Journal
METABOLIC ENGINEERING
Volume 32, Issue -, Pages 30-38Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ymben.2015.09.003
Keywords
Phenazine-1-carboxylic acid; Phenazine biosynthetic pathway; Chorismate-utilizing enzymes; Secondary metabolism; Pseudomonas aeruginosa PA1201
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Funding
- National High Technology Research and Development Program (863Program) of China [2012AA022107]
- National Key Basic Research Program of China (973 Program) [2012CB721000]
- National Key Technology R D Program [2012BAD19B01]
- National Natural Science Foundation of China [31270084]
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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 overproduction 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 fermenation, 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. (C) 2015 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.
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