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Development of new surfactants for protein extraction using reversed micelles

使用反胶束开发用于蛋白质提取的新型表面活性剂

基本信息

批准号：
05555213
负责人：
NAKASHIO Fumiyuki
金额：
$ 5.25万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research (B)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1994
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-05555213/
关键词：
Reversed Micelles Microemulsion Surfactant Protein Separation Extraction Molecular aggregation Biotechnology 酵素反応

项目摘要

Solvent extraction using reversed micelles provides an attractive to conventional procedures for the separation and purification of proteins from fermentation media and from very dilute solution. In the separation process, it is well known that a surfactant plays an important role, because solubilized proteins are stabilized by a surfactant layr. However, little attention has been paid to the design and synthesis of optimal surfactants for the protein extraction. In this study, new surfactants have been synthesized for potential use in reversed micellar protein extraction operations, and the following results were obtained :1.Preferential solubility of the surfactant in an aliphatic solvent such as hexane, heptane or iso-octane, and the formation of reversed micelles with solubilization of significant quantities of water can be achieved by using strongly hydrophobic, twin alkyl chains as the hydrophobic moiety.2.Different surfactants having identical water solubilizing capacities can have significantly different behavior in protein extractions, where extraction efficiency appears to be governed by the nature of the interfacial complex that forms between surfactants and proteins.3.Bulky surfactant chains provide a steric hindrance to the adsorption of the surfactant to the protein surface, thus inhibiting solvation of the protein/surfactant complex, and hence protein extraction. Under these conditions, a precipitate forms either in the bulk aqueous phase or at the interface. Surfactants that can from a close packed complex with the protein are excellent protein solubilizing agents. Dioleyl phosphoric acid (DOLPA) appears to be one of the best surfactants currently available for protein extraction.Now, we are studying more detail about extraction mechanism of proteins and novel application of newly developed reversed micelar sysytem.

使用反胶束的溶剂萃取为从发酵介质和非常稀的溶液中分离和纯化蛋白质提供了一种有吸引力的传统方法。在分离过程中，众所周知，表面活性剂起着重要的作用，因为溶解的蛋白质被表面活性剂层稳定。然而，对蛋白质提取最佳表面活性剂的设计和合成却很少关注。在本研究中，合成了新的表面活性剂，有望用于反胶束蛋白提取操作，并获得了以下结果：表面活性剂在脂肪族溶剂如己烷、庚烷或异辛烷中具有优先的溶解性，并且通过使用强疏水的双烷基链作为疏水基团，可以形成与大量水溶解的反胶束。具有相同水溶性的不同表面活性剂在蛋白质提取中可能有明显不同的行为，其中提取效率似乎由表面活性剂和蛋白质之间形成的界面复合物的性质决定。庞大的表面活性剂链为表面活性剂在蛋白质表面的吸附提供了空间位阻，从而抑制了蛋白质/表面活性剂复合物的溶剂化，从而抑制了蛋白质的提取。在这些条件下，沉淀要么在整体水相中形成，要么在界面处形成。表面活性剂能与蛋白质形成紧密的络合物，是极好的蛋白质增溶剂。二油基磷酸（DOLPA）是目前用于蛋白质提取的最佳表面活性剂之一。目前，我们正在对蛋白质的提取机理和新开发的反胶束体系的新应用进行更详细的研究。