Claims
- 1. An isolated gene coding for a protein having an N-terminal amino acid sequence of SEQ ID NO: 5, which has the enzymatic ability of hydroxylating the 3-position of L-proline end which acts on free L-proline in the presence of 2-ketoglutaric acid and divalent iron ions to produce cis-3-hydroxy-L-proline.
- 2. The gene according to claim 1, wherein the gene codes for a protein having the amino acid sequence selected from SEQ ID NOs : 1, 2, 15, 16 and 17.
- 3. The gene according to claim 1, wherein the gene is selected from the nucleotide sequences of SEQ ID NOs: 3, 4, 13 and 14.
- 4. The gene according to claim 1, wherein the gene is derived from microorganisms belonging to the genus Streptomyces or Bacillus.
- 5. The gene according to claim 4, wherein the microorganisms are selected from those of Streptomyces canus ATCC12647, Streptomyces canus ATCC12646, Streptomyces sp. TH1 (FERM BP-4399), Bacillus sp. TH2 (FERM BP-4397) and Bacillus sp. TH3 (FERM BP-4398).
- 6. A recombinant DNA constructed by inserting into a vector a DNA fragment that codes for a protein having an N-terminal amino acid sequence of SEQ ID NO: 5, which has the enzymatic ability of hydroxylating the 3-position of L-proline 5 and which acts on free L-proline in the presence of 2-ketoglutaric acid and divalent iron ions to produce cis-3-hydroxy-L-proline.
- 7. A recombinant DNA according to claim 6, wherein the gene codes for a protein having the amino acid sequence selected from the sequences of Sequence Nos. 1, 2, 15, 16 and 17.
- 8. The recombinant DNA according to claim 6, wherein the gene is selected from the nucleotide sequences of SEQ ID NOs: 3, 4, 13 and 14.
- 9. The recombinant DNA according to claim 6, wherein the gene is derived from microorganisms belonging to the genus Streptomyces or Bacillus.
- 10. The recombinant DNA according to claim 9, wherein the microorganisms are selected from those of Streptomyces canus ATCC12647, Streptomyces canus ATCC12646, Streptomyces sp. TH1 (FERM BP-4399), Bacillus sp. TH2 (FERM BP-4397) and Bacillus sp. TH3 (FERM BP-4398).
- 11. A transformant having a recombinant DNA constructed by inserting into a vector a DNA fragment that codes for a protein having the N-terminal amino acid sequence of SEQ ID NO: 5, which has the enzymatic ability of hydroxylating the 3-position of L-proline and which acts on free L-proline in the presence of 2-ketoglutaric acid and divalent iron ions to produce cis-3-hydroxy-L-proline.
- 12. The transformant according to claim 11, wherein the gene codes for a protein having the amino acid sequence selected from the sequences of Sequence Nos. 1, 2, 15, 16 and 17.
- 13. The transformant according to claim 11, wherein the gene is selected from nucleotide sequences of SEQ ID NOs: 3, 4, 13 and 14.
- 14. The transformant according to claim 11, wherein the gene is derived from microorganisms belonging to the genus Streptomyces or Bacillus.
- 15. The transformant according to claim 14, wherein the microorganisms are selected from those of Streptomyces canus ATCC12647, Streptomyces canus ATCC12646, Streptomyces sp. TH1 (FERM BP-4399), Bacillus sp. TH2 (FERM BP-4397) and Bacillus sp. TH3 (FERM BP-4398).
- 16. The transformant according to claim 11, which is selected from Escherichia coli XL2-Blue/pTH30 (FERM BP-5026) and from Escherichia coli XL2-Blue/pTH75 (FERM BP-5409).
- 17. A recombinant method for producing L-proline-3-hydroxylase, which comprises culturing in a medium a transformant, prepared by inserting into a vector a DNA fragment that codes for the hydroxylase having the N-terminal amino acid sequence of SEQ ID NO: 5, which has the enzymatic ability of hydroxylating the 3-position of L-proline and which acts on free L-proline in the presence of 2-ketoglutaric acid and divalent iron ions to produce cis-3-hydroxy-L-proline, thereby producing and accumulating L-proline-3-hydroxylase, followed by collecting the L-proline-3-hydroxylase from the resulting culture.
- 18. The method for producing L-proline-3-hydroxylase according to claim 17, wherein L-proline is added to the medium.
- 19. The method for producing L-proline-3-hydroxylase according to claim 17 or 18, wherein the gene codes for a protein having the amino acid sequence selected from SEQ ID NOs: 1, 2, 15, 16 and 17.
- 20. The method for producing an L-proline-3-hydroxylase according to claim 17 or 18, wherein the gene is selected from nucleotide sequences of SEQ ID NOs: 3, 4, 13 and 14.
- 21. The method for producing L-proline-3-hydroxylase according to claim 17 or 18, wherein the gene is derived from microorganisms belonging to the genus Streptomyces or Bacillus.
- 22. The method for producing L-proline-3-hydroxylase according to claim 21, wherein the microorganisms are selected from Streptomyces canus ATCC12647, Streptomyces canus ATCC12646, Streptomyces sp. TH1 (FERM BP-4399), Bacillus sp. TH2 (FERM BP-4397) and Bacillus sp. TH3 (FERM BP-4398).
- 23. An isolated gene encoding L-proline-3-hydroxylase from Streptomyces or Bacillus selected from SEQ ID NOs: 1, 2, 15, 16 and 17.
Priority Claims (1)
Number |
Date |
Country |
Kind |
5-221941 |
Sep 1993 |
JPX |
|
Parent Case Info
This application is a division of Ser. No. 08/708,856, which is a continuation-in-part of Ser. No. 08/474,135, filed Jul. 6, 1995, now abandoned, which is continuation-in-part of Ser. No. 08/301,654 filed Sep. 7, 1994, now abandoned.
Divisions (1)
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Number |
Date |
Country |
Parent |
708856 |
Sep 1996 |
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Continuation in Parts (2)
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Number |
Date |
Country |
Parent |
474135 |
Jul 1995 |
|
Parent |
301654 |
Sep 1994 |
|