表皮短杆菌ZJB-07021产R-酰胺酶培养条件的优化  

Optimization of Culture Conditions of R-Amidase-Producing Brevibacterium epidermidis Strain ZJB-07021

作　　者：金少军[1] 梁璐怡[1] 郑仁朝[1] 郑裕国[1] 沈寅初[1] 

机构地区：[1]浙江工业大学生物与环境工程学院生物工程研究所,浙江杭州310014

出　　处：《微生物学杂志》2008年第1期1-7,共7页Journal of Microbiology

基　　金：国家重点基础研究发展规划项目(973项目;2003CB716005);国家高技术研究发展计划(863项目;2006AA02Z241)

摘　　要：采用响应面分析法(RSM)对R-酰胺酶产生菌Brevibacterium epidermidis ZJB-07021的发酵培养基进行了优化。首先运用了单因子试验筛选出了发酵培养的最佳pH与温度,在此基础上采用Plackett-Burman(PB)设计法,对8种影响产酶的因素进行评价,实验结果表明,葡萄糖、酵母粉与乙酰胺含量对菌株产酰胺酶的活力具有显著的影响。通过旋转中心组合实验考察了葡萄糖、酵母粉和乙酰胺这三个主要因素对菌株所产酰胺酶活力的影响。发酵培养基优化结果为葡萄糖17.00g/L,酵母粉15.74g/L,乙酰胺7.05g/L,采用优化后的发酵培养条件进行摇瓶发酵培养,酰胺酶的酶活达到72.14U/L,比优化前的初始发酵培养条件下的酶活提高了73.3%。Fermentation medium of Brevibacterium epidermidis ZJB-07021 producing R-amidase was optimized by using response surface methodology （RSM）. The optimum pH and temperature were firstly screened with single factor experiment. Based on this, eight factors that affect the enzyme production were evaluated adopting Plackett-Burman （PB） designing method. The results showed that the contents of glucose, yeast extract, and acetamide possessed strong effects on the activity of R-amidase by the strain. The effects of three factors of glucose, yeast extract, and acetamide on the enzyme activity produced by the strain were observed by rotational center combination experiment. The result of fermentation medium optimization was as follow ： glucose 17. 00 g/L, yeast extract 15.74 g/L, and acetamide 7.05 g/L, and the enzymatic activity of R-amidase after adopting the optimized fermentation culture conditions in shaking flasks reached up to 72.14 U/L, increased by 73.3% as compared with initial fermentation culture conditions before the ontimization.

关 键 词：响应面分析法 R-酰胺酶 培养基优化 二甲基环丙甲酰胺 

分 类 号：Q939[生物学—微生物学]