枯草芽孢杆菌产角蛋白酶的发酵过程优化  

Optimization of Culture Process Conditions and Feeding Strategy for Keratinase by Recombinant Bacillus subtilis

作　　者：刘文涛[1] 刘柏宏[1,2] 张娟[1,2] 李江华[1,2] 陈坚[3,2] 堵国成[1,2] 

机构地区：[1]江南大学生物工程学院,江苏无锡214122 [2]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [3]江南大学粮食发酵工艺与技术国家工程实验室,江苏无锡214122

出　　处：《食品与生物技术学报》2014年第5期466-471,共6页Journal of Food Science and Biotechnology

基　　金：国家863计划项目(2011AA100905);国家"十二五"科技支撑计划项目(2011BAK10B03);江苏省自然科学基金项目(BK2012553)

摘　　要：为了实现重组角蛋白酶在基因工程菌Bacillus subtilis WB600中的高效生产,在3L发酵罐中对发酵条件和补料策略进行了优化,建立了补料分批发酵工艺.研究发现,两阶段控制pH,前6小时控制pH 7.0,后期控制pH 8.0、DO 20％,发酵过程以葡萄糖为流加碳源,7～11 h以μ为0.15 h-1指数流加、12～15 h以μ为0.1 h-1指数流加、16～27 h恒速流加葡萄糖3 g/（L·h）,在30 L发酵罐水平上,27 h酶活达到864 U/mL,生产强度为31.8 U/（mL·h）,国外报道的酶活达到2 900 mL,酶活在国内属领先水平,为重组枯草芽孢杆菌产角蛋白酶的工业化生产打下了坚实的基础.In order to achieve the high production of keratinase by recombinant Bacillus subtilis WB600,fermentation conditions and feeding strategy were optimized in 3 L fermentor,and a fedbatch fermentation was establishde.The results showed that,using a two-stage pH control strategy in which pH was controlled at 7.0 for the first 4 h and then 8.0 for remaining time and a mixed feeding strategy in which keeping the exponential feeding from 6 h to 15 h （6～11 h μ was 0.15 h-1,11～15 h μ was 0.1 h-1） and then constant-fed fermentation for remaining time,the keratinase activity reached 864 U/mL.The keratinase activity reached a leading level at home,and the keratinase reached 2 900 U/mL overseas.This work reached a high keratinase productivity and it is a stable theroret-ics for the fermention sacle-up process of keratinase.

关 键 词：重组枯草芽孢杆菌 流加策略 发酵优化 角蛋白酶 

分 类 号：TQ925.2[轻工技术与工程—发酵工程]