利用GS基因构建茶氨酸生物合成工程菌的研究

doi:10.13305/j.cnki.jts.2008.04.012

茶叶科学 ›› 2008, Vol. 28 ›› Issue (4): 242-248.doi: 10.13305/j.cnki.jts.2008.04.012

利用GS基因构建茶氨酸生物合成工程菌的研究

朱文娴^1,2, 黎星辉¹, 王丽鸳², 房婉萍^1,*, 成浩^1,2,*

1. 南京农业大学茶叶科学研究所,江苏南京 210095;
2. 中国农业科学院茶叶研究所,浙江杭州 310008

收稿日期:2008-05-05 出版日期:2008-08-15 发布日期:2019-09-12
作者简介:朱文娴(1982— ),女,硕士研究生,研究方向：茶叶生物技术;通讯作者：fangwp@njau.edu.cn
基金资助:
浙江省科技项目(2007C22074)、江苏省科技攻关计划(BE2007301)、常州市科技项目（CE2007210）

Construction of E. coli Recombinant Engineered Strain for Theanine Biosynthesis with GS Gene Embedded

ZHU Wen-xian^1,2, LI Xing-hui¹, WANG Li-yuan², FANG Wan-ping^1,*, CHENG Hao^1,2,*

1. Tea Research Institute, Nanjing Agricultural University, Nanjing 210095, China;
2. Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou 310008, China

Received:2008-05-05 Online:2008-08-15 Published:2019-09-12

摘要/Abstract

摘要： 为了高效合成茶氨酸,本研究通过将荧光假单胞菌GS基因转接入pET32a质粒中,再将重组质粒转化到E. coli BL21中,构建了一种生物合成茶氨酸的基因工程菌。工程菌株经0.1βmmol/L IPTG,28℃诱导表达,湿菌体的酶活达到41.79 U/mg prot,大约是出发菌株E. coli BL21的126.64倍。工程菌催化L-谷氨酸钠和盐酸乙胺反应生成茶氨酸的产量达到6.2 g/L,其催化L-谷氨酰胺和盐酸乙胺反应生成茶氨酸的能力较出发菌株E. coli BL21有显著提高。

关键词: 谷氨酰胺合成酶, 茶氨酸, 工程菌, 构建

Abstract: An experiment on the construction of E. coli recombinant strain for theanine biosynthesis with GS gene embedded was reported. In this experiment, Glutamine Synthetase gene in E. coli BL21 was ligated with the pET32a vector in order to produce theanine. The ligation product was transformed to E. coli BL21 and the engineered E. coli strain was constructed successfully. The glutamine Synthetase was expressed with induction of 0.1 mmol/L IPTG in 28℃. The activity of γ-glutamyl transpeptidase of the engineered strain of fresh E. coil engineered strain reached 41.79 U /mg prot, and was 126.64 times higher than that of the original strain one and the yield of theanine from L-Gln and ethylamine was 6.3 mg/ml. The ability of theanine biosynthesis of recombinant was enhanced obviously compared with that of original E. coli BL21.

Key words: glutamine synthetase, theanine, engineered strain, construction

中图分类号:

S571.1
Q523

朱文娴, 黎星辉, 王丽鸳, 房婉萍, 成浩. 利用GS基因构建茶氨酸生物合成工程菌的研究[J]. 茶叶科学, 2008, 28(4): 242-248. doi: 10.13305/j.cnki.jts.2008.04.012.

ZHU Wen-xian, LI Xing-hui, WANG Li-yuan, FANG Wan-ping, CHENG Hao. Construction of E. coli Recombinant Engineered Strain for Theanine Biosynthesis with GS Gene Embedded[J]. Journal of Tea Science, 2008, 28(4): 242-248. doi: 10.13305/j.cnki.jts.2008.04.012.

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利用GS基因构建茶氨酸生物合成工程菌的研究

Construction of E. coli Recombinant Engineered Strain for Theanine Biosynthesis with GS Gene Embedded

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