Watkins, Ella J. and Almhjell, Patrick J. and Arnold, Frances H. (2020) Direct enzymatic synthesis of a deep-blue fluorescent noncanonical amino acid from azulene and serine. ChemBioChem, 21 (1-2). pp. 80-83. ISSN 1439-4227. PMCID PMC6980211. doi:10.1002/cbic.201900497. https://resolver.caltech.edu/CaltechAUTHORS:20190912-124018949
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Abstract
We report a simple, one‐step enzymatic synthesis of the blue fluorescent noncanonical amino acid β‐(1‐azulenyl)‐l‐alanine (AzAla). Using an engineered tryptophan synthase β‐subunit (TrpB), stereochemically pure AzAla can be synthesized at scale starting from commercially available azulene and l‐serine. Mutation of a universally conserved catalytic glutamate in the active site to glycine has only a modest effect on native activity with indole but abolishes activity on azulene, suggesting that this glutamate activates azulene for nucleophilic attack by stabilization of the aromatic ion.
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| Additional Information: | © 2019 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim. Manuscript received: August 8, 2019; Accepted manuscript online: September 12, 2019; Version of record online: November 18, 2019. We thank Giovanni Tomaleri for his experimental assistance, Nathaniel Goldberg and Dr. Ben Levin for insightful discussions, and Dr. Tina Boville and Dr. David Romney for valuable contributions to experimental design. We thank Dr. Scott Virgil and the Center for Catalysis and Synthesis. This work was supported by the Rothenberg Innovation Initiative and by the National Institute of General Medical Sciences of the National Institutes of Health under Award Number R01GM125887. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Conflict of Interest: Enzyme variants in this manuscript and enzymatic synthesis of AzAla are the subject of a patent application (P.J.A. inventor). | |||||||||
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| Subject Keywords: | protein engineering; Amino acids; Biocatalysis; Sustainable Chemistry; Aromatic Ions | |||||||||
| Issue or Number: | 1-2 | |||||||||
| PubMed Central ID: | PMC6980211 | |||||||||
| DOI: | 10.1002/cbic.201900497 | |||||||||
| Record Number: | CaltechAUTHORS:20190912-124018949 | |||||||||
| Persistent URL: | https://resolver.caltech.edu/CaltechAUTHORS:20190912-124018949 | |||||||||
| Official Citation: | Direct enzymatic synthesis of a deep-blue fluorescent noncanonical amino acid from azulene and serine. E. J. Watkins, P. J. Almhjell, F. H. Arnold, ChemBioChem 2020, 21, 80; doi: 10.1002/cbic.201900497 | |||||||||
| Usage Policy: | No commercial reproduction, distribution, display or performance rights in this work are provided. | |||||||||
| ID Code: | 98611 | |||||||||
| Collection: | CaltechAUTHORS | |||||||||
| Deposited By: | Tony Diaz | |||||||||
| Deposited On: | 12 Sep 2019 20:17 | |||||||||
| Last Modified: | 09 Feb 2022 23:47 |
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