Applied and Environmental Microbiology, May 1999, p. 2078-2083, Vol. 65, No. 5
0099-2240/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Department of Microbiology and National Food Biotechnology Centre, University College Cork, Cork, Ireland,1 and Department of Food Science, Wageningen Agricultural University, Wageningen, The Netherlands2
Received 30 November 1998/Accepted 5 March 1999
The trimethylammonium compound glycine betaine
(N,N,N-trimethylglycine) can be
accumulated to high intracellular concentrations, conferring enhanced
osmo- and cryotolerance upon Listeria monocytogenes. We
report the identification of betL, a gene encoding a
glycine betaine uptake system in L. monocytogenes,
isolated by functional complementation of the betaine uptake mutant
Escherichia coli MKH13. The betL gene is
preceded by a consensus
B-dependent promoter and is
predicted to encode a 55-kDa protein (507 amino acid residues)
with 12 transmembrane regions. BetL exhibits significant sequence
homologies to other glycine betaine transporters, including OpuD from
Bacillus subtilis (57% identity) and BetP from
Corynebacterium glutamicum (41% identity). These high-affinity secondary transporters form a subset of the
trimethylammonium transporter family specific for glycine betaine,
whose substrates possess a fully methylated quaternary ammonium group.
The observed Km value of 7.9 µM for glycine
betaine uptake after heterologous expression of betL in
E. coli MKH13 is consistent with values obtained for
L. monocytogenes in other studies. In addition, a betL knockout mutant which is significantly affected in
its ability to accumulate glycine betaine in the presence or absence of
NaCl has been constructed in L. monocytogenes. This
mutant is also unable to withstand concentrations of salt as high as
can the BetL+ parent, signifying the role of the
transporter in Listeria osmotolerance.
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