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Metabolic engineering v.44, 2017년, pp.89 - 99   SCIE
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Engineering a bacterial platform for total biosynthesis of caffeic acid derived phenethyl esters and amides

Wang, Jian (College of Engineering, The University of Georgia, Athens, GA 30602, USA ) ; Mahajani, Monika (BiotecEra Inc., 220 Riverbend Rd., Athens, GA 30602, USA ) ; Jackson, Sheneika L. (Department of Chemistry, The University of Georgia, Athens, GA 30602, USA ) ; Yang, Yaping (College of Engineering, The University of Georgia, Athens, GA 30602, USA ) ; Chen, Mengyin (BiotecEra Inc., 220 Riverbend Rd., Athens, GA 30602, USA ) ; Ferreira, Eric M. (Department of Chemistry, The University of Georgia, Athens, GA 30602, USA ) ; Lin, Yuheng (BiotecEra Inc., 220 Riverbend Rd., Athens, GA 30602, USA ) ; Yan, Yajun (College of Engineering, The University of Georgia, Athens, GA 30602, USA ) ;
  • 초록  

    Abstract Caffeic acid has been widely recognized as a versatile pharmacophore for synthesis of new chemical entities, among which caffeic acid derived phenethyl esters and amides are the most extensively-investigated bioactive compounds with potential therapeutical applications. However, the natural biosynthetic routes for caffeic acid derived phenethyl esters or amides remain enigmatic, limiting their bio-based production. Herein, product-directed design of biosynthetic schemes allowed the development of thermodynamically favorable pathways for these compounds via acyltransferase (ATF) mediated trans-esterification. Production based screening identified a microbial O-ATF from Saccharomyces cerevisiae and a plant N-ATF from Capsicum annuum capable of forming caffeic acid derived esters and amides, respectively. Subsequent combinatorial incorporation of caffeic acid with various aromatic alcohol or amine biosynthetic pathways permitted the de novo bacterial production of a panel of caffeic acid derived phenethyl esters or amides in Escherichia coli for the first time. Particularly, host strain engineering via systematic knocking out endogenous caffeoyl-CoA degrading thioesterase and pathway optimization via titrating co-substrates enabled production enhancement of five caffeic acid derived phenethyl esters and amides, with titers ranging from 9.2 to 369.1mg/L. This platform expanded the capabilities of bacterial production of high-value natural aromatic esters and amides from renewable carbon source via tailoring non-natural biosynthetic pathways. Highlights Acyltransferases for synthesis of caffeic acid esters and amides were identified. Caffeic acid phenethyl esters and amides were produced from glucose in E. coli . Production improvement was made by up to 3-fold via host and pathway optimization.


  • 주제어

    Caffeic acid .   Esters and amides .   Acyltransferase .   Host engineering .   Promoter engineering.  

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