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Antimicrobial Agents and Chemotherapy, March 2008, p. 1094-1100, Vol. 52, No. 3
0066-4804/08/$08.00+0     doi:10.1128/AAC.01569-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Isolation and Purification of Enterocin E-760 with Broad Antimicrobial Activity against Gram-Positive and Gram-Negative Bacteria{triangledown}

J. E. Line,1* E. A. Svetoch,2 B. V. Eruslanov,2 V. V. Perelygin,2 E. V. Mitsevich,2 I. P. Mitsevich,2 V. P. Levchuk,2 O. E. Svetoch,2 B. S. Seal,1 G. R. Siragusa,1 and N. J. Stern1

USDA-ARS, Russell Research Center, Poultry Microbiological Safety Research Unit, Athens, Georgia,1 State Research Center for Applied Microbiology and Biotechnology, Obolensk, Russian Federation2

Received 18 December 2006/ Returned for modification 10 April 2007/ Accepted 9 December 2007

Strain NRRL B-30745, isolated from chicken ceca and identified as Enterococcus durans, Enterococcus faecium, or Enterococcus hirae, was initially identified as antagonistic to Campylobacter jejuni. The isolate produced a 5,362-Da bacteriocin (enterocin) that inhibits the growth of Salmonella enterica serovar Enteritidis, S. enterica serovar Choleraesuis, S. enterica serovar Typhimurium, S. enterica serovar Gallinarum, Escherichia coli O157:H7, Yersinia enterocolitica, Citrobacter freundii, Klebsiella pneumoniae, Shigella dysenteriae, Pseudomonas aeruginosa, Proteus mirabilis, Morganella morganii, Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, Campylobacter jejuni, and 20 other Campylobacter species isolates. The enterocin, E-760, was isolated and purified by cation-exchange and hydrophobic-interaction chromatographies. The proteinaceous nature of purified enterocin E-760 was demonstrated upon treatment with various proteolytic enzymes. Specifically, the antimicrobial peptide was found to be sensitive to beta-chymotrypsin, proteinase K, and papain, while it was resistant to lysozyme and lipase. The enterocin demonstrated thermostability by retaining activity after 5 min at 100°C and was stable at pH values between 5.0 and 8.7. However, activity was lost below pH 3.0 and above pH 9.5. Administration of enterocin E-760-treated feed significantly (P < 0.05) reduced the colonization of young broiler chicks experimentally challenged and colonized with two strains of C. jejuni by more than 8 log10 CFU. Enterocin E-760 also significantly (P < 0.05) reduced the colonization of naturally acquired Campylobacter species in market age broiler chickens when administered in treated feed 4 days prior to analysis.

* Corresponding author. Mailing address: PMSRU, USDA-ARS, Russell Research Center, 950 College Station Rd., Athens, GA 30604. Phone: (706) 546-3522. Fax: (706) 546-3771. E-mail: eric.line{at}ars.usda.gov

{triangledown} Published ahead of print on 17 December 2007.

Antimicrobial Agents and Chemotherapy, March 2008, p. 1094-1100, Vol. 52, No. 3
0066-4804/08/$08.00+0     doi:10.1128/AAC.01569-06
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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