Active site loop engineering abolishes water capture in hydroxylating sesquiterpene synthases

Srivastava, PrabhakarL. ORCID: https://orcid.org/0000-0002-8219-6419, Johns, Sam T., Walters, Rebecca, Miller, David J., Van der Kamp, Marc W. and Allemann, Rudolf K. ORCID: https://orcid.org/0000-0002-1323-8830 2023. Active site loop engineering abolishes water capture in hydroxylating sesquiterpene synthases. ACS Catalysis 13 (21) , pp. 14199-14204. 10.1021/acscatal.3c03920

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Abstract

Terpene synthases (TS) catalyze complex reactions to produce a diverse array of terpene skeletons from linear isoprenyl diphosphates. Patchoulol synthase (PTS) from Pogostemon cablin converts farnesyl diphosphate into patchoulol. Using simulation-guided engineering, we obtained PTS variants that eliminate water capture. Further, we demonstrate that modifying the structurally conserved Hα-1 loop also reduces hydroxylation in PTS, as well as in germacradiene-11-ol synthase (Gd11olS), leading to cyclic neutral intermediates as products, including α-bulnesene (PTS) and isolepidozene (Gd11olS). Hα-1 loop modification could be a general strategy for engineering sesquiterpene synthases to produce complex cyclic hydrocarbons without the need for structure determination or modeling.

Item Type:	Article
Date Type:	Published Online
Status:	Published
Schools:	Chemistry
Additional Information:	License information from Publisher: LICENSE 1: URL: https://creativecommons.org/licenses/by/4.0/, Start Date: 2023-10-20
Publisher:	American Chemical Society
ISSN:	2155-5435
Funders:	BBSRC
Date of First Compliant Deposit:	7 November 2023
Date of Acceptance:	12 October 2023
Last Modified:	16 Jan 2024 02:21
URI:	https://orca.cardiff.ac.uk/id/eprint/163700

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