L-天冬酰胺酶的补料分批发酵  

Fed-Batch Fermentation of L-Asparaginase

作　　者：王云龙 刘松[1,2] 冯岳[1,2] 堵国成[1,2] 陈坚[1,2] WANG Yunlong;LIU Song;FENG Yue;DU Guocheng;CHEN Jian(Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2020年第3期1-8,共8页Journal of Food Science and Biotechnology

基　　金：国家自然基金面上项目(31771913);江苏省重点研发计划社会发展项目(BE2016629)。

摘　　要：L-天冬酰胺酶(L-Asparaginase,EC 3.5.1.1,L-ASNase)广泛应用于食品和医药领域。为实现重组L-ASNase的高产,在3 L罐水平研究了搅拌转速与补料分批发酵条件对Bacillus subtilis/ASNΔ25/B2菌体生长与产酶的影响。通过优化确定补料分批发酵条件如下:发酵0~8 h搅拌转速:700 r/min;发酵8 h后搅拌转速:900 r/min;发酵16~28 h:恒速流加(18.75 mL/h)蔗糖(800 g/L),恒速流加(32 mL/h)酵母蛋白胨(200 g/L)和玉米浆(80 g/L)混合氮源。基于以上发酵条件,L-ASNase酶活在48 h可达到1413.6 U/mL,较分批发酵提高了66.2%,生产强度提高了24.6%。研究结果为重组L-ASNase工业化生产提供了基础数据。L-asparaginase(EC 3.5.1.1,L-ASNase)has been widely used in the fields of food and medicine.In order to achieve the efficient production of L-ASNase,the effects of stirring speed and fed-batch fermentation conditions on the cell growth and enzyme production of Bacillus subtilis/ASNΔ25/B2 were investigated in 3 L fermenter.After the optimization,conditions used for fed-batch fermentation were determined as follows:the stirring speed was set at 700 r/min for the first 8 h and shifted to 900 r/min till the end of the fermentation;for fermentation at 16~28 h,sucrose solution(800 g/L)and nitrogen sources(yeast peptone:200 g/L;corn syrup:80 g/L)were fed at 18.75 mL/h and 32 mL/h,respectively.Based on this fermentation condition,the L-ASNase yield reached 1413.6 U/mL at 48 h,which was 66.2%higher than that of batch fermentation.The final production intensity obtained here was 24.6%higher than that of the latter.The results provide the basic data for the industrial production of the recombinant L-ASNase.

关 键 词：枯草芽孢杆菌 L-天冬酰胺酶 发酵优化 分批发酵 补料分批发酵 

分 类 号：Q815[生物学—生物工程]