Journal
ACS CATALYSIS
Volume 8, Issue 7, Pages 5698-5707Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acscatal.8b00863
Keywords
biocatalysis; catalytic mechanism; stereoselective epoxide hydrolysis; free energy landscape; substrate specificity; QM/MM and MD calculations; internal dynamics; binding affinity
Categories
Funding
- SusChemLombardia: prodotti e processi sostenibili per l'industria lombarda project, Accordo Quadro Regione Lombardia-CNR [18096]
- AIRC [18216]
- EPSRC [EP/M022609/1]
- BBSRC [BB/M000354/1]
- BBSRC [BB/M026280/1, BB/K016601/1, BB/L018756/1] Funding Source: UKRI
- EPSRC [EP/J010588/1] Funding Source: UKRI
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Limonene-1,2-epoxide hydrolases (LEHs), a subset of the epoxide hydrolase family, present interesting opportunities for the mild, regio- and stereo-selective hydrolysis of epoxide substrates. However, moderate enantioselectivity for non-natural ligands, combined with narrow substrate specificity, has so far limited the use of LEHs as general biocatalytic tools. A detailed molecular understanding of the structural and dynamic determinants of activity may complement directed evolution approaches to expand the range of applicability of these enzymes. Herein, we have combined quantum mechanics/molecular mechanics (QM/MM) free energy calculations for the reaction with MD simulations of the enzyme internal dynamics, and the calculation of binding affinities (using the WaterSwap method) for various representatives of the enzyme conformational ensemble, to show that the presence of natural or non-natural substrates differentially modulates the dynamic and catalytic behavior of LEH. The cross-talk between the protein and the ligands favors the selection of specific substrate-dependent interactions in the binding site, priming reactive complexes to select different preferential reaction pathways. The knowledge gained via our combined approach provides a molecular rationale for LEH substrate preferences. The comprehensive strategy we present here is general and broadly applicable to other cases of enzyme substrate selectivity and reactivity.
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