Taming the reactivity of monoterpene synthases to guide regioselective product hydroxylation

Research output: Contribution to journalArticle

  • External authors:
  • Nicole Leferink
  • Kara E Ranaghan
  • Jaime Battye
  • Linus Johannissen
  • Marc W. van der Kamp
  • Adrian J. Mulholland

Abstract

Monoterpenoids are industrially important natural products with applications in the flavours, fragrances, fuels and pharmaceutical industries. Most monoterpenoids are produced by plants, but recently two bacterial monoterpene synthases have been identified, including a cineole synthase (bCinS). Unlike plant cineole synthases, bCinS is capable of producing nearly pure cineole from geranyl diphosphate in a complex cyclisation cascade that is tightly controlled. Here we have used a multidisciplinary approach to show that Asn305 controls water attack on the α‐terpinyl cation and subsequent cyclisation and depro‐tonation of the α‐terpineol intermediate, key steps in the cyclisation cascade which direct product formation towards cineole. Mutation of Asn305 results in variants that no longer produce α‐terpineol or cineole. MD simulations revealed that water coordination is disrupted in all variants tested. QM calculations indicate that Asn305 is most likely a (transient) proton acceptor for the final deprotonation step. Our synergistic approach gives unique insights into how a single residue, Asn305, tames the promiscuous chemistry of monoterpene synthase cyclisation cascades. It does this by tightly controlling the final steps in cineole formation catalysed by bCinS to form a single hydroxylated monoterpene product.

Bibliographical metadata

Original languageEnglish
Pages (from-to)1-7
Number of pages7
JournalCHEMBIOCHEM
Volume20
Early online date3 Dec 2019
DOIs
Publication statusPublished - 2020

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