Mosquitocidal xenorhabdus, lipopeptide and methods

Patent metadata

Field	Value
Publication number	US-10494407-B2
Application number	US-201615279119-A
Country	US
Kind code	B2
Filing date	Sep 28, 2016
Priority date	Sep 20, 2010
Publication date	Dec 3, 2019
Grant date	Dec 3, 2019

Abstract

Provided is a bacterial strain which produces a family of mosquitocidal toxins, Xenorhabdus MT, on deposit with the American Type Culture Collection, PTA-6826, insecticidal compositions comprising the mosquitocidal toxin(s) produced by Xenorhabdus MT, a mosquitocidal toxin preparation prepared from spent culture medium, whole culture or cells or a mixture thereof, of Xenorhabdus MT and method of insect control, especially mosquito control. Also provided are microbial compounds (same as mosquitocidal toxins) compositions comprising them and use in formulating therapeutic and other antimicrobial compositions, and methods of use for inhibiting microbial growth and for treating infection.

First claim

The invention claimed is:

1. A method of controlling a mosquito of the genus Culex, Aedes or Anopheles comprising the step of feeding the mosquito an effective amount of a lipopeptide toxin having oral toxin activity against the mosquito, wherein the lipopeptide toxin comprises one or more lipopeptides each having a peptide of eight amino acids wherein the peptide contains two histidines, three asparatic acids and/or asparagines, one 2,3-diaminobutryic acid, one glycine and one serine and each peptide has one fatty acid group selected from a saturated straight chain fatty acid, a 2 -oxo fatty acid, a 3-oxo fatty acid or a 4-oxo fatty acid, wherein the fatty acid has from 8 to 20 carbon atoms, wherein the lipopeptide toxin comprises lipopeptide molecules ranging in molecular weight from 1182 to 1478 daltons, and wherein the lipopeptide toxin is not contained in a nematode.

2. The method of claim 1 , wherein the fatty acid group has from 10 to 18 carbon atoms.

3. The method of claim 1 , wherein the lipopeptide toxin is in the form of a composition which further comprises an additional insecticidal component.

4. The method of claim 3 , wherein the additional insecticidal component is a chemical or a biological insecticide.

5. The method of claim 4 , wherein the biological insecticide is a bacterial insecticidal toxin, spinosyn, a plant insecticidal toxin or an insect virus.

6. The method of claim 5 , wherein the biological insecticide is a bacterial insecticidal toxin produced by a Bacillus thuringiensis or a Bacillus thuringiensis israeliensis strain.

7. The method of claim 1 , wherein the fatty acid group is selected from 2-oxo-fatty acid groups having 8 to 20 carbon atoms.

8. The method of claim 1 , wherein the fatty acid group is selected from 2-oxo-fatty acid groups having 10 to 18 carbon atoms.

9. The method of claim 1 , wherein said lipopeptide toxin is produced by a pure bacterial culture of Xenorhabdus MT deposited with the ATCC as PTA-6826.

10. The method of claim 1 , wherein the lipopeptide is contained within a composition comprising dried whole culture of Xenorhabdus innexi , concentrated whole culture of Xenorhabdus innexi , dried cell-free medium from a culture of Xenorhabdus innexi , or concentrated cell-free medium from a culture of Xenorhabdus innexi.

11. The method of claim 10 , wherein the lipopeptide is contained within a composition comprising dried whole culture of Xenorhabdus innexi , dried cell-free medium from a culture of Xenorhabdus innexi , or concentrated cell-free medium from a culture of Xenorhabdus innexi.

12. The method of claim 10 , wherein the lipopeptide is contained within a composition comprising dried whole culture of Xenorhabdus innexi , or dried cell-free medium from a culture of Xenorhabdus innexi.

13. The method of claim 1 , wherein the lipopeptide toxin is a mixture of lipopeptides ranging in molecular weight from 1182 to 1478 daltons.

14. The method of clam 1 , wherein the lipopeptide toxin consists of a mixture of lipopeptide molecules ranging in molecular weight from 1182 to 1478 daltons.

15. The method of claim 1 , wherein the lipopeptide toxin is produced by a Xenorhabdus innexi culture.

16. The method of claim 15 , wherein the lipopeptide toxin is isolated from cell-free growth liquor of said culture by:
precipitation from the liquor with acetone, removal of acetone from the acetone precipitate, forming a water solution of the precipitate from which acetone is removed; applying the water solution on an anion exchange column and collecting the flow through fraction from the column.

