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Modulation of the competition between renaturation and aggregation of lysozyme by additive mixtures.
Biotechnology and Applied Biochemistry ( IF 2.8 ) Pub Date : 2019-11-23 , DOI: 10.1002/bab.1864
Zeinab Takalloo 1 , Forouzan Niknaddaf 1 , S Shirin Shahangian 2 , Akbar Heydari 3 , Saman Hosseinkhani 1 , Reza H Sajedi 1
Affiliation  

The effects of 17 kinds of additive mixtures have been studied on refolding and aggregation of a model protein, lysozyme. Most of the prepared mixtures were efficient in inhibiting aggregation of the protein, and, surprisingly, four novel additive mixtures, i.e., lactic acid: l ‐arginine, lactic acid: l ‐glutamine, choline chloride: lactic acid, and imidazolium salt: β‐cyclodextrin as well as choline chloride: urea exhibited a more remarkable efficacy in suppressing aggregation. Among these, lactic acid: l ‐arginine was identified as the most efficient additive, and lactic acid: l ‐glutamine and choline chloride: lactic acid were inefficient to recover the enzyme activity. In contrast, choline chloride: ethylene glycol: imidazole, choline chloride: glycerol: imidazole, imidazole: betaine: ethylene glycol were found to be less effective mixtures in preventing enzyme aggregation. Totally, it was demonstrated that the protective effects of the mixtures were improved as their concentrations increased. The improvement was more remarkable for imidazolium salt: β‐cyclodextrin and choline chloride: urea, where the denatured lysozyme was reactivated and recovered up to 85% of its initial activity by enhancing their concentrations from 1 to 5% (V/V). It is suggested that such solution additives may be further employed as artificial chaperones to assist protein folding and stability.

中文翻译:

添加剂混合物对溶菌酶复性和聚集之间竞争的调节。

研究了17种添加剂混合物对模型蛋白溶菌酶的重折叠和聚集的影响。多数制备的混合物可有效抑制蛋白质的聚集,令人惊讶的是,四种新型添加剂混合物,即乳酸:l-精氨酸,乳酸:l-谷氨酰胺,氯化胆碱:乳酸和咪唑鎓盐:β环糊精和氯化胆碱:尿素在抑制聚集方面表现出更显着的功效。其中,乳酸:l-精氨酸被认为是最有效的添加剂,而乳酸:l谷氨酰胺和氯化胆碱:乳酸不能有效地恢复酶的活性。相反,发现胆碱氯化物:乙二醇:咪唑,胆碱氯化物:甘油:咪唑,咪唑:甜菜碱:乙二醇是防止酶聚集的不太有效的混合物。总的来说,证明了混合物的保护作用随着其浓度的增加而提高。咪唑鎓盐:β-环糊精和氯化胆碱:尿素的改善更为显着,其中变性的溶菌酶通过将其浓度从1增加到5%(V / V)而被重新活化并恢复到其初始活性的85%。建议将这种溶液添加剂进一步用作人工伴侣,以帮助蛋白质折叠和稳定。
更新日期:2019-11-23
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