Study on the effect of 1-octyl-3-methylimidazole bromide aggregation state on protein crystallization

doi:10.11949/0438-1157.20210214

Abstract

Abstract:

The effect of the aggregation state of the ionic liquid 1-octyl-3-methylimidazole bromide([C₈mim]Br) additives on the protein crystallization process was studied. Hen egg white lysozyme was chosen as the model protein and [C₈mim]Br was applied as the additive. It was found that the presence of [C₈mim]Br could improve the crystal morphology as well as affect the number and size of crystals. The influence mechanism of ionic liquid [C₈mim]Br on the crystallization of lysozyme was revealed by measuring the solubility, crystallization kinetics, aggregation state and zeta-potential at different [C₈mim]Br concentrations. The results showed that the interaction mechanism between lysozyme and [C₈mim]Br varied with the aggregation states of [C₈mim]Br. When the concentration of [C₈mim]Br was low (< 0.1 mol/L), there was hydrophobic interactions between [C₈mim]+ and lysozyme molecules, which promoted the aggregation and crystallization process. When the concentration of [C₈mim]Br was high (> 0.1 mol/L), [C₈mim]+ self-aggregated into micelle and formed aggregate complex with lysozyme molecules. The results from DLS further showed that the lysozyme-micelle complex was the basic unit in nucleation and hence, a slower crystallization rate and a higher crystal quality were obtained.

Key words: ionic liquids, aggregation, protein, crystallization

摘要：

研究了离子液体溴化1-辛基-3-甲基咪唑（［C₈mim］Br）添加剂的聚集状态对蛋白质结晶过程的影响。实验发现［C₈mim］Br可以改善溶菌酶晶体形貌，影响晶体数量和晶体尺寸。通过测定不同浓度［C₈mim］Br下溶菌酶的溶解度、结晶动力学、聚集状态和ζ-电势，揭示了离子液体［C₈mim］Br对溶菌酶结晶的影响机理。结果表明，［C₈mim］Br与溶菌酶的作用方式随［C₈mim］Br的聚集状态而改变。当［C₈mim］Br浓度较低（<0.1 mol/L）时，［C₈mim］⁺与溶菌酶分子之间存在疏水作用，可以促进蛋白质分子聚集，促进结晶过程。当［C₈mim］Br浓度较高（>0.1 mol/L）时，［C₈mim］⁺自发聚集成胶束，并与蛋白质分子相互作用形成溶菌酶-胶束复合物；动态光散射（DLS）结果显示结晶过程以溶菌酶-胶束复合物为基本单元聚集，因此结晶过程减缓，晶体质量提高。

关键词: 离子液体, 聚集, 蛋白质, 结晶

CLC Number:

TQ 423.9

Xiaoxi YU, Zhenzhen YAN, Qihui JIANG, Xia WU, Yuxiao ZHANG, Xiaojuan WANG, Fang HUANG. Study on the effect of 1-octyl-3-methylimidazole bromide aggregation state on protein crystallization[J]. CIESC Journal, 2021, 72(9): 4854-4860.

于筱溪, 闫真真, 蒋其辉, 吴霞, 张余晓, 王晓娟, 黄方. 溴化1-辛基-3-甲基咪唑聚集状态对蛋白质结晶的影响研究[J]. 化工学报, 2021, 72(9): 4854-4860.

Figures/Tables 8

Fig.1 Microgscope photos of lysozyme crystals (scale bars：100 μm)

Fig.2 Numbers of lysozyme crystals under different [C8mim]Br concentration

Fig.3 Lysozyme activity ratio under different [C8mim]Br concentration

Fig.4 Solubility of lysozyme under different [C8mim]Br concentration

Fig.5 Lysozyme crystallization kinetics under different [C8mim]Br concentration

Fig.6 The ζ-potential of lysozyme under different [C8mim]Br concentration

Fig.7 Particle size distribution of aggregates in [C8mim]Br solution (a); Particle size distribution of aggregates in crystallization solution with protein and [C8mim]Br (b)

Fig.8 Aggregates change during induction period

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