4.3 Article

Chromosomal editing of Corynebacterium glutamicum ATCC 13032 to produce gamma-aminobutyric acid

Journal

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
Volume 70, Issue 1, Pages 7-21

Publisher

WILEY
DOI: 10.1002/bab.2324

Keywords

Corynebacterium glutamicum; gamma-aminobutyric acid; glutamate dehydrogenase; glutamic acid decarboxylase; metabolic engineering

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A high GABA-producing strain of Corynebacterium glutamicum was constructed by chromosomal editing, without the need for antibiotics or inducers. The strategy of enhancing l-glutamic acid synthesis and optimizing the copy numbers of GABA-related genes resulted in increased GABA production.
Corynebacterium glutamicum has been used as a sustainable microbial producer for various bioproducts using cheap biomass resources. In this study, a high GABA-producing C. glutamicum strain was constructed by chromosomal editing. Lactobacillus brevis-derived gadB2 was introduced into the chromosome of C. glutamicum ATCC 13032 to produce gamma-aminobutyric acid and simultaneously blocked the biosynthesis of lactate and acetate. GABA transport and degradation in C. glutamicum were also blocked to improve GABA production. As precursor of GABA, l-glutamic acid synthesis in C. glutamicum was enhanced by introducing E. coli gdhA encoding glutamic dehydrogenase, and the copy numbers of gdhA and gadB2 were also optimized for higher GABA production. The final C. glutamicum strain CGY705 could produce 33.17 g/L GABA from glucose in a 2.4-L bioreactor after 78 h fed-batch fermentation. Since all deletion and expression of genes were performed using chromosomal editing, fermentation of the GABA-producing strains constructed in this study does not need supplementation of any antibiotics and inducers. The strategy used in this study has potential value in the development of GABA-producing bacteria.

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