Redesigning the Molecular Choreography to Prevent Hydroxylation in Germacradien-11-ol Synthase Catalysis.
| Autor: | Srivastava PL; School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, United Kingdom., Escorcia AM; School of Biochemistry, University of Bristol, University Walk, Bristol BS8 1TD, United Kingdom., Huynh F; School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, United Kingdom., Miller DJ; School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, United Kingdom., Allemann RK; School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff CF10 3AT, United Kingdom., van der Kamp MW; School of Biochemistry, University of Bristol, University Walk, Bristol BS8 1TD, United Kingdom. |
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| Jazyk: | angličtina |
| Zdroj: | ACS catalysis [ACS Catal] 2021 Feb 05; Vol. 11 (3), pp. 1033-1041. Date of Electronic Publication: 2021 Jan 07. |
| DOI: | 10.1021/acscatal.0c04647 |
| Abstrakt: | 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. Competing Interests: The authors declare no competing financial interest. (© 2021 American Chemical Society.) |
| Databáze: | MEDLINE |
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