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Journal of Bacteriology, June 1999, p. 3419-3426, Vol. 181, No. 11
Laboratoire de Génétique
Microbienne, Université de Technologie de Compiègne,
Centre de Recherches, 60205 Compiègne, France
Received 21 December 1998/Accepted 6 April 1999
Transposon mutagenesis of Mycobacterium smegmatis
mc2155 enabled the isolation of a mutant strain (called
LGM1) altered in the regulation of piperidine and pyrrolidine
utilization. The complete nucleotide sequence of the gene inactivated
in mutant LGM1 was determined from the wild-type strain. This gene
(pipR) encoded a member of the GntR family of bacterial
regulatory proteins. An insertion element (IS1096),
previously described for M. smegmatis, was
detected downstream of the gene pipR. Three additional open reading frames were found downstream of IS1096. The first
open reading frame (pipA) appeared to encode a protein
identified as a cytochrome P450 enzyme. This gene is the first member
of a new family, CYP151. By a gene replacement experiment,
it was demonstrated that the cytochrome P450 pipA gene is
required for piperidine and pyrrolidine utilization in M. smegmatis mc2155. Genes homologous to
pipA were detected by hybridization in several, previously
isolated, morpholine-degrading mycobacterial strains. A gene encoding a
putative [3Fe-4S] ferredoxin (orf1) and a truncated
gene encoding a putative glutamine synthetase (orf2') were
found downstream of pipA.
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Cloning and Characterization of the Genes Encoding a Cytochrome
P450 (PipA) Involved in Piperidine and Pyrrolidine Utilization and
Its Regulatory Protein (PipR) in Mycobacterium
smegmatis mc2155

and
*
Corresponding author. Mailing address: Laboratoire de
Génétique Microbienne, Université de
Technologie de Compiègne, B.P. 20529, 60205 Compiègne, France. Phone: 33 3 44 23 44 52. Fax: 33 3 44 20 48 13. E-mail: nicole.truffaut{at}utc.fr.
Present address: Marine Biotechnology Institute, 3-75-1 Heita,
Kamaishi, Iwate 026, Japan.
Present address: CIRSEE, 78230 Le Pecq, France.
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