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 . A method of controlling an insect comprising, the step of applying to an environment of the insect an effective amount of a lipopeptide toxin having functional activity against an insect, wherein said lipopeptide toxin is produced by a pure bacterial culture of the  Xenorhabdus  deposited with the ATCC as PTA-6826. 
     
     
         2 . The method of  claim 1 , wherein the lipopeptide toxin is applied to an environment of the insect. 
     
     
         3 . The method of  claim 1  or  2 , wherein the lipopeptide is contained within a composition comprising (i) dried whole culture of the  Xenorhabdus , (ii) concentrated whole culture of the  Xenorhabdus , (iii) dried cell free medium from a culture of the  Xenorhabdus , (iv) concentrated whole cell free medium from a culture of the  Xenorhabdus  or (v) purified lipopeptide toxin. 
     
     
         4 . The method of any one of  claims 1  to  3 , wherein the insect is a Dipteran insect. 
     
     
         5 . The method of  claim 4 , wherein the insect is a member of the Culicidae. 
     
     
         6 . The method of  claim 5 , wherein the insect is a mosquito of the genus  Culex, Aedes  or  Anopheles.    
     
     
         7 . The method of  claim 1 , further comprising an insecticidal toxin having functional activity against an insect in a mixture of one or more toxins produced from a purified culture of  Xenorhabdus  MT, deposited with American Type Culture Collection Accession No. PTA-6826. 
     
     
         8 . The method of any one of  claims 3  to  7 , wherein the composition comprises an additional insecticidal component. 
     
     
         9 . The method of  claim 8 , wherein the additional insecticidal component is a chemical or a biological insecticide. 
     
     
         10 . The method of  claim 9 , wherein the biological insecticide is a bacterial toxin, spinosyn, a plant insecticidal toxin or an insect virus. 
     
     
         11 . The method of  claim 9 , wherein the biological insecticide is a bacterial toxin produced by a  Bacillus thuringiensis  or a  Bacillus thuringiensis israeliensis  strain. 
     
     
         12 . The method of  claim 11 , wherein the  Xenorhabdus  protein is native to  Xenorhabdus  MT, American Type Culture Collection Accession No. PTA 6826. 
     
     
         13 . An insecticidal composition comprising the mosquitocidal toxin produced by  Xenorhabdus  MT, American Type Culture Collection Accession No. PTA 6826. 
     
     
         14 . The composition of  claim 13 , comprising whole growth liquor of  Xenorhabdus  deposited with the American Type Culture Collection as PTA-6826. 
     
     
         15 . The composition of  claim 14 , comprising dried or concentrated whole growth liquor of  Xenorhabdus  deposited with the American Type Culture Collection as PTA-6826. 
     
     
         16 . The composition of  claim 14 , comprising cells of  Xenorhabdus  deposited with the American Type Culture Collection as PTA-6826. 
     
     
         17 . The composition of  claim 14 , comprising cell-free spent medium of a culture of  Xenorhabdus  deposited with the American Type Culture Collection as PTA-6826. 
     
     
         18 . A composition comprising a mosquitocidal toxin produced by the  Xenorhabdus  deposited with the American Type Culture Collection as PTA-6826, said toxin comprising histidine, diaminobutyric acid, asparagine and/or aspartate, serine and glycine, and at least one saturated straight chain fatty acid, a 2-oxo fatty acid, 3-oxo fatty acid or a 4-oxo fatty acid, wherein the fatty acid has a chain length of from C8 to C20 or of from C10 to C18. 
     
     
         19 . A mosquitocidal and/or antimicrobial composition comprising a lipopeptide toxin produced by  Xenorhabdus innexi scapertisci  (ATCC PTA-6826), said toxin having a molecular weight, as determined by MALDI-TOF mass spectrophotometric analyses, of from about 1182 to about 1431 daltons, and comprising at least one fatty acid moiety covalently linked to the peptide, wherein said fatty acid moiety is a saturated fatty acid moiety of from C8 to C20, a straight chain saturated fatty acid moiety of from C10 to C18, or a 2-oxo-fatty acid moiety of from C8 to C20 or C10 to C18, a monounsaturated fatty acid moiety of from C8 to C20 or from Cl 0 to C18, a 3-oxo-fatty acid moiety of from C8 to C20 or C10 to C18, or a 4-oxo-fatty acid moiety of from C8 to C20 or from C10 to C18. 
     
