Production of hydroxycinnamoyl anthranilates from glucose in Escherichia coli

Microbial Cell Factories

Research

F William Collins¹  Edward E K Baidoo²  Darmawi Juminaga³  Aymerick Eudes⁴  Dominique Loqué⁴  Jay D Keasling⁵ 

[1] Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food, K1A 0C5, Ottawa, ON, Canada;Joint BioEnergy Institute, 94608, Emeryville, CA, USA;Joint BioEnergy Institute, 94608, Emeryville, CA, USA;California Institute for Quantitative Biosciences and the Synthetic Biology Institute at UC Berkeley, 94720, Berkeley, CA, USA;Joint BioEnergy Institute, 94608, Emeryville, CA, USA;Physical Biosciences Division, Lawrence Berkeley National Laboratory, 94720, Berkeley, CA, USA;Joint BioEnergy Institute, 94608, Emeryville, CA, USA;Physical Biosciences Division, Lawrence Berkeley National Laboratory, 94720, Berkeley, CA, USA;California Institute for Quantitative Biosciences and the Synthetic Biology Institute at UC Berkeley, 94720, Berkeley, CA, USA;Department of Bioengineering, Department of Chemical & Biomolecular Engineering, University of California, 94720, Berkeley, CA, USA;

关键词: Avenanthramide;    Tranilast;    BAHD;    Antioxidant;    Anti-inflammatory;    Tyrosine;    Anthranilate;    Hydroxycinnamate;    Biological synthesis;    Escherichia coli;   

DOI  :  10.1186/1475-2859-12-62

 received in 2013-05-25, accepted in 2013-06-18,  发布年份 2013

来源: Springer

【 摘 要 】

BackgroundOats contain hydroxycinnamoyl anthranilates, also named avenanthramides (Avn), which have beneficial health properties because of their antioxidant, anti-inflammatory, and antiproliferative effects. The microbial production of hydroxycinnamoyl anthranilates is an eco-friendly alternative to chemical synthesis or purification from plant sources. We recently demonstrated in yeast (Saccharomyces cerevisiae) that coexpression of 4-coumarate: CoA ligase (4CL) from Arabidopsis thaliana and hydroxycinnamoyl/benzoyl-CoA/anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT) from Dianthus caryophyllus enabled the biological production of several cinnamoyl anthranilates upon feeding with anthranilate and various cinnamates. Using engineering strategies to overproduce anthranilate and hydroxycinnamates, we describe here an entire pathway for the microbial synthesis of two Avns from glucose in Escherichia coli.ResultsWe first showed that coexpression of HCBT and Nt4CL1 from tobacco in the E. coli anthranilate-accumulating strain W3110 trpD9923 allowed the production of Avn D [N-(4′-hydroxycinnamoyl)-anthranilic acid] and Avn F [N-(3′,4′-dihydroxycinnamoyl)-anthranilic acid] upon feeding with p-coumarate and caffeate, respectively. Moreover, additional expression in this strain of a tyrosine ammonia-lyase from Rhodotorula glutinis (Rg TAL) led to the conversion of endogenous tyrosine into p-coumarate and resulted in the production of Avn D from glucose. Second, a 135-fold improvement in Avn D titer was achieved by boosting tyrosine production using two plasmids that express the eleven genes necessary for tyrosine synthesis from erythrose 4-phosphate and phosphoenolpyruvate. Finally, expression of either the p-coumarate 3-hydroxylase Sam5 from Saccharothrix espanensis or the hydroxylase complex HpaBC from E. coli resulted in the endogenous production of caffeate and biosynthesis of Avn F.ConclusionWe established a biosynthetic pathway for the microbial production of valuable hydroxycinnamoyl anthranilates from an inexpensive carbon source. The proposed pathway will serve as a platform for further engineering toward economical and sustainable bioproduction of these pharmaceuticals and other related aromatic compounds.

【 授权许可】

Unknown   
© Eudes et al.; licensee BioMed Central Ltd. 2013. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.