A computationally designed panel of diverse and selective peroxygenases for terpene oxyfunctionalization
Listed in
This article is not in any list yet, why not save it to one of your lists.Abstract
Enzyme engineering has a critical role in the transition to economical, low-energy and environmentally friendly chemical production. Current approaches relying on costly iterations of mutation and selection are limited to reactions with a straightforward experimental readout and struggle to address mutational epistasis. We focus on unspecific peroxygenases (UPOs), prized engineering targets due to their ability to oxyfunctionalize diverse organic molecules of industrial and environmental value. To address the lack of scalable screening for UPO functions, we applied enzyme-design calculations to focus experiments. Starting from an AlphaFold2 model of Mth UPO, the automated FuncLib algorithm generated 50 diverse active-site multipoint designs—all of which were functional. Screening against nine diverse terpenes revealed large improvements and new oxyfunctionalization products, resulting in molecules of high pharmaceutical and industrial value. This work demonstrates that computational design can overcome the shortcomings of traditional enzyme engineering and accelerate the urgently needed green transition of the chemical industry.
BIGGER PICTURE
Enzymes enable energy- and resource-efficient chemical reactions and are key players in the drive to a sustainabil chemical industry. But natural enzymes are seldom optimized for industrial use, demanding optimization by enzyme engineers. Traditional enzyme-engineering approaches, however, are typically time-consuming, costly and struggle to address the complexity of implementing multiple mutations in an enzyme active site. To address these challenges, we apply AI-based structure modeling and computational design calculations to the active site of an enzyme belonging to the unspecific peroxygenase (UPO) family, which are highly prized in the synthesis of valuable flavor, fragrance, and medicinal molecules. Strikingly, the 50 active-site designs we tested were all highly functional and many exhibited different activity profiles and the production of commercially valuable molecules.
This work demonstrates that computational design can complement traditional methods for enzyme engineering to accelerate the urgently needed green transition of the chemical industry.
HIGHLIGHTS
-
One-shot computational design of 50 functional UPO designs
-
Diverse oxyfunctionalization products of different terpene substrates
-
Remarkable improvement in activity, regio-, chemo- and enantioselectivity
-
Valuable production of commercially relevant molecules
