Manipulating crystallization in lysozyme and supramolecular self-arrangement in solution using ionic liquids

Abstract

Manipulating crystallization for macromolecule using ionic liquids (ILs) as an additive has received great attention, but how ILs accurately control their formation and crystallization pathway remains unclear. Here, we aim to illuminate the role of ILs on the solution chemistry of macromolecules by investigating their effects on the crystallization of lysozyme. The solubility findings show ILs changed intermolecular contacts, leading to lysozyme supramolecular repulsive interactions being induced or attractive interactions enhanced. Lysozyme was crystallized in the form of tetragonal crystals in the presence of [C₄mim]BF₄ (1-butyl-3-methylimidazolium tetrafluoroborate), [C₄mim]Cl (1-butyl-3-methylimidazolium chloride) and [C₄mim]Br (1-butyl-3-methylimidazolium bromide), while it was crystallized in the form of monoclinic crystals in the presence of [dmim]I (1,3-dimethylimidazolium iodine). Compared with that in the absence of ILs, lysozyme in the presence of [C₄mim]Cl and [dmim]I contained a decreased α-helix content, but increased β-sheet content. The presence of [dmim]I also caused the β-turn content to increase and random coil content to decrease. This demonstration shows the formation of crystal nucleus is involved in transition at the protein secondary structure level, which may contribute to a better understanding of the role of ILs in the crystallization of bio-macromolecular materials and to developing a strategy to produce new bio-macromolecular materials using ILs.