响应面法优化菌株Bacillus sp.W77酯酶发酵条件  

Optimization of Fermentation Conditions for Bacillus sp. W77 Eaterase by Response Surface Methodology

作　　者：张敏文[1] 刘悦[1] 李荷[1] 

机构地区：[1]广东药学院基础学院生物化学与分子系,广州510006

出　　处：《应用与环境生物学报》2012年第6期993-998,共6页Chinese Journal of Applied and Environmental Biology

基　　金：国家自然科学基金项目(No.30570 0 55);广东省科技计划项目(No.2009B020313005);广东省自然科学基金项目(No.94510224010 03873)资助~~

摘　　要：为开发适应性好的工业用酶,从深圳福田红树林自然保护区土壤中分离纯化获得一株产耐高温嗜碱性酯酶的菌株,通过形态学和分子生物学初步鉴定该菌株为芽孢杆菌属(Bacillus sp.),命名为Bacillus sp.W77;对所产的粗酶液进行酶活测定,发现该菌产的酯酶的最适反应温度是50℃,最适pH为8.0,且在碱性条件下较稳定;采用响应面法(Response surface methodology)对土壤菌株Bacillus sp.W77的产酯酶条件进行了优化,酵母提取物6.67 g/L,磷酸氢二钾0.56 g/L,硫酸镁0.36 g/L时,此时预测的最大酯酶活力的D405 nm值为1.266,在最佳产酶条件下,优化后酯酶活力的D405 nm值由0.829提高到1.235,实际值达到理论预测值的97.5%.A thermostable basophilic esterase-producing bacterial strain was isolated from soil sample in Futian Mangrove Natural Reserve, Shenzhen, China. This strain was identified as Bacillus sp. W77 according to the morphological and molecular characterization. The results of the ester ase activity showed that the optimal temperature and pH for the activity of the crude enzyme was 50 ℃ and 8.0, respectively. The response surface methodology was applied to study the optimal conditions of fermentation of esterase-producing Bacillus sp. W77. When the yeast extracts, K2HPO4 and MgSO4 were 6.67 g/L, 0.56 g/L and 0.36 g/L, respectively, the maximum predicted D40~ nm was verified as 1.266. However, the D405 nm was practically improved from 0.829 to 1.235 after the optimization. The practical measured value approached almost 97.5% of the theoretical predicted value. This research provided theoretical basis for industrial application of microbial esterase.

关 键 词：芽孢杆菌 菌株筛选 酯酶 酶学性质 响应面法(RSM) 发酵条件优化 

分 类 号：TQ925[轻工技术与工程—发酵工程]