title: The stachydrine catabolism region in Sinorhizobium meliloti encodes a multi-enzyme complex similar to the xenobiotic degrading systems in other bacteria.
abstract: Stachydrine (proline betaine) can be used by Sinorhizobium meliloti as a source of carbon and nitrogen. Catabolism depends on an initial N-demethylation, after which the resultant N-methyl proline enters general metabolism. Deletion and insertion mutagenesis demonstrated that the information necessary for catabolism is carried on the symbiotic plasmid (pSym) distal to nodD2 and the nod-nif cluster. Sequencing of an 8.5kb fragment spanning this region revealed four open reading frames with functional homology to known proteins, including a putative monooxygenase and a putative NADPH-FMN-reductase, which were shown by insertional and frame-shift mutagenesis to be necessary for stachydrine catabolism. Other open reading frames, encoding a putative flavoprotein and a repressor, were judged not to be required for stachydrine catabolism, since they were not included in a fragment capable of complementing a deletion of the entire stc region. Sequence and mutagenesis data suggest that stachydrine is demethylated by an iron-sulfur monooxygenase of the Rieske type with a requirement for a specific reductase. The stc catabolic cluster, therefore, resembles xenobiotic degradation in other bacteria and recalls rhizopine catabolism in S. meliloti. Stachydrine appears to have multiple roles in osmoprotection, nutrition and nodulation. Genes involved in stachydrine catabolism are also necessary for carnitine degradation; thus, they could be important in the catabolism of a variety of root exudates and mediate other relationships.
authors.affiliation: Laboratoire de Biologie de la Rhizosphère, INRA, 78026, Versailles, France. michael.burnet@aguk.zeneca.com
authors.forename: M W
authors.initials: MW
authors.lastname: Burnet
authors.forename: A
authors.initials: A
authors.lastname: Goldmann
authors.forename: B
authors.initials: B
authors.lastname: Message
authors.forename: R
authors.initials: R
authors.lastname: Drong
authors.forename: A
authors.initials: A
authors.lastname: El Amrani
authors.forename: O
authors.initials: O
authors.lastname: Loreau
authors.forename: J
authors.initials: J
authors.lastname: Slightom
authors.forename: D
authors.initials: D
authors.lastname: Tepfer
mesh_terms: D000595:Amino Acid Sequence; D001419:Bacteria / Q000378:metabolism*; D001673:Biodegradation, Environmental; D002250:Carbon Radioisotopes; D002331:Carnitine / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D005420:Flavoproteins / Q000235:genetics; D005816:Genetic Complementation Test; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000235:genetics* / Q000378:metabolism; D016254:Mutagenesis, Insertional; D009586:Nitrogen Fixation / Q000235:genetics; D016366:Open Reading Frames; D010088:Oxidoreductases / Q000235:genetics; D010105:Oxygenases / Q000235:genetics; D018517:Plant Roots / Q000382:microbiology; D010957:Plasmids / Q000235:genetics; D011392:Proline / Q000031:analogs & derivatives* / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D016962:Sinorhizobium meliloti / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D013559:Symbiosis; D015262:Xenobiotics / Q000378:metabolism*
chemical_list: D002250:Carbon Radioisotopes; D004269:DNA, Bacterial; D005420:Flavoproteins; D009097:Multienzyme Complexes; D012097:Repressor Proteins; D015262:Xenobiotics; D011392:Proline; D010088:Oxidoreductases; D010105:Oxygenases; C003342:stachydrine; D002331:Carnitine