|本期目录/Table of Contents|

[1]成成,李兆丰,李彬,等.利用重组大肠杆菌生产α-环糊精葡萄糖基转移酶[J].生物加工过程,2009,7(03):56-63.
 CHENG Cheng,LI Zhao-feng,LI Bin,et al.Production of α-cyclodextrin glycosyltransferase in recombinant Escherichia coli[J].Chinese Journal of Bioprocess Engineering,2009,7(03):56-63.
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利用重组大肠杆菌生产α-环糊精葡萄糖基转移酶
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《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
7
期数:
2009年03期
页码:
56-63
栏目:
出版日期:
2009-05-30

文章信息/Info

Title:
Production of α-cyclodextrin glycosyltransferase in recombinant Escherichia coli
文章编号:
1672-3678(2009)03-0056-08
作者:
成成12李兆丰12李彬12刘花12陈坚12吴敬12
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
Author(s):
CHENG Cheng12 LI Zhao-feng12 LI Bin12 LIU Hua12 CHEN Jian12 WU Jing12
1. State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi 214122, China;
2. School of Biotechnology, Key Laboratory of Industrial Biotechnology of the Ministry of Education,
Jiangnan University, Wuxi 214122, China
关键词:
α-环糊精葡萄糖基转移酶cgt基因' target="_blank" rel="external">">cgt基因大肠杆菌克隆
分类号:
Q555+.4
文献标志码:
A
摘要:
将来源于软化类芽孢杆菌(Paenibacillus macerans)的α-环糊精葡萄糖基转移酶(α-CGT)基因插入含pelB信号肽的质粒pET-20b(+)中,构建了表达载体pET-20b(+)/cgt,并将其转化表达宿主E.coli BL21(DE3)。对重组菌E.coli BL21/ pET-cgt进行摇瓶发酵条件的优化,确定了其胞外表达α-CGT酶的最适条件:葡萄糖8 g/L,乳糖 0.5 g/L,蛋白胨12 g/L,酵母膏24 g/L,K2HPO4 72 mmol/L,KH2PO4 17 mmol/L,CaCl2 2.5 mmol/L;初始pH为7.0,诱导温度为25 ℃。在该条件下培养90 h后最终α-CGT酶的胞外比活达到22.1 U/mL,与来源菌P. macerans所产天然酶比活相比提高了42倍,实现了α-CGT酶的高效生产。将基因工程菌在上述条件下于3 L发酵罐中发酵,90 h后胞外酶比活达到22.6 U/mL,证实了工业化放大的可能性。

参考文献/References:

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[12]陈坚, 吴敬, 李兆丰, 等. 一种α环糊精葡萄糖基转移酶基因的克隆与表达:中国,101294149[P]. 2008-10-29.
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备注/Memo

备注/Memo:
收稿日期:2008-12-26
基金项目:国家杰出青年基金资助项目(20625619);食品科学与技术国家重点实验室科研基金资助项目(SKLF-MB-200802);江苏省自然科学基金资助项目(BK2007019)
作者简介:成成(1985—),男,江苏泰州人,硕士研究生,研究方向:发酵工程及酶工程;吴敬(联系人),教授,E-mail:jingwu@jiangnan.edu.cn
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