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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (10): 50-56    
研究报告     
重组大肠杆菌产γ-环糊精葡萄糖基转移酶的摇瓶发酵优化
纪丽萍1,2, 吴丹1,2, 吴敬1,2, 陈坚1,2
1. 江南大学食品科学与技术国家重点实验室 无锡 214122;
2. 江南大学生物工程学院工业生物技术教育部重点实验室 无锡 214122
Optimization of Fermentation in Shake Flasks for Recombinant γ-CGTase Expression in E. coli
JI Li-ping1,2, WU Dan1,2, WU Jing1,2, CHEN Jian1,2
1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
2. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi 214122, China
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摘要:

为了实现来源于碱性芽孢杆菌Alkalophilic Bacillus clarkii 7364的γ-环糊精葡萄糖基转移酶的高效胞外表达,对OmpA信号肽介导的E.coli BL21(DE3)/pET20b(+)-γcgt基因工程菌进行发酵培养基及发酵条件的优化,并进行正交试验,获得最优培养基:甘油5g/L、蛋白胨6g/L、酵母膏24g/L、钙离子6mmol/L、镁离子2mmol/L、甘氨酸0.75%、PO43- 0.1mol/L;在此基础上最适发酵条件:pH6.5、25℃培养、装液量30ml/250ml、转速220r/min、0.02%SDS、在发酵10h时利用5g/L乳糖进行诱导,使得酶活从初始的5189.2U/ml提高到20268.8U/ml。研究结果得到高效表达的培养条件,为实现该酶的工业化应用打下了基础。

关键词: 重组大肠杆菌环糊精葡萄糖基转移酶发酵优化    
Abstract:

In order to achieve efficient extracellular production of γ-cyclodextrin glycosyltransferase(γ-CGTase), E. coli BL21(DE3) carrying the γ-cgt gene of Alkalophilic Bacillus clarkii 7364 was investigated. By comparison of different culture media and conditions, got the optimized medium: 5g/L glycerol, 6g/L peptone, 24g/L yeast extract, 6mmol/L Ca2+, 2mmol/L Mg2+, 0.75% glycine, the optimized conditions: cultivated in 25℃, 220r/min, 30ml/250ml, pH 6.5, induced by 5g/L lactose at 10h of culture, which contained 0.02% SDS, the enzyme activity rised from 5189.2U/ml to 14683.4U/ml.

Key words: E. coli    γ-CGTase    Fermentation optimization
收稿日期: 2011-05-09 出版日期: 2011-10-25
ZTFLH:  819  
基金资助:

中央高校基本科研业务费专项资金(JUSRP20917)资助项目

通讯作者: 吴敬     E-mail: jingwu@jiangnan.edu.cn
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引用本文:

纪丽萍, 吴丹, 吴敬, 陈坚. 重组大肠杆菌产γ-环糊精葡萄糖基转移酶的摇瓶发酵优化[J]. 中国生物工程杂志, 2011, 31(10): 50-56.

JI Li-ping, WU Dan, WU Jing, CHEN Jian. Optimization of Fermentation in Shake Flasks for Recombinant γ-CGTase Expression in E. coli. China Biotechnology, 2011, 31(10): 50-56.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I10/50


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