8EUO
Hydroxynitrile Lyase from Hevea brasiliensis with Seven Mutations
Summary for 8EUO
| Entry DOI | 10.2210/pdb8euo/pdb |
| Descriptor | (S)-hydroxynitrile lyase (2 entities in total) |
| Functional Keywords | engineered protein, alpha/beta hydrolase fold, esterase, catalytic promiscuity, lyase |
| Biological source | Hevea brasiliensis (rubber tree) |
| Total number of polymer chains | 1 |
| Total formula weight | 30258.74 |
| Authors | Greenberg, L.R.,Walsh, M.E.,Kazlauskas, R.J.,Pierce, C.T.,Shi, K.,Aihara, H.,Evans, R.L. (deposition date: 2022-10-19, release date: 2022-11-30, Last modification date: 2025-09-17) |
| Primary citation | Pierce, C.T.,Greenberg, L.R.,Walsh, M.E.,Shi, K.,Magee, D.J.,Aihara, H.,Gordon, W.,Evans 3rd, R.L.,Kazlauskas, R.J. Crystal structure of a seven-substitution mutant of hydroxynitrile lyase from rubber tree. Acta Crystallogr.,Sect.F, 81:398-405, 2025 Cited by PubMed Abstract: The α/β-hydrolase fold superfamily includes esterases and hydroxynitrile lyases which, despite catalyzing different reactions, share a Ser-His-Asp catalytic triad. We report a 1.99 Å resolution crystal structure of HNL6V, an engineered variant of hydroxynitrile lyase from Hevea brasiliensis (HbHNL) containing seven amino-acid substitutions (T11G, E79H, C81L, H103V, N104A, G176S and K236M). The structure reveals that HNL6V maintains the characteristic α/β-hydrolase fold while exhibiting systematic shifts in backbone and catalytic atom positions. Compared with wild-type HbHNL, the C positions in HNL6V differ by a mean of 0.2 ± 0.1 Å, representing a statistically significant displacement. Importantly, the catalytic triad and oxyanion-hole atoms have moved 0.2-0.8 Å closer to their corresponding positions in SABP2, although they remain 0.3-1.1 Å from fully achieving the configuration of SABP2. The substitutions also increase local flexibility, particularly in the lid domain covering the active site. This structural characterization demonstrates that targeted amino-acid substitutions can systematically shift catalytic geometries towards those of evolutionarily related enzymes. PubMed: 40864147DOI: 10.1107/S2053230X25007034 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.99 Å) |
Structure validation
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