茶氨酸生物合成工程菌构建

doi:10.13305/j.cnki.jts.2007.01.009

Abstract

Abstract: γ-ggt was cloned by PCR from E.coli DH5α. Then, the PCR product of γ-ggt digested with two restriction enzymes, Kpn I and Xho I, was purified and ligated with the pET-32a vector digested with the same enzymes by T4 DNA ligase. Then the ligation product was transformed to E.coli BL21 and the engineered Escherichia coli strain was constructed successfully. The γ-Glutamyltranspeptidase was expressed with induction of 0.05βmmol/L IPTG in 32℃ incubation. The activity of 1βg fresh cells of engineered strain was 2.0βU/g,it is about 15 times higher than that of E.coli DH5α．Under the catalysis of cells of the engineered strain induced with IPTG, the yield of theanine from L-Gln and ethylamine was 29.40βg/L and the conversion rate of L-Gln as to theanine being 48.22%; it is about 100 times higher than that of E.coli DH5α．

Key words: γ-Glutamyltranspeptidase;, theanine, engineered strain, construction

CLC Number:

S571.1
Q523

WANG Xian-bo, WANG Li-yuan, CHENG Hao, ZHOU Jian, LIN Zhi. The Construction of the Engineered Escherichia coli Strain for the Biosynthesis of Theanine[J]. Journal of Tea Science, 2007, 27(1): 61-66.

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The Construction of the Engineered Escherichia coli Strain for the Biosynthesis of Theanine

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