A Smart Library of Epoxide Hydrolase Variants and the Top Hits for Synthesis of (<i>S</i>)‐β‐Blocker Precursors

Kong, Xiangxing; Ma, Qian; Zhou, Jiahai; Zeng, Bu‐Bing; Xu, Jian‐He

doi:10.1002/ange.201402653

Cited by 12 publications

(7 citation statements)

References 30 publications

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“…Interestingly, it contains a "U" shape substrate binding pocket (SBP) with the active site residues placed in the center, giving rise to two narrow tunnels (tunnel 1 and tunnel 2), whose spatial properties have an effect on the substrate preference. 28,39 Tunnel 1 was constituted of W 102 , I 105 , V 126 , P 127 , L 129 , T 137 , A 143 , M 144 , I 151 , V 264 , S 267 , L 268 , and M 270 , whereas tunnel 2 was constituted of F 33 , P 34 , I 176 , T 179 , K 181 , G 183 , P 185 , N 197 , M 263 , and F 298 (Figure S3). Both tunnels of VrEH3 SBP are interlinked with each other and also lead eventually to the exterior of the protein, which dramatically differs with those SBPs of other α/β hydrolase fold EHs, such as StEH (PDB: 2CJP), 27 AnEH LCP (PDB: 3G0I), 40 BmEH (PDB: 4G02), 41 and MtEHB (PDB: 2ZJF), 42 whose tunnels closed at the internal region.…”

Section: Journal Of Agricultural and Food Chemistrymentioning

confidence: 99%

Stereoselective Hydrolysis of Epoxides by reVrEH3, a Novel Vigna radiata Epoxide Hydrolase with High Enantioselectivity or High and Complementary Regioselectivity

Hu¹,

Tang

Li³

et al. 2017

J. Agric. Food Chem.

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To provide more options for the stereoselective hydrolysis of epoxides, an epoxide hydrolase (VrEH3) gene from Vigna radiata was cloned and expressed in Escherichia coli. Recombinant VrEH3 displayed the maximum activity at pH 7.0 and 45 °C and high stability at pH 4.5-7.5 and 55 °C. Notably, reVrEH3 exhibited high and complementary regioselectivity toward styrene oxides 1a-3a and high enantioselectivity (E = 48.7) toward o-cresyl glycidyl ether 9a. To elucidate these interesting phenomena, the interactions of the three-dimensional structure between VrEH3 and enantiomers of 1a and 9a were analyzed by molecular docking simulation. Using E. coli/vreh3 whole cells, gram-scale preparations of (R)-1b and (R)-9a were performed by enantioconvergent hydrolysis of 100 mM rac-1a and kinetic resolution of 200 mM rac-9a in the buffer-free water system at 25 °C. These afforded (R)-1b with >99% ee and 78.7% overall yield after recrystallization and (R)-9a with >99% ee, 38.7% overall yield, and 12.7 g/L/h space-time yield.

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Section: Journal Of Agricultural and Food Chemistrymentioning

confidence: 99%

Stereoselective Hydrolysis of Epoxides by reVrEH3, a Novel Vigna radiata Epoxide Hydrolase with High Enantioselectivity or High and Complementary Regioselectivity

Hu¹,

Tang

Li³

et al. 2017

J. Agric. Food Chem.

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“…Recently, protein engineering has emerged as a powerful tool in tuning enzyme properties, 21 including catalytic activity, 22 substrate spectrum, 23 selectivity, 24,25 thermostability, 26 and new reaction types. 27 The same strategy has also been applied in the engineering of IDOs aiming to improve their catalytic activities.…”

Section: ■ Introductionmentioning

confidence: 99%

Engineering Bacillus subtilis Isoleucine Dioxygenase for Efficient Synthesis of (2S,3R,4S)-4-Hydroxyisoleucine

Yan

Qian

et al. 2020

J. Agric. Food Chem.

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Isoleucine dioxygenase (IDO)-catalyzed hydroxylation of isoleucine is a promising method for the synthesis of the diabetic drug (2S,3R,4S)-4-hydroxyisoleucine [(2S,3R,4S)-4-HIL]. However, the low activity of IDO significantly limits its practical application. In this work, a high-throughput screening method was developed and directed evolution was performed on the IDO from Bacillus subtilis, resulting in a double mutant with improvements in specific activity, protein expression level, and fermentation titer of 3.2-, 2.8-, and 9.4-fold, respectively. l-Isoleucine (228 mM) was completely converted to (2S,3R,4S)-4-HIL by the best variant with a space-time yield of up to 80.8 g L–1 d–1, which is the highest record reported so far. With a further increase of the substrate loading to 1 M, a high conversion of 91% could also be achieved. At last, enzymatic synthesis of (2S,3R,4S)-4-HIL was successfully carried out on a 3 L scale, indicating tremendous potential of the IDO variant I162T/T182N for green and efficient production of (2S,3R,4S)-4-HIL.

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“…Epoxides are known versatile intermediates giving rise to drugs such as pioglitazone, metoprolol and levofloxacin; or chemical probes such as wiskostatin [ 23 , 31 , 37 , 38 ]. Epoxide installation is typically by direct coupling of an epihalohydrin [ 37 – 40 ] or through m -chloroperoxybenzoic acid, hydrogen peroxide or tert -butyl hydroperoxide olefin epoxidation [ 41 – 43 ].…”

Section: Introductionmentioning

confidence: 99%

In situ epoxide generation by dimethyldioxirane oxidation and the use of epichlorohydrin in the flow synthesis of a library of β-amino alcohols

Cossar¹,

Baker²,

Cain³

et al. 2018

R. Soc. open sci.

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The flow coupling of epichlorohydrin with substituted phenols, while efficient, limits the nature of the epoxide available for the development of focused libraries of β-amino alcohols. This limitation was encountered in the production of analogues of 1-(4-nitrophenoxy)-3-((2-((4-(trifluoromethyl)pyrimidin-2-yl)amino)ethyl)amino)propan-2-ol 1, a potential antibiotic lead. The in situ (flow) generation of dimethyldoxirane (DMDO) and subsequent flow olefin epoxidation abrogates this limitation and afforded facile access to structurally diverse β-amino alcohols. Analogues of 1 were readily accessed either via (i) a flow/microwave hybrid approach, or (ii) a sequential flow approach. Key steps were the in situ generation of DMDO, with olefin epoxidation in typically good yields and a flow-mediated ring opening aminolysis to form an expanded library of β-amino alcohols 1 and 10a–18g, resulting in modest (11a, 21%) to excellent (12g, 80%) yields. Alternatively flow coupling of epichlorohydrin with phenols 4a–4m (22%–89%) and a Bi(OTf)3 catalysed microwave ring opening with amines afforded a select range of β-amino alcohols, but with lower levels of aminolysis regiocontrol than the sequential flow approach.

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A Smart Library of Epoxide Hydrolase Variants and the Top Hits for Synthesis of (S)‐β‐Blocker Precursors

Cited by 12 publications

References 30 publications

Stereoselective Hydrolysis of Epoxides by reVrEH3, a Novel Vigna radiata Epoxide Hydrolase with High Enantioselectivity or High and Complementary Regioselectivity

Stereoselective Hydrolysis of Epoxides by reVrEH3, a Novel Vigna radiata Epoxide Hydrolase with High Enantioselectivity or High and Complementary Regioselectivity

Engineering Bacillus subtilis Isoleucine Dioxygenase for Efficient Synthesis of (2S,3R,4S)-4-Hydroxyisoleucine

In situ epoxide generation by dimethyldioxirane oxidation and the use of epichlorohydrin in the flow synthesis of a library of β-amino alcohols

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