Summary
Rigidification of active site residues accelerates enzyme activity. This was shown in a Kemp eliminase where residues in the first and second shell surrounding the active site were mutated (1,2), as well as synthetic de novo designed enzymes (3).
1.
Broom A, Rakotoharisoa RV, Thompson MC, Zarifi N, Nguyen E, Mukhametzhanov N, et al. Ensemble-based enzyme design can recapitulate the effects of laboratory directed evolution in silico. Nature Communications. 2020;11(1). Available from: https://doi.org/10.1038/s41467-020-18619-x
2.
Zarifi N, Asthana P, Doustmohammadi H, Klaus C, Sanchez J, Hunt SE, et al. Distal mutations enhance catalysis in designed enzymes by facilitating substrate binding and product release. Nature Communications. 2025;16(1). Available from: https://doi.org/10.1038/s41467-025-63802-7
3.
Hou K, Huang W, Qi M, Tugwell TH, Alturaifi TM, Chen Y, et al. De novo design of porphyrin-containing proteins as efficient and stereoselective catalysts. Science. 2025;388(6747):665–70. Available from: https://doi.org/10.1126/science.adt7268