9FD7
Re-engineered peroxygenase variant of 2-deoxy-D-ribose-5-phosphate aldolase in substrate-free state
Summary for 9FD7
| Entry DOI | 10.2210/pdb9fd7/pdb |
| Descriptor | Deoxyribose-phosphate aldolase, GLYCEROL (3 entities in total) |
| Functional Keywords | protein engineering, directed evolution, aldolase, peroxygenase, schiff base, lyase |
| Biological source | Escherichia coli |
| Total number of polymer chains | 2 |
| Total formula weight | 57891.94 |
| Authors | Thunnissen, A.M.W.H.,Zhou, H.,Frietema, H.O.T.,Poelarends, G.J. (deposition date: 2024-05-16, release date: 2025-03-12, Last modification date: 2025-05-21) |
| Primary citation | Zhou, H.,Kunzendorf, A.,Xu, G.,Frietema, H.O.T.,Thunnissen, A.W.H.,Poelarends, G.J. Engineering 2-Deoxy-D-ribose-5-phosphate Aldolase for anti- and syn-Selective Epoxidations of alpha , beta-Unsaturated Aldehydes. Angew.Chem.Int.Ed.Engl., 64:e202503054-e202503054, 2025 Cited by PubMed Abstract: The enzyme 2-deoxy-D-ribose-5-phosphate aldolase (DERA) naturally catalyzes the reversible aldol addition between acetaldehyde and D-glyceraldehyde-3-phosphate to yield 2-deoxy-D-ribose-5-phosphate. Herein we describe the redesign of DERA into a proficient non-natural peroxygenase that promotes the asymmetric epoxidation of various α,β-unsaturated aldehydes. This repurposed aldolase, named DERA-EP, is able to utilize HO to accomplish both anti- and syn-selective epoxidations of various α,β-unsaturated aldehydes to give the corresponding epoxides with moderate to high diastereoselectivity (diastereomeric ratio up to 99 : 1) and excellent enantioselectivity (enantiomeric ratio up to 99 : 1). Crystallographic analysis of DERA-EP in a substrate-free and substrate-bound state provides a structural context for the evolved activity, a clear explanation for the high enantioselectivity, and compelling evidence for catalysis via enzyme-bound iminium ion intermediates. The unprecedented anti-selectivity of DERA-EP with multiple α,β-unsaturated aldehydes is complementary to the syn-selectivity of previously reported enzyme-, metal- and organo-catalysts, making DERA-EP an attractive new asset to the toolbox of epoxidation catalysts. PubMed: 39993220DOI: 10.1002/anie.202503054 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.4 Å) |
Structure validation
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