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Journal of Tea Science ›› 2008, Vol. 28 ›› Issue (4): 242-248.doi: 10.13305/j.cnki.jts.2008.04.012

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

ZHU Wen-xian1,2, LI Xing-hui1, WANG Li-yuan2, FANG Wan-ping1,*, CHENG Hao1,2,*   

  1. 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

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

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