Ene‐reductases and their Applications

Knaus, Tanja; Toogood, Helen S.; Scrutton, Nigel S.

doi:10.1002/9781118828083.ch18

Cited by 31 publications

(30 citation statements)

References 72 publications

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“…9 As the 2 H on C3 has higher chiral priority than the 1 H, the product may be assigned as [2R, 3R-2 H2]-(5R)-dihydrocarvone. 10 Hence, the two 2 H nuclei are added in an antirelationship across C2 and C3, which is in accordance with previous mechanistic studies of similar ene reductase enzymes. 9,10 Finally…”

Section: S363 Results A: [2 3-2 H2]-(2r 5r)-dihydrocarvone (5a)supporting

confidence: 88%

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Bringing Biocatalysis into the Deuteration Toolbox

Rowbotham¹,

Lenz²,

Reeve³

et al. 2019

Preprint

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Chemicals labelled with the heavy hydrogen isotope deuterium (2H) have long been used in chemical and biochemical mechanistic studies, spectroscopy, and as analytical tracers. More recently, demonstration of selectively deuterated drug candidates that exhibit advantageous pharmacological traits has spurred innovations in metal-catalysed 2H insertion at targeted sites, but asymmetric deuteration remains a key challenge. Here we demonstrate an easy-to-implement biocatalytic deuteration strategy, achieving high chemo-, enantio- and isotopic selectivity, requiring only 2H2O (D2O) and unlabelled dihydrogen under ambient conditions. The vast library of enzymes established for NADH-dependent C=O, C=C, and C=N bond reductions have yet to appear in the toolbox of commonly employed 2H-labelling techniques due to requirements for suitable deuterated reducing equivalents. By facilitating transfer of deuterium atoms from 2H2O solvent to NAD+, with H2 gas as a clean reductant, we open up biocatalysis for asymmetric reductive deuteration as part of a synthetic pathway or in late stage functionalisation. We demonstrate enantioselective deuteration via ketone and alkene reductions and reductive amination, as well as exquisite chemo-control for deuteration of compounds with multiple unsaturated sites.

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Section: S363 Results A: [2 3-2 H2]-(2r 5r)-dihydrocarvone (5a)supporting

confidence: 88%

“…10 Hence, the two 2 H nuclei are added in an antirelationship across C2 and C3, which is in accordance with previous mechanistic studies of similar ene reductase enzymes. 9,10 Finally…”

Section: S363 Results A: [2 3-2 H2]-(2r 5r)-dihydrocarvone (5a)supporting

confidence: 88%

Bringing Biocatalysis into the Deuteration Toolbox

Rowbotham¹,

Lenz²,

Reeve³

et al. 2019

Preprint

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“…Thus, ene reductases offer a valuable access route to asymmetric compounds, which is complementary to the chemical cis-hydrogenation catalyzed by chiral rhodium or ruthenium phosphine catalysts [10,11]. Today, ene reductases are classified into five enzyme groups, which differ in structure, reaction mechanism, substrate spectrum, and stereoselectivity ( Figure 1) [12]. While enoate reductases, medium-and short-chain dehydrogenases/reductases (MDR and SDR), as well as the recently discovered quinone Figure 1.…”

Section: Introductionmentioning

confidence: 99%

Characterization of the Novel Ene Reductase Ppo-Er1 from Paenibacillus Polymyxa

2020

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Ene reductases enable the asymmetric hydrogenation of activated alkenes allowing the manufacture of valuable chiral products. The enzymes complement existing metal- and organocatalytic approaches for the stereoselective reduction of activated C=C double bonds, and efforts to expand the biocatalytic toolbox with additional ene reductases are of high academic and industrial interest. Here, we present the characterization of a novel ene reductase from Paenibacillus polymyxa, named Ppo-Er1, belonging to the recently identified subgroup III of the old yellow enzyme family. The determination of substrate scope, solvent stability, temperature, and pH range of Ppo-Er1 is one of the first examples of a detailed biophysical characterization of a subgroup III enzyme. Notably, Ppo-Er1 possesses a wide temperature optimum (Topt: 20–45 °C) and retains high conversion rates of at least 70% even at 10 °C reaction temperature making it an interesting biocatalyst for the conversion of temperature-labile substrates. When assaying a set of different organic solvents to determine Ppo-Er1′s solvent tolerance, the ene reductase exhibited good performance in up to 40% cyclohexane as well as 20 vol% DMSO and ethanol. In summary, Ppo-Er1 exhibited activity for thirteen out of the nineteen investigated compounds, for ten of which Michaelis–Menten kinetics could be determined. The enzyme exhibited the highest specificity constant for maleimide with a kcat/KM value of 287 mM−1 s−1. In addition, Ppo-Er1 proved to be highly enantioselective for selected substrates with measured enantiomeric excess values of 92% or higher for 2-methyl-2-cyclohexenone, citral, and carvone.

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“…Ene-reductases (ERs) are a large class of enzymes characterized by their capacity to reduce activated olefins including enones, enals, unsaturated carboxylic acids, esters and cyclic imides. 17 Enantioenriched amino acid derivatives, important polymer precursors, and chiral building blocks have been synthesized using ERs as a key synthetic step. 17 The Old Yellow Enzyme (OYE) family of flavoproteins belong to this class and have been employed in a large number of reaction cascades and industrially-relevant processes due to their substrate promiscuity and excellent stereoselectivity.…”

Section: Introductionmentioning

confidence: 99%

“…17 Enantioenriched amino acid derivatives, important polymer precursors, and chiral building blocks have been synthesized using ERs as a key synthetic step. 17 The Old Yellow Enzyme (OYE) family of flavoproteins belong to this class and have been employed in a large number of reaction cascades and industrially-relevant processes due to their substrate promiscuity and excellent stereoselectivity. 18,19 The mechanism of OYEs in the asymmetric reduction of olefins has been well-studied, 20 and is understood to involve the delivery of a hydride from flavin mononucleotide (FMNH 2 , 6 ) to the β-position of the substrate which is coordinated by a strictly conserved histidine/asparagine or histidine/histidine pair (Figure 3A).…”

Section: Introductionmentioning

confidence: 99%