产顺式环氧琥珀酸水解酶的红球菌M1菌株的分离鉴定及其产酶条件优化

Isolation, Identification and Phylogenetic Analysis of Rhodococcus sp. Strain M1 Producing cis-epoxysuccinate Hydrolase and Optimization of Production Conditions

从土壤中筛选到一株产顺式环氧琥珀酸水解酶 (ESH)的菌株 ,经生理生化、Biolog碳源利用试验和 1 6SrDNA序列分析系统发育研究 ,菌株可能为赤红球菌 (Rhodococcusruber)M1。摇瓶试验确定了最佳碳源、氮源、顺式环氧琥珀酸二钠添加时间和添加量。正交优化试验的最佳培养基和培养时间为 :葡萄糖 1 2 % ,硫酸铵 0 6 % ,酵母膏0 5 % ;顺式环氧琥珀酸二钠投加时间为 30h ,投加量为 3 5 8% ;菌体培养时间 70h。摇瓶试验ESH酶活达 75 0U g湿细胞。目前该菌株已经应用于固定化细胞连续生产L(+)

Strain M1, capable of producing cis epoxysuccinate hydrolase (ESH) with high yield,was isolated from the soil on campus of Zhejiang University,and identified as Rhodococcus ruber based on physiological experiment,Biolog carbon source utilization experiment and phylogenetic analysis of 16S rDNA sequence. The conditions for enzyme formation of strain M1 were optimized. The optimum conditions were glucose,1 2%,(NH 4) 2SO 4,0 6%,and yeast extract 0 5%,the time and quantity of adding disodium cis epoxysuccinate was 32h and 3 58%,respectively. The ESH activity was about 750U/g wet cells when the organism was cultured under above optimized conditions. This is the first report dealing with strain producing the cis epoxysuccinate hydrolase in Rhodococcus.