Yeast has been an indispensable host for synthesizing complex plant-derived naturalcompounds, yet the yields remained largely constrained. This limitation mainly arises from overlookingthe importance of cell and pathw...Yeast has been an indispensable host for synthesizing complex plant-derived naturalcompounds, yet the yields remained largely constrained. This limitation mainly arises from overlookingthe importance of cell and pathway suitability during the optimization of enzymes and pathways. Herein,beyond conventional enzyme engineering, we dissected metabolic suitability with a framework forsimultaneously augmenting cofactors and carbon flux to enhance the biosynthesis of heterogenoustriterpenoids. We further developed phospholipid microenvironment engineering strategies, dramaticallyimproving yeast’s suitability for the high performance of endoplasmic reticulum (ER)-localized, ratelimitingplant P450s. Combining metabolic and microenvironment suitability by manipulating only threegenes, NHMGR (NADH-dependent HMG-CoA reductase), SIP4 (a DNA-binding transcription factor)andGPP1 (Glycerol-1-phosphate phosphohydrolase 1), we enabled the high-level production of 4.92 g/L rarelicorice triterpenoids derived from consecutive oxidation of b-amyrin by two P450 enzymes afterfermentation optimization. This production holds substantial commercial value, highlighting the criticalrole of establishing cell suitability in enhancing triterpenoid biosynthesis and offering a versatileframework applicable to various plant natural product biosynthetic pathways.展开更多
基金grants from the National Key Research and Development Program of China(2021YFC2100800)the National Natural Science Foundation of China(No.22108154,No.22138006,No.32171430,No.22278240)+1 种基金the Natural Science Foundation of Beijing Municipality(M21010,China)the Shuimu Tsinghua Scholar Program(2020SM097,China).
文摘Yeast has been an indispensable host for synthesizing complex plant-derived naturalcompounds, yet the yields remained largely constrained. This limitation mainly arises from overlookingthe importance of cell and pathway suitability during the optimization of enzymes and pathways. Herein,beyond conventional enzyme engineering, we dissected metabolic suitability with a framework forsimultaneously augmenting cofactors and carbon flux to enhance the biosynthesis of heterogenoustriterpenoids. We further developed phospholipid microenvironment engineering strategies, dramaticallyimproving yeast’s suitability for the high performance of endoplasmic reticulum (ER)-localized, ratelimitingplant P450s. Combining metabolic and microenvironment suitability by manipulating only threegenes, NHMGR (NADH-dependent HMG-CoA reductase), SIP4 (a DNA-binding transcription factor)andGPP1 (Glycerol-1-phosphate phosphohydrolase 1), we enabled the high-level production of 4.92 g/L rarelicorice triterpenoids derived from consecutive oxidation of b-amyrin by two P450 enzymes afterfermentation optimization. This production holds substantial commercial value, highlighting the criticalrole of establishing cell suitability in enhancing triterpenoid biosynthesis and offering a versatileframework applicable to various plant natural product biosynthetic pathways.
