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United States Department of Agriculture

Agricultural Research Service

Research Project: INTERVENTIONS AND METHODOLOGIES TO REDUCE HUMAN FOOD-BORNE BACTERIAL PATHOGENS IN CHICKENS Title: Inducer bacteria, unique signal peptides and nutrient limitation stimulate in-vitro bacteriocin production

Authors
item Svetoch, Edward -
item Eruslanov, Boris -
item Perelygin, Vladimir -
item Levchuk, Vladimir -
item Seal, Bruce
item Stern, Norman

Submitted to: International Symposium on Antimicrobial Peptides
Publication Type: Abstract Only
Publication Acceptance Date: April 15, 2009
Publication Date: June 17, 2009
Citation: Svetoch, E.A., Eruslanov, B.V., Perelygin, V.V., Levchuk, V.P., Seal, B.S., Stern, N.J. 2009. Inducer bacteria, unique signal peptides and nutrient limitation stimulate in-vitro bacteriocin production. International Symposium on Antimicrobial Peptides.

Technical Abstract: Bacteriocins (BCN) provide enormous potential for controlling bacterial infections in human and veterinary medicine, in feedstuffs and human foods, and in cosmetic applications. To successfully apply such antimicrobial proteins, adequate commercial quantities of these valuable BCN must be efficiently produced and harvested. We have previously described three Type IIa BCN produced by lactic acid bacteria (NRRL B-30514, NRRL B-30745 and NRRL B-30746) which in vitro are demonstrably effective against a wide array of pathogenic bacteria and when orally administered to infected adult broiler chickens, milligram quantities reduced six logs of pathogen colonization. To produce required commercial quantities of BCN we determined that a combination of adding signal peptides in the presence of both producer and inducer bacteria to a dilute fermentation medium enabled marked increase in output. The three signal peptides ranged from 24 to 30 amino acids in length and had a terminal carboxyl sequence of VKGLT. The inducer bacterial isolates added to the BCN fermentation were Lactobacillus acidophilus NRRL B-30510 and Lactobacillus crispatus NRRL B-30884. Although other starvation media would likely provide similar influence, we employed a 10% Brucella Broth to optimize our BCN production. Using the combination of the above three parameters enabled us to reproducibly harvest an excess of 200 mg BCN per liter of the spent fermentation broth for each of the three BCN studied. A commercial quantity of each of these three Type IIa BCN can now be reproducibly and efficiently created.

Last Modified: 11/25/2014
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