Structural insights into the ene-reductase synthesis of profens

Waller, John; Toogood, Helen S.; Karuppiah, V.; Rattray, Nicholas J. W.; Mansell, David; Leys, D.; Gardiner, John M.; Fryszkowska, Anna; Ahmed, Syed Thouheed; Bandichhor, Rakeshwar; Reddy, G. Panduranga; Scrutton, Nigel S.

doi:10.1039/c7ob00163k

Cited by 22 publications

(19 citation statements)

References 60 publications

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“…These KYE2‐catalyzed reduction reactions proceeded in a highly stereoselective manner, giving products with higher than 95 % ee in most cases. The configuration of the major bioreduction product was found to be R , the same as that of the products generated by other OYEs . As reported recently, OYE‐mutant‐catalyzed enantioselective radical dehalogenation of α‐bromoesters could potentially be used to access ( S )‐profens …”

Section: Resultssupporting

confidence: 67%

“…Recent studies have suggested that the R enantiomers of profens, in particular ( R )‐flurbiprofen, have potential to be developed as β‐amyloid‐lowering agents . Several OYEs, including YqjM, have been reported to catalyze the synthesis of profen derivatives with various efficiencies . In order to demonstrate the synthetic potential of KYE2, we sought to apply this enzyme to the chemoenzymatic synthesis of profen methyl esters.…”

Section: Resultsmentioning

confidence: 99%

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Identification of an Ene Reductase from Yeast Kluyveromyces Marxianus and Application in the Asymmetric Synthesis of (R)‐Profen Esters

Wang

Meng

et al. 2018

Asian J Org Chem

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Given their ability to reduce activated alkenes thereby generating up to two stereogenic centers, ene reductases from the old yellow enzyme (OYE) family have received much attention as effective biocatalysts. Through genome mining, a" classical" OYE, KYE2, was identified from Kluyveromycesm arxianus CBS4857 and fully characterizedi n vitro. This NADPH-dependent enzyme displayed ab road substrate spectrumf or the bioreduction of activated alkenes. The highly stereoselective synthesis (> 95 % ee)o fv arious (R)-profen methyl esters, including the synthesis of (R)-flurbiprofen methyl ester on asemi-preparative scale further demonstrates the potential of KYE2 to be developed as ag eneral biocatalyst for chemical synthesis.

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Section: Resultssupporting

confidence: 67%

Section: Resultsmentioning

confidence: 99%

Identification of an Ene Reductase from Yeast Kluyveromyces Marxianus and Application in the Asymmetric Synthesis of (R)‐Profen Esters

Wang

Meng

et al. 2018

Asian J Org Chem

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“…The isolated enzyme screening approach was applied to the bioreduction of the 2-arylpropenoic acids and their esters in the chemoenzymatic synthesis of the pharmacologically-relevant profen class 48. Classically, the reduction of α,β-unsaturated carboxylic acids and esters by ERs is challenging, requiring the presence of a second conjugated electron withdrawing group.…”

Section: Individual and Cascading Biocatalytic Reactionsmentioning

confidence: 99%

Discovery, Characterization, Engineering, and Applications of Ene-Reductases for Industrial Biocatalysis

2018

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Recent studies of multiple enzyme families collectively referred to as ene-reductases (ERs) have highlighted potential industrial application of these biocatalysts in the production of fine and speciality chemicals. Processes have been developed whereby ERs contribute to synthetic routes as isolated enzymes, components of multi-enzyme cascades, and more recently in metabolic engineering and synthetic biology programmes using microbial cell factories to support chemicals production. The discovery of ERs from previously untapped sources and the expansion of directed evolution screening programmes, coupled to deeper mechanistic understanding of ER reactions, have driven their use in natural product and chemicals synthesis. Here we review developments, challenges and opportunities for the use of ERs in fine and speciality chemicals manufacture. The ER research field is rapidly expanding and the focus of this review is on developments that have emerged predominantly over the last 4 years.

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“…Synthetic applications of ene reductases are manifold and range from the preparation of profens [25][26][27] and chiralγ-amino acids [28][29][30] to the synthesis of chiral phosphonates [31] and nitroalkanes [32], precursors in the synthesis of pharmaceutically active ingredients. To further promote an off-the-shelve synthetic use of ene reductases, which can reduce the time and cost of the implementation of a biocatalytic step into a process significantly, we set out to expand the available biocatalytic toolbox [15].…”

Section: Isolated In 1932 Bymentioning

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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Structural insights into the ene-reductase synthesis of profens

Cited by 22 publications

References 60 publications

Identification of an Ene Reductase from Yeast Kluyveromyces Marxianus and Application in the Asymmetric Synthesis of (R)‐Profen Esters

Identification of an Ene Reductase from Yeast Kluyveromyces Marxianus and Application in the Asymmetric Synthesis of (R)‐Profen Esters

Discovery, Characterization, Engineering, and Applications of Ene-Reductases for Industrial Biocatalysis

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

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