title: Engineering the central biosynthetic and secondary metabolic pathways of Pseudomonas aeruginosa strain PA1201 to improve phenazine-1-carboxylic acid production.
abstract: The secondary metabolite phenazine-1-carboxylic acid (PCA) is an important component of the newly registered biopesticide Shenqinmycin. We used a combined method involving gene, promoter, and protein engineering to modify the central biosynthetic and secondary metabolic pathways in the PCA-producing Pseudomonas aeruginosa strain PA1201. The PCA yield of the resulting strain PA-IV was increased 54.6-fold via the following strategies: (1) blocking PCA conversion and enhancing PCA efflux pumping; (2) increasing metabolic flux towards the PCA biosynthetic pathway through the over-production of two DAHP synthases and blocking the synthesis of 21 secondary metabolites; (3) increasing the PCA precursor supply through the engineering of five chorismate-utilizing enzymes; (4) engineering the promoters of two PCA biosynthetic gene clusters. Strain PA-IV produced 9882 mg/L PCA in fed-batch fermentation, which is twice as much as that produced by the current industrial strain. Strain PA-IV was also genetically stable and comparable to Escherichia coli in cytotoxicity.
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Kaiming
authors.initials: K
authors.lastname: Jin
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Lian
authors.initials: L
authors.lastname: Zhou
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Haixia
authors.initials: H
authors.lastname: Jiang
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Sun
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Yunling
authors.initials: Y
authors.lastname: Fang
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
authors.forename: Xuehong
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Microbial Metabolism, School of Life Sciences & Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address: yawenhe@sjtu.edu.cn.
authors.forename: Ya-Wen
authors.initials: YW
authors.lastname: He
mesh_terms: D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D053898:Biosynthetic Pathways / Q000235:genetics*; D002826:Chorismate Mutase / Q000235:genetics / Q000378:metabolism; D003470:Culture Media; D004331:Drosophila melanogaster; D005285:Fermentation; D060847:Metabolic Engineering / Q000379:methods*; D053858:Metabolic Networks and Pathways / Q000235:genetics*; D005810:Multigene Family / Q000235:genetics; D009154:Mutation / Q000235:genetics; D010619:Phenazines / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism* / Q000472:pathogenicity
chemical_list: D003470:Culture Media; D010619:Phenazines; C037165:1-phenazinecarboxylic acid; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D002826:Chorismate Mutase
keywords: Chorismate-utilizing enzymes; Phenazine biosynthetic pathway; Phenazine-1-carboxylic acid; Pseudomonas aeruginosa PA1201; Secondary metabolism