Existence of aa3-type ubiquinol oxidase as a terminal oxidase in sulfite oxidation of Acidithiobacillus thiooxidans.
It was found that Acidithiobacillus thiooxidans has sulfite:ubiquinone oxidoreductase and ubiquinol oxidase activities in the cells. Ubiquinol oxidase was purified from plasma membranes of strain NB1-3 in a nearly homogeneous state. A purified enzyme showed absorption peaks at 419 and 595 nm in the oxidized form and at 442 and 605 nm in the reduced form. Pyridine ferrohaemochrome prepared from the enzyme showed an alpha-peak characteristic of haem a at 587 nm, indicating that the enzyme contains haem a as a component. The CO difference spectrum of ubiquinol oxidase showed two peaks at 428 nm and 595 nm, and a trough at 446 nm, suggesting the existence of an aa(3)-type cytochrome in the enzyme. Ubiquinol oxidase was composed of three subunits with apparent molecular masses of 57 kDa, 34 kDa, and 23 kDa. The optimum pH and temperature for ubiquinol oxidation were pH 6.0 and 30 degrees C. The activity was completely inhibited by sodium cyanide at 1.0 mM. In contrast, the activity was inhibited weakly by antimycin A(1) and myxothiazol, which are inhibitors of mitochondrial bc(1) complex. Quinone analog 2-heptyl-4-hydoroxyquinoline N-oxide (HOQNO) strongly inhibited ubiquinol oxidase activity. Nickel and tungstate (0.1 mM), which are used as a bacteriostatic agent for A. thiooxidans-dependent concrete corrosion, inhibited ubiquinol oxidase activity 100 and 70% respectively.
Sugio
Tsuyoshi
T
Division of Science and Technology for Energy Conversion, Graduate School of Natural Science and Technology, Okayama University, Okayama 700-8530, Japan. sugio@cc.okayama-u.ac.jp
Hisazumi
Tomohiro
T
Kanao
Tadayoshi
T
Kamimura
Kazuo
K
Takeuchi
Fumiaki
F
Negishi
Atsunori
A
D013856:Acidithiobacillus thiooxidans / Q000201:enzymology*; D000968:Antimycin A / Q000737:chemistry; D002462:Cell Membrane / Q000201:enzymology; D003576:Electron Transport Complex IV / Q000737:chemistry; D006418:Heme / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D006912:Hydroxyquinolines / Q000737:chemistry; D008689:Methacrylates / Q000737:chemistry; D009532:Nickel / Q000737:chemistry; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D021122:Protein Subunits / Q000737:chemistry; D012966:Sodium Cyanide / Q000737:chemistry; D013447:Sulfites / Q000737:chemistry*; D013844:Thiazoles / Q000737:chemistry; D017973:Tungsten Compounds / Q000737:chemistry; D014451:Ubiquinone / Q000031:analogs & derivatives / Q000737:chemistry
D006912:Hydroxyquinolines; D008689:Methacrylates; D021122:Protein Subunits; D013447:Sulfites; D013844:Thiazoles; D017973:Tungsten Compounds; D014451:Ubiquinone; C001333:2-(n-heptyl)-4-hydroxyquinoline N-oxide; D006418:Heme; D000968:Antimycin A; C030517:myxothiazol; D009532:Nickel; D010088:Oxidoreductases; C432586:ubiquinol oxidase; D003576:Electron Transport Complex IV; C003741:ubiquinol; D012966:Sodium Cyanide; C045951:tungstate