离子液体对溶菌酶晶体形态和活性的影响

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摘要以鸡蛋清溶菌酶为模型蛋白，采用冷却结晶法结晶以及浊度法测定酶活性，研究了分别添加4种亲水性离子液体（1-丁基-3-甲基咪唑四氟硼酸盐（[C₄mim]BF₄）、1-丁基-3-甲基咪唑氯盐（[C₄mim]Cl）、1-丁基-3-甲基咪唑溴盐（[C₄mim]Br）和1，3-二甲基咪唑碘化物（[dmim]I））条件下溶菌酶晶体形态和晶体活性的变化。利用光学显微镜对溶菌酶结晶过程的晶体形态进行分析，结果表明添加离子液体会影响溶菌酶的晶体形态和尺寸。离子液体[C₄mim]BF₄介导的溶菌酶形成棒状晶体。与未添加离子液体相比，[C₄mim]Cl和[C₄mim]Br均未改变溶菌酶晶体形态，均呈块状晶体，但[C₄mim]Cl和[C₄mim]Br促进溶菌酶晶体生长，晶体尺度均增加。而添加[dmim]I，溶菌酶呈现针状晶体。采用浊度法测定溶菌酶晶体活性，结果表明不同的离子液体对溶菌酶晶体的活性影响不同。其中，[C₄mim]BF₄和[dmim]I的添加降低了溶菌酶晶体的活性，而[C₄mim]Cl和[C₄mim]Br的添加则提高了酶晶体的活性。

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	李娜
	王占忠

关键词：溶菌酶离子液体结晶酶活性

Abstract： In this study, egg white lysozyme was used as a model protein, crystals were prepared by cooling crystallization and the activity of lysozyme crystal was determined by the turbidity method, and changes in morphology and activity of lysozyme crystals were investigated under the condition of adding four kinds of hydrophilic ionic liquids (ILs) respectively. The four ILs are 1-butyl-3 methylimidazolium tetrafluoroborate ([C₄mim]BF₄), 1-butyl-3 methylimidazolium chloride ([C₄mim]Cl), 1-butyl-3 methylimidazolium bromide ([C₄mim]Br) and 1,3 dimethylimidazolium iodide ([dmim]I). The crystal morphology of the lysozyme during the crystallization process was analyzed by optical microscopy, and the results showed that the addition of IL affects the morphology and size of the lysozyme crystal.[C₄mim]BF₄ mediates lysozyme to form rod-like crystals. Both[C₄mim]Cl and[C₄mim]Br do not change the morphology of lysozyme crystals but promote the growth of lysozyme crystals compared with no IL addition. In the presence of[C₄mim]Cl and[C₄mim]Br, lysozyme crystals are square and increase in size. When[dmim]I was added, lysozyme shows needle-like crystals. The activity of the lysozyme crystal was determined by turbidimetry. The results show that different ILs have different effects on the activity of lysozyme crystals. Among them, the addition of[C₄mim]BF₄ and[dmim]I decreased the activity of lysozyme crystals, while the addition of[C₄mim]Cl and[C₄mim]Br increased the activity of enzyme crystals.

Keywords： lysozyme ionic liquid crystallization enzymatic activity

Received 2022-07-23;

Fund:国家自然科学基金（21978206）。

Corresponding Authors: 王占忠,副教授,E-mail:wzz7698@tju.edu.cn。 Email: wzz7698@tju.edu.cn

About author: 李娜(1998-),女,硕士研究生,现从事蛋白质结晶方面的研究。

引用本文:

李娜, 王占忠.离子液体对溶菌酶晶体形态和活性的影响[J]. 化学工业与工程, 2023,40(1): 112-119

LI Na, WANG Zhanzhong.Influences of ionic liquids on morphology and activity of lysozyme crystal[J]. Chemcial Industry and Engineering, 2023,40(1): 112-119

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