Department of Chemical Engineering, University of California at Berkeley, Berkeley, California 94720, USA.
Biotechnol Bioeng. 1992 Jun 5;40(1):91-102. doi: 10.1002/bit.260400114.
Solubilization properties of alpha-chymotrypsin and alcohol dehydrogenase (LADH) in reverse micelles are reported for three different solubilization techniques. The solubilization properties for these two proteins depend on the method used for protein addition. The addition of a dry protein powder to a reverse-micelle-containing organic phase does not appreciably solubilize the protein until the diameter of the reverse micelle is similar to that of the protein. However, when an aqueous protein solution is injected an organic phase, protein solubilization is not strongly dependent on micelle size. For chymotrypsin, multiple protein occupancy occurs at large micelle size, with as many as 11 chymotrypsin molecules solubilized in one reverse micelle. The solubilization of chymotrypsin using a phase-transfer technique with a positively charged surfactant follows the expected trend based on protein-surfactant electrostatic interactions. When a negatively charged surfactants is used for phase transfer, at low pH the solubilization data do not fit this electrostatic interaction mechanism. In this case, protein-surfactant aggregation may be occurring at the aqueous-organic interface.
报道了三种不同增溶技术中α-糜蛋白酶和醇脱氢酶（LADH）在反胶束中的增溶性质。这两种蛋白质的增溶性质取决于用于添加蛋白质的方法。将干燥的蛋白质粉末添加到含有反胶束的有机相中，直到反胶束的直径与蛋白质相似，否则不会明显增溶蛋白质。然而，当将蛋白质水溶液注入有机相时，蛋白质的增溶并不强烈依赖于胶束的大小。对于糜蛋白酶，在大胶束尺寸下会发生多个蛋白质占据，多达 11 个糜蛋白酶分子溶解在一个反胶束中。使用带正电荷的表面活性剂进行相转移技术增溶糜蛋白酶，基于蛋白质-表面活性剂静电相互作用，符合预期趋势。当使用带负电荷的表面活性剂进行相转移时，在低 pH 值下，增溶数据不符合这种静电相互作用机制。在这种情况下，蛋白质-表面活性剂的聚集可能发生在水-有机界面处。
