Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds
Abstract Enantio-pure α-hydroxy amides are valuable intermediates for the synthesis of chiral pharmaceuticals. The asymmetric reduction of α-keto amides to generate chiral α-hydroxy amides is a difficult and challenging task in biocatalysis. In this study, iolS, an aldo–keto reductase from Bacillus...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-10-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-023-02225-9 |
| _version_ | 1797450974657249280 |
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| author | Ruixuan Bai Baoling Chen Liangyu Zheng |
| author_facet | Ruixuan Bai Baoling Chen Liangyu Zheng |
| author_sort | Ruixuan Bai |
| collection | DOAJ |
| description | Abstract Enantio-pure α-hydroxy amides are valuable intermediates for the synthesis of chiral pharmaceuticals. The asymmetric reduction of α-keto amides to generate chiral α-hydroxy amides is a difficult and challenging task in biocatalysis. In this study, iolS, an aldo–keto reductase from Bacillus subtilis 168 was exhibited as a potential biocatalyst, which could catalyze the reduction of diaryl α-keto amide such as 2-oxo-N, 2-diphenyl-acetamide (ONDPA) with moderate S-selectivity (76.1%, ee) and 60.5% conversion. Through semi-rational engineering, two stereocomplementary variants (I57F/F126L and N21A/F126A) were obtained with ee value of 97.6% (S) and 99.9% (R) toward ONDPA (1a), respectively, delivering chiral α-hydroxy amide with > 98% conversions. Moreover, the excellent S- and R-preference variants displayed improved stereoselectivities toward the other α-keto amide compounds. Molecular dynamic and docking analysis revealed that the two key residues at 21 and 126 were identified as the “switch”, which specifically controlled the stereopreference of iolS by regulating the shape of substrate binding pocket as well as the substrate orientation. Our results offer an effective strategy to obtain α-hydroxy amides with high optical purity and provide structural insights into altering the stereoselectivity of AKRs. Graphical Abstract |
| first_indexed | 2024-03-09T14:48:15Z |
| format | Article |
| id | doaj.art-5a9d1e5bf76d475eab62430ee452e4f8 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-03-09T14:48:15Z |
| publishDate | 2023-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-5a9d1e5bf76d475eab62430ee452e4f82023-11-26T14:38:18ZengBMCMicrobial Cell Factories1475-28592023-10-0122111410.1186/s12934-023-02225-9Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compoundsRuixuan Bai0Baoling Chen1Liangyu Zheng2Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin UniversityKey Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin UniversityKey Laboratory for Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin UniversityAbstract Enantio-pure α-hydroxy amides are valuable intermediates for the synthesis of chiral pharmaceuticals. The asymmetric reduction of α-keto amides to generate chiral α-hydroxy amides is a difficult and challenging task in biocatalysis. In this study, iolS, an aldo–keto reductase from Bacillus subtilis 168 was exhibited as a potential biocatalyst, which could catalyze the reduction of diaryl α-keto amide such as 2-oxo-N, 2-diphenyl-acetamide (ONDPA) with moderate S-selectivity (76.1%, ee) and 60.5% conversion. Through semi-rational engineering, two stereocomplementary variants (I57F/F126L and N21A/F126A) were obtained with ee value of 97.6% (S) and 99.9% (R) toward ONDPA (1a), respectively, delivering chiral α-hydroxy amide with > 98% conversions. Moreover, the excellent S- and R-preference variants displayed improved stereoselectivities toward the other α-keto amide compounds. Molecular dynamic and docking analysis revealed that the two key residues at 21 and 126 were identified as the “switch”, which specifically controlled the stereopreference of iolS by regulating the shape of substrate binding pocket as well as the substrate orientation. Our results offer an effective strategy to obtain α-hydroxy amides with high optical purity and provide structural insights into altering the stereoselectivity of AKRs. Graphical Abstracthttps://doi.org/10.1186/s12934-023-02225-9α-keto amidesα-hydroxyl amidesAldo–keto reductaseStereocomplementarySemi-rational engineeringSwitch |
| spellingShingle | Ruixuan Bai Baoling Chen Liangyu Zheng Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds Microbial Cell Factories α-keto amides α-hydroxyl amides Aldo–keto reductase Stereocomplementary Semi-rational engineering Switch |
| title | Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds |
| title_full | Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds |
| title_fullStr | Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds |
| title_full_unstemmed | Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds |
| title_short | Semi-rational engineering an aldo–keto reductase for stereocomplementary reduction of α-keto amide compounds |
| title_sort | semi rational engineering an aldo keto reductase for stereocomplementary reduction of α keto amide compounds |
| topic | α-keto amides α-hydroxyl amides Aldo–keto reductase Stereocomplementary Semi-rational engineering Switch |
| url | https://doi.org/10.1186/s12934-023-02225-9 |
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