5-Substituted hydantoin racemase, DNA encoding the same, recombinant DNA, transformed cell, and process for production of optically active N-carbamylamino acid or optically active amino acid

1. An isolated polypeptide comprising the amino acid sequence of SEQ ID NO: 1.

2. An isolated DNA encoding a polypeptide having hydantoin racemase activity,

said DNA selected from the group consisting of: (1) a DNA that encodes a polypeptide comprising the amino acid sequence of SEQ ID NO: 1; (2) a DNA consisting of the polynucleotide sequence of SEQ ID NO: 2; and (3) a DNA consisting of the polynucleotide sequence of SEQ ID NO: 2 in which 1 to 8 bases are substituted, deleted, inserted and/or added.

3. An isolated polypeptide encoded by a DNA selected from the group consisting of: a DNA consisting of the polynucleotide sequence of SEQ ID NO: 2 and; a DNA consisting of the polynucleotide sequence of SEQ ID NO: 2 in which 1 to 8 bases are substituted, deleted, inserted and/or added.

4. A recombinant plasmid comprising the DNA of claim 2 .

5. The recombinant plasmid of claim 4 , wherein said recombinant plasmid is a plasmid pBHR001 separatable from Escherichia coli HB101 (pBHR001) (FERM BP-10476).

6. A transformant obtained by transforming a host microorganism with the recombinant plasmid of claim 4 .

7. The transformant of claim 6 , wherein the host microorganism is Escherichia coli.

8. The transformant of claim 7 , wherein the transformant is Escherichia coli HB101 (pBHR001) (FERM BP-10476).

9. An isolated microorganism that produces the polypeptide of claim 1 , wherein said microorganism belongs to the Genus Bacillus.

10. The isolated microorganism of claim 9 , wherein the microorganism is Bacillus sp. KNK519HR (FERM BP-10477), or a mutant strain thereof.

11. A process for producing hydantoin racemase, comprising culturing the transformant of claim 6 , accumulating a polypeptide having hydantoin racemase activity in a culture of the transformant, and collecting the polypeptide.

12. A process for producing hydantoin racemase, comprising culturing the microorganism of claim 9 , accumulating said polypeptide in a culture of the microorganism, and collecting the polypeptide.

13. A process for racemizing optically active 5-substituted hydantoin, comprising causing an optically active 5-substituted hydantoin compound to be acted upon by a polypeptide of claim 1 .

14. A process for racemizing optically active 5-substituted hydantoin as set forth in claim 13 , wherein the optically active 5-substituted hydantoin compound is optically active 5-substituted hydantoin represented by General Formula (1)

where R represents an alkyl group having 1 to 20 carbon atoms with or without a substituent, an aralkyl group having 7 to 20 carbon atoms with or without a substituent, or an aryl group having 6 to 20 carbon atoms with or without a substituent.

15. A process for producing optically active N-carbamylamino acid, comprising causing a 5-substituted hydantoin compound to be acted upon by a polypeptide of claim 1 .

16. A process for producing optically active N-carbamylamino acid as set forth in claim 15 ,

wherein the optically active 5-substituted hydantoin compound is 5-substituted hydantoin represented by General Formula (1)

where R represents an alkyl group having 1 to 20 carbon atoms with or without a substituent, an aralkyl group having 7 to 20 carbon atoms with or without a substituent, or an aryl group having 6 to 20 carbon atoms with or without a substituent, and

wherein the optically active N-carbamylamino acid is optically active N-carbamylamino acid represented by General Formula (2)

where R represents an alkyl group having 1 to 20 carbon atoms with or without a substituent, an aralkyl group having 7 to 20 carbon atoms with or without a substituent, or an aryl group having 6 to 20 carbon atoms with or without a substituent.

17. A process as set forth in claim 16 , wherein the optically active N-carbamylamino acid is one of N-carbamyl-D-leucine, N-carbamyl-D-isoleucine, N-carbamyl-D-valine, N-carbamyl-D-norleucine, N-carbamyl-D-norvaline, N-carbamyl-D-methionine, N-carbamyl-D-cysteine, N-carbamyl-D-penicillamine, N-carbamyl-D-phenylalanine, N-carbamyl-D-phenylglycine, and N-carbamyl-D-4-hydroxyphenylglycine.

18. A process for producing an optically active amino acid, comprising causing a 5-substituted hydantoin compound to be acted upon by hydantoinase and N-carbamylamino acid amidohydrolase, together with the polypeptide of claim 1 .

19. A process for producing an optically active amino acid as set forth in claim 18 ,

wherein the 5-substituted hydantoin compound is 5-substituted hydantoin represented by General Formula (1)

where R represents an alkyl group having 1 to 20 carbon atoms with or without a substituent, an aralkyl group having 7 to 20 carbon atoms with or without a substituent, or an aryl group having 6 to 20 carbon atoms with or without a substituent, and

wherein the optically active amino acid is represented by General Formula (3)

where R represents an alkyl group having 1 to 20 carbon atoms with or without a substituent, an aralkyl group having 7 to 20 carbon atoms with or without a substituent, or an aryl group having 6 to 20 carbon atoms with or without a substituent.

20. A process as set forth in claim 19 , wherein the optically active amino acid is one of D-leucine, D-isoleucine, D-valine, D-norleucine, D-norvaline, D-methionine, D-cysteine, D-penicillamine, D-phenylalanine, D-phenylglycine, and D-4-hydroxyphenylglycine.