毛细管区带电泳用于酶微反应器酶解条件的研究

将氨基酰化酶通过戊二醛固定在毛细管内壁,制备毛细管酶微反应器,用毛细管区带电泳对毛细管酶微反应器的酶解产物进行分离,以生成物的峰面积优化底物N-乙酰-DL-蛋氨酸的酶解条件。实验结果表明,在温度37℃的条件下,10μg/mL N-乙酰-DL-蛋氨酸磷酸盐缓冲溶液(pH7.5)以4μL/min的速度通过15 cm长的毛细管酶微反应器,具有良好的酶解效果。利用毛细管酶微反应器对底物N-乙酰-DL-蛋氨酸进行酶解,每天酶解5次,10天后酶活仅下降了8.66%,说明制备的毛细管酶微反应器具有良好的稳定性。

Study on the enzymatic hydrolysis in capillary enzyme micro-reactor with capillary zone electrophoresis

The capillary enzyme micro-reactor was prepared by immobilizing aminoacylase on the inner wall of capillary with glutaraldehyde as a cross-linking agent. The enzymatic hydrolysis of N-acetyl-DL-methioine in the prepared micro-reactor was evaluated by capillary zone electrophoresis. The results showed that high efficiency of the enzymatic hydrolysis could be obtained at pH 7.5 and 37 ℃ for 10 μg/mL N-acetyl-DL-methioine with a flow rate of 4 μL/min in a 15 cm capillary enzyme micro-reactor. The enzyme micro-reactor was stable while its activity decreased less than 8.66% within 10 days.