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Appl Environ Microbiol, April 1998, p. 1359-1365, Vol. 64, No. 4
Institut für Lebensmitteltechnologie,
Universität Hohenheim, 70593 Stuttgart, Germany
Received 19 August 1997/Accepted 20 January 1998
The catalase gene katA of Lactobacillus
sakei LTH677 was cloned and expressed in Escherichia
coli UM2, Lactobacillus casei LK1, and
Lactobacillus curvatus LTH1432. The last host is a
catalase-deficient plasmid-cured derivative of a starter organism used
in meat fermentation. The regulation of katA expression was
found to be the same in L. sakei LTH677 and the recombinant
strains. The addition of H2O2 to anaerobic
cultures, as well as a switch to aerobic conditions, resulted in a
strong increase in KatA activity. The expression was investigated in
more detail with L. sakei LTH677 and L. curvatus LTH4002. The recombinant strain LTH4002 did not
accumulate H2O2 under glucose-limited aerobic
conditions and remained viable in the stationary phase. Under inductive
conditions, the katA-specific mRNA and the apoenzyme were
synthesized de novo. Deletion derivatives of the katA
promoter were produced, and the regulatory response was investigated by
fusion to the
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Copyright © 1998, American Society for Microbiology. All rights reserved.
Oxygen-Dependent Regulation of the Expression of
the Catalase Gene katA of Lactobacillus
sakei LTH677
-glucuronidase reporter gene gusA and
expression in L. sakei LTH677. The fact that gene
expression was subject to induction was confirmed at the level of
transcription and protein synthesis. A small putative regulatory
sequence of at least 25 bp was identified located upstream of the
35
site. Competition experiments performed with L. sakei
LTH677 harboring the fusion constructs consisting of the
katA promoter and gusA revealed that an
activator protein is involved in the transcriptional induction of
katA.
*
Corresponding author. Mailing address: Institut
für Lebensmitteltechnologie, Universität Hohenheim,
Garbenstrasse 28, 70599 Stuttgart, Germany. Phone: 49 711 459 4255. Fax: 49 711 459 4199. E-mail: hertel{at}uni-hohenheim.de.
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