Crystal Structures and Catalytic Mechanism of l‐erythro‐3,5‐Diaminohexanoate Dehydrogenase and Rational Engineering for Asymmetric Synthesis of β‐Amino Acids
- 24 February 2021
- journal article
- research article
- Published by Wiley in Angewandte Chemie-International Edition
- Vol. 60 (18), 10203-10210
- https://doi.org/10.1002/anie.202017225
Abstract
Amino acid dehydrogenases (AADHs) have shown considerable potential as biocatalysts in the asymmetric synthesis of chiral amino acids. However, compared to the widely studied α‐AADHs, limited knowledge is available about β‐AADHs that enable the synthesis of β‐amino acids. Herein, we report the crystal structures of a l‐erythro‐3,5‐diaminohexanoate dehydrogenase and its variants, the only known member of β‐AADH family. Crystal structure analysis, site‐directed mutagenesis studies and quantum chemical calculations revealed the differences in the substrate binding and catalytic mechanism from α‐AADHs. A number of rationally engineered variants were then obtained with improved activity (by 110–800 times) toward various aliphatic β‐amino acids without an enantioselectivity trade‐off. Two β‐amino acids were prepared by using the outstanding variants with excellent enantioselectivity (>99 % ee) and high isolated yields (86–87 %). These results provide important insights into the molecular mechanism of 3,5‐DAHDH, and establish a solid foundation for further design of β‐AADHs for the asymmetric synthesis of β‐amino acids.Keywords
Funding Information
- National Natural Science Foundation of China (21778072 and 92056101)
- National Key Research and Development Program of China (2019YFA09005000 and 2018YFA0901900)
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