工程酵母菌发酵过程中因氧化胁迫、蛋白质错误折叠等压力致使生产能力降低。该研究发现,在甲羟戊酸途径增强的酵母工程菌中,过表达人源细胞凋亡调控因子B细胞淋巴瘤/白血病-2(B-cell lymphoma-2,BCL-2)和还原型谷胱甘肽合成酶(glutathione synthase,GSH1)编码基因可以促进橙花叔醇的合成,橙花叔醇摇瓶产量分别提高77.7%和32.7%,达到594.1和446.9 mg/L。对表达和未表达BCL-2蛋白的菌株进行代谢组学分析,发现了182种差异代谢物,主要涉及脂肪酸代谢、氨基酸代谢以及辅酶A和泛酸的生物合成等。此外,发现连接有内质网定位信号肽的BCL-2蛋白对工程菌产橙花叔醇的促进作用更强,橙花叔醇摇瓶产量进一步提高了29.2%,达到767.6 mg/L。该研究为提高外源萜类化合物在工程酿酒酵母中的产量提供了一个有效的策略。
Production capacity of engineered yeast decreases due to some pressures, such as oxidative stress and protein misfolding. Results of this study showed that, the engineered yeast with enhanced mevalonate pathway, overexpression of human apoptosis regulator BCL-2 and reduced glutathione synthase GSH1 promoted the production of nerolidol by 77.7% and 32.7% in shake flasks, reached 594.1 and 446.9 mg/L, respectively. Metabolome analysis between strains with and without BCL-2 overexpression showed that there were 182 differential metabolites between these two strains. It mainly involved the metabolic pathways of fatty acids and amino acids, and biosynthetic pathways of coenzyme A and pantothenic acid. In addition, BCL-2 protein linked with endoplasmic reticulum localization signal peptide had a stronger promoting effect on nerolidol production, which was further increased by 29.2%, reaching 767.6 mg/L. This study provides an effective strategy for increasing the yield of exogenous terpenoids in engineered Saccharomyces cerevisiae.
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