Sephadex G-50纯化生姜蛋白酶的研究

采用Sephadex G-50为填料对粗提生姜蛋白酶进行了纯化,充分利用了尺寸排阻层析简便、高效、重复性好的特点,以酶得率、纯化倍数、柱效分析等参数为指标,研究了凝胶柱床高度、洗脱速度、上样量等条件对纯化效果的影响。确定了最佳纯化方案为:柱床高度50cm,上样量3mL粗酶,流速45cm/h,洗脱液总用量150mL。纯化后的生姜蛋白酶比活性是姜汁的4.192倍,粗酶的2.113倍,而且还去除了色素、不溶物等杂质,纯化效果极佳。

Study on purification of ginger protease by Sephadex G-50

Crude ginger protease was purified by Sephadex G-50, and the optimal purification method was obtained through researches about the gel bed length, flow rate, sample application amount, and was measured by ginger protease yield, purified folds, and the efficiency of chromatography column.The parameters were:gel bed length-50cm,sample application amount-crude ginger protease 3mL (extracted from 10g ginger), flow rate-45cm/h,and the total elution length was 150mL.After purifying,not only the specific activity of purified ginger protease increased to 4.192 folds of ginger juice, and 2.113 folds of crude ginger protease, but also pigment, insoluble impurities were removed,which indicts a good effect of purification.