A Novel Unspecific Peroxygenase from Galatian marginata for Biocatalytic Oxyfunctionalization Reactions.

• A novel peroxygenase from Galerina marginata (GmaUPO) was identified by database mining • GmaUPO was recombinatly expressed in Pichia pastoris initially characertized • GmaUPO catalyzes the H2O2-driven oxyfunctionalisation of alkanes, alkenes, alcohols and thioethers • Molecular modelling puts the basis for a molecular understanding of factors influencing GmaUPO's activity and selectivity Unspecific peroxygenases (UPOs, EC 1.11.2.1) are promising oxyfunctionalization catalysts because of their unique stereoselectivity. However, so far only a few UPOs have been reported. In this study, gene mining was used to identify a gene from Galerina marginata that coded for a novel UPO (Gma UPO). Gma UPO was expressed in Pichia pastoris X-33 by scale-up fermentation (the UPO activity of the culture supernatant was 118 U/L). Gma UPO exhibited a molecular weight of 40 kDa and exhibited highest activity at 35°C and pH 9, respectively. Furthermore, Gma UPO was demonstrated to catalyze the epoxidation, sulfoxidation, and hydroxylation of common substrates, particularly fatty acids such as tridecanoic acid. The molecular basis for Gma UPO regioselectivity for fatty acid hydroxylation was explored by molecular modelling. The regioselectivity was mostly governed by the architecture of the enzyme's active site. [Display omitted] [ABSTRACT FROM AUTHOR]