响应面法优化重组枯草芽孢杆菌生产纤维素酶EGA的研究  

Research on Cellulase EGA Production through Optimization and Recombination of Bacillus Subtilis Based on Response Surface Methodology

作　　者：陈涛[1] 张漫莉[1] 李晶虹 吴程雨[1] 赵辅昆[1] 陈玮[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）》2014年第4期451-455,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：浙江省大学生科技创新项目(2013R406021)

摘　　要：通过响应面优化法(response surface methodology,RSM)对分泌表达纤维素酶EGA重组枯草芽孢杆菌的发酵条件进行优化。利用Plackett-Burman实验设计对淀粉、酵母粉、蛋白胨等8个因素对枯草芽孢杆菌产纤维素酶EGA的显著影响与否进行评估分析,筛选得到淀粉、酵母粉和MgSO4·7H2O为3个显著影响的因素,再利用Box-Behnken实验设计对这3个因素进行进一步优化,确定最佳培养基的配比为淀粉21g/L,酵母粉11.26g/L,蛋白胨25g/L,MgSO4·7H2O 1.52g/L,KH2PO40.395g/L,NaCl 3.25g/L,CaCl20.1g/L,FeSO4·7H2O 0.005g/L。在最优条件下,利用小型发酵罐对重组菌进行扩大培养,其最高酶活力达到1 264U/L。Response Surface Methodology was used to optimize the fermentation conditions for secretory expression cellulose EGA and recombination of bacillus subtilis.Plackett-Burman experimental design was adopted to evaluate and analyze obvious effects of 8 factors（including starch,yeast powder and peptone）on producing cellulase EGA by bacillus subtilis.3 factors with obvious effects were screened out：starch,yeast powder and MgSO4·7H2O.Then,Box-Behnken experimental design was used to further optimize the 3 factors.Finally,the ratio of optimal culture medium was confirmed as follows：starch 21g/L,culture medium11.26g/L,peptone 25g/L,MgSO4·7H2O 1.52g/L,KH2PO40.395g/L,NaCl 3.25g/L,CaCl20.1g/L and FeSO4·7H2O 0.005g/L.Under the optimal conditions,a small fermentation tank was used to enlarge cultivation of recombinant bacteria.The highest enzyme activity reached 1 264U/L.

关 键 词：纤维素酶EGA 枯草芽孢杆菌 响应面优化 发酵 

分 类 号：Q786[生物学—分子生物学]