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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,
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.
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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