TABLE 3

Kinetic parameters of native EcAAO and evolved AAO (FX7 mutant)a

SubstrateKinetic constantEcAAOFX7
MeanSEMMeanSEM
p-Methoxybenzyl alcoholKm (mM)0.0350.0010.0340.001
kcat (s−1)1056544
kcat/Km (s−1 mM−1)297966156244
Veratryl alcoholKm (mM)0.5040.0430.3880.023
kcat (s−1)662281
kcat/Km (s−1 mM−1)1318713
Benzyl alcoholKm (mM)0.7700.0110.5100.001
kcat (s−1)221191
kcat/Km (s−1 mM−1)282363
2,4-Hexadien-1-olKm (mM)0.0870.0010.0590.004
kcat (s−1)1363521
kcat/Km (s−1 mM−1)15556786653
  • a Steady-state constants were estimated in 100 mM sodium phosphate buffer (pH 6) at 24°C. All reactions were performed in triplicate.