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Applied and Environmental Microbiology, March 2003, p. 1589-1597, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1589-1597.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Identification and Characterization of Two Novel Clostridial Bacteriocins, Circularin A and Closticin 574

Robèr Kemperman,1 Anneke Kuipers,1 Harma Karsens,1 Arjen Nauta,2 Oscar Kuipers,1 and Jan Kok1*

Department of Molecular Genetics, Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen,1 FCDF Corporate Research, 7400 AB Deventer, The Netherlands2

Received 28 June 2002/ Accepted 1 November 2002

Two novel antibacterial peptides of clostridial species were purified, N-terminally sequenced, and characterized. Moreover, their structural genes were identified. Closticin 574 is an 82-amino-acid bacteriocin produced by Clostridium tyrobutyricum ADRIAT 932. The supernatant of the producing strain showed a high level of activity against the indicator strain C. tyrobutyricum. The protein is synthesized as a preproprotein that is possibly secreted via the general secretion pathway, after which it is hydrolyzed at an Asp-Pro site. Circularin A is produced by Clostridium beijerinckii ATCC 25752 as a prepeptide of 72 amino acids. Cleavage of the prepeptide between the third leucine and fourth valine residues followed by a head-to-tail ligation between the N and C termini creates a circular antimicrobial peptide of 69 amino acids. The unusually small circularin A leader peptide of three amino acids is cleaved off in this process. The supernatant of C. beijerinckii ATCC 25752 showed a broad antibacterial activity range.

* Corresponding author. Mailing address: Department of Molecular Genetics, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands. Phone: 31-503 63 21 11. Fax: 31-503 63 23 48. E-mail: J.kok{at}biol.rug.nl.

Applied and Environmental Microbiology, March 2003, p. 1589-1597, Vol. 69, No. 3
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.3.1589-1597.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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