8JHE
Hyper-thermostable ancestral L-amino acid oxidase 2 (HTAncLAAO2)
Summary for 8JHE
| Entry DOI | 10.2210/pdb8jhe/pdb |
| Descriptor | Hyper thermostable ancestral L-amino acid oxidase, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total) |
| Functional Keywords | l-amino acid oxidases, fad-dependent, ancestral sequence reconstruction, oxidoreductase |
| Biological source | synthetic construct |
| Total number of polymer chains | 4 |
| Total formula weight | 253824.36 |
| Authors | Kawamura, Y.,Ishida, C.,Miyata, R.,Miyata, A.,Hayashi, S.,Fujinami, D.,Ito, S.,Nakano, S. (deposition date: 2023-05-23, release date: 2023-10-04) |
| Primary citation | Kawamura, Y.,Ishida, C.,Miyata, R.,Miyata, A.,Hayashi, S.,Fujinami, D.,Ito, S.,Nakano, S. Structural and functional analysis of hyper-thermostable ancestral L-amino acid oxidase that can convert Trp derivatives to D-forms by chemoenzymatic reaction. Commun Chem, 6:200-200, 2023 Cited by PubMed Abstract: Production of D-amino acids (D-AAs) on a large-scale enables to provide precursors of peptide therapeutics. In this study, we designed a novel L-amino acid oxidase, HTAncLAAO2, by ancestral sequence reconstruction, exhibiting high thermostability and long-term stability. The crystal structure of HTAncLAAO2 was determined at 2.2 Å by X-ray crystallography, revealing that the enzyme has an octameric form like a "ninja-star" feature. Enzymatic property analysis demonstrated that HTAncLAAO2 exhibits three-order larger k/K values towards four L-AAs (L-Phe, L-Leu, L-Met, and L-Ile) than that of L-Trp. Through screening the variants, we obtained the HTAncLAAO2(W220A) variant, which shows a > 6-fold increase in k value toward L-Trp compared to the original enzyme. This variant applies to synthesizing enantio-pure D-Trp derivatives from L- or rac-forms at a preparative scale. Given its excellent properties, HTAncLAAO2 would be a starting point for designing novel oxidases with high activity toward various amines and AAs. PubMed: 37737277DOI: 10.1038/s42004-023-01005-1 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.201 Å) |
Structure validation
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