Purification and characterization of histamine dehydrogenase from Nocardioides simplex IFO 12069.
Histamine dehydrogenase from Nocardioides simplex IFO 12069 was purified to homogeneity. The enzyme had a molecular mass of 170 kDa and was suggested to be a dimer of subunits that had a molecular mass of 84 kDa. The enzyme showed highest activity toward histamine and produced ammonia in its oxidative deamination to imidazole acetaldehyde. The K(m) and V(max) values for histamine were 0.075 mM and 4.76 micromol min(-1) mg(-1), respectively. The enzyme was sensitive to the carbonyl reagent iproniazid and a structurally similar compound, tryptophan. The enzyme showed absorption maxima at 442 and 280 nm. Reduction with histamine under anaerobic conditions resulted in a different absorption maximum at 360 nm instead of 442 nm. The enzyme was most active at pH 8.5 in Tris-HCl buffer and most stable at pH 7.0 in potassium phosphate buffer. The E(1%) value of the enzyme was 8.6 at 280 nm.
Siddiqui
J A
JA
Department of Bioscience and Biotechnology, Faculty of Agriculture, Shinshu University, Minamiminowa, 399-4598, Nagano-ken, Japan.
Shoeb
S M
SM
Takayama
S
S
Shimizu
E
E
Yorifuji
T
T
D006631:Amine Oxidase (Copper-Containing) / Q000302:isolation & purification* / Q000378:metabolism*; D002384:Catalysis; D009618:Nocardiaceae / Q000201:enzymology*; D013379:Substrate Specificity
D006631:Amine Oxidase (Copper-Containing)