Cascade Biocatalysis Designed for the Allylic Oxidation of α-Pinene

Abstract

A biocatalytic cascade system using a cocktail of oxidoreductase enzymes (2-1B peroxidase and M120 laccase) was designed for the allylic oxidation of (+)-α-pinene into value-added products (e.g., verbenol and verbenone). The oxidative transformation involved a two-step process as follows: (+)-α-pinene was (i) oxidized on the allylic position with H2O2 mainly assisted by 2-1B peroxidase leading to verbenol as the principal reaction product, and (ii) directed to verbenone in the presence of M120 laccase responsible for further oxidation of verbenol to verbenone. The reaction environment was ensured by the acetate buffer (0.1 M, pH = 5). Optimum values for the experimental parameters (e.g., concentration of 2-1B peroxidase, M120 laccase, and H2O2) were set up. The biocatalytic cascade process was monitored for 24 h in order to evaluate the process pathway. Maximum performance under optimum conditions was reached after 5 h incubation time (e.g., 80% (+)-α-pinene conversion and 70% yield in verbenol). Therefore, the developed biocatalytic cascade system offered promising perspectives for (+)-α-pinene valorization.