Projects per year
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.
|Number of pages||7|
|Early online date||3 Dec 2019|
|Publication status||Published - 2020|
Research Beacons, Institutes and Platforms
- Manchester Institute of Biotechnology
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- 3 Finished
Manchester Synthetic Biology Research Centre for Fine and Speciality Chemicals
Scrutton, N., Azapagic, A., Balmer, A., Barran, P., Breitling, R., Delneri, D., Dixon, N., Faulon, J., Flitsch, S., Goble, C., Goodacre, R., Hay, S., Kell, D., Leys, D., Lloyd, J., Lockyer, N., Martin, P., Micklefield, J., Munro, A., Pedrosa Mendes, P., Randles, S., Salehi Yazdi, F., Shapira, P., Takano, E., Turner, N. & Winterburn, J.
14/11/14 → 13/05/20
Innovative routes to monoterpene hydrocarbons and their high value derivatives
Scrutton, N., Breitling, R., Gardiner, J., Hay, S., Leys, D., Pedrosa Mendes, P. & Takano, E.
1/11/14 → 30/04/20
TERPENOSOME: Engineered compartments for monoterpenoid production using synthetic biology
1/07/14 → 30/06/17