5NX4

Crystal structure of Linalool/Nerolidol synthase from Streptomyces clavuligerus

Summary for 5NX4

• Entry DOI: 10.2210/pdb5nx4/pdb
• Descriptor: Pentalenene synthase, GLYCEROL (3 entities in total)
• Functional Keywords: terpene synthase, linalool, nerolidol, monoterpenoid, ligase
• Biological source: Streptomyces clavuligerus
• Total number of polymer chains: 2
• Total formula weight: 73452.67

Authors

Karuppiah, V.,Leys, D.,Scrutton, N.S. (deposition date: 2017-05-09, release date: 2017-09-20, Last modification date: 2024-01-17)

Primary citation

Karuppiah, V.,Ranaghan, K.E.,Leferink, N.G.H.,Johannissen, L.O.,Shanmugam, M.,Ni Cheallaigh, A.,Bennett, N.J.,Kearsey, L.J.,Takano, E.,Gardiner, J.M.,van der Kamp, M.W.,Hay, S.,Mulholland, A.J.,Leys, D.,Scrutton, N.S.
Structural Basis of Catalysis in the Bacterial Monoterpene Synthases Linalool Synthase and 1,8-Cineole Synthase.
ACS Catal, 7:6268-6282, 2017

PubMed 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 flavors and fragrances. We report here structures for the recently discovered monoterpene synthases linalool synthase (bLinS) and 1,8-cineole synthase (bCinS), and we show that these are active biocatalysts for monoterpene production using biocatalysis and metabolic engineering platforms. In metabolically engineered monoterpene-producing 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 characterized 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 toward 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.

PubMed: 28966840
DOI: 10.1021/acscatal.7b01924

Experimental method

X-RAY DIFFRACTION (2.38 Å)