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Applied and Environmental Microbiology, January 2007, p. 168-178, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.02023-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Isolation and Identification of a Paenibacillus polymyxa Strain That Coproduces a Novel Lantibiotic and Polymyxin ^,

Zengguo He,¹ Duygu Kisla,^1, Liwen Zhang,³ Chunhua Yuan,⁴ Kari B. Green-Church,³ and Ahmed E. Yousef^1,2*

Department of Food Science & Technology,¹ Department of Microbiology,² Mass Spectrometry and Proteomics Facility, Campus Chemical Instrument Center,³ Nuclear Magnetic Resonance Facility, Campus Chemical Instrument Center, The Ohio State University, Columbus, Ohio 43210⁴

Received 25 August 2006/ Accepted 30 October 2006

A new bacterial strain, displaying potent antimicrobial properties against gram-negative and gram-positive pathogenic bacteria, was isolated from food. Based on its phenotypical and biochemical properties as well as its 16S rRNA gene sequence, the bacterium was identified as Paenibacillus polymyxa and it was designated as strain OSY-DF. The antimicrobials produced by this strain were isolated from the fermentation broth and subsequently analyzed by liquid chromatography-mass spectrometry. Two antimicrobials were found: a known antibiotic, polymyxin E1, which is active against gram-negative bacteria, and an unknown 2,983-Da compound showing activity against gram-positive bacteria. The latter was purified to homogeneity, and its antimicrobial potency and proteinaceous nature were confirmed. The antimicrobial peptide, designated paenibacillin, is active against a broad range of food-borne pathogenic and spoilage bacteria, including Bacillus spp., Clostridium sporogenes, Lactobacillus spp., Lactococcus lactis, Leuconostoc mesenteroides, Listeria spp., Pediococcus cerevisiae, Staphylococcus aureus, and Streptococcus agalactiae. Furthermore, it possesses the physico-chemical properties of an ideal antimicrobial agent in terms of water solubility, thermal resistance, and stability against acid/alkali (pH 2.0 to 9.0) treatment. Edman degradation, mass spectroscopy, and nuclear magnetic resonance were used to sequence native and chemically modified paenibacillin. While details of the tentative sequence need to be elucidated in future work, the peptide was unequivocally characterized as a novel lantibiotic, with a high degree of posttranslational modifications. The coproduction of polymyxin E1 and a lantibiotic is a finding that has not been reported earlier. The new strain and associated peptide are potentially useful in food and medical applications.

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* Corresponding author. Mailing address: Parker Food Science Building, The Ohio State University, 2015 Fyffe Road, Columbus, OH 43210. Phone: (614) 292-7814. Fax: (614) 292-0218. E-mail: yousef.1{at}osu.edu.

Published ahead of print on 27 October 2006.

Supplemental material for this article may be found at http://aem.asm.org/.

Present address: Ege University, Faculty of Fisheries, Fish Processing Technology Department, Bornova, Izmir, Turkey.

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Applied and Environmental Microbiology, January 2007, p. 168-178, Vol. 73, No. 1
0099-2240/07/$08.00+0     doi:10.1128/AEM.02023-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.