文献类型：期刊文献

英文题名：Engineered ethanol-driven biosynthetic system for improving production of acetyl-CoA derived drugs in Crabtree-negative yeast

作者：Liu, Yiqi[1];Bai, Chenxiao[1];Liu, Qi[1];Xu, Qin[1];Qian, Zhilan[1];Peng, Qiangqiang[1];Yu, Jiahui[1];Xu, Mingqiang[1];Zhou, Xiangshan[1,2];Zhang, Yuanxing[1];Cai, Menghao[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：54

起止页码：275

外文期刊名：METABOLIC ENGINEERING

收录：;EI(收录号：20192006930218);WOS:【SCI-EXPANDED(收录号:WOS:000470680100025)】；

基金：This work was supported by the National Natural Science Foundation of China [grant number 31870073], Shanghai Rising-Star Program, China [grant number 19QA1402700], Fundamental Research Funds for the Shanghai Science and Technology Innovation Action Plan, China [grant number 17JC1402400], the 111 Project of China [grant number B18022], Fundamental Research Funds for the Central Universities, China [grant number 22221818014] and Research Program of State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Ethanol; Crabtree-negative yeast; Acetyl-CoA; Monacolin J; Metabolic engineering; Coculture

摘要：Many natural drugs use acetyl-CoA as the key biosynthetic precursor. While in eukaryotic chassis host like yeast, efficient biosynthesis of these drugs is often hampered by insufficient acetyl-CoA supply because of its compartmentalized metabolism. Reported acetyl-CoA engineering commonly modifies central carbon metabolism to pull and push acetyl-CoA into cytosol from sugars or redirects biosynthetic pathways in organelles, involving complicated metabolic engineering strategies. We constructed a new biosynthetic system based on a Crabtree-negative yeast, which grew exceptionally on ethanol and assimilated ethanol directly in cytosol to acetyl-CoA (3 steps). A glucose-repressed and ethanol-induced transcriptional signal amplification device (ESAD) with 20-fold signal increase was constructed by rewiring native transcriptional regulation circuits. This made ethanol the sole and fast-growing substrate, acetyl-CoA precursor, and strong biosynthetic pathway inducer simultaneously. The ESAD was used for biosynthesis of a commercial hypolipidemic drug intermediate, monacolin J. A strain producing dihydromonacolin L was firstly constructed and systematically engineered. We further developed a co-culture system equipped with this upstream strain and a downstream strain with dihydromonacolin L-to-monacolin J module controlled by a synthetic constitutive transcriptional signal amplification device (CSAD). It produced a high monacolin J titre of 2.2 g/L on ethanol in bioreactor. Engineering glucose-supported and ethanol-repressed fatty acids biosynthesis in the upstream strain contributed more acetyl-CoA for monacolin J and improved its titre to 3.2 g/L, far surpassing other reported productions in yeasts. This study provides a new paradigm for facilitating the high-yield production of acetyl-CoA derived pharmaceuticals and value-added molecules.