Open AccessPurification and characterization of histamine dehydrogenase from Nocardioides simplex IFO 12069
Author(s) -
Siddiqui Jahan Ara,
Shoeb Syed Mohammed,
Takayama Shigeo,
Shimizu Eiichi,
Yorifuji Takamitsu
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09227.x
Subject(s) - oxidative deamination , chemistry , enzyme , nuclear chemistry , chromatography , biochemistry , stereochemistry
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 μmol 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.