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Applied and Environmental Microbiology, September 1998, p. 3140-3146, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Isolation, Characterization, and Heterologous Expression of the Novel Lantibiotic Epicidin 280 and Analysis of Its Biosynthetic Gene Cluster

Christoph Heidrich,1 Ulrike Pag,1 Michaele Josten,1 Jörg Metzger,2,dagger Ralph W. Jack,2 Gabriele Bierbaum,1 Günther Jung,2 and Hans-Georg Sahl1,*

Institut für Medizinische Mikrobiologie und Immunologie der Universität Bonn, D-53105 Bonn,1 and Institut für Organische Chemie, Universität Tübingen, D-72076 Tübingen,2 Germany

Received 22 December 1997/Accepted 19 June 1998

Epicidin 280 is a novel type A lantibiotic produced by Staphylococcus epidermidis BN 280. During C18 reverse-phase high-performance liquid chromatography two epicidin 280 peaks were obtained; the two compounds had molecular masses of 3,133 ± 1.5 and 3,136 ± 1.5 Da, comparable antibiotic activities, and identical amino acid compositions. Amino acid sequence analysis revealed that epicidin 280 exhibits 75% similarity to Pep5. The strains that produce epicidin 280 and Pep5 exhibit cross-immunity, indicating that the immunity peptides cross-function in antagonization of both lantibiotics. The complete epicidin 280 gene cluster was cloned and was found to comprise at least five open reading frames (eciI, eciA, eciP, eciB, and eciC, in that order). The proteins encoded by these open reading frames exhibit significant sequence similarity to the biosynthetic proteins of the Pep5 operon of Staphylococcus epidermidis 5. A gene for an ABC transporter, which is present in the Pep5 gene cluster but is necessary only for high yields (G. Bierbaum, M. Reis, C. Szekat, and H.-G. Sahl, Appl. Environ. Microbiol. 60:4332-4338, 1994), was not detected. Instead, upstream of the immunity gene eciI we found an open reading frame, eciO, which could code for a novel lantibiotic modification enzyme involved in reduction of an N-terminally located oxopropionyl residue. Epicidin 280 produced by the heterologous host Staphylococcus carnosus TM 300 after introduction of eciIAPBC (i.e., no eciO was present) behaved homogeneously during reverse-phase chromatography.

* Corresponding author. Mailing address: Institut für Medizinische Mikrobiologie und Immunologie, Sigmund-Freud-Str. 25, 53105 Bonn, Germany. Phone: 49 228 287 5704. Fax: 49 228 2876763. E-mail: sahl{at}mibi03.meb.uni-bonn.de.

dagger Present address: Institut für Siedlungswasserbau, Universität Stuttgart, Stuttgart, Germany.

Applied and Environmental Microbiology, September 1998, p. 3140-3146, Vol. 64, No. 9
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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