     
         20 . The composition of  claim 19 , further comprising a pharmaceutically acceptable carrier. 
     
     
         21 . The composition of  claim 19  or  20 , having antimicrobial activity against a microorganism selected from the group consisting of a bacterium, a fungus, a virus, and a protozoan. 
     
     
         22 . The composition of  claim 21 , wherein the microorganism is a bacterium. 
     
     
         23 . The composition of  claim 22 , wherein the bacterium is  Pseudomonas aeruginosa, Escherichia coli, Salmonella enteriditis, Salmonella typhimurium, Salmonella agona, Listeria monocytogenes, Micrococcus luteus , or  Bacillus cereus.    
     
     
         24 . The composition of  claim 21 , wherein the fungus is  Candida albicans.    
     
     
         25 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 13 - 19 . 
     
     
         26 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 13 - 19 , wherein said microorganism is selected from the group consisting of a gram-negative bacterium, a gram-positive bacterium and a fungus. 
     
     
         27 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 13 - 19 , wherein said controlling is by inhibiting growth, replication, or infectivity of said microorganism. 
     
     
         28 . A mosquitocidal and antimicrobial composition produced by reverse phase chromatography of a culture of by  Xenorhabdus innexi scapertisci  (ATCC PTA-6826), wherein said material is eluted during the chromatography using 25-40% (vol/vol) acetonitrile/0.1% trifluoroacetic acid (vol/vol) as solvent for elution. 
     
     
         29 . The composition of  claim 32 , wherein there are two successive reverse phase chromatography steps. 
     
     
         30 . The composition of  claim 32 , where there are three successive reverse phase chromatography steps. 
     
     
         31 . The composition of any one of  claims 32  to  34 , wherein the composition comprises lipopeptides of molecular weights of 1195 plus or minus 3, 1306 plus or minus 3, 1349 plus or minus 3, 1363 plus or minus 3, and 1478 plus or minus 3. 
     
     
         32 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 32 - 34 . 
     
     
         33 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 32 - 34 , wherein said microorganism is selected from the group consisting of a gram-negative bacterium, a gram-positive bacterium and a fungus. 
     
     
         34 . A method of controlling a microorganism, said method comprising the step of administering an effective amount of the composition of any one of  claims 32 - 34 , wherein said controlling is by inhibiting growth, replication, or infectivity of said microorganism. 
     
     
         35 . A pure  Xenorhabdus innexi  culture deposited with the American Type Culture Collection as PA-6826. 
     
     
         36 . A method for treating an infection caused by a microorganism in a subject, said method comprising the step of administering a therapeutically effective amount of the composition of any one of  claim 13 - 9  or  32 - 34  to a subject in need thereof. 
     
     
         37 . A method for treating an infection caused by a microorganism in a subject, said method comprising the step of administering a therapeutically effective amount of the composition of any one of  claim 13 - 9  or  32 - 34  to a subject in need thereof, wherein the microorganism is selected from the group consisting of a gram-positive bacterium, a gram-negative bacterium and a fungus. 
     
     
         38 . A method for treating an infection caused by a microorganism in a subject, said method comprising the step of administering a therapeutically effective amount of the composition of any one of  claim 13 - 9  or  32 - 34  to a subject in need thereof, wherein the gram-negative bacterium is  Pseudomonas aeruginosa, Escherichia coli, Salmonella enteriditis, Salmonella typhimurium, Salmonella agona, Listeria monocytogenes, Micrococcus luteus , or  Bacillus cereus.    
     
     
         39 . A method for treating an infection caused by a microorganism in a subject, said method comprises the step of administering a therapeutically effective amount of the composition of any one of  claim 13 - 9  or  32 - 34  to a subject in need thereof, wherein the fungus is  Candida albicans.