Publication
Article Metrics
Citations
Online attention
Structural basis of catalysis in the bacterial monoterpene synthases linalool synthase and 1,8-cineole synthase
Authors:
Vijaykumar
Karuppiah
(Manchester Institute of Biotechnology, University of Manchester)
,
Kara
Ranaghan
(University of Bristol)
,
Nicole G. H.
Leferink
(Manchester Institute of Biotechnology, University of Manchester)
,
Linus O.
Johannissen
(Manchester Institute of Biotechnology, University of Manchester)
,
Muralidharan
Shanmugam
(Manchester Institute of Biotechnology, University of Manchester)
,
Aisling
Ni Cheallaigh
(Manchester Institute of Biotechnology, University of Manchester)
,
Nathan
Bennett
(Manchester Institute of Biotechnology, University of Manchester)
,
Lewis
Kearsey
(Manchester Institute of Biotechnology, University of Manchester)
,
Eriko
Takano
(Manchester Institute of Biotechnology, University of Manchester)
,
John
Gardiner
(Manchester Institute of Biotechnology, University of Manchester)
,
Marc
Van Der Kamp
(University of Bristol)
,
Sam
Hay
(Manchester Institute of Biotechnology, University of Manchester)
,
Adrian J.
Mulholland
(University of Bristol)
,
David
Leys
(Manchester Institute of Biotechnology, University of Manchester)
,
Nigel S.
Scrutton
(Manchester Institute of Biotechnology, University of Manchester)
Co-authored by industrial partner:
No
Type:
Journal Paper
Journal:
Acs Catalysis
State:
Published (Approved)
Published:
August 2017
Diamond Proposal Number(s):
12788

Abstract: Terpenoids form the largest and stereochemically most diverse class of natural products, and there is considerable interest in producing these by biocatalysis with whole cells or purified enzymes, and by metabolic engineering. The monoterpenes are an important class of terpenes and are industrially important as flavours and fragrances. We report here structures for the recently discovered Streptomyces clavuligerus monoterpene synthases linalool synthase (bLinS) and 1,8-cineole synthase (bCinS) and show that these are active biocatalysts for monoterpene production using biocatalysis and metabolic engineering platforms. In metabolically engineered monoterpene-producing E. coli strains use of bLinS leads to 300-fold higher linalool production compared with the corresponding plant monoterpene synthase. With bCinS, 1,8-cineole is produced with 96% purity compared to 67% from plant species. Structures of bLinS and bCinS, and their complexes with fluorinated substrate analogues, show that these bacterial monoterpene synthases are similar to previously characterised sesquiterpene synthases. Molecular dynamics simulations suggest that these monoterpene synthases do not undergo large-scale conformational changes during the reaction cycle making them attractive targets for structured-based protein engineering to expand the catalytic scope of these enzymes towards alternative monoterpene scaffolds. Comparison of the bLinS and bCinS structures indicates how their active sites steer reactive carbocation intermediates to the desired acyclic linalool (bLinS) or bicyclic 1,8-cineole (bCinS) products. The work reported here provides the analysis of structures for this important class of monoterpene synthase. This should now guide exploitation of the bacterial enzymes as gateway biocatalysts for the production of other monoterpenes and monoterpenoids.
Journal Keywords: monoterpene synthase; monoterpenoids; sesquiterpene synthase; terpenes; protein crystallography; molecular dynamics simulations
Diamond Keywords: Bacteria; Enzymes
Subject Areas:
Chemistry,
Biology and Bio-materials
Instruments:
I04-1-Macromolecular Crystallography (fixed wavelength)
,
I04-Macromolecular Crystallography
Added On:
14/08/2017 08:57
Documents:
acscatal4.pdf
Discipline Tags:
Biotechnology
Biochemistry
Catalysis
Chemistry
Structural biology
Engineering & Technology
Life Sciences & Biotech
Technical Tags:
Diffraction
Macromolecular Crystallography (MX)