Substrate, solvent, and multiple KIE on AAO steady-state constants for p-anisyl alcohol and 2,4-hexadien-1-ol oxidation
Assays were performed in normal and deuterated 100 mm phosphate, pH 6 (at 25 °C, under air saturation conditions) using the α-protiated and α-bideuterated alcohol substrates. Different KIE estimated: Dk, substrate KIE in water (i.e. k for α-protiated substrate in water/k for α-bideuterated substrate in water); Dk(D2O), substrate KIE in D2O (i.e. k for α-protiated substrate in deuterium oxide/k for α-bideuterated substrate in D2O); D2Ok, solvent KIE using α-protiated substrate (i.e. k for α-protiated substrate in water/k for α-protiated substrate in D2O); D2Ok(D), solvent KIE using α-bideuterated substrate (i.e. k for α-bideuterated substrate in water/k for α-bideuterated substrate in D2O);D,D2Ok, simultaneous substrate and solvent KIE (i.e. k for α-protiated substrate in water/k for α-bideuterated substrate in D2O). The experimental kinetic parameters for Vmax and Km used to calculate the KIE are reported in supplemental Table S1.