Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (R)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis

Su, Bing-Mei; Xu, Liang; Xu, Xinqi; Wang, Lichao; Li, Aipeng; Lin, Juan; Ye, Lidan; Yu, Hongwei

doi:10.1016/j.gresc.2020.09.003

Cited by 35 publications

(21 citation statements)

References 34 publications

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“…Molecular dynamics simulations : The complex treatment before the MD simulation, including the construction of force fields for micro molecules, topology files for system and the three‐stage energy minimization were performed as previous described [9a] . After the minimization, the system was heated from 0 to 300 K at constant volume in 50 ps, equilibrated at constant pressure in 50 ps, and equilibrated for 500 ps.…”

Section: Methodsmentioning

confidence: 99%

Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Wang

et al. 2021

Chemistry A European J

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l‐Threonine aldolase from Actinocorallia herbida (AhLTA) is an ideal catalyst for producing l‐threo‐4‐methylsulfonylphenylserine [(2S,3R)‐1 b], a key chiral precursor for florfenicol and thiamphenicol. The moderate Cβ stereoselectivity is the main obstacle to the industrial application of AhLTA. To address this issue, a combinatorial active‐site saturation test (CAST) together with sequence conservatism analysis was applied to engineer the AhLTA toward improved Cβ stereoselectivity. The optical mutant Y314R could asymmetrically synthesize l‐threo‐4‐methylsulfonylphenylserine with 81 % diastereomeric excess (de), which is 23 % higher than wild‐type AhLTA. Molecular dynamic (MD) simulations revealed that the mechanism for the improvement in Cβ stereoselectivity of Y314R is due to the acylamino group of residues Arg314 controlling the orientation of substrate 4‐methylsulfonyl benzaldehyde (1 a) in the active pocket by directed interaction with the methylsulfonyl group; this leads to asymmetric synthesis of l‐threo‐4‐methylsulfonylphenylserine. The success in this study demonstrates that direct control of substrates in an active pocket is an attract strategy to address the Cβ stereoselectivity problem of LTA and contribute to the industrial application of LTA.

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Section: Methodsmentioning

confidence: 99%

Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Wang

et al. 2021

Chemistry A European J

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“…To alleviate this inconvenience, use of KREDs recombinantly overexpressed in E. coli should be a viable option. Due to their excellent stereoselectivity, broad substrate spectrum, good stability, and high volumetric productivity, such over-expressed KREDs either in isolated form (purified enzyme or crude-cell lysate) or in whole-cell form, have been widely adopted in the synthesis of pharmaceuticals and bioactive molecules 43,45,[70][71][72] . To the best of our knowledge, recombinantly over-expressed KRED-catalyzed stereoselective reduction has not yet been reported for PG synthesis.…”

Section: Resultsmentioning

confidence: 99%

Unified Strategy to Prostaglandins: Chemoenzymatic Total Synthesis of Cloprostenol, Bimatoprost, PGF2α, Fluprostenol, and Travoprost Guided by Biocatalytic Retrosynthesis

Zhu

Jiang

et al. 2021

Preprint

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Development of efficient and stereoselective synthesis of prostaglandins (PGs) is of utmost importance, owing to their valuable medicinal applications and unique chemical structures. We report here a unified synthesis of PGs cloprostenol, bimatoprost, PGF2α, fluprostenol, and travoprost from the readily available dichloro-containing bicyclic ketone 6a guided by biocatalytic retrosynthesis, in 11-12 steps with 2.9-6.5% overall yields. A Baeyer-Villiger monooxygenase (BVMO)-catalyzed stereoselective oxidation of 6a (99% ee), and a ketoreductase (KRED)-catalyzed diastereoselective reduction of enones 12 (87 : 13 to 99 : 1 dr) were utilized in combination for the first time to set the critical stereochemical configurations under mild conditions. Another key transformation was the copper (II)-catalyzed regioselective p-phenylbenzoylation of the secondary alcohol of diol 10 (3.8 : 1 rr). This study not only provides an alternative route to the highly stereoselective synthesis of PGs, but also showcases the usefulness and great potential of biocatalysis in construction of complex molecules.

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“…In the same line, Yu and co‐workers recently have significantly improved the stereoselectivity of the ketoreductase EbSDR in the synthesis of ( R )‐HPBE ( 17 ) by using a computation‐based design approach [22] . According to per‐residue free energy decomposition and sequence conservation analysis, the authors identified key residues involved in the enantioselectivity control.…”

Section: Creation Of One Stereocenter Through Kred‐catalyzed Reductionmentioning

confidence: 99%

Application of Ketoreductase in Asymmetric Synthesis of Pharmaceuticals and Bioactive Molecules: An Update (2018–2020)

Yang

Liu

et al. 2021

The Chemical Record

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With the rapid development of genomic DNA sequencing, recombinant DNA expression, and protein engineering, biocatalysis has been increasingly and widely adopted in the synthesis of pharmaceuticals, bioactive molecules, fine chemicals, and agrochemicals. In this review, we have summarized the most recent advances achieved (2018–2020) in the research area of ketoreductase (KRED)‐catalyzed asymmetric synthesis of chiral secondary alcohol intermediates to pharmaceuticals and bioactive molecules. In the first part, synthesis of chiral alcohols with one stereocenter through the bioreduction of four different ketone classes, namely acyclic aliphatic ketones, benzyl or phenylethyl ketones, cyclic aliphatic ketones, and aryl ketones, is discussed. In the second part, KRED‐catalyzed dynamic reductive kinetic resolution and reductive desymmetrization are presented for the synthesis of chiral alcohols with two contiguous stereocenters.

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Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (R)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis

Cited by 35 publications

References 34 publications

Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Unified Strategy to Prostaglandins: Chemoenzymatic Total Synthesis of Cloprostenol, Bimatoprost, PGF2α, Fluprostenol, and Travoprost Guided by Biocatalytic Retrosynthesis

Application of Ketoreductase in Asymmetric Synthesis of Pharmaceuticals and Bioactive Molecules: An Update (2018–2020)

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Redesign of a short-chain dehydrogenase/reductase for asymmetric synthesis of ethyl (R)-2-hydroxy-4-phenylbutanoate based on per-residue free energy decomposition and sequence conservatism analysis

Cited by 35 publications

References 34 publications

Improving the Cβ Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Improving the Cβ Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Unified Strategy to Prostaglandins: Chemoenzymatic Total Synthesis of Cloprostenol, Bimatoprost, PGF2α, Fluprostenol, and Travoprost Guided by Biocatalytic Retrosynthesis

Application of Ketoreductase in Asymmetric Synthesis of Pharmaceuticals and Bioactive Molecules: An Update (2018–2020)

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Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance

Improving the C_β Stereoselectivity of l‐Threonine Aldolase for the Synthesis of l‐threo‐4‐Methylsulfonylphenylserine by Modulating the Substrate‐Binding Pocket To Control the Orientation of the Substrate Entrance