Insights in the kinetic mechanism of the eukaryotic Baeyer-Villiger monooxygenase BVMOAf1 from Aspergillus fumigatus Af293

MASCOTTI, ML; KURINA-SANZ, MB; JURI AYUB, M; FRAAIJE, MW

BIOCHIMIE

ELSEVIER FRANCE-EDITIONS SCIENTIFIQUES MEDICALES ELSEVIER

Lugar: Paris; Año: 2014 vol. 107 p. 270 - 276

0300-9084

This work reports a detailed kinetic study of the recently discovered BVMOAf1 from Aspergillus fumigatusAf293. By performing steady state and pre-steady state kinetic analyses, it was demonstrated that therate of catalysis is partially limited by the NADPH-mediated reduction of the flavin cofactor, a uniquehallmark of BVMOAf1. In addition, the oxygenating C4a-(hydro)peroxyflavin intermediate could bespectrophotometrically detected and it was found to be the most stable among all analyzed BVMOs. Toassess the possible influence of some residues on the kinetic features, model-inspired site-directedmutagenesis was performed. Among the mutants, the Q436A variant showed a slightly broader substratescope and a better catalytic efficiency. In summary, this study describes for the first time the kineticparameters for an eukaryotic BVMO.