Metabolic engineering of fast-growing Vibrio natriegens for efficient pyruvate production

Pyruvate is a widely used value-added chemical which also serves as a hub of various metabolic pathways. The fastest-growing bacterium Vibrio natriegens is a promising chassis for synthetic biology applications with high substrate uptake rates. The aim of this study was to investigate if the high su...

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Bibliographic Details
Published in:Microbial cell factories 2023-09, Vol.22 (1), p.1-172, Article 172
Main Authors: Wu, Fengli, Wang, Shucai, Peng, Yanfeng, Guo, Yufeng, Wang, Qinhong
Format: Article
Language:English
Subjects:
Bacteria
Biosynthesis
Carbon
Cell growth
Dehydrogenases
E coli
Engineering
Ethanol
Fermentation
Fine-tuning gene expression
Gene clusters
Gene expression
Genes
Genomes
Glucose
Kinases
Metabolic engineering
Metabolic pathways
Metabolism
Metabolites
Pathway engineering
Phosphoenolpyruvate carboxylase
Ppc gene
Productivity
Pyruvate
Pyruvate dehydrogenase (lipoamide)
Pyruvic acid
Substrates
Sucrose
Synthetic biology
Tricarboxylic acid cycle
Vibrio natriegens
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Summary:Pyruvate is a widely used value-added chemical which also serves as a hub of various metabolic pathways. The fastest-growing bacterium Vibrio natriegens is a promising chassis for synthetic biology applications with high substrate uptake rates. The aim of this study was to investigate if the high substrate uptake rates of V. natriegens enable pyruvate production at high productivities. Two prophage gene clusters and several essential genes for the biosynthesis of byproducts were first deleted. In order to promote pyruvate accumulation, the key gene aceE encoding pyruvate dehydrogenase complex E1 component was down-regulated to reduce the carbon flux into the tricarboxylic acid cycle. Afterwards, the expression of ppc gene encoding phosphoenolpyruvate carboxylase was fine-tuned to balance the cell growth and pyruvate synthesis. The resulting strain PYR32 was able to produce 54.22 g/L pyruvate from glucose within 16 h, with a yield of 1.17 mol/mol and an average productivity of 3.39 g/L/h. In addition, this strain was also able to efficiently convert sucrose or gluconate into pyruvate at high titers. A novel strain of V. natriegens was engineered which was capable to provide higher productivity in pyruvate synthesis. This study lays the foundation for the biosynthesis of pyruvate and its derivatives in fast-growing V. natriegens.
ISSN:1475-2859
1475-2859
DOI:10.1186/s12934-023-02185-0