Enzymatic synthesis of γ-glutamylglutamine, a stable glutamine analogue, by γ-glutamyltranspeptidase from Escherichia coli K-12
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The optimal reaction conditions for the synthesis of γ-glutamylglutamine using γ-glutamyltranspeptidase from Escherichia coli were determined. The maximum yield of γ-glutamylglutamine (110 mM) was obtained using 250 mM l-glutamine and 1.1 U γ-glutamyltranspeptidase/ml at pH 10.5 and at 37°C for 7 h; the conversion of glutamine to γ-glutamylglutamine was 88%.
KeywordsGlutamine γ-Glutamylglutamine γ-Glutamyltranspeptidase Transpeptidation
We would like to thank Dr. Bunnta Watanabe and Prof. Jun Hiratake, Institute for Chemical Research, Kyoto University, for the NMR and MS analyses and for their productive discussion. This work was supported by a Grant-in-Aid for Scientific Research (no. 21380059) to HS from the Ministry of Education, Culture, Sports, Science and Technology of Japan. CY was supported by the 21st Century COE Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan.
- Hara T, Yokoo Y, Furukawa T (1992) Potential of γ-glutamyl-l-cystine and bis-γ-l-glutamyl-l-cystine as a cystine-containing peptide for parental nutrition. In: Takai K (ed) Frontiers and new horizons in amino acid research. Elsevier, Amsterdam, pp 607–611Google Scholar
- Inoue M, Hiratake J, Suzuki H, Kumagai H, Sakata K (2000) Identification of catalytic nucleophile of Escherichia coli γ-glutamyltranspeptidase by γ-monofluorophosphono derivative of glutamic acid: N-terminal Thr-391 in small subunit is the nucleophile. Biochemistry 39:7764–7771CrossRefPubMedGoogle Scholar