Glucose-6-phosphate dehydrogenase
AssigneeKyowa Hakko Kogyo Co., Ltd.
InventorsHaruhiko Yokoi, Seiko Ando, Keiko Ochiai, Yoshiyuki Yonetani, Shin-ichi Hashimoto
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Abstract
The present invention relates to a novel glucose-6-phosphate dehydrogenase (hereinafter referred to as “G6PD”) derived from a bacterium belonging to the genus Corynebacterium, a DNA encoding the enzyme, a recombinant DNA comprising the DNA, a transformant comprising the recombinant DNA, a transformant comprising the DNA on its chromosome, and a process for producing L-amino acid or G6PD which comprises culturing the transformant. According to the present invention, a modified G6PD and a DNA encoding the G6PD are obtained, and the productivity of L-amino acid by a microorganism can be improved by using the modified G6PD.
Claims17
An isolated or purified DNA which encodes a polypeptide selected from the group consisting of (a) to (d):
- (a) a polypeptide which consists of SEQ ID NO:2,
- (b) a polypeptide which consists of a variant of SEQ ID NO:2, wherein only Ala at position 213 in SEQ ID NO:2 is replaced with another amino acid, and a variant of SEQ ID NO:2, wherein glucose-6-phosphate dehydrogenase activity,
- (c) a polypeptide which consists of SEQ ID NO:12, and
- (d) a polypeptide which consists of a variant of SEQ ID NO:12, wherein up to ten amino acids other than the amino acid residue at position 213 in SEQ ID NO:12 are deleted, substituted or added, and a variant of SEQ ID NO:12, has glucose-6-phosphate dehydrogenase activity.
An isolated or purified DNA which consists of the nucleotide sequence SEQ ID NO:1.
An isolated or purified DNA which consists of a variant of SEQ ID NO:1, wherein only positions 637 to 639 in the nucleotide sequence of SEQ ID NO:1 is replaced with a codon encoding an amino acid other than Ala and wherein said variant encodes a polypeptide with glucose-6-phospahte dehydrogenase activity.
An isolated or purified DNA which consists of the nucleotide sequence SEQ ID NO:11.
An isolated or purified DNA which hybridizes with a DNA comprising the nucleotide sequence of SEQ ID NO:1 at 65° C. in the presence of 0.7 to 1.0 mol/l of sodium chloride using a filter on which colony- or plaque-derived DNA is immobilized, followed by washing at 65° C. with 0.1 to 2×SSC solution, and encodes a polypeptide having glucose-6-phosphate dehydrogenase activity,
- wherein a nucleotide sequence corresponding to the nucleotide sequence of positions 637 to 639 encoding in the nucleotide sequence SEQ ID NO:1 is replaced with a codon encoding an amino acid other than Ala and an amino acid sequence of the polypeptide encoded by the hybridizable DNA has homology of 95% or more compared with the amino acid sequence of SEQ ID NO:12.
An isolated or purified DNA which hybridizes with a DNA comprising the nucleotide sequence of SEQ ID NO:1 at 65° C. in the presence of 0.7 to 1.0 mol/l of sodium chloride using a filter on which colony- or plaque-derived DNA is immobilized, followed by washing at 65° C. with 0.1 to 2×SSC solution, and encodes a polypeptide having glucose-6-phosphate dehydrogenase activity,
- wherein a nucleotide sequence corresponding to the nucleotide of position 637 in the nucleotide sequence SEQ ID NO:1 is replaced with adenine and an amino acid sequence of the polypeptide encoded by the hybridizable DNA has homology of 95% or more compared with the amino acid sequence of SEQ ID NO:12.
A plasmid pCRBzwfM found in Escherichia coli TOP10 (FERM BP-7135).
Dependencies are inferred from the claim text.