CTAB/正丁醇/异辛烷反胶团法纯化α-淀粉酶

Purification of α-amylase by using CTAB/n-butane/isooctane reversed micelles system

以CTAB/正丁醇/异辛烷构成反胶团系统,通过反胶团萃取方式纯化精制α-淀粉酶。最佳反应条件为:萃取温度40℃,水相组成为NaCl 0.03 mol/L,pH12.0,有机相∶无机相=1∶2,振荡时间10 min;反萃取最佳条件为:温度60℃,水相组成为KCl 3 mol/L,pH4.0,有机相∶无机相=2∶1,反萃取振荡时间10 min。在上述条件下,经过一个萃取与反萃取循环后,α-淀粉酶的萃取率最高可达90.78%。

Purification of roughcast α-amylase was carried out by using reversed micellar extraction with CTAB, n-butane and isooctane reversed micellar solution as the extractant. The optimal experimental conditions to purify α-amylase were found： the compositions of initial aqueous phase is NaCl 0.03 mol/L buffered solution,extraction time is 40 ℃, pH = 12.0 containing crude enzyme, the proportion of the organic substance and the buffered solution is 1：2, the forward extraction time is 10 min;the composition of stripping solution is KCl 3 mol/L of reversed micellar in buffered solution, extraction temperature is 60 ℃, pH = 4.0, the proportion of the organic substance and the buffered solution is 2 ： 1, the backward extraction time is 10 min. Under such conditions,the extraction rate of α-amylase is 90.78% after a full forward and backward extraction cycle.