title: Pseudomonas putida A ATCC 12633 oxidizes trimethylamine aerobically via two different pathways.
abstract: The present study examined the aerobic metabolism of trimethylamine in Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine. In both conditions, the trimethylamine was used as a nitrogen source and also accumulated in the cell, slowing the bacterial growth. Decreased bacterial growth was counteracted by the addition of AlCl(3). Cell-free extracts prepared from cells grown aerobically on tetradecyltrimethylammonium bromide exhibited trimethylamine monooxygenase activity that produced trimethylamine N-oxide and trimethylamine N-oxide demethylase activity that produced dimethylamine. Cell-free extracts from cells grown on trimethylamine exhibited trimethylamine dehydrogenase activity that produced dimethylamine, which was oxidized to methanal and methylamine by dimethylamine dehydrogenase. These results show that this bacterial strain uses two enzymes to initiate the oxidation of trimethylamine in aerobic conditions. The apparent K(m) for trimethylamine was 0.7 mM for trimethylamine monooxygenase and 4.0 mM for trimethylamine dehydrogenase, but both enzymes maintain similar catalytic efficiency (0.5 and 0.4, respectively). Trimethylamine dehydrogenase was inhibited by trimethylamine from 1 mM. Therefore, the accumulation of trimethylamine inside Pseudomonas putida A ATCC 12633 grown on tetradecyltrimethylammonium bromide or trimethylamine may be due to the low catalytic efficiency of trimethylamine monooxygenase and trimethylamine dehydrogenase.
authors.affiliation: Departamento de Biología Molecular, Facultad de Ciencias Exactas, Fisico-Químicas y Naturales, Universidad Nacional de Río Cuarto, CPX5804BYA, Río Cuarto, Córdoba, Argentina.
authors.forename: Andrés S
authors.initials: AS
authors.lastname: Liffourrena
authors.forename: Mario A
authors.initials: MA
authors.lastname: Salvano
authors.forename: Gloria I
authors.initials: GI
authors.lastname: Lucchesi
mesh_terms: D000332:Aerobiosis; D004123:Dimethylamines / Q000378:metabolism; D007700:Kinetics; D008744:Methylamines / Q000378:metabolism*; D010084:Oxidation-Reduction; D010089:Oxidoreductases, N-Demethylating / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010105:Oxygenases / Q000378:metabolism*; D016958:Pseudomonas putida / Q000254:growth & development / Q000378:metabolism*; D050337:Trimethyl Ammonium Compounds / Q000378:metabolism
chemical_list: D004123:Dimethylamines; D008744:Methylamines; D050337:Trimethyl Ammonium Compounds; C034516:dimethylamine; C027451:methylamine; D010105:Oxygenases; C043996:tertiary amine monooxygenase; D010089:Oxidoreductases, N-Demethylating; C023186:trimethylamine dehydrogenase; C005855:trimethyloxamine; C023336:trimethylamine; C009410:tetradecyltrimethylammonium