Grogan, Gideon James orcid.org/0000-0003-1383-7056, Gilio, Amelia, Steflik, Jeremy et al. (4 more authors) (2023) Engineering of a Reductive Aminase to Enable the Synthesis of a Key Intermediate to a CDK 2/4/6 Inhibitor. ACS Catalysis. ISSN 2155-5435
Abstract
Biocatalytic reductive amination reactions with reductive aminases (RedAms) are emerging transformations with a high potential value for pharmaceutical synthesis. Here, we report the identification and engineering of a RedAm to catalyze a reductive amination reaction, making a key intermediate in the synthesis of an investigational cyclin-dependent kinase (CDK) inhibitor, using the relatively bulky benzylamine as a nucleophile. The engineered enzyme contains six mutations with respect to the wild-type and displays high productivity at high substrate concentrations (50-fold improved over the wild-type). After the optimized enzyme variant was identified, crystal structures of both the wild-type and mutant enzymes were solved and used to rationalize how structural changes to the RedAm improved its performance under process conditions. Results suggest that mutations affecting both substrate binding and enzyme thermostability contribute to improved enzyme performance. By enabling the multikilogram-scale synthesis of the chiral intermediate, this work highlights the versatility and industrial utility of RedAm-catalyzed reductive amination.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | ©2023 American Chemical Society. This is an author-produced version of the published paper. Uploaded in accordance with the University’s Research Publications and Open Access policy. |
Dates: |
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Institution: | The University of York |
Academic Units: | The University of York > Faculty of Sciences (York) > Chemistry (York) |
Funding Information: | Funder Grant number BBSRC (BIOTECHNOLOGY AND BIOLOGICAL SCIENCES RESEARCH COUNCIL) BB/T017805/1 |
Depositing User: | Pure (York) |
Date Deposited: | 25 Jul 2023 13:10 |
Last Modified: | 07 Feb 2025 00:36 |
Published Version: | https://doi.org/10.1021/acscatal.3c01534 |
Status: | Published online |
Refereed: | Yes |
Identification Number: | 10.1021/acscatal.3c01534 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:201825 |
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