17. The method of claim 16 , wherein the flow through fraction of the anion exchange column is applied to a cation exchange column and eluted with 1.0 M aqueous NaCl.

18. The method of claim 10 , wherein the Xenorhabdus innexi is Xenorhabdus MT deposited with the ATCC as PTA-6826.

19. A method of controlling a mosquito of the genus Culex, Aedes or Anopheles comprising the step of feeding the mosquito an effective amount of a lipopeptide toxin having oral toxin activity against the mosquito, wherein the lipopeptide toxin is produced by a pure bacterial culture of the Xenorhabdus MT deposited with the ATCC as PTA-6826, and isolated from cell-free growth liquor of said culture by:
precipitation from the liquor with acetone, removal of acetone from the acetone precipitate, forming a water solution of the precipitate from which acetone is removed; applying the water solution on an anion exchange column and collecting the flow through fraction from the column.

20. The method of claim 19 , wherein the flow through fraction of the anion exchange column is applied to a cation exchange column and eluted with 1.0 M aqueous NaCl.

21. The method of claim 20 , wherein the fraction eluted from the cation exchange column with 1.0 M aqueous NaCl is separated by reverse phase high pressure liquid chromatography with an acetonitrile/0.1% trifluoroacetic acid gradient.

22. The method of claim 21 , wherein the lipopeptide toxin comprises peptide molecules ranging in molecular weight from 1182 to 1478 daltons.

23. The method of claim 22 , wherein the lipopeptide toxin comprises one or more lipopeptides each having a peptide of eight amino acids wherein the peptide contains two histidines, three asparatic acids and/or asparagines, one 2,3-diaminobutryic acid, one glycine and one serine and each peptide has one fatty acid group selected from a saturated straight chain fatty acid, a 2-oxo fatty acid, a 3-oxo fatty acid or a 4-oxo fatty acid, wherein the fatty acid has from 8 to 20 carbon atoms.

24. The method of claim 22 , wherein the lipopeptide toxin is a mixture of lipopeptides each having a peptide of eight amino acids wherein the peptide contains two histidines, three asparatic acids and/or asparagines, one 2,3-diaminobutryic acid, one glycine and one serine and each peptide has one fatty acid group selected from a 2-oxo fatty acid group having from 8 to 20 carbon atoms.

25. The method of claim 22 , wherein the lipopeptide toxin is a mixture of lipopeptides each having a peptide of eight amino acids wherein the peptide contains two histidines, three asparatic acids and/or asparagines, one 2,3-diaminobutryic acid, one glycine and one serine and each peptide has one fatty acid group selected from a 2-oxo fatty acid group having from 10 to 18 carbon atoms.

26. A method of controlling a mosquito of the genus Culex, Aedes or Anopheles comprising the step of feeding the mosquito an effective amount of a lipopeptide toxin having oral toxin activity against the mosquito, wherein the lipopeptide toxin comprises one or more lipopeptides each having a peptide of eight amino acids wherein the peptide contains two histidines, three asparatic acids and/or asparagines, one 2,3-diaminobutryic acid, one glycine and one serine and each peptide has one fatty acid group selected from a saturated straight chain fatty acid, a 2-oxo fatty acid, a 3-oxo fatty acid or a 4-oxo fatty acid, wherein the fatty acid has from 8 to 20 carbon atoms, wherein the lipopeptide toxin is not contained in a nematode, and wherein the lipopeptide toxin is produced by a Xenorhabdus innexi culture and is isolated from cell-free growth liquor of said culture by:
precipitation from the liquor with acetone, removal of acetone from the acetone precipitate, forming a water solution of the precipitate from which acetone is remo

Assignees

• Wisconsin Alumni Res Found

Inventors

• Ensign Jerald Coleman
• Lan Que
• Dyer David

Classifications

• A61P31/00

  Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics · CPC title

• A61K35/74

  Bacteria (therapeutic use of a bacterial protein A61K38/00) · CPC title

• A01N37/46

  N-acyl derivatives · CPC title

• A61K38/03

  Peptides having up to 20 amino acids in an undefined or only partially defined sequence; Derivatives thereof · CPC title

• C07K14/24 Primary

  from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia · CPC title