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- Title
Efficient synthesis of γ-glutamyl compounds by co-expression of γ-glutamylmethylamide synthetase and polyphosphate kinase in engineered Escherichia coli.
- Authors
Pan, Xinru; Yu, Jinhai; Du, Qinglin; Zeng, Shuiyun; Liu, Junzhong; Jiao, Qingcai; Zhang, Hongjuan
- Abstract
γ-Glutamyl compounds have unveiled their importance as active substances or precursors of pharmaceuticals. In this research, an approach for enzymatic synthesis of γ-glutamyl compounds was developed using γ-glutamylmethylamide synthetase (GMAS) from Methylovorus mays and polyphosphate kinase (PPK) from Corynebacterium glutamicum. GMAS and PPK were co-recombined in pETDuet-1 plasmid and co-expressed in E. coli BL21 (DE3), and the enzymatic properties of GMAS and PPK were investigated, respectively. Under the catalysis of the co-expression system, l-theanine was synthesized with 89.8% conversion when the substrate molar ratio of sodium glutamate and ethylamine (1:1.4) and only 2 mM ATP were used. A total of 14 γ-glutamyl compounds were synthesized by this one-pot method and purified by cation exchange resin and isoelectric point crystallization with a yield range from 22.3 to 72.7%. This study provided an efficient approach for the synthesis of γ-glutamyl compounds by GMAS and PPK co-expression system.
- Subjects
ESCHERICHIA coli; MONOSODIUM glutamate; CORYNEBACTERIUM glutamicum; ISOELECTRIC point; YIELD strength (Engineering); ION exchange resins; GAMMA-glutamyltransferase
- Publication
Journal of Industrial Microbiology & Biotechnology, 2020, Vol 47, Issue 8, p573
- ISSN
1367-5435
- Publication type
Article
- DOI
10.1007/s10295-020-02305-4