Broad Spectrum Bacteriocin for Control of Unwanted Bacteria

Abstract

A composition containing a newly identified bacteriocin produced by a Lactobacillus plantarum strain, isolated from natural corn mash used in commercial fermentation process for production of ethanol. The bacteriocin composition is effective in broad range killing against the vast majority of the lactic acid bacteria (LAB) isolates in ethanol fermentation mash, which include multiple species of Lactobacillus, Lactococcus, Weissella, Leuconostoc, Pediococcus, Enterococcus , and Streptococcus as well as killing activity against non-LAB isolates, such as strains of Staphylococcus, Enterobacter, Bacillus , and Clostridium . The bacteriocin is also combined with broad range bacteriophage to provide synergistic effectiveness again unwanted bacteria in industrial biofuel fermentation process and in human and animal bacterial infections.

1 . A composition comprising a two-peptide lantibiotic bacteriocin isolated from the indigenous lactic acid bacteria  L. plantarum  contained in corn mash from an ethanol fermentation process. 
     
     
         2 . The composition of  claim 1  in an aqueous solution having a pH range of 4-10 and maintained below 65° C. 
     
     
         3 . The composition of  claim 1  wherein the bacteriocin is GP-15cin as defined in the specification. 
     
     
         4 . The composition of  claim 1  derived from over-producing system mutants of  L. plantarum  by natural mutation or genetic manipulation. 
     
     
         5 . A method of control of unwanted bacteria in a fermentation process by adding to the feed to the process or to the fermentation vessel a composition comprising a two-peptide lantibiotics bacteriocin produced by  Lactobacillus plantarum , isolated from the indigenous lactic acid bacteria from an ethanol fermentation process. 
     
     
         6 . The method of  claim 5  comprising contacting unwanted bacteria with  Lactobacillus  bacteria GP-15, as defined in the specification, capable of producing bacteriocin virulent for the unwanted bacteria in a sufficient amount and for sufficient time for the bacteriocin bacteria to produce an effective amount of bacteriocin virulent for unwanted bacteria. 
     
     
         7 . The method of  claim 5  also comprising contacting unwanted bacteria containing material with other bacteriocin(s) capable of killing the bacteria GP-15 from which the GP-15cin bacteriocin is produced. 
     
     
         8 . The method of  claim 5  also comprising adding to the bacteriocin a bacteriophage that is capable of acting synergistically with the bacteriocin in broadening the scope and increasing the efficiency of treating unwanted bacteria. 
     
     
         9 . The method of  claim 5  also comprising adding to the bacteriocin produced by  Lactobacillus plantarum  the bacteriophage LferInf as described in the specification. 
     
     
         10 . The method of  claim 5  wherein the bacteriocin producing GP-15 is added to the fermentation reactor after at least an hour of fermentation. 
     
     
         11 . A method for treating unwanted bacteria in humans or animals by application of a composition comprising a two-peptide lantibiotics bacteriocin produced by  L. plantarum  isolated from lactic acid bacteria contained in corn mash from an ethanol fermentation process. 
     
     
         12 . The method of  claim 11  also comprising contacting unwanted bacteria containing material with other bacteriocin(s) capable of killing the bacteria GP-15 from which GP-15cin is produced. 
     
     
         13 . The method of  claim 11  also comprising adding to the bacteriocin a bacteriophage that is capable of acting synergistically with the bacteriocin in broadening the scope and increasing the efficiency of treating unwanted bacteria. 
     
     
         14 . The method of  claim 11  also comprising adding to the bacteriocin produced by  Lactobacillus plantarum , GP-15cin, the bacteriophage LferInf as described in the specification. 
     
     
         15 . The method of  claim 14  wherein the bacteriocin produced by  Lactobacillus plantarum , GP-15cin, and the bacteriophage LferInf as described in the specification are formulated into a cocktail effective for reduction of unwanted bacteria.