Mapping selective oxidations of unspecific peroxygenases

Abstract

Several unspecific peroxygenases (UPOs) have been identified that perform a broad range of selective oxyfunctionalizations and hence represent a pivotal addition to the biocatalysis “toolbox”. To make these “oxidation tools” broadly applicable it is crucial to provide a detailed “user manual” for their substrate preference, chemo- and regioselectivity. We therefore selected 11 different substrates with a panel of 15 diverse UPOs and mapped their preferences. Various UPOs proved to be highly selective — discriminating based on either position or chemical properties of the substrate — with up to 99% chemo- and regioselectivity while achieving turnover numbers (TONs) of a few hundred up to multiple thousands. ChimeraUPO-II was found to oxidize small alkanes selectively at the 2-position with regioselectivity values up to 99%. This map of UPO selectivity shall serve as a starting point for new chemoenzymatic routes and starting points for protein engineering endeavors.