Systematic Engineering of Saccharomyces cerevisiae for the De Novo Biosynthesis of Genistein and Glycosylation Derivatives
文献类型： 外文期刊
作者： Wang, Yongtong 1 ; Xiao, Zhiqiang 1 ; Zhang, Siqi 1 ; Tan, Xinjia 1 ; Zhao, Yifei 1 ; Liu, Juan 1 ; Jiang, Ning 2 ; Shan, Yang 1 ;
作者机构： 1.Hunan Univ, Coll Biol, Longping Branch, Changsha 410125, Peoples R China
2.Hunan Acad Agr Sci, Agr Prod Proc Inst, Changsha 410125, Peoples R China
3.Hunan Agr Prod Proc Inst, Hunan Key Lab Fruits & Vegetables Storage Proc Qua, Changsha 410125, Peoples R China
4.Chalmers Univ Technol, Dept Life Sci, SE-41296 Gothenburg, Sweden
关键词： isoflavones; genistein; heme; malonyl-CoA; glycosylation
期刊名称：JOURNAL OF FUNGI （ 影响因子：4.7； 五年影响因子：5.2 ）
ISSN：
年卷期： 2024 年 10 卷 3 期
页码：
收录情况： SCI
摘要： Isoflavones are predominantly found in legumes and play roles in plant defense and prevention of estrogen-related diseases. Genistein is an important isoflavone backbone with various biological activities. In this paper, we describe how a cell factory that can de novo synthesize genistein was constructed in Saccharomyces cerevisiae. Different combinations of isoflavone synthase, cytochrome P450 reductase, and 2-hydroxyisoflavone dehydratase were tested, followed by pathway multicopy integration, to stably de novo synthesize genistein. The catalytic activity of isoflavone synthase was enhanced by heme supply and an increased intracellular NADPH/NADP(+) ratio. Redistribution of the malonyl-CoA flow and balance of metabolic fluxes were achieved by adjusting the fatty acid synthesis pathway, yielding 23.33 mg/L genistein. Finally, isoflavone glycosyltransferases were introduced into S. cerevisiae, and the optimized strain produced 15.80 mg/L of genistin or 10.03 mg/L of genistein-8-C-glucoside. This is the first de novo synthesis of genistein-8-C-glucoside in S. cerevisiae, which is advantageous for the green industrial production of isoflavone compounds.
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