Practical method for enzymatically synthesizing cyclic di-GMP
AssigneeYamasa Corporation
InventorsKaori Tanabe, Kazuya Ishige
Life of this patent
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Abstract
A practical method for enzymatically synthesizing c-di-GMP with excellent productivity is provided. A diguanylate cyclase having physical and chemical characteristics (A) to (F): (A) catalytic action on reaction “2 GTP→c-di-GMP”; (B) a molecular weight of 19800±2000; (C) an optimum pH of 7.3 to 9.4; (D) an optimum temperature of 35 to 60° C.; (E) thermal stability as the remaining activity of 90% or higher after heated for 60 minutes under conditions of 50° C. and pH7.8; and (F) the presence of GGDEF (SEQ ID NO:26) domain and the lack of amino acid sequence KXXD (SEQ ID NO:23) in the i-site.
Claims10
A method for producing c-di-GMP from two GTP molecules using an enzyme, wherein the enzyme is a diguanylate cyclase having a molecular weight of 19,800±2,000 Da and comprising an amino acid sequence selected from the group consisting of:
- (A) the amino acid sequence of SEQ ID NO:6;
- and
- (B) an amino acid sequence having at least 90% identity to the amino acid sequence of SEQ ID NO:6, wherein glycine at position 54 in the amino acid of SEQ ID NO:6 is conserved and the amino acid sequence comprises a GGDEF (SEQ ID NO:26) domain or GGEEF (positions 83-87 of SEQ ID NO:6) domain comprising i-site, but lacks the amino acid sequence KXXD (SEQ ID NO:
- 23) in the i-site;
- the method comprising contacting two GTP molecules with said enzyme.
Dependencies are inferred from the claim text.