Patent ID: 8039232

Claim:
A method for producing a dipeptide comprising: culturing a transformed cell in a medium for a time and under conditions suitable to produce a peptide-forming enzyme in a culture, and mixing the culture with a carboxy component and an amine component to synthesize a dipeptide by enzymatic catalysis facilitated by a peptide-forming enzyme encoded by said DNA; wherein said carboxy component is an amino acid ester or an amino acid amide; wherein said amine component is selected from the group consisting of an amino acid and a C-protected amino acid; and wherein said transformed cell comprises a recombinant DNA comprising a polynucleotide encoding a protein selected from the group consisting of (E), (F), (Q), and (R), wherein said protein has an amino acid sequence defined as follows: (E) the amino acid sequence consisting of amino acid residues numbers 23 to 625 of SEQ ID NO:18, (F) an amino acid sequence which includes substitution, deletion, insertion, and/or addition, of one to 30 amino acids in the amino acid sequence consisting of amino acid residues numbers 23 to 625 of SEQ ID NO:18, and has at least 50% of the peptide-forming activity of a protein corresponding to unmutated amino acid residue numbers 23 to 625 of SEQ ID NO:18 at 50° C. and a pH of 8, (Q) the amino acid sequence consisting of SEQ ID NO:18, (R) an amino acid sequence which includes substitution, deletion, insertion, and/or addition, of one to 30 amino acids in the amino acid sequence consisting of SEQ ID NO:18, and has at least 50% of the peptide-forming activity of a protein corresponding to unmutated SEQ ID NO:18 at 50° C. and a pH of 8, or a polynucleotide selected from the group consisting of (e), (e2), and (q), wherein said polynucleotide has a nucleotide sequence defined as follows: (e) a nucleotide sequence consisting of nucleotide numbers 127 to 1935 of SEQ ID NO:17 (e2) a nucleotide sequence that hybridizes under stringent conditions with a DNA having a nucleotide sequence complementary to a nucleotide sequence consisting of nucleotide numbers 127 to 1935 of SEQ ID NO:17 under stringent conditions, wherein said stringent conditions are 0.1×SSC and 0.1% SDS at 65° C., and encodes a protein that has at least 50% of the peptide-forming activity at 50° C. and a pH of 8 of a protein encoded by unmutatetd nucleotide numbers 127 to 1935 of SEQ ID NO:17, (q) a nucleotide sequence consisting of nucleotide numbers 61 to 1935 of SEQ ID NO:17.