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Journal of Bacteriology, October 1998, p. 5218-5226, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Carbon-Source-Dependent Expression of the
PalkB Promoter from the Pseudomonas oleovorans
Alkane Degradation Pathway
Luis
Yuste,
Inés
Canosa, and
Fernando
Rojo*
Departamento de Biotecnología
Microbiana, Centro Nacional de Biotecnología, CSIC, Campus de
la Universidad Autónoma de Madrid, Cantoblanco, 28049-Madrid,
Spain
Received 30 March 1998/Accepted 20 July 1998
Pseudomonas oleovorans GPo1 can metabolize
medium-chain-length alkanes by means of an enzymatic system whose
induction is regulated by the AlkS protein. In the presence of alkanes,
AlkS activates the expression of promoter PalkB, from which
most of the genes of the pathway are transcribed. In addition,
expression of the first enzyme of the pathway, alkane hydroxylase, is
known to be influenced by the carbon source present in the growth
medium, indicating the existence of an additional overimposed level of regulation associating expression of the alk genes with the
metabolic status of the cell. Reporter strains bearing
PalkB-lacZ transcriptional fusions were constructed to
analyze the influence of the carbon source on induction of the
PalkB promoter by a nonmetabolizable inducer. Expression
was most efficient when cells grew at the expense of citrate,
decreasing significantly when the carbon source was lactate or
succinate. When cells were grown in Luria-Bertani rich medium,
PalkB was strongly down-regulated. This effect was partially relieved when multiple copies of the gene coding for the AlkS
activator were present and was not observed when the promoter was moved
to Escherichia coli, a heterologous genetic background.
Possible mechanisms responsible for PalkB regulation are
discussed.
*
Corresponding author. Mailing address: Centro Nacional
de Biotecnología, CSIC, Campus de la Universidad Autónoma
de Madrid, Cantoblanco, 28049-Madrid, Spain. Phone: (34) 91 585 45 39. Fax: (34) 91 585 45 06. E-mail: frojo{at}cnb.uam.es.
Journal of Bacteriology, October 1998, p. 5218-5226, Vol. 180, No. 19
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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