不同破壁方法对丙酮酸激酶活性的影响

Influence of Different Wall-breaking Methods on Activity of Pyruvate Kinase

目的研究4种不同处理方法对纤维堆囊菌中丙酮酸激酶酶活性的影响。方法分别采用液氮研磨法、石英砂研磨法、超声波法和溶菌酶法将纤维堆囊菌细胞破壁,然后分别测定丙酮酸激酶的酶活性,比较酶活性及操作难易度。结果石英砂研磨条件下酶活性最小,超声波破碎和液氮研磨酶活性相近,溶菌酶法最高;溶菌酶法耗时间最长,其他3种相近。结论超声波破碎法成本低,效率高,是首选的方法。

Objective To study the effects of 4 kinds of processing methods on the activity of pyruvate kinase from Sorangium cellulosum. Methods The liquid nitrogen grinding, quartz sand grinding, ultrasonic breaking and enzymolysis using lysozyme were respectively used for cell disruption of Sorangium cellulosum, then the activity of pyruvate kinase obtained by different wall-breaking methods was determined in order to compare the activity and operation complexity. Results Of the activity of pyruvate kinase obtained by different methods, the lowest was that obtained by quartz sand grinding and the highest was that obtained by enzymolysis; the activity of pyruvate kinase obtained by ultrasonic breaking and liquid nitrogen grinding was equivalent. The enzymolysis was most time consuming, and the efficiency was similar for the other three methods. Conclusion The ultrasonic breaking is the preferred method because of low cost and high efficiency.