产青霉素酶枯草芽孢杆菌发酵条件的优化  

Optimization of Bacillus subtilis DB104/pWB-pemp Fermentation Conditions for Efficient Biosynthesis of Penicillinase

作　　者：黄琳[1] 刘逸寒[1] 王春霞[1] 王建玲[1] 杜连祥[1] 路福平[1] 

机构地区：[1]工业发酵微生物教育部重点实验室工业酶国家工程实验室天津市工业微生物重点实验室天津科技大学生物工程学院,天津300457

出　　处：《生物技术》2011年第6期77-80,共4页Biotechnology

基　　金：国家"十二五"农村领域科技计划课题(No.2011AA100905);国家自然科学基金项目(31101219)资助

摘　　要：目的:对一株产青霉素酶的重组枯草芽孢杆菌(Bacillus subtilis DB104/pWB-pemp)的摇瓶发酵条件进行优化。方法:利用单因素实验及正交实验等方法考查重组菌摇瓶发酵条件,研究不同浓度的碳源、氮源、金属离子、磷酸盐及不同pH、接种量、装液量和温度等条件对产青霉素酶的影响。结果:重组菌株摇瓶最适发酵条件为pH 7.0、接种量3%、装液量14%、发酵温度37℃,培养基成分为2%蔗糖、3.5%酵母膏、0.1mmol/L镁离子、0.4%磷酸盐。结论:最优条件下,发酵36h,产酶活力达到2227.8U/mL,为先前研究结果(1 580 U/mL)的1.41倍,为重组菌的大规模发酵生产奠定了基础。Objective: The fermentation conditions in shaking flasks of Bacillus subtilis DB104∕pWB-pemp producing Penicillinase were investigated.Method:The fermentation in shaking flasks was studied by Single-factor comparative experiments and orthogonal experiments.The effects of different concentration of carbon source,nitrogen source,metal ion,phosphate and different initial pH,inoculums,medium volume and temperature on Penicillinase synthesis were studied.Result:The optimal fermentation conditions in shaking flasks were as followed:initial pH 7.0,inoculums size 3%,medium volume 14%,temperature 37℃,sucrose 2%,yeast extract 3.5%,magnesium ions 0.1mmol/L,phosphate 0.4%.Conclusion:Under these conditions,enzyme activities of Penicillinase reached 2 227.8U/mL,which was 1.41 times of before result(1 580 U/mL).These laid a foundation for large scale fermentation production of recombinant bacterium.

关 键 词：青霉素酶 产酶条件 摇瓶培养 发酵 酶活力 

分 类 号：Q81[生物学—生物工程]