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Redesigning the molecular choreography to prevent hydroxylation in Germacradien-11-ol synthase catalysis

Srivastava, Prabhakar L., Escorcia, Andrés M., Huynh, Florence, Miller, David J., Allemann, Rudolf K. and van der Kamp, Marc W. 2021. Redesigning the molecular choreography to prevent hydroxylation in Germacradien-11-ol synthase catalysis. ACS Catalysis 11 (3) , 1033–1041. 10.1021/acscatal.0c04647

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Abstract

Natural sesquiterpene synthases have evolved to make complex terpenoids by quenching reactive carbocations either by proton transfer or by hydroxylation (water capture), depending on their active site. Germacradien-11-ol synthase (Gd11olS) from Streptomyces coelicolor catalyzes the cyclization of farnesyl diphosphate (FDP) into the hydroxylated sesquiterpene germacradien-11-ol. Here, we combine experiment and simulation to guide the redesign of its active site pocket to avoid hydroxylation of the product. Molecular dynamics simulations indicate two regions between which water molecules can flow that are responsible for hydroxylation. Point mutations of selected residues result in variants that predominantly form a complex nonhydroxylated product, which we identify as isolepidozene. Our results indicate how these mutations subtly change the molecular choreography in the Gd11olS active site and thereby pave the way for the engineering of terpene synthases to make complex terpenoid products.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Chemistry
Additional Information: Made available through a Creative Commons CC-BY License
Publisher: American Chemical Society
ISSN: 2155-5435
Funders: BBSRC
Date of First Compliant Deposit: 9 February 2021
Date of Acceptance: 7 January 2021
Last Modified: 18 Mar 2021 12:06
URI: http://orca.cardiff.ac.uk/id/eprint/138382

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