Reprogramming the Metabolism of Yeast for High-Level Production of Miltiradiene

作者：Xue Bai, Shuling Wang, Qin Zhang, Yuhan Hu, Jiawei Zhou, Lianhui Men, Dengyu Li, Jing Ma, Qiuhui Wei, Mengdie Xu, Xiaopu Yin, Tianyuan Hu · 发表于：Journal of Agricultural and Food Chemistry · 年份：2024 · DOI：10.1021/acs.jafc.4c01203 · 被引用次数：14 · 研究领域：Plant biochemistry and biosynthesis、Microbial Natural Products and Biosynthesis、Microbial Metabolic Engineering and Bioproduction

Miltiradiene serves as a crucial precursor in the synthesis of various high-value abietane-type diterpenes, exhibiting diverse pharmacological activities. Previous efforts to enhance miltiradiene production have primarily focused on the mevalonate acetate (MVA) pathway. However, limited emphasis has been placed on optimizing the supply of acetyl-CoA and NADPH. In this study, we constructed a platform yeast strain for miltiradiene production by reinforcing the biosynthetic pathway of geranylgeranyl diphosphate (GGPP) and acetyl-CoA, and addressing the imbalance between the supply and demand of the redox cofactor NADPH within the cytoplasm, resulting in an increase in miltiradiene yield to 1.31 g/L. Furthermore, we conducted modifications to the miltiradiene synthase fusion protein t Sm KSL1- Cf TPS1. Finally, the comprehensive engineering strategies and protein modification strategies culminated in 1.43 g/L miltiradiene in the engineered yeast under shake flask culture conditions. Overall, our work established efficient yeast cell factories for miltiradiene production, providing a foothold for heterologous biosynthesis of abietane-type diterpenes.