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Applied and Environmental Microbiology, October 2003, p. 5839-5848, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5839-5848.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Functional Analysis of the Gene Cluster Involved in Production of the Bacteriocin Circularin A by Clostridium beijerinckii ATCC 25752

Robèr Kemperman,1 Marnix Jonker,1 Arjen Nauta,2 Oscar P. Kuipers,1 and Jan Kok1*

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

Received 31 March 2003/ Accepted 1 August 2003

A region of 12 kb flanking the structural gene of the cyclic antibacterial peptide circularin A of Clostridium beijerinckii ATCC 25752 was sequenced, and the putative proteins involved in the production and secretion of circularin A were identified. The genes are tightly organized in overlapping open reading frames. Heterologous expression of circularin A in Enterococcus faecalis was achieved, and five genes were identified as minimally required for bacteriocin production and secretion. Two of the putative proteins, CirB and CirC, are predicted to contain membrane-spanning domains, while CirD contains a highly conserved ATP-binding domain. Together with CirB and CirC, this ATP-binding protein is involved in the production of circularin A. The fifth gene, cirE, confers immunity towards circularin A when expressed in either Lactococcus lactis or E. faecalis and is needed in order to allow the bacteria to produce bacteriocin. Additional resistance against circularin A is conferred by the activity of the putative transporter consisting of CirB and CirD.

* 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, October 2003, p. 5839-5848, Vol. 69, No. 10
0099-2240/03/$08.00+0     DOI: 10.1128/AEM.69.10.5839-5848.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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