Microorganism having improved ability to produce N-acetylglucosamine as a result of modulating glycolytic flux
AssigneeKorea University Research and Business Foundation
InventorsMin-Kyu Oh, Sang-Woo Lee
Life of this patent
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Abstract
The present invention relates to a mutant microorganism in which a gene that encodes phosphofructokinase-2 is disrupted or deleted to reduce glycolytic flux to thereby improve the ability of the microorganism to produce N-acetylglucosamine, and to a method of producing N-acetylglucosamine using the mutant microorganism. The mutant microorganism according to the present invention has advantages in that it has high resistance to various chemical substances, grows rapidly, is easily cultured, and produces N-acetylglucosamine with high efficiency, indicating that it is useful for production of a large amount of N-acetylglucosamine.
Claims9
A mutant Saccharomyces cerevisiae BY4742 that overproduces N-acetylglucosamine when cultured in the presence of galactose as a sole carbon source, said mutant Saccharomyces cerevisiae BY4742 comprising a gene encoding a mutant enzyme comprising the amino acid sequence of SEQ ID NO:
- 4, SEQ ID NO:
- 5, or SEQ ID NO:
- 6,
- wherein genes pfk26 and pfk27 which encode phosphofructokinase-2 (PFK-2) are disrupted or deleted,
- wherein the gene pfk26 has the nucleotide sequence of SEQ ID NO:
- 1 and the gene pfk27 has the nucleotide sequence of SEQ ID NO:
- 2, and
- wherein expression of the genes encoding phosphofructokinase-1 (PFK-1) and/or pyruvate kinase (Pyk1p) is inhibited, but not completely inactivated,
- said mutant Saccharomyces cerevisiae BY4742 comprising a Cas protein and/or a vector for expression of Cas protein, and guide RNA for reducing expression of said genes encoding phosphofructokinase-1 (PFK-1) and/or pyruvate kinase (Pyk1p),
- wherein said guide RNA comprises gRNA of SEQ ID NO:
- 10 for reducing expression of the gene encoding phosphofructokinase-1 (PFK-1) and/or gRNA of SEQ ID NO:
- 11 for reducing expression of the gene encoding pyruvate kinase (Pyk1p).
Dependencies are inferred from the claim text.