Polypeptides having peroxygenase activity

Patent metadata

Field	Value
Publication number	US-9670468-B2
Application number	US-201615370027-A
Country	US
Kind code	B2
Filing date	Dec 6, 2016
Priority date	Oct 12, 2012
Publication date	Jun 6, 2017
Grant date	Jun 6, 2017

Abstract

The present invention relates to isolated polypeptides having peroxygenase activity, and polynucleotides encoding the polypeptides. The invention also relates to nucleic acid constructs, vectors, and host cells comprising the polynucleotides as well as methods of producing and using the polypeptides.

First claim

The invention claimed is:

1. A method for hydroxylation in position 2 or 3 of either end of a substituted or unsubstituted, linear or branched, alkane or saturated fatty acid having at least 3 carbons and having a hydrogen attached to the carbon in position 2 or 3, comprising contacting said alkane or said saturated fatty acid with hydrogen peroxide and an isolated polypeptide having peroxygenase activity and comprising an amino acid sequence having at least 95% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2, wherein said alkane or said saturated fatty acid is optionally substituted with one or two substituents selected from the group consisting of halogen, hydroxyl, carboxyl, amino, nitro, cyano, thiol, sulphonyl, formyl, acetyl, methoxy, ethoxy, phenyl, benzyl, xylyl, carbamoyl and sulfamoyl.

2. The method of claim 1 , wherein said alkane is hydroxylated in the method.

3. The method of claim 2 , wherein the alkane is pentane, hexane, heptane, octane, nonane, decane, or an isomer thereof.

4. The method of claim 2 , wherein the alkane is unsubstituted.

5. The method of claim 2 , wherein the alkane is linear.

6. The method of claim 2 , wherein the alkane is converted to a diol by introduction of two hydroxy groups.

7. The method of claim 1 , wherein said saturated fatty acid is hydroxylated in the method.

8. The method of claim 7 , wherein the saturated fatty acid is selected from the group consisting of butanoic acid, pentanoic acid, hexanoic acid, heptanoic acid, octanoic acid, nonanoic acid, decanoic acid, dodecanoic acid, tetradecanoic acid, hexadecanoic acid, octadecanoic acid, and eicosanoic acid.

9. The method of claim 7 , wherein the saturated fatty acid is unsubstituted.

10. The method of claim 7 , wherein the saturated fatty acid is linear.

11. The method of claim 7 , wherein the saturated fatty acid is converted to a diol by introduction of two hydroxy groups.

12. The method of claim 2 , wherein the polypeptide comprises an amino acid sequence that has at least 97% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

13. The method of claim 2 , wherein the polypeptide comprises the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

14. The method of claim 7 , wherein the polypeptide comprises an amino acid sequence that has at least 97% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

15. The method of claim 7 , wherein the polypeptide comprises the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

16. A method for introducing a keto group at the second or third carbon of at least two ends of a substituted or unsubstituted, linear or branched, alkane or saturated fatty acid having at least five carbons and having at least one hydrogen attached to said second or third carbon, comprising contacting said alkane or said saturated fatty acid with hydrogen peroxide and an isolated polypeptide having peroxygenase activity and comprising an amino acid sequence having at least 95% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2, wherein said alkane or said saturated fatty acid is optionally substituted with one or two substituents selected from the group consisting of halogen, hydroxyl, carboxyl, amino, nitro, cyano, thiol, sulphonyl, formyl, acetyl, methoxy, ethoxy, phenyl, benzyl, xylyl, carbamoyl and sulfamoyl.

17. The method of claim 16 , wherein the polypeptide comprises an amino acid sequence that has at least 97% sequence identity to the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

18. The method of claim 16 , wherein the polypeptide comprises the sequence of amino acids 1 to 244 of SEQ ID NO: 2.

19. The method of claim 16 , wherein the alkane or saturated fatty acid is unsubstituted.

20. The method of claim 16 , wherein the alkane or saturated fatty acid is linear.

Assignees

• Novozymes As

Inventors

• Landvik Sara
• Oestergaard Lars Henrik
• Kalum Lisbeth

Classifications

• C12P17/00

  Preparation of heterocyclic carbon compounds with only O, N, S, Se or Te as ring hetero atoms (C12P13/04 - C12P13/24 take precedence) · CPC title

• C12Y111/02004

  Fatty-acid peroxygenase (1.11.2.4) · CPC title

• C12Y111/02001

  Unspecific peroxygenase (1.11.2.1) · CPC title

• C12N2310/141

  MicroRNAs, miRNAs · CPC title

• C11D3/38654

  containing oxidase or reductase · CPC title