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| Construction and Optimization of p-coumaric Acid Producing Saccharomyces cerevisiae |
| ZHANG Wei1,2, LIU Duo1,2, LI Bing-zhi1,2, YUAN Ying-jin1,2 |
1. Department of Pharmaceutical Engineering, School of Chemical Engineering and Technology, Tianjin University;Key Laboratory of System Bioengineering(Ministry of Education), Tianjin 300072, China;
2. SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering(Tianjin), School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China |
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Abstract p-coumaric acid is widely used in bio-pharmaceutical, cosmetics and food industry as a very important precursor compound of phenylpropanoids, stilbenes and flavonoids. Saccharomyces cerevisiae was used as the host cell to synthesize p-coumaric acid with the methods of synthetic biology. It was demonstrated that the multi-copies plasmid harbored TAL showed a larger accumulation of p-coumaric acid; meanwhile, to eliminate the feedback inhibition of L-tyrosine, specific amino acid mutation Aro4pK229L and Aro7pG141S was obtained. The relevant mutated gene was integrated into yeast genome using delta site integration. 24 strains were picked out to verify the production of p-coumaric acid, the different yield between the highest strain and the lowest strain was 28.87mg/L. To strength the metabolic flux to p-coumaric, gene ARO10 and PDC5 which involved in the biosynthesis of byproduct aromatic alcohols were knocked out. Production of p-coumaric acid in strain with two gene knock out was improved to 87.56mg/L, 2.05-fold to the control one. Furthermore, when 1mmol/L L-tyrosine was added, the production of p-coumaric acid arrived the peak, about (174.57±0.30)mg/L. The p-coumaric acid over-producing S. cerevisiae using the synthetic biology method as well as lay a foundation of the biosynthesis of subsequent stilbenes and flavonoids has been successfuly constructed.
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Received: 08 March 2017
Published: 25 September 2017
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