Ancestral L-amino acid oxidases for deracemization and stereoinversion of amino acids
Open Access
- 4 December 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Communications Chemistry
- Vol. 3 (1), 1-11
- https://doi.org/10.1038/s42004-020-00432-8
Abstract
L-amino acid oxidases (LAAOs) can be applied to convert racemic amino acids to D-isomers, which are potential precursors of pharmaceuticals. However, this application is hampered by the lack of available stable and structure-determined LAAOs. In this study, we attempt to address this limitation by utilizing two ancestral LAAOs: AncLAAO-N4 and AncLAAO-N5. AncLAAO-N4 has the highest thermal and temporal stabilities among the designed LAAOs that can be used for deracemization and stereoinversion. AncLAAO-N5 can provide X-ray crystal structures, which are helpful to reveal substrate recognition and reaction mechanisms of LAAOs at the molecular level. Next, we attempted to improve activity of AncLAAO-N4 toward L-Val through a semi-rational protein engineering method. Three variants with enhanced activity toward L-Val were obtained. Taken together, we believe that the activity and substrate selectivity of AncLAAOs give them the potential to be key enzymes in various chemoenzymatic reactions.Keywords
Funding Information
- MEXT | Japan Society for the Promotion of Science (18K14391, 17H06169, 17K06931)
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