Assessing the stereoselectivity of carbonyl reductases toward the reduction of OPBE and docking analysis

Chen, Rong; Deng, Jiao; Lin, Jian; Yin, Xia; Xie, Tian; Yang, Shengli; Wei, Dongzhi

doi:10.1002/bab.1397

Cited by 8 publications

(1 citation statement)

References 35 publications

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“…The generated model was geometrically refined by the Princeton TIGRESS 2.0 ( Khoury et al, 2017 ). The final model was evaluated using Procheck program, and the lowest energy conformational model was chosen for docking studies ( Chen et al, 2016 ).…”

Section: Methodsmentioning

confidence: 99%

Protein Engineering of a Germacrene A Synthase From Lactuca sativa and Its Application in High Productivity of Germacrene A in Escherichia coli

et al. 2022

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Germacrene A (GA) is a key intermediate for the synthesis of medicinal active compounds, especially for β-elemene, which is a broad-spectrum anticancer drug. The production of sufficient GA in the microbial platform is vital for the precursors supply of active compounds. In this study, Escherichia coli BL21 Star (DE3) was used as the host and cultivated in SBMSN medium, obtaining a highest yield of FPP. The GA synthase from Lactuca sativa (LTC2) exhibited the highest level of GA production. Secondly, two residues involved in product release (T410 and T392) were substituted with Ser and Ala, respectively, responsible for relatively higher activities. Next, substitution of selected residues S243 with Asn caused an increase in activity. Furthermore, I364K-T410S and T392A-T410S were created by combination with the beneficial mutation, and they demonstrated dramatically enhanced titers with 1.90-fold and per-cell productivity with 5.44-fold, respectively. Finally, the production titer of GA reached 126.4 mg/L, and the highest productivity was 7.02 mg/L.h by the I364K-T410S mutant in a shake-flask batch culture after fermentation for 18 h. To our knowledge, the productivity of the I364K-T410S mutant is the highest level ever reported. These results highlight a promising method for the industrial production of GA in E. coli, and lay a foundation for pathway reconstruction and the production of valuable natural sesquiterpenes.

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Section: Methodsmentioning

confidence: 99%

Protein Engineering of a Germacrene A Synthase From Lactuca sativa and Its Application in High Productivity of Germacrene A in Escherichia coli

et al. 2022

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Multi‐faceted Set‐up of a Diverse Ketoreductase Library Enables the Synthesis of Pharmaceutically‐relevant Secondary Alcohols

et al. 2021

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Enzymes are valuable tools to introduce chirality into small molecules. Especially, ketoreductase (KRED)-catalyzed transformations of ketones to yield chiral secondary alcohols have become an established biocatalytic process step in the pharmaceutical and fine chemical industry. Development time, however, remains a critical factor in chemical process development and thus, the competitiveness of a biocatalytic reaction step is often governed by the availability of off-the-shelf enzyme libraries. To expand the biocatalytic toolbox with additional ketoreductases, we established a multi-faceted screening procedure to capture KRED diversity from different sources, such as literature, available genome data, and uncharacterized microbial strains. Overall, we built a library consisting of 51 KRED enzymes, 29 of which have never been described in literature before. Notably, 18 of the newly described enzymes exhibited anti-Prelog preference complementing the majority of ketoreductases which generally follow Prelog's rule. Analysis of the library's catalytic activity toward a chemically diverse ketone substrate set of pharmaceutical interest further highlighted the broad substrate scope and the complementing enantio-preference of the individual KREDs. Using the generated sequence-function data of the included short chain dehydrogenases in a bioinformatic analysis led to the identification of possible sequence determinants of the stereospecificity exhibited by these enzymes.

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Regenerated coenzyme-based preparation of bienzyme-polymer nanoconjugates and their applications for the synthesis of ethyl (R)-2-hydroxy-4-phenylbutyrate

Lü

Dai

Cheng

et al. 2021

Korean J. Chem. Eng.

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Assessing the stereoselectivity of carbonyl reductases toward the reduction of OPBE and docking analysis

Cited by 8 publications

References 35 publications

Protein Engineering of a Germacrene A Synthase From Lactuca sativa and Its Application in High Productivity of Germacrene A in Escherichia coli

Protein Engineering of a Germacrene A Synthase From Lactuca sativa and Its Application in High Productivity of Germacrene A in Escherichia coli

Multi‐faceted Set‐up of a Diverse Ketoreductase Library Enables the Synthesis of Pharmaceutically‐relevant Secondary Alcohols

Regenerated coenzyme-based preparation of bienzyme-polymer nanoconjugates and their applications for the synthesis of ethyl (R)-2-hydroxy-4-phenylbutyrate

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