大肠杆菌BL21(DE3)生产β-环状糊精转移酶发酵条件的优化  

Optimization of the culture conditions of E.coli BL21(DE3) producing β-cyclodextrin glucanotransferase

作　　者：宋拓[1] 李俊俊[1] 唐湘华[1] 谢振荣[1] 张学林[1] 王秋云 许波[1] 周峻沛[1] 慕跃林[1] 黄遵锡[1] 

机构地区：[1]生物能源持续开发利用教育部工程研究中心云南省酶资源开发与应用工程研究中心云南省生物质能与环境生物技术重点实验室云南师范大学酶工程重点实验室云南师范大学生命科学学院,云南昆明650500

出　　处：《中国酿造》2013年第4期52-57,共6页China Brewing

基　　金：科技部农业科技成果转化资金项(2010GB2F300436);国家自然科学基金委员会资助项目(31160229)

摘　　要：采用正交试验确定大肠杆菌BL21(DE3)产β-环糊精转移酶的发酵条件。结果表明,生产β-环状糊精转移酶的最适条件为:接种量1%,100mL三角瓶装液量15mL,初始pH值为7.0,Ca2+浓度为1.25mmol/L,Mg2+浓度为2.50mmol/L,温度控制在37℃,转速控制为150r/min。当OD600达到1.4时,加入终浓度5.0g/L的α-乳糖,转速提高至170r/min,37℃诱导12h后过滤,β-CGTase酶活最高可达16.3μ/mL。The culture condition for producing β-cyclodextrin glucanotransferase by E.coli BL21 （DE3） was primarily established by orthogonal experimental design. The results showed that the optimal fermentation conditions were as follows： the inoculum size was 1%; The load of 100ml-flask was 15ml; initial pH value was 7.0; the concentration of Ca^2＋ was 1.25mmol/L, the concentration of Mg^2＋ was 2.50mmol/L, incubation temperature was 37~C and the speed was 150r/rain. When the value of OD600 reached 1.4, u-lactose was added so as to the final concentration to 5.0g/L, the speed was increased to 170r/min, and induced 12h at 37℃ then filtered, the β-CGTase activity reached the maximum value as 16.3μ/ml.

关 键 词：发酵条件 β-环状糊精转移酶 重组大肠杆菌 优化 

分 类 号：TS05[轻工技术与工程]