TABLE 1

Steady-state kinetic parameters hAOX1 wild type and the L438V variant with different substrates

Steady-state kinetic parameters were corrected to a molybdenum saturation of 100%. Kinetic parameters were recorded in 50 mM Tris-HCl, 200 mM NaCl, and 1 mM EDTA (pH 8.0) in the presence of 250 µM O2 as electron acceptor using 200–400 nM enzyme. Substrate concentration were varied using 5–200 μM for benzaldehyde, 5–120 μM for phthalazine, and 75–600 μM for zoniporide. Data are mean values from three independent measurements (±S.D.).

EnzymeBenzaldehydePhthalazineZoniporide
hAOX1-WTkcat (min−1)14.87 ± 1.42123.11 ± 1.020.43 ± 0.03
KM (µM)104.98 ± 19.96201.56 ± 31.98623.82 ± 9.09
hAOX1-L438Vkcat (min−1)10.58 ± 1.0789.28 ± 1.140.42 ± 0.02
KM (µM)101.48 ± 21.01117.06 ± 28.92573.41 ± 37.51