Food Chem. 2026 Jul 7;525(Pt 1):150333. doi: 10.1016/j.foodchem.2026.150333. Online ahead of print.

ABSTRACT

To overcome the activity-stability trade-off of enzymes in organic solvents, we propose a “Tunnel dynamics guided” engineering strategy. MD simulations revealed that solvents induce a defensive compaction in lipases, creating pathological, narrow tunnels that obstruct catalysis in a dynamic perspective. By targeting critical tunnel-gating sites, we engineered mutants that effectively mitigated activity-stability trade-off, exhibiting 7.53- and 5.56-fold higher tolerance to 66.7% DMSO and 10% pyridine, respectively. Mechanistic analysis identified a newly formed D285-K290 salt bridge that rigidifies the tunnel entrance and shortens the substrate pathway by ∼40%, significantly alleviating mass transfer resistance. Validated via kilogram-scale sucrose laurate synthesis (96.5% conversion), this study establishes tunnel dynamics remodeling as a robust paradigm for designing next-generation industrial biocatalysts.

PMID:42462309 | DOI:10.1016/j.foodchem.2026.150333