A Novel Unspecific Peroxygenase from Galatian marginata for Biocatalytic Oxyfunctionalization Reactions
Yunjian Ma (Macau University of Science and Technology, South China University of Technology)
Hongjing Liang (South China University of Technology)
Zexin Zhao (Hubei University of Technology)
Bin Wu (South China University of Technology)
Dongming Lan (South China University of Technology)
Frank Hollmann (TU Delft - Applied Sciences)
Yonghua Wang (South China University of Technology, Guangdong Youmei Institute of Inteligent Bio-manufacturing Co, Guangdong)
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Abstract
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 (GmaUPO). GmaUPO was expressed in Pichia pastoris X-33 by scale-up fermentation (the UPO activity of the culture supernatant was 118 U/L). GmaUPO exhibited a molecular weight of 40 kDa and exhibited highest activity at 35°C and pH 9, respectively. Furthermore, GmaUPO was demonstrated to catalyze the epoxidation, sulfoxidation, and hydroxylation of common substrates, particularly fatty acids such as tridecanoic acid. The molecular basis for GmaUPO regioselectivity for fatty acid hydroxylation was explored by molecular modelling. The regioselectivity was mostly governed by the architecture of the enzyme's active site.
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