PUB - Publikationen an der Universität Bielefeld

The interconversion of monoterpenes is facilitated by a complex network of carbocation rearrangement pathways. Controlling these isomerization pathways is challenging when using common Bronsted and Lewis acid catalysts, which often produce product mixtures that are difficult to separate. In contrast, natural monoterpene cyclases exhibit high control over the carbocation rearrangement reactions but are reliant on phosphorylated substrates.In this study, we present engineered squalene-hopene cyclases fromAlicyclobacillus acidocaldarius(AacSHC) that catalyze the challenging isomerization of monoterpenes with unprecedented precision.Starting from a promiscuous isomerization of (+)-beta-pinene, we first demonstrate noticeable shifts in the product distribution solely by introducing single point mutations. Furthermore, we showcase the tuneable cation steering by enhancing (+)-borneol selectivity from 1% to >90% (>99%de) aided by iterative saturation mutagenesis. Our combined experimental and computational datasuggeststhat the reorganization of key aromatic residues leads to the restructuring of the water network that facilitates the selective termination of the secondary isobornyl cation. This work expands our mechanistic understanding of carbocation rearrangements and sets the stage for target-oriented skeletal reorganization of broadly abundant terpenes. © 2024 Wiley-VCH GmbH.