Construction Fermentation of Engineered Strain and Properties of Recombinantγ-glutamyltranspeptidase

China Biotechnology

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Construction Fermentation of Engineered Strain and Properties of Recombinantγ-glutamyltranspeptidase

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Abstract The γ-glutamyltranspeptidase encoding gene(ggt) from Bacillus subtilis SYU 20016 was amplified by PCR.The ggt gene was inserted in pBV220 to yield the recombinant expression vector pBV220-ggt.Over-expression of ggt in E.coli JM109 was achieved with pBV220-ggt. SDS-PAGE analysis showed an over-expressed recombinant product at about 65kD,consistent with the molecular weight predicted from gene sequence.The characteristics of γ-glutamyltranspeptidase were also discussed. The optimum temperature and pH for the enzyme were determined as 30℃ and 7.2 respectively.The cultures were incubated at 42℃ for 4h with broth volume 20mL/250mL flask and the yield of 6U/mL was obtained, enzyme activity of B. subtilis NX-2 was only 3.2 U/mL.

Key words： biotransformation fermentation condition plasmid stability γ-glutamyltranspeptidase theanine

Received: 21 June 2006 Published: 25 November 2006

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. Construction Fermentation of Engineered Strain and Properties of Recombinantγ-glutamyltranspeptidase. China Biotechnology, 2006, 26(11): 48-53.

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https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2006/V26/I11/48

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