酿酒酵母过氧化物酶体脂肪酸代谢调控高效合成脂肪醇

Microbial synthesis is an alternative and important route toward the sustainable production of chemicals and liquid biofuels. However, the complex of cellular metabolism make it challenge to rational optimize the biosynthetic pathways. Especially, the eukaryotic microbial metabolism is functionally distributed in different sub-organelles, which limited the foreseeability of biosynthetic pathway engineering, and thus requires to develop more rational and controllable regulation strategy. This project aims to engineering the model eukaryotic microbe Saccharomyces cerevisiae for production high-value oleo-chemical fatty alcohol by harnessing the peroxisome, a sub-organelle with active fatty acid metabolism. We will investigate the dynamic regulation of peroxisome biogenesis and its fatty acid metabolism by using external fatty acid and H2O2 stresses, and reveal the responsive genes and its regulation elements by using transcriptomic analysis. With the learned knowledge and characterized genetic elements, we will construct and optimize dynamic and controllable metabolic pathways, smartly responding to the fatty acid metabolism and external stresses, for enhancing cell robustness and the production of fatty alcohols. This study will reveal dynamic spatiotemporal fatty acid metabolism in peroxisome and also its adaptation mechanism with downstream oleo-chemical biosynthetic pathways, and develop the spatiotemporal metabolic engineering strategy for improving biosynthesis. The resulting data and knowledge will provide theoretical and technical basis for bio-manufacturing of oleo-chemicals and other chemicals.

微生物合成是实现化学品及液体燃料可持续生产的重要路径。然而，细胞的代谢复杂性给代谢途径优化带来挑战，特别是真核微生物中代谢网络动态分布于多个亚细胞器中，制约了生物合成途径优化的可预见性。因而迫切需要发展更理性、可控的代谢调控策略。本项目以油脂化学品脂肪醇高效生物合成为目标，以真核模式微生物酿酒酵母（Saccharomyces cerevisiae）为宿主，理性改造脂肪酸代谢活跃的亚细胞器-过氧化物酶体提高脂肪醇合成效率。通过脂肪酸与过氧化氢（H2O2）胁迫，探索过氧化物酶体脂肪酸代谢动态分布规律；通过转录组学挖掘响应脂肪酸与H2O2胁迫的基因元件；利用相关元件设计适配脂肪酸代谢以及响应外界刺激的智能生物合成途径，提高脂肪醇生物合成效率及细胞性能。研究成果将揭示过氧化物酶体脂肪酸代谢动态调控规律及其与下游生物合成途径适配规律，发展新颖时空代谢调控策略，为化学品高效生物合成提供理论及技术支持。

本项目以油脂化学品脂肪醇高效生物合成为目标，以真核模式微生物酿酒酵母（Saccharomyces cerevisiae）为宿主，理性改造脂肪酸代谢活跃的亚细胞器-过氧化物酶体，建立了时空代谢调控策略提高脂肪醇合成效率。通过转录组学挖掘响应脂肪酸的基因元件并利用相关元件设计适配脂肪酸代谢以及响应外界刺激的智能生物合成途径，提高脂肪醇生物合成效率及细胞性能；通过脂肪酸与过氧化氢（H2O2）胁迫，进一步提高了脂肪酸生物合成效率。项目系统探索了过氧化物酶体和胞浆脂肪酸代谢规律，建立了时空代谢调控策略，为脂肪酸衍生物生物合成提供了指导。特别是鉴定了响应脂肪酸与酯酰-CoA的启动子库，为构建酵母细胞内时空平衡生物合成途径提供了可靠元件。本项目设计的时空调控策略实现了脂肪醇代谢与细胞初级代谢耦合，有望用于其他产物高效合成。