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.
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.
Poupin
P
P
Laboratoire de Génétique Microbienne, Université de Technologie de Compiègne, Centre de Recherches, 60205 Compiègne, France.
Ducrocq
V
V
Hallier-Soulier
S
S
Truffaut
N
N
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004251:DNA Transposable Elements / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D029968:Escherichia coli Proteins; D005798:Genes, Bacterial / Q000235:genetics; D005947:Glucose / Q000378:metabolism; D007700:Kinetics; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D020102:Mycobacterium smegmatis / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D016366:Open Reading Frames / Q000235:genetics; D010880:Piperidines / Q000378:metabolism*; D011759:Pyrrolidines / Q000378:metabolism*; D012097:Repressor Proteins / Q000737:chemistry / Q000235:genetics* / Q000502:physiology; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014157:Transcription Factors
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; C081598:GntR protein, E coli; D010880:Piperidines; D011759:Pyrrolidines; D012097:Repressor Proteins; D014157:Transcription Factors; C032727:piperidine; D003577:Cytochrome P-450 Enzyme System; D005947:Glucose; C032519:pyrrolidine