A Study on Protein Separation with AOT-Isooctane Reverse Micellar System

AOT-Isooctane 역마이셀계를 이용한 단백질 분리에 관한 연구

  • Published : 1989.08.01


For the selective separation of proteins, the solubilization and desolubilization of proteins in sodium-di-2-ethylhexyl sulfosuccinate (AOT)-isooctane reverse micellar system were investigated. Protein solubilization increased with increasing the concentration of AOT to 200 mM and then decreased above that concentration. Protein was solubilized into reverse micelles in the pH range below the isoelectric Point of each protein, pH 4-10 for lysozyme and pH 5-6 for trypsin and ${\alpha}-chymotrypsin$, Lysozyme, trypsin and ${\alpha}-chymotrypsin$ were efficiently extracted in the precence of KCl and NaCl while larger molecular weight proteins such as pepsin and BSA had high solubilization with $CaCl_2$. At higher ionic strength all proteins exhibited murk less tendency to solubilize and the increase of ionic strength resulted in the decrease of micelle size. Lysozyme was successfully back transfered at pH 12.2 and 1.0M KCl; trypsin at pH 12.6 and 0.5M KCl; and ${\alpha}-chymotrypsin$ at pH 6.7 and 0.5M KCl. In a test group separation experiments, complete separation of lysozyme from BSA could be obtained.