Dehydrogenase and a gene encoding the same
View Patent ↗The object of the present invention is to provide a novel dehydrogenase having a property which is different from that of known dehydrogenases. The present invention provides a dehydrogenase having the following physicochemical properties: (1) effect: to produce N-alkyl-L-alanine from pyruvic acid and alkylamine or dialkylamine using NADPH and/or NADH as coenzyme; (2) substrate specificity: to show activity to alkylamine or dialkylamine but not to ammonium; (3) optimal pH when using phenylpyruvic acid and methylamine as substrates is around 10; and (4) when treated at 30° C. for 30 minutes, the enzyme is stable at around pH 5 to 10.5.
1. An isolated dehydrogenase isolated from Pseudomonas putida having the following physicochemical properties:
(1) effect: to produce N-alkyl-L-alanine from pyruvic acid and alkylamine or dialkylamine using NADPH and/or NADH as coenzyme;
(2) substrate specificity: to show activity to alkylamine or dialkylamine but not to ammonium;
(3) optimal pH when using phenylpyruvic acid and methylamine as substrates is around 10;
(4) when treated at 30° C. for 30 minutes, the enzyme is stable at around pH 5 to 10.5; and
(5) a molecular weight of approximately 36 kiloDaltons in SDS-polyacrylamide electrophoresis.
2. An isolated polypeptide as in (1) or (2) below
(1) a polypeptide comprising SEQ ID NO: 1; or
(2) a polypeptide having an amino acid sequence wherein 1 to 20 amino acids are deleted, substituted and/or added in the amino acid sequence comprising SEQ ID No. 1 and having N-methyl-L-amino acid dehydrogenase activity.
3. A polypeptide according to claim 2 having an amino acid sequence which shows 95% or more homology to the amino acid sequence comprising SEQ ID No. 1, and having dehydrogenase activity.
4. A polypeptide according to claim 2 having an amino acid sequence which shows 97% or more homology to the amino acid sequence comprising SEQ ID No. 1, and having N-methyl-L-amino acid dehydrogenase activity.
5. A polypeptide according to claim 2 having an amino acid sequence wherein 1 to 15 amino acids are deleted, substituted and/or added in the amino acid sequence comprising SEQ ID No. 1, and having N-methyl-L-amino acid dehydrogenase activity.
6. A polypeptide according to claim 2 having an amino acid sequence wherein 1 to 10 amino acids are deleted, substituted and/or added in the amino acid sequence comprising SEQ ID No. 1, and having N-methyl-L-amino acid dehydrogenase activity.
7. A polypeptide according to claim 2 having an amino acid sequence wherein 1 to 7 amino acids are deleted, substituted and/or added in the amino acid sequence comprising SEQ ID No. 1, and having N-methyl-L-amino acid dehydrogenase activity.
8. A polypeptide according to claim 2 having an amino acid sequence wherein approximately 1 to 5 amino acids are deleted, substituted and/or added in the amino acid sequence comprising SEQ ID No. 1, and having N-methyl-L-amino acid dehydrogenase activity.