Bt培养基配比及伴孢晶体纯化条件的优化  

Optimization of Culture Condition and Purification of Bacillus thuringiensis HD-1

作　　者：王善利[1] 马景芝 袁勤生[1] 

机构地区：[1]华东理工大学生物工程学院,上海200237

出　　处：《华东理工大学学报（自然科学版）》2006年第3期285-289,共5页Journal of East China University of Science and Technology

摘　　要：苏云金芽孢杆菌〔B t(HD-1)〕发酵中培养基的配方影响发酵水平,还决定发酵产物杀虫毒力的效果。本实验在基础参数试验基础上,采用均匀设计法对B t发酵中培养基组分、杀虫晶体蛋白纯化条件进行优化,并直接采用发酵中伴孢晶体量作为最终评价指标。结果表明:B t培养基成分对发酵的主要影响因素依次为:M gSO4、蛋白胨、可溶性淀粉,最佳用量为(g/L):1.3、2.6、4.5;晶体蛋白双液相纯化的主要影响因素依次为:K2HPO4/KH2PO4质量比、PEG浓度、沉淀量、离心速度,最佳条件为:3∶1、110 g/L、1 g、1 440 r/m in,其中离心速度若选500 r/m in,可能对晶体纯度提高更有利,但低速对回收率有负影响。The medium components of Bacillus thuringiensis subsp, kurstaki HD-1 and aqueous two phase distribution purification condition of HD-1 crystal were optimized by uniform-designed method. Crystal amount was used directly as an evaluation standard in both processes. Results show that the main confluence factors in the fermentation of Bacillus thuringiensis HD-1 are MgSO4, peptone, soluble starch with the concentration of 1.3 g/L,2.6 g/L,4.5 g/L,receptively. And the main confluence factors in the process of purification of HD-1 crystal are the ratio of K2HPO4/KH2PO4,PEG4000 concentration, precipitate amount and centrifugation speed. The optimum condition of purification is as follows： m （K2HPO4）/m （KH2PO4） 3 ： 1, PEG4000 110g/L, precipitate amount 1 gandcentrifugationspeed 1 440r/min. Lowering of the centrifugation speed is useful to get high purity crystal, while at the cost of the recovery rate.

关 键 词：苏云金芽孢杆菌 培养基优化 晶体 提纯 均匀设计 

分 类 号：Q503[生物学—生物化学] TQ92[轻工技术与工程—发酵工程]