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Journal of Bacteriology, June 1999, p. 3419-3426, Vol. 181, No. 11
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

Pascal Poupin, Véronique Ducrocq,dagger Sylvie Hallier-Soulier,Dagger and Nicole Truffaut*

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.

* 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.

dagger Present address: Marine Biotechnology Institute, 3-75-1 Heita, Kamaishi, Iwate 026, Japan.

Dagger Present address: CIRSEE, 78230 Le Pecq, France.

Journal of Bacteriology, June 1999, p. 3419-3426, Vol. 181, No. 11
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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