The dipeptide γ‐glutamylcysteine (γ‐GC), the first intermediate of glutathione (GSH) synthesis, is considered as a promising drug to reduce or prevent plethora of age‐related disorders such as Alzheimer and Parkinson diseases. The unusual γ‐linkage between the two constitutive amino acids, namely cysteine and glutamate, renders its chemical synthesis particularly challenging. Herein, we report on the metabolic engineering of the non‐conventional yeast Yarrowia lipolytica for efficient γ‐GC synthesis. The yeast was first converted into a γ‐GC producer by disruption of gene GSH2 encoding GSH synthase and by constitutive expression of GSH1 encoding glutamylcysteine ligase. Subsequently genes involved in cysteine and glutamate anabolism, namely MET4, CYSE, CYSF, and GDH1 were overexpressed with the aim to increase their intracellular availability. With such a strategy, a γ‐GC titer of 464 nmol mg−1 protein (93 mg gDCW−1) was obtained within 24 h of cell growth.

中文翻译：

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利用工程解脂耶氏酵母生物合成抗氧化剂 γ-谷氨酰-半胱氨酸

二肽γ-谷氨酰半胱氨酸（γ-GC）是谷胱甘肽（GSH）合成的第一个中间体，被认为是一种有前途的药物，可以减少或预防多种与年龄相关的疾病，例如阿尔茨海默病和帕金森病。两种组成氨基酸（即半胱氨酸和谷氨酸）之间不寻常的 γ 连接使其化学合成特别具有挑战性。在此，我们报告了非常规酵母的代谢工程解脂耶氏酵母高效的γ-GC合成。酵母首先通过基因破坏转化为 γ-GC 生产者谷胱甘肽2编码 GSH 合酶并通过组成型表达还原型谷胱甘肽1编码谷氨酰半胱氨酸连接酶。随后涉及半胱氨酸和谷氨酸合成代谢的基因，即MET4,CYSE,CYSF， 和GDH1被过度表达，目的是增加它们的细胞内可用性。采用这种策略，γ-GC 滴度为 464 nmol mg−1蛋白质（93 毫克 gDCW−1）是在细胞生长24小时内获得的。