Wang Yongtong, Xiao Zhiqiang, Zhang Siqi, Tan Xinjia, Zhao Yifei, Liu Juan, Jiang Ning, Shan Yang
Longping Branch, College of Biology, Hunan University, Changsha 410125, China.
Agriculture Product Processing Institute, Hunan Academy of Agricultural Sciences, Changsha 410125, China.
J Fungi (Basel). 2024 Feb 26;10(3):176. doi: 10.3390/jof10030176.
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 . 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 , and the optimized strain produced 15.80 mg/L of genistin or 10.03 mg/L of genistein-8--glucoside. This is the first de novo synthesis of genistein-8--glucoside in , which is advantageous for the green industrial production of isoflavone compounds.
异黄酮主要存在于豆类中，在植物防御和预防雌激素相关疾病中发挥作用。染料木黄酮是一种具有多种生物活性的重要异黄酮骨架。在本文中，我们描述了如何在……构建一个能够从头合成染料木黄酮的细胞工厂。测试了异黄酮合酶、细胞色素P450还原酶和2-羟基异黄酮脱水酶的不同组合，随后进行途径多拷贝整合，以稳定地从头合成染料木黄酮。通过血红素供应和增加细胞内NADPH/NADP比率提高了异黄酮合酶的催化活性。通过调整脂肪酸合成途径实现了丙二酰辅酶A流的重新分配和代谢通量的平衡，产生了23.33mg/L的染料木黄酮。最后，将异黄酮糖基转移酶引入……，优化后的菌株产生了15.80mg/L的染料木苷或10.03mg/L的染料木黄酮-8-O-葡萄糖苷。这是在……中首次从头合成染料木黄酮-8-O-葡萄糖苷，这有利于异黄酮化合物的绿色工业化生产。
