产顺式环氧琥珀酸水解酶的博德特氏菌BK-52的筛选、鉴定及其产酶条件优化  

Isolation and identification of a novel cis-epoxysuccinate hydrolase-producing Bordetella sp.BK-52 and optimization of enzyme production

作　　者：潘海峰[1] 谢志鹏[1] 鲍文娜 张建国[1] 

机构地区：[1]浙江大学生物化学研究所,杭州310058 [2]杭州宝晶生物化工有限公司,杭州311106

出　　处：《微生物学报》2008年第8期1075-1081,共7页Acta Microbiologica Sinica

基　　金：funded by Hangzhou Bioking Biochemical Engineering Co.,Ltd.

摘　　要：【目的】产顺式环氧琥珀酸水解酶(CESH)新型菌株的筛选、鉴定及其产酶条件优化。【方法】通过电镜、Biolog GN、(G+C)含量和16S rDNA序列研究,对筛选菌株进行分类鉴定。通过红外光谱、核磁共振氢谱和碳谱、质谱以及旋光度实验,鉴定纯化产物的结构并优化CESH产酶条件。【结果】本文筛选出一株产CESH的新型菌株,该菌株可将顺式琥珀酸盐转化为D(-)-酒石酸,属于博德特氏菌属,并将其命名为博德特氏菌BK-52。最佳发酵条件为:最佳温度30°C,最佳pH7.0,最佳发酵时间36h,最佳碳源蔗糖,最佳无机氮源硫酸铵,最佳酶诱导剂顺式环氧琥珀酸二钠。在此最佳条件下,CESH酶活最高达764U/g生物量。【结论】本文筛选的新菌株博德特氏菌BK-52为D(-)-酒石酸的生产提供了一种新的方法。[Objective] To isolate and identify a novel strain with cis-epoxysuccinate hydrolase （CESH） activity and to optimize its enzyme production. [Methods] The isolated strain was identified by electron microscopy, Biolog gram negative （GN） test, G＋C （guanine plus cytosine） content measurement and 16S rDNA sequence. The purified enzymatic biotransformed product was identified by IR, ^1H-NMR, ^13C-NMR, MS and optical rotation analysis. Then the fermentation conditions for CESH production were optimized. [Results] A novel CESH-producing strain was isolated for biotransforming cis-epoxysuccinate to D（-）-tartaric acid. It was assigned to genus Bordetella and named Bordetella sp. BK-52. The optimal conditions were found to be 30℃, pH 7.0, fermentation time 36 h, carbon source of saccharose, inorganic nitrogen source of ammonium sulfate and enzyme inducer of disodium cis-epoxysuccinate. Under these conditions, the maximum CESH activity reached 764 U/g biomass. [Conelusion] The isolated Bordetella sp. BK-52 provided a new alternative for biosynthesis of D（-）-tartaric acid from cis-epo x ysuccinate.

关 键 词：博德特氏菌 顺式环氧琥珀酸水解酶 D(-)-酒石酸 筛选 鉴定 

分 类 号：TQ920.1[轻工技术与工程—发酵工程]