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Journal of Bacteriology, December 2000, p. 6724-6731, Vol. 182, No. 23
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Inducible Metabolism of Phenolic Acids in
Pediococcus pentosaceus Is Encoded by an Autoregulated
Operon Which Involves a New Class of Negative Transcriptional
Regulator
Lise
Barthelmebs,
Bruno
Lecomte,
Charles
Divies, and
Jean-François
Cavin*
Laboratoire de Microbiologie UMR-INRA,
ENSBANA, Université de Bourgogne, F-21000 Dijon, France
Received 19 April 2000/Accepted 21 September 2000
Pediococcus pentosaceus displays a substrate-inducible
phenolic acid decarboxylase (PAD) activity on p-coumaric
acid. Based on DNA sequence homologies between the three PADs
previously cloned, a DNA probe of the Lactobacillus plantarum
pdc gene was used to screen a P. pentosaceus genomic
library in order to clone the corresponding gene of this bacteria. One
clone detected with this probe displayed a low PAD activity. Subcloning
of this plasmid insertion allowed us to determine the part of the
insert which contains a 534-bp open reading frame (ORF) coding for a
178-amino-acid protein presenting 81.5% of identity with L. plantarum PDC enzyme. This ORF was identified as the
padA gene. A second ORF was located just downstream of the
padA gene and displayed 37% identity with the product of
the Bacillus subtilis yfiO gene. Subcloning,
transcriptional analysis, and expression studies with Escherichia
coli of these two genes under the padA gene promoter,
demonstrated that the genes are organized in an autoregulated
bicistronic operonic structure and that the gene located upstream of
the padA gene encodes the transcriptional repressor of the
padA gene. Transcription of this pad operon in
P. pentosaceus is acid phenol dependent.
*
Corresponding author. Mailing address: Laboratoire de
Microbiologie UMR-INRA, ENSBANA, 1 Esplanade Erasme, F-21000 Dijon, France. Phone: (33) 03.80.39.66.72. Fax: (33) 03.80.39.66.40. E-mail:
cavinjf{at}u-bourgogne.fr.
Journal of Bacteriology, December 2000, p. 6724-6731, Vol. 182, No. 23
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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