Chan, Kam KhongKam KhongChanSundaram, VidyaVidyaSundaramTan, JullyJullyTanHo, Yong KuenYong KuenHoRamanan, Ramakrishnan NagasundaraRamakrishnan NagasundaraRamananOoi, Chien WeiChien WeiOoi2025-08-312025-08-312024-01-0110.1080/07391102.2023.22625902-s2.0-85173475836http://repository.iitgn.ac.in/handle/IITG2025/2648337787564As a class of ionic liquids with higher biocompatibility, cholinium aminoates ([Cho][AA]) hold potential as solvation media for enzymatic bioprocessing. Herein, solvation effect of [Cho][AA] on structural stability and enzymatic activity of Candida antarctica lipase B (CALB) was evaluated using experimental and computational approaches. Influence of [Cho][AA] on CALB stability was investigated using amino acid anions ([AA]^-) with varying hydrophobicity levels. Choline phenylalaninate ([Cho][Phe]) resulted in 109.1% and 110.4% of relative CALB activity to buffer medium at 25 °C and 50 °C, respectively. Simulation results revealed the improvement of CALB’s enzymatic activities by [AA]^- with a strong hydrophobic character. Shielding of CALB from water molecules by [AA]^- was observed. The level of CALB activity was governed by accumulation level of [AA]^- at CALB’s first hydration layer. The stronger interaction between His224 and Asp187 was postulated to be driven by [Cho][AA], resulting in the activity enhancement of CALB. The slight improvement of CALB activity in 0.05 M [Cho][Phe] at 50 °C could be due to the larger size of entrance to the catalytic site and the stronger interaction between the catalytic residues. The promising effect of [Cho][Phe] on CALB activation may stimulate research efforts in designing a ‘fully green’ bioreaction for various industrial applications. Communicated by Ramaswamy H. Sarma.falseCALB | choline phenylalaninate | conformational changes | enzyme activation | molecular dynamic simulationArticle1538025411351-1136520246arJournal6