Optimization of the l-tyrosine metabolic pathway in Saccharomyces cerevisiae by analyzing p-coumaric acid production

• Published in: 3 Biotech Vol. 10; no. 6; p. 258
• Main Authors: Li, Yuanzi, Mao, Jiwei, Song, Xiaofei, Wu, Yuzhen, Cai, Miao, Wang, Hesuiyuan, Liu, Quanli, Zhang, Xiuming, Bai, Yanling, Xu, Haijin, Qiao, Mingqiang
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.06.2020; Springer Nature B.V
• Subjects: Acid production; Acids; Agriculture; Ammonia; Bioinformatics; Biomaterials; Biotechnology; Cancer Research; Chemistry; Chemistry and Materials Science; Chorismate mutase; Coumaric acid; Feedback; Feedback inhibition; Food irradiation; L-Tyrosine; Metabolic engineering; Metabolic pathways; Metabolism; Optimization; Original; Original Article; p-Coumaric acid; Phenylpyruvate decarboxylase; Phosphopyruvate hydratase; Pyruvate decarboxylase; Pyruvic acid; Saccharomyces cerevisiae; Stem Cells; Transaldolase; Tyrosine; Tyrosine; Coumaric acid; Metabolic engineering; p-Coumaric acid; l-Tyrosine; Saccharomyces cerevisiae
• ISSN: 2190-572X; 2190-5738; 2190-5738
• DOI: 10.1007/s13205-020-02223-3

In this study, we applied a series of genetic modifications to wild-type S. cerevisiae strain BY4741 to address the bottlenecks in the l -tyrosine pathway. A tyrosine ammonia-lyase (TAL) gene from Rhodobacter capsulatus , which can catalyze conversion of l -tyrosine into p -coumaric acid, was overexpressed to facilitate the analysis of l -tyrosine and test the strain’s capability to synthesize heterologous derivatives. First, we enhanced the supply of precursors by overexpressing transaldolase gene TAL1 , enolase II gene ENO2 , and pentafunctional enzyme gene ARO1 resulting in a 1.55-fold increase in p -coumaric acid production. Second, feedback inhibition of 3-deoxy- d -arabino-heptulosonate-7-phosphate synthase and chorismate mutase was relieved by overexpressing the mutated feedback-resistant ARO4 K229L and ARO7 G141S , and a 3.61-fold improvement of p -coumaric acid production was obtained. Finally, formation of byproducts was decreased by deleting pyruvate decarboxylase gene PDC5 and phenylpyruvate decarboxylase gene ARO10 , and p -coumaric acid production was increased 2.52-fold. The best producer—when TAL1 , ENO2 , ARO1 , ARO4 K229L , ARO7 G141S , and TAL were overexpressed, and PDC5 and ARO10 were deleted—increased p -coumaric acid production by 14.08-fold (from 1.4 to 19.71 mg L −1 ). Our study provided a valuable insight into the optimization of l -tyrosine metabolic pathway.