Harnessing natural deep eutectic solvents Pickering emulsion droplets for biocatalytic production of diacylglycerols via glycerolysis: Elucidating the underlying mechanism of lipase thermostability.

Glycerolysis is a crucial approach for the preparation of diacylglycerols (DAG). The challenges such as enzyme stability and interfacial mass transfer have been insufficiently addressed. To validate the Pickering interfacial biocatalysis (PIB) for the desired target, a boosting system containing Pickering emulsion droplets based on natural deep eutectic solvents (NADES) was developed as a reaction medium for enhancing the catalysis efficiency of CALB-catalyzed (Candida antarctica lipase B, CALB) glycerolysis. The effects of types of oils and NADES on the properties of Pickering emulsions and catalytic efficiency were evaluated. Comparative analyses within the system revealed a 3.20-fold enhancement in catalytic efficiency for the NADES-based Pickering emulsion compared to its non-emulsion control, and an independent 9.13-fold enhancement for the glycerol-based Pickering emulsion relative to corresponding non-emulsion control. Additionally, molecular dynamics simulations offered insights into the temperature-dependent conformational behavior of CALB across various polar media. This study highlights the potential applicability of NADES Pickering emulsions as an efficient and sustainable platform for biocatalysis of lipids.