Oxidation of Coniferyl Alcohol Catalyzed by Laccases from Trametes versicolor

Oxidation of coniferyl alcohol catalyzed by commercial laccase and crude laccase produced during the submerged cultivation of Trametes versicolor MZKI G-99 in a medium containing the waste from paper industry was investigated. pH of 6.6 and temperature of 35 °C was found to be optimal for coniferyl alcohol oxidation catalyzed by commercial laccase. Based on the initial reaction rate measurements, apparent Michaelis-Menten kinetic parameters for commercial laccase were determined in an aqueous media (Vm = 4.387 U mg–1, Km = 0.025 mmol dm–3), as well as in 1:1 (v/v) methanol: phosphate buffer mixture (Vm = 0.979 U mg–1, Km = 0.019 mmol dm–3). Inhibition of substrate was found for crude laccase and the following apparent kinetic parameters Vm = 9.272 U mg–1, Km = 0.045 mmol dm–3 and Ki = 0.002 mmol dm–3 were estimated. Mathematical model of batch process, which includes double-substrate Michaelis-Menten kinetics with oxygen as the second substrate and mass balances, has been developed and validated in experiments with or without additional aeration. 100 % conversions of up to 0.8 mmol dm–3 of coniferyl alcohol in batch experiment due to the high operational stability of enzymes was realized with both laccases.