文献类型：期刊文献

英文题名：Complete Biosynthesis of Anthocyanins Using E. coli Polycultures

作者：Jones, J. Andrew[1,4];Vernacchio, Victoria R.[1];Collins, Shannon M.[1];Shirke, Abhijit N.[3];Xiu, Yu[1,5,6];Englaender, Jacob A.[2];Cress, Brady F.[1];McCutcheon, Catherine C.[4];Linhardt, Robert J.[1,2,3];Gross, Richard A.[3];Koffas, Mattheos A. G.[1,2]

第一作者：Jones, J. Andrew

通讯作者：Koffas, MAG[1];Koffas, MAG[2]

机构：[1]Rensselaer Polytech Inst, Dept Chem & Biol Engn, Troy, NY 12180 USA;[2]Rensselaer Polytech Inst, Dept Biol Sci, Troy, NY 12180 USA;[3]Rensselaer Polytech Inst, Dept Chem, Troy, NY USA;[4]Hamilton Coll, Dept Chem, Clinton, NY 13323 USA;[5]Beijing Univ Chem Technol, State Key Lab Chem Resource Engn, Beijing, Peoples R China;[6]Beijing Union Univ, Beijing Key Lab Bioact Subst & Funct Food, Beijing, Peoples R China

第一机构：Rensselaer Polytech Inst, Dept Chem & Biol Engn, Troy, NY 12180 USA

通讯机构：[1]corresponding author), Rensselaer Polytech Inst, Dept Chem & Biol Engn, Troy, NY 12180 USA;[2]corresponding author), Rensselaer Polytech Inst, Dept Biol Sci, Troy, NY 12180 USA.

年份：2017

卷号：8

期号：3

外文期刊名：MBIO

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000404733300046)】；

语种：英文

外文关键词：Escherichia coli; anthocyanins; coculture; de novo; flavonoids; pelargonidin 3-O-glucoside; polyculture; recombinant production

摘要：Fermentation-based chemical production strategies provide a feasible route for the rapid, safe, and sustainable production of a wide variety of important chemical products, ranging from fuels to pharmaceuticals. These strategies have yet to find wide industrial utilization due to their inability to economically compete with traditional extraction and chemical production methods. Here, we engineer for the first time the complex microbial biosynthesis of an anthocyanin plant natural product, starting from sugar. This was accomplished through the development of a synthetic, 4-strain Escherichia coli polyculture collectively expressing 15 exogenous or modified pathway enzymes from diverse plants and other microbes. This synthetic consortium-based approach enables the functional expression and connection of lengthy pathways while effectively managing the accompanying metabolic burden. The de novo production of specific anthocyanin molecules, such as calistephin, has been an elusive metabolic engineering target for over a decade. The utilization of our polyculture strategy affords milligram-per-liter production titers. This study also lays the groundwork for significant advances in strain and process design toward the development of cost-competitive biochemical production hosts through nontraditional methodologies. IMPORTANCE To efficiently express active extensive recombinant pathways with high flux in microbial hosts requires careful balance and allocation of metabolic resources such as ATP, reducing equivalents, and malonyl coenzyme A (malonyl-CoA), as well as various other pathway-dependent cofactors and precursors. To address this issue, we report the design, characterization, and implementation of the first synthetic 4-strain polyculture. Division of the overexpression of 15 enzymes and transcription factors over 4 independent strain modules allowed for the division of metabolic burden and for independent strain optimization for module-specific metabolite needs. This study represents the most complex synthetic consortia constructed to date for metabolic engineering applications and provides a new paradigm in metabolic engineering for the reconstitution of extensive metabolic pathways in nonnative hosts.