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.

γ-谷氨酰胺化合物已经显示出它们作为药物的活性物质或前体的重要性。在这项研究中，开发了一种酶法合成γ-谷氨酰基化合物的方法，该方法使用了来自甲基甲虫的γ-谷氨酰甲基酰胺合成酶（GMAS）和来自谷氨酸棒杆菌的多磷酸激酶（PPK）。GMAS和PPK在pETDuet-1质粒中共重组并在大肠杆菌BL21（DE3）中共表达，并分别研究了GMAS和PPK的酶学性质。下的共表达系统的催化，升当使用谷氨酸钠和乙胺的底物摩尔比（1：1.4）和仅使用2 mM ATP时，β-茶氨酸以89.8％的转化率合成。通过一锅法合成了14种γ-谷氨酰基化合物，并通过阳离子交换树脂和等电点结晶进行纯化，收率范围为22.3％至72.7％。该研究为GMAS和PPK共表达系统合成γ-谷氨酰胺化合物提供了有效途径。