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Journal of Bacteriology, March 1999, p. 1610-1616, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
The PalkBFGHJKL Promoter Is under Carbon
Catabolite Repression Control in Pseudomonas oleovorans but
Not in Escherichia coli alk+
Recombinants
Ivo E.
Staijen,
Rosanna
Marcionelli, and
Bernard
Witholt*
Institut für Biotechnologie, Swiss
Federal Institute of Technology (ETH), ETH Hönggerberg, HPT,
8093 Zürich, Switzerland
Received 8 July 1998/Accepted 21 December 1998
The alk genes are located on the OCT plasmid of
Pseudomonas oleovorans and encode an inducible pathway for
the utilization of n-alkanes as carbon and energy sources.
We have investigated the influence of alternative carbon sources on the
induction of this pathway in P. oleovorans and
Escherichia coli alk+ recombinants. In doing
so, we confirmed earlier reports that induction of alkane hydroxylase
activity in pseudomonads is subject to carbon catabolite repression.
Specifically, synthesis of the monooxygenase component AlkB is
repressed at the transcriptional level. The alk genes have
been cloned into plasmid pGEc47, which has a copy number of about 5 to
10 per cell in both E. coli and pseudomonads.
Pseudomonas putida GPo12 is a P. oleovorans
derivative cured of the OCT plasmid. Upon introduction of pGEc47 in
this strain, carbon catabolite repression of alkane hydroxylase
activity was reduced significantly. In cultures of recombinant E. coli HB101 and W3110 carrying pGEc47, induction of AlkB and
transcription of the alkB gene were no longer subject to
carbon catabolite repression. This suggests that carbon catabolite
repression of alkane degradation is regulated differently in
Pseudomonas and in E. coli strains. These
results also indicate that PalkBFGHJKL, the
Palk promoter, might be useful in attaining
high expression levels of heterologous genes in E. coli
grown on inexpensive carbon sources which normally trigger carbon
catabolite repression of native expression systems in this host.
*
Corresponding author. Mailing address: Institut
für Biotechnologie, Swiss Federal Institute of Technology (ETH),
ETH Hönggerberg, HPT, 8093 Zürich, Switzerland. Phone:
41.1.633.32.86. Fax: 41.1.633.10.51. E-mail:
bw{at}biotech.biol.ethz.ch.
Journal of Bacteriology, March 1999, p. 1610-1616, Vol. 181, No. 5
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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