Abstract:In this study, an unspecific peroxygenase AbvbUPO derived from Agaricusbisporus var. bisporus was used as the target protein to achieve heterologous expression in Pichia pastoris GS115. Western blot results showed that the molecular mass of AbvbUPO was 35 ku. Biochemical characterization of the enzyme revealed that the optimal reaction temperature and pH of the unspecific peroxygenase AbvbUPO were 35 ℃ and 3.0, respectively. Hydrogen peroxide tolerance experiments revealed that hydrogen peroxide above 2.0 mM deactivated AbvbUPO. In order to maintain the activity of AbvbUPO in the reaction process, the insitu production of H2O2 occurring in the AbvbUPO enzyme cascade catalytic reaction was used for the identification of its catalytic ability. After the 4 h AbvbUPO enzyme cascade catalytic reaction with ethylbenzene as the substrate, the yield of the product β-phenylethanol reached 14.40%. The above research results show that AbvbUPO was a mesophilic enzyme that could not tolerate high temperature and high concentrations of H2O2, but can maintain good catalytic performance in the cascade reaction of insitu H2O2 production. This study provides a reference for the heterologous expression and biocatalytic application of unspecific peroxygenase.
