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Applied and Environmental Microbiology, September 2009, p. 5734-5738, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.01070-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Potential Applicability of Chymotrypsin-Susceptible Microcin J25 Derivatives to Food Preservation{triangledown}

María Fernanda Pomares,1 Raúl A. Salomón,1 Olga Pavlova,2 Konstantin Severinov,2,3 Ricardo Farías,1 and Paula A. Vincent1*

Departamento de Bioquímica de la Nutrición, Instituto Superior de Investigaciones Biológicas (Consejo Nacional de Investigaciones Científicas y Técnicas-Universidad Nacional de Tucumán) and Instituto de Química Biológica Dr. Bernabé Bloj, Chacabuco 461, 4000 San Miguel de Tucumán, Tucumán, Argentina,1 Department of Molecular Biology and Biochemistry, Waksman Institute, Rutgers, The State University of New Jersey, Piscataway, New Jersey 08854,2 Institutes of Molecular Genetics and Gene Biology, Russian Academy of Sciences, Moscow, Russia3

Received 11 May 2009/ Accepted 3 July 2009

Microcin J25 (MccJ25) is a 21-residue ribosomally synthesized lariat peptide antibiotic. MccJ25 is active against such food-borne disease-causing pathogens as Salmonella spp., Shigella spp., and Escherichia coli, including E. coli O157:H7 and non-O157 strains. MccJ25 is highly resistant to digestion by proteolytic enzymes present in the stomach and intestinal contents. MccJ25 would therefore remain active in the gastrointestinal tract, affecting normal intestinal microbiota, and this limits the potential use of MccJ25 as a food preservative. In the present paper, we describe a chymotrypsin-susceptible MccJ25 derivative with a mutation of Gly12 to Tyr that retained almost full antibiotic activity and efficiently inhibited the growth of pathogenic Salmonella enterica serovar Newport and Escherichia coli O157:H7 in skim milk and egg yolk. However, unlike the wild-type MccJ25, the MccJ25(G12Y) variant was inactivated by digestive enzymes both in vitro and in vivo. To our knowledge, our results represent the first example of a rational modification of a microcin aimed at increasing its potential use in food preservation.

* Corresponding author. Mailing address: Departamento de Bioquímica de la Nutrición, INSIBIO, Chacabuco 461, 4000 San Miguel de Tucumán, Tucumán, Argentina. Phone: (54) (381) 4248921. Fax: (54) (381) 4248921. E-mail: pvincent{at}fbqf.unt.edu.ar

{triangledown} Published ahead of print on 10 July 2009.

Applied and Environmental Microbiology, September 2009, p. 5734-5738, Vol. 75, No. 17
0099-2240/09/$08.00+0     doi:10.1128/AEM.01070-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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