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Journal of Bacteriology, September 2000, p. 4738-4743, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
The Membrane-Bound H+-ATPase Complex Is
Essential for Growth of Lactococcus lactis
Brian J.
Koebmann,1,2
Dan
Nilsson,2
Oscar P.
Kuipers,3,
and
Peter
R.
Jensen1,*
Department of Microbiology, Technical
University of Denmark, DK-2800 Lyngby,1
and Department of Physiology, Chr. Hansen A/S, DK-2970
Hørsholm,2 Denmark, and Microbial
Ingredients Section, NIZO Food Research, NL-6710 BA Ede, The
Netherlands3
Received 10 January 2000/Accepted 13 June 2000
The eight genes which encode the (F1Fo)
H+-ATPase in Lactococcus lactis subsp.
cremoris MG1363 were cloned and sequenced. The genes were
organized in an operon with the gene order atpEBFHAGDC; i.e., the order of atpE and atpB is reversed
with respect to the more typical bacterial organization. The deduced
amino acid sequences of the corresponding H+-ATPase
subunits showed significant homology with the subunits from other
organisms. Results of Northern blot analysis showed a transcript at
approximately 7 kb, which corresponds to the size of the
atp operon. The transcription initiation site was mapped by
primer extension and coincided with a standard promoter sequence. In
order to analyze the importance of the H+-ATPase for
L. lactis physiology, a mutant strain was constructed in
which the original atp promoter on the chromosome was
replaced with an inducible nisin promoter. When grown on GM17 plates
the resulting strain was completely dependent on the presence of nisin for growth. These data demonstrate that the H+-ATPase is
essential for growth of L. lactis under these conditions.
*
Corresponding author. Mailing address: Department of
Microbiology, Technical University of Denmark, Building 301, DK-2800 Lyngby, Denmark. Phone: 45 45252510. Fax: 45 45932809. E-mail: imprj{at}pop.dtu.dk.

Present address: Molecular Genetics, Groningen Biomolecular
Sciences and Biotechnology Institute, University of Groningen,
NL-9750
AA Haren, The
Netherlands.
Journal of Bacteriology, September 2000, p. 4738-4743, Vol. 182, No. 17
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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