文献类型：期刊文献

英文题名：Engineered dynamic distribution of malonyl-CoA flux for improving polyketide biosynthesis in Komagataella phaffii

作者：Wen, Jiao[1];Tian, Lin[1];Liu, Qi[1];Zhang, Yuanxing[1,2];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：2020

卷号：320

起止页码：80

外文期刊名：JOURNAL OF BIOTECHNOLOGY

收录：;EI(收录号：20202708884009);WOS:【SCI-EXPANDED(收录号:WOS:000548643800012)】；

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

语种：英文

外文关键词：Komagataella phaffii; FapR/fapO; Malonyl-CoA; Dynamic control; Synthetic biology

摘要：Malonyl-CoA is a basic but limited precursor for the biosynthesis of various bioactive compounds and life-supporting fatty acids in cells. This study develops a biosynthetic system to dynamically redirect malonyl-CoA flux and improve production of malonyl-CoA derived polyketide (6-MSA) in Komagataella phaffii. A synthetic regulatory protein fusing a yeast activator Prm1 with a bacterial repressor FapR was proved to work with a hybrid promoter (-7)fapO-cP(AOX1) and activate gene expression. Expression mode by the Prm1-FapR/(-7)fapO-cP(AOX1) device was not affected by intracellular malonyl-CoA levels. Further, 9 promoter variants of P-GAP with insertion of fapO at various sites were tested with the Prm1-FapR. It generated a biosensor of Prm1-FapR/P-GAP-(+2)fapO with regulation behavior of malonyl-CoA-low-level repression/high-level derepression. Both devices were subsequently integrated into a single cell, for which fatty acid synthesis module was driven by Prm1-FapR/ P-GAP-(+2)fapO but 6-MSA synthesis module was expressed by Prm1-FapR/(-7)fapO-cP(AOX1). The integrated system allowed continuous polyketide synthesis but malonyl-CoA-high-level "on"/low-level "off" fatty acid synthesis. This design finally increased 6-MSA production capacity by 260 %, proving the positive effects of dynamic malonyl-CoA distribution to the target compounds. It provides a new strategy for synthesis of malonyl-CoA derived compounds in eukaryotic chassis hosts.