Optimization of the l -tyrosine metabolic pathway in Saccharomyces cerevisiae by analyzing p -coumaric acid production
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ORIGINAL ARTICLE
Optimization of the l‑tyrosine metabolic pathway in Saccharomyces cerevisiae by analyzing p‑coumaric acid production Yuanzi Li1 · Jiwei Mao2 · Xiaofei Song1 · Yuzhen Wu1 · Miao Cai1 · Hesuiyuan Wang1 · Quanli Liu2 · Xiuming Zhang1 · Yanling Bai1 · Haijin Xu1 · Mingqiang Qiao1 Received: 31 July 2019 / Accepted: 24 April 2020 / Published online: 18 May 2020 © King Abdulaziz City for Science and Technology 2020
Abstract 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 ARO4K229L and ARO7G141S, 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, ARO4K229L, ARO7G141S, 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. Keywords l-Tyrosine · Metabolic engineering · p-Coumaric acid · Saccharomyces cerevisiae
Introduction The plant secondary metabolites flavonoids, stilbenoids and alkaloids have attracted increasing attention due to their pharmaceutical and nutritional applications (Akinwumi et al. 2018; Chougule et al. 2011; Yao et al. 2004). They are Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02223-3) contains supplementary material, which is available to authorized users. * Haijin Xu [email protected] * Mingqiang Qiao [email protected] 1
The Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, College of Life Sciences, Nankai University, No. 94 Weijin Road, Nankai District, Tianjin 300071, People’s Republic of China
Department of Biology and Biological Engineering, Chalmers University of Technology, 41296 Gothenburg, Sweden
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mainly obtained by extraction from plants and the extraction is energy intensive, inefficient and not environmentally friendly (Donnez et al. 2009; Sato et al. 2007; Silva et al. 2017). Heterologous biosynthesis in engineered microbes may be a good choice to achieve low consumption of energy and high yield of t
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