纤维素酶的CMC酶活测定条件的研究

Study on the optimum conditions of determination of cellulase activity

研究和分析了羧甲基纤维素酶(CMC酶)活性测定法的实验条件。通过单因素实验对酶促反应时间、酶促反应温度、pH、测定波长、底物浓度、粗酶液和底物添加量六个影响因素进行了研究。最后结合正交实验、方差分析和多重比较,确定了纤维素酶活力测定的最佳条件组合。结果表明,在pH6.2、粗酶液和底物添加量各为2mL的情况下,影响纤维素酶活力测定的主次因素依次为酶促反应时间、酶促反应温度、测定波长和底物浓度。纤维素酶活力测定的最佳条件为:在酶促反应温度为40℃,pH6.2,底物浓度为10g/L,粗酶液和底物添加量各为2mL,酶促反应时间30min,测定波长为520nm。

The experimental conditions of CMC enzyme activity assay were researched. Six influencing factors： enzymatic reaction time,reaction-temperature,pH,detective wavelength,substrate concentration,crude enzyme and substrate addition were studied by a single factor test. Finally,orthogonal experiment,variance analysis and multiple comparisons were combined together,and the optimum conditions of determination of CMC enzyme activity were determined. The result showed that,when the volume of crude enzyme and substrate were 2mL and pH6.2,the primary and secondary factors which affected cellulase activity were enzymatic reaction time , reaction temperature , enzymatic determination of wavelength and substrate concentration. Experiments showed that：the enzymatic reaction temperature 40℃ ,pH6.2,substrate concentration 10g/L,the volume of crude enzyme and substrate were 2mL,enzymatic reaction time 30min,and the detective wavelength at 520nm were optimum conditions for the determination.