TABLE 3

Steady-state kinetics of hAOX1-WT and SNP variant S1271L

Steady-state kinetic parameters were calculated and related to a molybdenum saturation of 100% for hAOX1-WT and hAOX1-S1271L under assay conditions using molecular oxygen (phenanthridine), 1 mM ferricyanide, and 100 μM 2,6 dichlorophenolindophenol as electron acceptors. Data are mean values from three independent measurements (±S.D.).

EnzymeSubstrateElectron AcceptorkcatKMkcat/KM
min−1μMmin−1 μM−1
hAOX1-WT codon-optimizedPhenanthridineMolecular oxygen306.5 ± 11.715.91 ± 2.919.3
Ferricyanide220.5 ± 5.624.85 ± 3.58.8
DCPIP27.5 ± 5.01.8 ± 0.515.3
PhthalazineFerricyanide271.7 ± 13.9196.2 ± 10.81.4
DCPIP27.4 ± 3.722.14 ± 4.11.2
BenzaldehydeFerricyanide187.8 ± 13.980.16 ± 8.42.3
DCPIP56.9 ± 8.612.1 ± 2.74.7
hAOX1-WT native sequence (pSS130)PhenanthridineMolecular oxygen284.8 ± 11.326.53 ± 6.510.7
Ferricyanide292.1 ± 15.825.5 ± 4.811.5
DCPIP26.8 ± 1.40.78 ± 0.134.4
PhthalazineFerricyanide223.3 ± 10.6125.7 ± 4.11.8
DCPIP31.62 ± 3.98.96 ± 0.83.5
BenzaldehydeFerricyanide203.7 ± 12.145.18 ± 6.24.5
DCPIP29.8 ± 1.14.6 ± 0.36.5
hAOX1-S1271LPhenanthridineMolecular oxygen233.36 ± 13.612.03 ± 0.919.4
Ferricyanide124.96 ± 2.515.98 ± 3.97.8
DCPIP15.65 ± 1.31.56 ± 0.110.0
PhthalazineFerricyanide175.6 ± 8.2177.4 ± 12.40.9
DCPIP17.44 ± 0.712.16 ± 0.81.4
BenzaldehydeFerricyanide95.7 ± 5.267.8 ± 10.91.4
DCPIP15.46 ± 1.27.83 ± 0.71.9