Chemoenzymatic synthesis of the alarm pheromone (+)-verbenone from geranyl diphosphate

Yoosuf-Aly, Zulfa; Faraldos, Juan A.; Miller, David J.; Allemann, Rudolf Konrad

doi:10.1039/c2cc32883f

Cited by 11 publications

(3 citation statements)

References 28 publications

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“…Terpene synthases generate many high‐value products with applications, for instance, as drugs, agrochemicals or fragrances. Understanding the intricate details of their catalytic mechanism will lead to improved methods for the production of naturally occurring terpenes and help the development of designer products with new or improved properties …”

Section: Resultsmentioning

confidence: 99%

Nucleophilic Water Capture or Proton Loss: Single Amino Acid Switch Converts δ‐Cadinene Synthase into Germacradien‐4‐ol Synthase

et al. 2017

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Abstractδ‐Cadinene synthase is a sesquiterpene cyclase that utilises the universal achiral precursor farnesyl diphosphate (FDP) to generate predominantly the bicyclic sesquiterpene δ‐cadinene and about 2 % germacradien‐4‐ol, which is also generated from FDP by the cyclase germacradien‐4‐ol synthase. Herein, the mechanism by which sesquiterpene synthases discriminate between deprotonation and reaction with a nucleophilic water molecule was investigated by site‐directed mutagenesis of δ‐cadinene synthase. If W279 in δ‐cadinene synthase was replaced with various smaller amino acids, the ratio of alcohol versus hydrocarbon product was directly proportional to the van der Waals volume of the amino acid side chain. DCS‐W279A is a catalytically highly efficient germacradien‐4‐ol synthase (k cat/K M=1.4×10−3 μm s−1) that produces predominantly germacradien‐4‐ol in addition to 11 % δ‐cadinene. Water capture is not achieved through strategic positioning of a water molecule in the active site, but through a coordinated series of loop movements that allow bulk water access to the final carbocation in the active site prior to product release.

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Section: Resultsmentioning

confidence: 99%

Nucleophilic Water Capture or Proton Loss: Single Amino Acid Switch Converts δ‐Cadinene Synthase into Germacradien‐4‐ol Synthase

et al. 2017

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“…Stereospecific carbon–carbon bond formation is of high importance in synthetic organic chemistry and biocatalysis. , Terpene cyclases are of high synthetic interest − as they enable stereospecific polycyclizations that are challenging to achieve by synthetic chemistry . Therefore, an enhanced understanding of the reaction mechanism displayed by these fascinating enzymes would be beneficial for chemical and biotechnological industries.…”

Section: Introductionmentioning

confidence: 99%

MD Simulations Reveal Complex Water Paths in Squalene–Hopene Cyclase: Tunnel-Obstructing Mutations Increase the Flow of Water in the Active Site

et al. 2017

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Squalene–hopene cyclase catalyzes the cyclization of squalene to hopanoids. A previous study has identified a network of tunnels in the protein, where water molecules have been indicated to move. Blocking these tunnels by site-directed mutagenesis was found to change the activation entropy of the catalytic reaction from positive to negative with a concomitant lowering of the activation enthalpy. As a consequence, some variants are faster and others are slower than the wild type (wt) in vitro under optimal reaction conditions for the wt. In this study, molecular dynamics (MD) simulations have been performed for the wt and the variants to investigate how the mutations affect the protein structure and the water flow in the enzyme, hypothetically influencing the activation parameters. Interestingly, the tunnel-obstructing variants are associated with an increased flow of water in the active site, particularly close to the catalytic residue Asp376. MD simulations with the substrate present in the active site indicate that the distance for the rate-determining proton transfer between Asp376 and the substrate is longer in the tunnel-obstructing protein variants than in the wt. On the basis of the previous experimental results and the current MD results, we propose that the tunnel-obstructing variants, at least partly, could operate by a different catalytic mechanism, where the proton transfer may have contributions from a Grotthuss-like mechanism.

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“…Allemann and co-workers have shown that high-value terpenoids can be easily prepared in a one-pot chemoenzymatic approach. 28 Geranyl diphosphate was converted to (+)-apinene using the Mg 2+ -dependent monoterpene cyclase, a-pinene synthase (APS) (Scheme 4). Allylic oxidation then completed the one-pot process giving (+)-verbenone in isolated yields of up to 50% over the two steps.…”

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Hill

Sutherland

2012

Nat. Prod. Rep.

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Chemoenzymatic synthesis of the alarm pheromone (+)-verbenone from geranyl diphosphate

Abstract: The enzyme-guided asymmetric synthesis of (+)-verbenone from geranyl diphosphate in a simple two-step, one pot transformation highlights the potential of chemoenzymatic procedures for the generation of high-value terpenoids.

Cited by 11 publications

References 28 publications

Nucleophilic Water Capture or Proton Loss: Single Amino Acid Switch Converts δ‐Cadinene Synthase into Germacradien‐4‐ol Synthase

Nucleophilic Water Capture or Proton Loss: Single Amino Acid Switch Converts δ‐Cadinene Synthase into Germacradien‐4‐ol Synthase

MD Simulations Reveal Complex Water Paths in Squalene–Hopene Cyclase: Tunnel-Obstructing Mutations Increase the Flow of Water in the Active Site

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