Development of Novel Molecular Assemblies that Enable High Selective Separation for Biomolecules

反胶束在有机介质中创造了独特的环境。它们能够在水中溶解亲水性生物分子(如蛋白质、氨基酸、DNA)。在第一个研究中，我们使用反胶束从水溶液中提取蛋白质到异辛烷中，并阐明了实现有效蛋白质转移的一些重要操作参数。利用阴离子表面活性剂在异辛烷中形成的反胶束成功地提取了目标蛋白。蛋白质的萃取度受表面活性剂的种类、有机溶剂和水相离子强度的影响很大。蛋白质转移的驱动力是阴离子表面活性剂与带正电的蛋白质之间的静电相互作用。具有两个长链烷基链的阴离子表面活性剂是蛋白质从水溶液成功转移到反胶束溶液的最佳候选者。在接下来的研究中，反胶束作为蛋白质复性介质的新功能被实现。在反胶束相中，变性蛋白质在纳米结构的反胶束溶液中被完全重新激活。这种新的复性技术有助于在高蛋白浓度下获得较高的复性产率。将这种反胶束技术应用于蛋白质浓度较高的溶液中，可以提高蛋白质复性的效率，而稀释法会导致蛋白质的聚集。反胶束技术不仅是蛋白质分离的有用工具，也是蛋白质复性的有用工具。

Reversed micelles create unique environment in organic media. They are capable of solubilizing hydrophilic biomolecules (e.g., proteins, amino acids, DNA) in their aqueous interior. In the first study, we conducted protein extraction from an aqueous solution into isooctane using reversed micelles, and some important operational parameters to achieve an efficient protein transfer are elucidated. Targeted proteins were successfully extracted by reversed micelles that are formed by an anionic surfactant in isooctane. The degree of extraction of proteins was strongly influenced by the type of surfactant ih the organic solvent and ionic strength in the aqueous phase. The driving force of the protein transfer was electrostatic interaction between anionic surfactants and the positively charged protein. Anionic surfactants possessing two long-alkyl chains are the best candidate for the successful transfer of proteins from the aqueous solution to the reversed micellar solution.In the next study, novel function of reversed micelles as a protein refolding medium was carried out. In the reversed micellar phase, denatured proteins were completely reactivated in a nano-structured reversed micellar solution. This novel refolding technique facilitates a high renaturation yield at high protein concentration. The efficiency of protein refolding was enhanced when this novel technique by reversed micelles is applied to the solution containing higher protein concentration which, in the case of dilution method, would lead to protein aggregation. The reversed micellar technique is found to be a useful tool not only for protein separation but also for protein refolding.