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Disruption of genes for the enhanced biosynthesis of α-ketoglutarate in Corynebacterium glutamicum

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Abstract:

The development of microbial strains for the enhanced production of α-ketoglutarate (α-KG) was investigated using a strain of Corynebacterium glutamicum that overproduces of l-glutamate, by disrupting three genes involved in the α-KG biosynthetic pathway. The pathways competing with the biosynthesis of α-KG were blocked by knocking out aceA (encoding isocitrate lyase, ICL), gdh (encoding glutamate dehydrogenase, l-gluDH), and gltB (encoding glutamate synthase or glutamate-2-oxoglutarate aminotransferase, GOGAT). The strain with aceA, gltB, and gdh disrupted showed reduced ICL activity and no GOGAT and l-gluDH activities, resulting in up to 16-fold more α-KG production than the control strain in flask culture. These results suggest that l-gluDH is the key enzyme in the conversion of α-KG to l-glutamate; therefore, prevention of this step could promote α-KG accumulation. The inactivation of ICL leads the carbon flow to α-KG by blocking the glyoxylate pathway. However, the disruption of gltB did not affect the biosynthesis of α-KG. Our results can be applied in the industrial production of α-KG by using C. glutamicum as producer.

Keywords: Corynebacterium glutamicum; biosynthesis; biosynthèse; gene disruption; interruption génique; α-cétoglutarate; α-ketoglutarate

Document Type: Research Article

DOI: https://doi.org/10.1139/w11-132

Affiliations: 1: Department of Bioscience and Biotechnology, Hankuk University of Foreign Studies, Yongin, Gyeonggi-Do, 449-791, Korea. 2: MH2 Biochemical, Eumseong, Chungcheongbuk-Do, 396-841, Korea.

Publication date: 2012-03-17

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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