Application of Reverse Micelles for the Extraction of Amino Acids and Proteins

Application of Reverse Micelles for the Extraction of Amino Acids and Proteins
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反胶束在氨基酸和蛋白质提取中的应用

DOI：
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发表时间：
1990
期刊：
Chimia (Basel)
影响因子：
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通讯作者：
P. Luisi
P. Luisi
中科院分区：
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文献类型：
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作者：
M. Leser;P. Luisi

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被引文献

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研究了使用液-液萃取和固-液萃取方法使用油包水微乳液溶解和提取氨基酸和蛋白质。小单体蛋白（如细胞色素 c、溶菌酶、 通过改变几个外部参数，例如 wo (wo = [H2O]/[AOT]）、有机相中的表面活性剂类型和浓度，以及 H2O 相中的 pH、离子强度、盐类型。在液-液萃取方法中，本发明的双相系统由水和上清液反胶束溶液组成。化合物 从水相转移到有机胶束相。还考虑到同时发生的 H2O 转移来研究该过程。转移动力学研究提供了有关 H2O 和蛋白质提取之间联系的信息。 在第二种技术中，化合物以固态提供并悬浮在含有已知量溶解水的有机胶束溶液中。通过液-液相转移方法，结果表明，在一定条件下，定量且相当 可以实现显着的转移，其中盐的类型及其在水相中的浓度以及水相 pH 值在决定传质程度方面起着最重要的作用。胶束和提取物之间静电相互作用的意义 通过比较 AOT 和 CTAB（十六烷基三甲基溴化铵；是一种阳离子表面活性剂）油包水微乳液的提取结果，也可以证明分子的存在。此外，一系列带正电、带负电和中性氨基酸衍生物的提取率为 研究了两种表面活性剂体系，以表征表面活性剂分子的静电和疏水相互作用的重要性。还用固相萃取法对这一系列氨基酸衍生物进行了研究。对于蛋白质转移的情况 从固态来看，溶解率受蛋白质和胶束液滴尺寸的影响。对于小蛋白质，即分子量低于约。 15 000 D，提取曲线作为 wo 的函数，令人惊讶地在小 wo 中显示出足够的峰值 区域（6 到 12 之间）。通过将 H2O 含量增加到超过此限制，H2O 可溶性蛋白质的转移效率会降低。对于分子量高于 20 000 D 的蛋白质，提取曲线为 wo 的函数 随着 wo 的增加而单调增加，没有任何尖锐的 wo 最大值。提供了一个定性模型来解释这种不同的行为。还表明市售蛋白粉的电离状态显着影响提取 行为。胶束排斥效应最终应用于从粗细菌蛋白质混合物中提取蛋白质。
The use of water-in-oil microemulsions for the solubilization and extraction of amino acids and proteins is investigated, using both the liquid-liquid extraction and the solid-liquid extraction method. The solubilization behaviour of small monomeric proteins such as cytochrome c, lysozyme, ribonuclease A, trypsin, ?-chymotrypsin, papain, pepsin, and bovine serum albumin (BSA) into mainly AOT (the anionic surfactant bis(2-ethylhexyl) sodium sulfosuccinate)/H2O/isooctane is studied by varying several external parameters, such as wo (wo = [H2O]/[AOT]), surfactant type and concentration in the organic phase, and, pH, ionic strength, salt type in the H2O phase. In the liquid-liquid extraction method, the present biphasic system consists of an aqueous and a supernatant reverse micellar solution. The compounds are transferred from the H2O phase into the organic micellar phase. This process is investigated also in view of the simultaneously occurring H2O transfer. Studies of transfer kinetics give an information about the connection between the H2O and protein extraction. In the second technique, the compounds are given in their solid state and suspended in the organic micellar solution which contains already a known amount of solubilized water.With the liquid-liquid phase-transfer method, it is shown that, under certain conditions, a quantitative and quite remarkable transfer can be achieved, whereby the salt type and its concentration in the H2O phase, as well as the aqueous pH, play the most important role in determining the extent mass transfer. The significance of electrostatic interactions between the micelles and the extracted molecules is evidenced also by comparing extraction results obtained with AOT and CTAB (cetyltrimethylammonium bromide; is a cationic surfactant) water-in-oil microemulsions. Moreover, the extraction yield of a series of positively, negatively charged, and neutral amino-acid derivatives is investigated in the two surfactant systems, in order to characterize the significance of electrostatic and hydrophobic interactions with the surfactant molecules. This series of amino-acid derivatives is also studied with the solid-phase extraction method. In the case of proteins transferred from the solid state, the solubilization yield is affected by the protein and micellar droplet size. For small proteins, i.e. with a molecular weight below ca. 15 000 D, the extraction curve as a function of wo shows surprisingly enough a peak in the small wo region (between 6 and 12). By increasing the H2O content above this wo limit, the transfer efficiency of the H2O-soluble proteins decreases. For proteins having a molecular weight above 20 000 D, the extraction curve as a function of wo shows a monotonous increase with increasing wo without any sharp wo maxima. A qualitative model is offered to explain this different behaviour. It is also shown that the ionization state of commercially available protein powders markedly affects the extraction behaviour. The micellar exclusion effect is finally applied in the extraction of proteins from a crude bacterial protein mixture.