Purification and characterization of a glucose-tolerant β-glucosidase from black plum seed and its structural changes in ionic liquids

The objective of this study was to characterize a plant origin β-glucosidase from black plum seeds and identify its conformational changes in twenty-six imidazolium- and amino acid-based ionic liquids (ILs). The results revealed that the purified 60 kDa enzyme was monomeric in nature, maximally active at 55 °C and pH 5.0, and nearly completely inhibited by Hg²⁺ and Ag⁺. Attractive peculiarities of the relative low kinetic and higher glucose inhibition constants (K_m = 0.58 mM [pNPG]; K_i = 193.5 mM [glucose]) demonstrated its potential applications in food industry. Circular dichroism studies showed that the secondary structural changes of the enzyme depended not only on the anions, but also on the cations of the assayed ILs. Interestingly, no corresponding relations were observed between the changes in enzyme structure induced by ILs and its catalytic activities, suggesting that the influences of ILs on enzymatic processes don't rely simply on enzyme conformational changes.