Chondroitin-producing bacterium and method of producing chondroitin
AssigneeSeikagaku Corporation
InventorsKiyoshi Suzuki, Kentaro Miyamoto, Hiromi Kaseyama
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Abstract
Chondroitin is produced by culturing a UDP-glucuronic acid-producing bacterium transfected with a kfoA gene derived from Escherichia coli K4 strain and a kfoC gene derived from Escherichia coli K4 strain and having chondroitin-producing ability. Chondroitin is collected from the bacterium.
Claims6
An isolated UDP-glucuronic acid-producing bacterium, which is introduced with a kfoA gene derived from the Escherichia coli K4 strain and a kfoC gene derived from Escherichia coli K4 strain, wherein said bacterium has chondroitin-producing ability, and wherein the kfoA gene encodes a protein selected from the group consisting of (A) and (B), and wherein the kfoC gene encodes a protein selected from the group consisting of (C), (D), (E) and (F):
- (A) a protein comprising the amino acid sequence of SEQ ID NO:
- 2;
- (B) a protein comprising the amino acid sequence of SEQ ID NO:
- 2 comprising substitution, deletion, insertion, or addition of 1-20 amino acids and having UDP-glucose-4-epimerase activity;
- (C) a protein comprising the amino acid sequence of SEQ ID NO:
- 4;
- (D) a protein comprising the amino acid sequence of SEQ ID NO:
- 4 comprising substitution, deletion, insertion, or addition of 1-20 amino acids and having chondroitin synthase activity;
- (E) a protein comprising the amino acid sequence of SEQ ID NO:
- 6;
- and
- (F) a protein comprising the amino acid sequence of SEQ ID NO:
- 6 comprising substitution, deletion, insertion, or addition of 1-20 amino acids and having chondroitin synthase activity.
A vector comprising a kfoA gene derived from Escherichia coli K4 strain and a kfoC gene derived from Escherichia coli K4 strain, wherein the kfoA gene is a DNA selected from the group consisting of (a) and (b), and wherein the kfoC gene is a DNA selected from the group consisting of (c), (d), (e) and (f), and wherein stringent conditions are conditions for hybridization at 42° C. in a solution containing 50% formamide, 4×SSC, 50 mM HEPES (pH 7.0), 10×Denhardt's solution, 100 μg/ml salmon sperm DNA and washing with 2×SSC, 0.1% SDS solution at room temperature and then 0.1×SSC, 0.1% SDS solution at 60° C.:
- (a) a DNA comprising the nucleotide sequence of SEQ ID NO:
- 1;
- (b) a DNA that hybridizes with a DNA comprising the nucleotide sequence complementary to SEQ ID NO:
- 1 under stringent conditions and encodes a protein having UDP-glucose-4-epimerase activity;
- (c) a DNA comprising the nucleotide sequence of SEQ ID NO:
- 3;
- (d) a DNA that hybridizes with a DNA comprising the nucleotide sequence complementary to SEQ ID NO:
- 3 under stringent conditions and encodes a protein having chondroitin synthase activity;
- (e) a DNA comprising the nucleotide sequence of SEQ ID NO:
- 5;
- and
- (f) a DNA that hybridizes with a DNA comprising the nucleotide sequence complementary to SEQ ID NO:
- 5 under stringent conditions and encodes a protein having chondroitin synthase activity.
Dependencies are inferred from the claim text.