The inducible trimethylamine N-oxide reductase of Escherichia coli K12: its localization and inducers

• Published in: Biochimica et biophysica acta Vol. 999; no. 2; pp. 208 - 216
• Main Authors: Silvestro, André, Pommier, Janine, Pascal, Marie-Claire, Giordano, Gérard
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 30.11.1989; Elsevier; North-Holland
• Subjects: Anaerobiosis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Dimethyl Sulfoxide - pharmacology; E. coli; Enzyme Induction; Enzymes and enzyme inhibitors; Escherichia coli - drug effects; Escherichia coli - enzymology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Genotype; Immune Sera; Immunodiffusion; Immunoelectrophoresis; Kinetics; Macromolecular Substances; Methylamines - pharmacology; NADH, NADPH Oxidoreductases - biosynthesis; NADH, NADPH Oxidoreductases - isolation & purification; NADH, NADPH Oxidoreductases - metabolism; Oxidoreductases; Oxidoreductases Acting on CH-NH Group Donors; Pyridines - pharmacology; Trimethylamine- N-oxide reductase; TMAO; PNO; LMS; DMSO; Enzyme induction; Trimethylamine- N-oxide reductase; THTO; Immunoelectrophoresis; E. coli; Enzyme inducer; Enzyme; Escherichia coli; Immunodiffusion; Enzymatic activity; Trimethylamine-N-oxide reductase; Spheroplast; Immunoblotting assay; Bacteria; FPLC chromatography; Localization; Escherichieae; Enterobacteriaceae
• ISSN: 0167-4838; 0006-3002; 1879-2588; 1878-2434
• DOI: 10.1016/0167-4838(89)90220-3

We used an anti-trimethylamine- N-oxide reductase (EC 1.6.6.9) serum and different immunological techniques (Ouchterlony, rocket immunoelectrophoresis, immunoblotting) to show that dimethylsulphoxide (DMSO), tetrahydrothiophene 1-oxide (THTO) and pyridine N-oxide (PNO) were effective inducers of the inducible form of trimethylamine N-oxide reductase. We confirmed this genetically and biochemically using a strain in which phage MudII 1734 carrying lacZ was inserted into torA, the structural gene for inducible trimethylamine- N-oxide reductase. By subcellular fractionation and quantitation with rocket immunoelectrophoresis, we showed that the enzyme was principally localized in the periplasmic fraction. Constitutive trimethylamine- N-oxide reductase was localized in the membrane fraction and, like the inducible enzyme showed a broad specificity with respect to various compounds such as DMSO, THTO and PNO. Apart from their immunological properties, the two enzymes could be clearly differentiated by their temperature stability.