Mosquitocidal xenorhabdus, lipopeptide and methods

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.

1 .- 33 . (canceled) 
     
     
         34 . A mosquitocidal composition, the composition comprising a lipopeptide toxin comprising one or more lipopeptides each having a peptide of eight amino acids in length, the amino acids being selected from one or more of histidine, aspartic acid, asparagine, 2,3-diaminobutryic acid, glycine, and serine, or one or more conservative amino acid substitutions thereof,
 wherein 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, the fatty acid having from 8 to 20 carbon atoms, and   wherein the lipopeptide toxin comprises lipopeptide molecules ranging in molecular weight from 1182 to 1478 daltons.   
     
     
         35 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises an alanine substituted for a glycine. 
     
     
         36 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises a glutamine substituted for an asparagine. 
     
     
         37 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises a threonine substituted for a serine. 
     
     
         38 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises a cysteine substituted for a serine. 
     
     
         39 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises a lysine substituted for a histidine. 
     
     
         40 . The composition of  claim 34 , wherein the one or more conservative amino acid substitutions comprises a lysine substituted for a 2,3-diaminobutryic acid. 
     
     
         41 . The composition of  claim 34 , wherein the fatty acid group has from 10 to 18 carbon atoms. 
     
     
         42 . A mosquitocidal composition, the composition comprising a lipopeptide toxin comprising one or more lipopeptides each having a peptide of eight amino acids in length, the amino acids being selected from one or more of histidine, aspartic acid, asparagine, 2,3-diaminobutryic acid, glycine, and serine, or one or more conservative amino acid substitutions thereof,
 wherein at least two amino acids are histidine or a conservative amino acid substitution thereof,   wherein 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, the fatty acid having from 8 to 20 carbon atoms, and   wherein the lipopeptide toxin comprises lipopeptide molecules ranging in molecular weight from 1182 to 1478 daltons.   
     
     
         43 . The composition of  claim 42 , wherein the one or more conservative amino acid substitutions comprises a lysine substituted for a histidine, a lysine substituted for a 2,3-diaminobutryic acid, a glutamine substituted for an asparagine, a threonine substituted for a serine, a cysteine substituted for a serine, an alanine substituted for a glycine, or a combination thereof. 
     
     
         44 . The composition of  claim 43 , wherein the fatty acid group has from 10 to 18 carbon atoms. 
     
     
         45 . The composition of  claim 44 , further comprising an additional insecticidal component selected from a bacterial insecticidal toxin, spinosyn, a plant insecticidal toxin or an insect virus. 
     
     
         46 . The composition of  claim 45 , wherein the bacterial insecticidal toxin is produced by a  Bacillus thuringiensis  or a  Bacillus thuringiensis  israeliensis strain. 
     
     
         47 . The composition of  claim 46 , wherein the lipopeptide toxin is produced by a pure bacterial culture of  Xenorhabdus  MT deposited with the ATCC as PTA-6826. 
     
     
         48 . The composition of  claim 47 , wherein the lipopeptide toxin 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.    
     
     
         49 . A method of controlling a mosquito, the method comprising applying an effective amount of a mosquitocidal composition to an environment of the mosquito, the mosquitocidal composition comprising a lipopeptide toxin comprising one or more lipopeptides each having a peptide of eight amino acids in length, the amino acids being selected from one or more of histidine, aspartic acid, asparagine, 2,3-diaminobutryic acid, glycine, and serine, or one or more conservative amino acid substitutions thereof,
 wherein 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, the fatty acid having from 8 to 20 carbon atoms, and   wherein the lipopeptide toxin comprises lipopeptide molecules ranging in molecular weight from 1182 to 1478 daltons.   
     
     
         50 . The method of  claim 49 , wherein the one or more conservative amino acid substitutions comprises an alanine substituted for a glycine, a glutamine substituted for an asparagine, a threonine substituted for a serine, a cysteine substituted for a serine, a lysine substituted for a histidine, a lysine substituted for a 2,3-diaminobutryic acid, or a combination thereof. 
     
     
         51 . The method of  claim 49 , wherein applying an effective amount of the mosquitocidal composition to an environment of the mosquito comprises feeding the mosquito an effective amount of the mosquitocidal composition. 
     
     
         52 . The method of  claim 49 , wherein the mosquito is from the genus  Culex, Aedes , or  Anopheles.    
     
     
         53 . The method of  claim 49 , wherein the lipopeptide toxin is not contained in a nematode.