响应面法优化重组枯草芽孢杆菌发酵生产青霉素G酰化酶  

Optimization of Penicillin G Acylase Production by Recombinant Bacillus Subtilisvia Response Surface Analysis

作　　者：仇晶晶[1] 陈玮[1] 丁明[1] 张漫莉[1] 赵辅昆[1] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2012年第6期868-873,共6页Journal of Zhejiang Sci-Tech University(Natural Sciences)

摘　　要：利用响应面法优化重组枯草芽孢杆菌pAUB-BmPGA/BS168发酵生产青霉素G酰化酶的条件。通过Plackett-Burman设计法对碳源﹑氮源﹑无机盐﹑温度等19个因素对发酵产青霉素G酰化酶的影响进行评价。筛选出发酵温度和酵母膏为影响产酶的显著因素。采用了中心组合设计实验对两种显著影响因素进行了优化,应用响应面分析确定了显著因素的最佳水平。结果表明,当温度为34℃,酵母膏为8.8g/L时,最终的酶活达到28.2U/mL,比初始发酵条件提高了1.19倍。在最佳条件利用15L的发酵罐对重组枯草芽孢杆菌进行扩大培养,最终酶活可达51.0U/mL,相对于摇瓶发酵又有大幅度的提高。Response surface methodology and design is used to optimize the fermentation conditions for Penicillin G acylase production by recombination bacillus subtilis. Initial screening via the Plackett-Burman method and design reveals that the fermentation temperature and yeast extract are the most significant a- mong 19 factors. Further optimization with central design and response surface analysis predicts that the final enzyme activity can reach 28.2 U/mL when the temperature is 34~C and the yeast extract is 8. 8 g/L. The enzyme activity of the optimization conditions is 2. 19 times more than the initial conditions. In accord- ance with the optimization conditions, we use the 15 L fermentor to expand fermenting and find that the highest PGA activity of the fermentation liquid can reach 51 U/mL, which is improved more significantly than when it is in the flask.

关 键 词：青霉素G酰化酶 枯草芽孢杆菌 响应面 发酵 

分 类 号：Q815[生物学—生物工程]