DFPase enzymes from aplysia californica

Patent metadata

Field	Value
Publication number	US-9447392-B2
Application number	US-201414209463-A
Country	US
Kind code	B2
Filing date	Mar 13, 2014
Priority date	May 6, 2009
Publication date	Sep 20, 2016
Grant date	Sep 20, 2016

Abstract

The present invention relates to isolated polypeptides having organophosphorous hydrolase activity and isolated 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 non-natural nucleic acid construct comprising a polynucleotide sequence that encodes a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2 operably linked to one or more control sequences that direct the production of the polypeptide in an expression host, wherein the polypeptide has organophosphorous hydrolase activity.

2. A recombinant expression vector comprising the nucleic acid construct of claim 1 .

3. A recombinant host cell comprising the nucleic acid construct of claim 1 .

4. A method of producing a polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2 comprising:
(a) cultivating a recombinant host cell comprising a nucleic acid construct comprising a polynucleotide sequence that encodes the polypeptide comprising an amino acid sequence having at least 90% sequence identity to the mature polypeptide of SEQ ID NO: 2, under conditions conducive for production of the polypeptide; and (b) recovering the polypeptide, wherein the polypeptide has organophosphorous hydrolase activity.

5. The method of producing a polypeptide in accordance with claim 4 , wherein the polynucleotide sequence that encodes the polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2, and wherein the polypeptide comprises an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.

6. A method of using a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2 for decontamination comprising: contacting the mature polypeptide with an area or a device contaminated with at least one harmful or undesired organophosphorous compound, wherein the at least one harmful or undesired organophosphorous compound is G-agents, V-agents or pesticides.

7. A method for removing organophosphorous compound comprising contacting the organophosphorous compound with: 1) a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2 or 2) a composition comprising a polypeptide comprising an amino acid sequence having at least 95% sequence identity to the mature polypeptide of SEQ ID NO: 2.

8. A method for removing an organophosphorous compound, comprising contacting the organophosphorous compound with a polypeptide having at least 90%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

9. The method of claim 8 , wherein the polypeptide has at least 95%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

10. The method of claim 8 , wherein the polypeptide has at least 96%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

11. The method of claim 8 , wherein the polypeptide has at least 97%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

12. The method of claim 8 , wherein the polypeptide has at least 98%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

13. The method of claim 8 , wherein the polypeptide has at least 99%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

14. The method of claim 8 , wherein the organophosphorous compound is a nerve gas, toxin or pesticide.

15. A method for decontaminating an organophosphorous compound, comprising contacting the organophosphorous compound with a polypeptide having at least 90%, sequence identity to the mature polypeptide of SEQ ID NO: 2, wherein the polypeptide has detoxification activity.

16. The method of claim 15 , wherein the polypeptide has at least 99%, sequence identity to the mature polypeptide of SEQ ID NO: 2.

17. The method of claim 15 , wherein the organophosphorous compound is a G-agent, V-agent or pesticide.

Assignees

• Novozymes As

Inventors

• Danielsen Steffen
• Skov Lars Kobberoee
• Leite Ricardo
• Laize Vincent
• Da Fonseca M Leonor Cancela

Classifications

• C12Y301/08001

  Aryldialkylphosphatase (3.1.8.1), i.e. paraoxonase · CPC title

• A62D2101/26

  containing nitrogen or phosphorus · CPC title

• C12Y301/08002

  Diisopropyl-fluorophosphatase (3.1.8.2) · CPC title

• A62D3/02

  by biological methods, i.e. processes using enzymes or microorganisms · CPC title

• A62D2101/04

  Pesticides, e.g. insecticides, herbicides, fungicides or nematocides · CPC title