Structure-guided evolution of carbonyl reductase for efficient biosynthesis of ethyl (<i>R</i>)-2-hydroxy-4-phenylbutyrate

Tang, Wen; Chen, Lulu; Deng, Jiao; Kuang, Yuyao; Chen, Chao; Yin, Biaolin; Wang, Hualei; Lin, Jian; Wei, Dongzhi

doi:10.1039/d0cy01411g

Cited by 5 publications

(3 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recent examples indicated the effectiveness of improving enantioselectivity through structure-oriented engineering of substrate-binding pockets. 15,32,33 Blast analysis predicted that the substrate binding pocket of the esterase pnbA-BS constituted 13 key residues (G105, G106, A107, E188, S189, A190, M193, T326, A330, L331, M358, A400, L403). Except for S189 as one of the catalytic triads, those residues were subjected to alanine scanning by site-directed mutagenesis.…”

Section: Resultsmentioning

confidence: 99%

Engineering aBacillus subtilisesterase for selective hydrolysis ofd,l-menthyl acetate in an organic solvent-free system

et al. 2023

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Engineering aBacillus subtilisesterase for selective hydrolysis ofd,l-menthyl acetate in an organic solvent-free system

et al. 2023

View full text Add to dashboard Cite

show abstract

“…For example, Lin and co‐workers previously disclosed that Go CR, a NADH‐dependent KRED from Gluconobacter oxydans , was able to catalyze this reaction, albeit with unsatisfactory specific activity (3.7 U/mg) and stereoselectivity (43.2 % ee) [20] . Recently, these authors have improved the catalytic performance of this enzyme and realized an efficient production of ( R )‐HPBE at high substrate loadings using a structure‐guided rational design strategy [21] . In short, three hotspots identified through computational techniques including molecular docking and the molecular mechanics generalized Born surface area (MM‐GBSA) method, were subjected to single‐site and cooperative mutation studies, ultimately yielding the best mutant mut‐W193L/C93I/I187L with simultaneously improved catalytic efficiency, stereoselectivity, and thermostability.…”

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

View full text Add to dashboard Cite

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.

show abstract

“…Ethyl (R)-2-hydroxy-4-phenylbutanoate ((R)-HPBE) is an important chiral alcohol intermediate for the synthesis of angiotensin-converting enzyme inhibitors such as enalapril and lisinopril. Various methods for preparing (R)-HPBE have been developed, including multistep chemical synthesis [1], asymmetric enzymatic reduction of 2-oxo-4phenyl-butyric acid ethyl ester (OPBE), kinetic resolution of racemates, and the enzymatic esterification of 2-hydroxy-4-phenylbutanoic acid [2][3][4]. Asymmetric catalysis is one of the most efficient methods to prepare (R)-HPBE [5,6].…”

Section: Introductionmentioning

confidence: 99%

Construction of Bi-Enzyme Self-Assembly Clusters Based on SpyCatcher/SpyTag for the Efficient Biosynthesis of (R)-Ethyl 2-hydroxy-4-phenylbutyrate

et al. 2023

View full text Add to dashboard Cite

Cascade reactions catalyzed by multi-enzyme systems are important in science and industry and can be used to synthesize drugs and nutrients. In this study, two types of macromolecules of bi-enzyme self-assembly clusters (BESCs) consisting of carbonyl reductase (CpCR) and glucose dehydrogenase (GDH) were examined. Stereoselective CpCR and GDH were successfully fused with SpyCatcher and SpyTag, respectively, to obtain four enzyme modules, namely: SpyCatcher-CpCR, SpyCatcher-GDH, SpyTag-CpCR, and SpyTag-GDH, which were covalently coupled in vitro to form two types of hydrogel-like BESCs: CpCR-SpyCatcher-SpyTag-GDH and GDH-SpyCatcher-SpyTag-CpCR. CpCR-SpyCatcher-SpyTag-GDH showed a better activity and efficiently converted ethyl 2-oxo-4-phenylbutyrate (OPBE) to ethyl(R)2-hydroxy-4-phenylbutanoate ((R)-HPBE), while regenerating NADPH. At 30 °C and pH 7, the conversion rate of OPBE with CpCR-SpyCatcher-SpyTag-GDH as a catalyst reached 99.9%, with the ee% of (R)-HPBE reaching above 99.9%. This conversion rate was 2.4 times higher than that obtained with the free bi-enzyme. The pH tolerance and temperature stability of the BESCs were also improved compared with those of the free enzymes. In conclusion, bi-enzyme assemblies were docked using SpyCatcher/SpyTag to produce BESCs with a special structure and excellent catalytic activity, improving the catalytic efficiency of the enzyme.

show abstract

Structure-guided evolution of carbonyl reductase for efficient biosynthesis of ethyl (R)-2-hydroxy-4-phenylbutyrate

Abstract: Ethyl (R)-2-hydroxy-4-phenylbutanoate ((R)-HPBE) is an important versatile intermediate for the synthesis of angiotensin-converting enzyme inhibitors. Herein, a structure-guided rational design was adopted to improve the catalytic performance of carbonyl reductase...

Cited by 5 publications

References 45 publications

Engineering aBacillus subtilisesterase for selective hydrolysis ofd,l-menthyl acetate in an organic solvent-free system

Engineering aBacillus subtilisesterase for selective hydrolysis ofd,l-menthyl acetate in an organic solvent-free system

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

Construction of Bi-Enzyme Self-Assembly Clusters Based on SpyCatcher/SpyTag for the Efficient Biosynthesis of (R)-Ethyl 2-hydroxy-4-phenylbutyrate

Contact Info

Product

Resources

About