title: Modification of flavin adenine dinucleotide in alcohol oxidase of the yeast Hansenula polymorpha.
abstract: Alcohol oxidase, a major peroxisomal protein of methanol-utilizing yeasts, may possess two different forms of flavin adenine dinucleotide, classical FAD and so-called modified FAD (mFAD). Conversion of FAD into mFAD was observed both in purified preparations of the enzyme and in cells grown in batch and continuous culture. The relative amount of mFAD in the enzyme varied from 5 to 95%, depending on the growth or storage conditions. The presence of mFAD led to a slight decrease in Vmax and a significant (about one order) decrease in the Km of alcohol oxidase with respect to methanol. The kinetics of modification measured in purified preparations of the enzyme obeyed first-order kinetics (k = 0.78 h-1). The modification process was strongly inhibited by methanol, formaldehyde or hydroxylamine. Modification observed in continuous culture under steady state conditions depended on the dilution rate and could also be described as a spontaneous first-order reaction (kapp = 0.27 h-1). FAD modification could only be detected in alcohol oxidase and not in other yeast peroxisomal flavoenzymes, such as D-amino acid oxidase from Candida boidinii.
authors.affiliation: Department of Microbiology, University of Groningen, Haren, The Netherlands.
authors.forename: L V
authors.initials: LV
authors.lastname: Bystrykh
authors.forename: L
authors.initials: L
authors.lastname: Dijkhuizen
authors.forename: W
authors.initials: W
authors.lastname: Harder
mesh_terms: D000429:Alcohol Oxidoreductases / Q000378:metabolism*; D002175:Candida / Q000378:metabolism; D005182:Flavin-Adenine Dinucleotide / Q000031:analogs & derivatives* / Q000378:metabolism*; D005557:Formaldehyde / Q000494:pharmacology; D005656:Fungal Proteins / Q000378:metabolism*; D019811:Hydroxylamine; D006898:Hydroxylamines / Q000494:pharmacology; D007700:Kinetics; D000432:Methanol / Q000378:metabolism / Q000494:pharmacology; D010843:Pichia / Q000187:drug effects / Q000378:metabolism*
chemical_list: D005656:Fungal Proteins; D006898:Hydroxylamines; C072368:modified FAD; D005182:Flavin-Adenine Dinucleotide; D005557:Formaldehyde; D019811:Hydroxylamine; D000429:Alcohol Oxidoreductases; C019643:alcohol oxidase; D000432:Methanol