文献类型：期刊文献

英文题名：Metabolic Engineering of Escherichia coli for De Novo Biosynthesis of the Platform Chemical Pelletierine

作者：Li, Wei[1];Zhao, Peng[2];Li, Ying[3];Wu, Shimin[1];Tian, Pingfang[1]

第一作者：Li, Wei

通讯作者：Tian, PF[1]

机构：[1]Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing Key Lab Bioproc, Beijing 100029, Peoples R China;[2]Beijing Univ Agr, Coll Biosci & Resources Environm, Beijing 102206, Peoples R China;[3]Beijing Union Univ, Coll Biochem Engn, Beijing 100023, Peoples R China

第一机构：Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing Key Lab Bioproc, Beijing 100029, Peoples R China

通讯机构：[1]corresponding author), Beijing Univ Chem Technol, Coll Life Sci & Technol, Beijing Key Lab Bioproc, Beijing 100029, Peoples R China.

年份：2025

卷号：13

期号：2

起止页码：778-790

外文期刊名：ACS SUSTAINABLE CHEMISTRY & ENGINEERING

收录：;WOS:【SCI-EXPANDED(收录号:WOS:001393345900001)】；

基金：This study was funded by grants from the National Key Research and Development Program of China (2018YFA0901800 and 2023YFA0914700) and National Natural Science Foundation of China (22278022).

语种：英文

外文关键词：huperzine A; pelletierine; Alzheimer'sdisease; Escherichia coli; metabolic engineering

摘要：Pelletierine is a versatile plant alkaloid having a C5N-C3 structure from which numerous chemicals can be derived. One notable derivative is huperzine A (HupA) which may alleviate the symptoms of Alzheimer's disease. Currently, industrial production of pelletierine relies primarily on chemical synthesis and plant extraction. However, chemical synthesis leads to analogues that complicate product separation, and plant extraction is constrained by limited resources. Herein, we report that pelletierine can be produced by recombinant Escherichia coli in which the engineered pelletierine biosynthesis pathway comprises four modules involving seven key genes native to E. coli, three genes from other bacteria, and three genes from plants. To overproduce pelletierine, the intrinsic l-lysine biosynthesis pathway in E. coli was simplified, and a clustered regularly interspaced short palindromic repeats (CRISPR) interference (CRISPRi) system was engineered to minimize the byproducts. Moreover, the transporter MatC was overexpressed to enhance the intracellular concentration of 3-oxoglutaryl ketide, which is another precursor of pelletierine. Based on the aforementioned manipulations, the resulting recombinant E. coli harboring the pelletierine biosynthesis pathway and CRISPRi system produced 3.40 and 8.23 mg/L pelletierine in a shake-flask and a 5 L bioreactor, respectively. This is the first report of microbial production of pelletierine, which represents a sustainable route to produce the precursor of HupA and beyond.