenzhuo;ZHANG Zhaohui(College of Biological Engineering, Zhejiang University of Technology, Hangzhou 310014, China)

机构地区：[1]浙江工业大学生物工程学院,杭州310014

出　　处：《工业微生物》2018年第6期19-24,共6页Industrial Microbiology

摘　　要：从活性污泥样品中分离得到一株能不对称水解(D,L)-N-癸酰草铵膦的革兰氏阴性菌。经过16S r DNA测序分子生物学鉴定,命名该菌株为Pseudomonas sp. zjut126。当(D,L)-N-癸酰草铵膦浓度为8 g/L,湿菌体(含水75%):N-癸酰草铵膦=1∶1(w/w),反应24 h,产物L-草铵膦得率为16. 3%,eep> 95%。该菌体催化的最适温度为30℃,在温度40℃以下催化稳定性较好。最适p H为7,pH在6～8之间催化稳定性较好。加入异丙醇有利于N-癸酰草铵膦的溶解,但对细胞催化活性有明显的抑制作用。建立了一条利用Pseudomonas sp. zjut126细胞作催化剂拆分(D,L)-N-癸酰草铵膦制备L-草铵膦的工艺路线,第一步先将(D,L)-草铵膦化学酰化得到(D,L)-N-癸酰草铵膦,然后用菌株选择性水解(D,L)-N-癸酰草铵膦生成L-草铵膦。将未反应的(D)-N-癸酰草铵膦与L-草铵膦进行分离,终产品L-草铵膦的含量是92. 0%,ee值为95. 2%。A gram-negative bacterium was isolated from active sludge samples,which had high activity for enantioselective hydrolysis of(D,L)-N-decanoyl-glufosinate.The stain was identified as Pseudomonas by 16S rDNA analysis.It was named Pseudomonas sp.zjut 126.At 30℃,8 g/L of the wet cells(75%water content)catalyzed(D,L)-N-decanoyl-glufosinate at the concentration of 8 g/L for 24 h,the yield of L-glufosinate reached 16.3%,its ee p value exceeded 95%.The optimum catalytic temperature and pH was 30℃and 7 respectively.The cells had stable catalytic activity below 40℃and at pH 6~8.Organic solvent(isopropanol)was conducive to the dissolution of N-decanoyl-glufosinate,but it had a significant inhibitory effect on the cells’activity.A new process for producing L-glufosinate from(D,L)-glufosinate was established by using Pseudomonas sp.zjut 126 cells as the resolution catalyst.In the first step,racemized glufosinate was chemically acylated to(D,L)-N-decanoyl-glufosinate,which was then enantioselective hydrolyzed by the strain.After separating L-glufosinate from the unreacted(D)-N-decanoyl-glufosinate,the final product of L-glufosinate with the purity of 92.0%and ee p value of 95.2%was obtained.

关 键 词：生物拆分 (D L)-N-癸酰草铵膦 L-草铵膦菌株筛选与鉴定 PSEUDOMONAS 

分 类 号：TQ457[化学工程—农药化工]