Biochemical and electrochemical characterization of quinohemoprotein amine dehydrogenase from Paracoccus denitrificans.
A new quinohemoprotein amine dehydrogenase from Paracoccus denitrificans IFO 12442 was isolated and characterized in views of biochemistry and electrochemistry. This enzyme exists in periplasm and catalyzes the oxidative deamination of primary aliphatic and aromatic amines. n-Butylamine or benzylamine as a carbon and energy source strongly induces the expression of the enzyme. Carbonyl reagents inhibit the enzyme activity irreversibly. This enzyme is a heterodimer constituted of alpha and beta subunits with the molecular mass of 59.5 and 36.5 kDa, respectively. UV-vis and EPR spectroscopy, and the quinone-dependent redox cycling and heme-dependent peroxidative stains of SDS-PAGE bands revealed that the alpha subunit contains one quinonoid cofactor and one heme c per molecule, while the beta subunit has no prosthetic group. The redox potential of the heme c moiety was determined to be 0.192 V vs NHE at pH 7.0 by a mediator-assisted continuous-flow column electrolytic spectroelectrochemical technique. The analysis of the substrate titration curve allowed the evaluation of the redox potential of the quinone/semiquinone and semiquinone/quinol redox couples as 0.19 and 0.11 V, respectively.
Takagi
K
K
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Japan.
Torimura
M
M
Kawaguchi
K
K
Kano
K
K
Ikeda
T
T
D004227:Dithionite / Q000737:chemistry; D004563:Electrochemistry; D004578:Electron Spin Resonance Spectroscopy; D006418:Heme / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D010084:Oxidation-Reduction; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D010231:Paracoccus denitrificans / Q000201:enzymology*; D019897:Periplasm / Q000201:enzymology; D011809:Quinones / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity; D017186:Titrimetry
D011809:Quinones; D004227:Dithionite; C031690:heme C; D006418:Heme; D000587:Oxidoreductases Acting on CH-NH Group Donors; C119497:quinohemoprotein amine dehydrogenase