Polypeptides having peroxygenase activity

Patent metadata

Field	Value
Publication number	US-9719072-B2
Application number	US-201314432416-A
Country	US
Kind code	B2
Filing date	Oct 8, 2013
Priority date	Oct 12, 2012
Publication date	Aug 1, 2017
Grant date	Aug 1, 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. A polynucleotide encoding a peroxygenase was isolated from Thielavia hyrcaniae.

First claim

The invention claimed is:

1. A method of producing a polypeptide having peroxygenase activity, comprising:
(a) cultivating a recombinant host cell under conditions conducive for production of the polypeptide, wherein
(i) the recombinant host cell comprises a nucleic acid construct or expression vector comprising a polynucleotide encoding the polypeptide;
(ii) the polypeptide has at least 80% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2; and
(iii) the polynucleotide is operably linked to one or more control sequences that direct the production of the polypeptide in the recombinant host cell; and

(b) recovering the polypeptide.

2. The method of claim 1 , wherein the polypeptide has at least 85% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.

3. The method of claim 1 , wherein the polypeptide has at least 90% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.

4. The method of claim 1 , wherein the polypeptide has at least 95% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.

5. The method of claim 1 , wherein the polypeptide has at least 97% sequence identity to the polypeptide of amino acids 1-241 of SEQ ID NO: 2.

6. The method of claim 1 , wherein the polypeptide is encoded by a polynucleotide that hybridizes under high stringency conditions with (i) the mature polypeptide coding sequence of SEQ ID NO: 1, (ii) the cDNA sequence thereof, or (iii) the full-length complement of (i) or (ii).

7. The method of claim 1 , wherein the polypeptide is encoded by a polynucleotide having at least 80% sequence identity to the mature polypeptide coding sequence of SEQ ID NO: 1 , or the cDNA sequence thereof.

8. The method of claim 1 , wherein the polypeptide is a variant of the polypeptide of amino acids 1-241 of SEQ ID NO: 2 comprising a substitution, deletion, and/or insertion at one or more positions.

9. The method of claim 1 , wherein the polypeptide is a fragment of the polypeptide of amino acids 1-241 of SEQ ID NO: 2 and wherein the fragment has peroxygenase activity.

10. The method of claim 1 , wherein the polypeptide comprises amino acids 1-241 of SEQ ID NO: 2.

11. The method of claim 1 , wherein the polypeptide consists of amino acids 1-241 of SEQ ID NO: 2.

12. The method of claim 1 , wherein the polypeptide comprises an amino acid sequence including the motif:
E-H-D-[G,A]-S-[L,I]-S-R, or
R-[G,A]-P-C-P-X-[M,L]-N-[S,T]-L.

13. The method of claim 1 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chlysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

14. The method of claim 2 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

15. The method of claim 3 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

16. The method of claim 4 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

17. The method of claim 5 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

18. The method of claim 9 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

19. The method of claim 10 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

20. The method of claim 12 , wherein the recombinant host cell is a filamentous fungal host cell selected from the group consisting of Acremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phiebia, Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium, Trametes , and Trichoderma.

Assignees

• Novozymes As

Inventors

• Landvik Sara
• Oestergaard Lars Henrik
• Kalum Lisbeth

Classifications

• C12P7/18

  polyhydric · CPC title

• C12N9/0065 Primary

  acting on hydrogen peroxide as acceptor (1.11) · CPC title

• C11D3/38654

  containing oxidase or reductase · CPC title

• C12P7/64

  Fats; Fatty oils; Ester-type waxes; Higher fatty acids, i.e. having at least seven carbon atoms in an unbroken chain bound to a carboxyl group; Oxidised oils or fats · CPC title

• C12Y111/02001 Primary

  Unspecific peroxygenase (1.11.2.1) · CPC title