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Journal of Bacteriology, July 2001, p. 4202-4209, Vol. 183, No. 14
Departamento de Biotecnología
Microbiana, Centro Nacional de Biotecnología, CSIC, Campus de
la Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid,
Spain,1 and Institute of
Biotechnology, ETH Hönggerberg, CH-8093 Zürich,
Switzerland2
Received 23 March 2001/Accepted 24 April 2001
In many microorganisms the first step for alkane degradation is the
terminal oxidation of the molecule by an alkane hydroxylase. We report
the characterization of a gene coding for an alkane hydroxylase in a
Burkholderia cepacia strain isolated from an oil-contaminated site. The protein encoded showed similarity to other
known or predicted bacterial alkane hydroxylases, although it clustered
on a separate branch together with the predicted alkane hydroxylase of
a Mycobacterium tuberculosis strain. Introduction of the
cloned B. cepacia gene into an alkane hydroxylase
knockout mutant of Pseudomonas fluorescens CHAO restored
its ability to grow on alkanes, which confirms that the gene analyzed
encodes a functional alkane hydroxylase. The gene, which was named
alkB, is not linked to other genes of the alkane
oxidation pathway. Its promoter was identified, and its expression was
analyzed under different growth conditions. Transcription was induced
by alkanes of chain lengths containing 12 to at least 30 carbon atoms
as well as by alkanols. Although the gene was efficiently expressed during exponential growth, transcription increased about fivefold when
cells approached stationary phase, a characteristic not shared by the
few alkane degraders whose regulation has been studied. Expression of
the alkB gene was under carbon catabolite repression when cells were cultured in the presence of several organic acids and
sugars or in a complex (rich) medium. The catabolic repression process
showed several characteristics that are clearly different from what has
been observed in other alkane degradation pathways.
0021-9193/01/$04.00+0 DOI: 10.1128/JB.183.14.4202-4209.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
The Alkane Hydroxylase Gene of Burkholderia
cepacia RR10 Is under Catabolite Repression Control

*
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.
Present address: IATE/P, EPFL, CH-1020 Lausanne, Switzerland.
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