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Originally published In Press as doi:10.1074/jbc.M207237200 on October 11, 2002
J. Biol. Chem., Vol. 278, Issue 1, 89-94, January 3, 2003
Function of Lactococcus lactis Nisin
Immunity Genes nisI and nisFEG after
Coordinated Expression in the Surrogate Host Bacillus
subtilis*
Torsten
Stein,
Stefan
Heinzmann,
Irina
Solovieva, and
Karl-Dieter
Entian
From the Institut für Mikrobiologie, Johann Wolfgang
Goethe-Universität, D-60439 Frankfurt am Main, Germany
Nisin-producing Lactococcus lactis
strains show a high degree of resistance to the action of nisin, which
is based upon expression of the self-protection (immunity) genes
nisI, nisF, nisE, and nisG. Different combinations of nisin immunity genes were
integrated into the chromosome of a nisin-sensitive Bacillus
subtilis host strain under the control of an inducible promoter.
For the recipient strain, the highest level of acquired nisin tolerance
was achieved after coordinated expression of all four nisin immunity
genes. But either the lipoprotein NisI or the ABC
transporter-homologous system NisFEG, respectively, were also able to
protect the Bacillus host cells. The acquired immunity was
specific to nisin and provided no tolerance to subtilin, a closely
related lantibiotic. Quantitative in vivo peptide release
assays demonstrated that NisFEG diminished the quantity of
cell-associated nisin, providing evidence that one role of NisFEG is to
transport nisin from the membrane into the extracellular space. NisI
solubilized from B. subtilis membrane vesicles and
recombinant hexahistidine-tagged NisI from Escherichia coli
interacted specifically with nisin and not with subtilin. This suggests
a function of NisI as a nisin-intercepting protein.
*
This work was supported by a grant from the Stipendien-Fonds
des Verbandes der Chemischen Industrie (to S. H.) and Deutscher Akademischer Austauschdienst (DAAD) Grant 325-A/02/06602 (to I. S.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Institut für
Mikrobiologie, Johann Wolfgang Goethe-Universität,
Marie-Curie-Str. 9, D-60439 Frankfurt am Main, Germany. Tel.:
49-69-7982-9526; Fax: 49-69-7982-9527; E-mail:
Entian@em.uni-frankfurt.de.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2003 by the American Society for Biochemistry and Molecular Biology.
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