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Applied and Environmental Microbiology, April 2000, p. 1305-1310, Vol. 66, No. 4
Department of Biology, Third University of
Rome, 00146 Rome, Italy,1 and Laboratory
of Microbial Gene Technology, Department of Biotechnological
Sciences, Agricultural University of Norway, 1432 Aas,2 and UNIGEN Center for
Molecular Biology and Laboratory of Biotechnology, Norwegian
University of Science and Technology, 7489 Trondheim,3 Norway
Received 23 September 1999/Accepted 11 January 2000
The effects of different carbon sources on expression of the
styrene catabolism genes in Pseudomonas fluorescens ST
were analyzed by using a promoter probe vector, pPR9TT, which contains
transcription terminators upstream and downstream of the
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Physiological Analysis of the Expression of the Styrene
Degradation Gene Cluster in Pseudomonas fluorescens
ST
-galactosidase reporter system. Expression of the promoter of the
stySR operon, which codes for the styrene two-component
regulatory system, was found to be constitutive and not subject to
catabolite repression. This was confirmed by the results of an analysis
of the stySR transcript in P. fluorescens
ST cells grown on different carbon sources. The promoter of the operon
of the upper pathway, designated PstyA, was induced by
styrene and repressed to different extents by organic acids or
carbohydrates. In particular, cells grown on succinate or lactate in
the presence of styrene started to exhibit -galactosidase activity
during the mid-exponential growth phase, before the preferred carbon
sources were depleted, indicating that there is a threshold succinate
and lactate concentration which allows induction of styrene catabolic
genes. In contrast, cells grown on glucose, acetate, or glutamate and
styrene exhibited a diauxic growth curve, and -galactosidase
activity was detected only after the end of the exponential growth
phase. In each experiment the reliability of the reporter system
constructed was verified by comparing the -galactosidase activity
and the activity of the styrene monooxygenase encoded by the first gene
of the styrene catabolic operon.
*
Corresponding author. Mailing address: Department of
Biology, Third University of Rome, Viale Marconi 446, 00146 Rome,
Italy. Phone: 39 0655176318. Fax: 39 0655176321. E-mail:
zennaro{at}bio.uniroma3.it.
Applied and Environmental Microbiology, April 2000, p. 1305-1310, Vol. 66, No. 4
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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