摘要
目的采用不同的检测方法测定重组人透明质酸酶活性,以确定不同条件下最适宜的检测方法。方法采用3,5-二硝基水杨酸法、对-二甲氨基苯甲醛法、浊度法测定重组毕赤酵母发酵上清液和凝胶过滤分离产物中透明质酸酶活性,并分析比较其结果。结果 3,5-二硝基水杨酸法与对-二甲氨基苯甲醛法的线性范围分别是50～450 U/mL,50～350 U/mL。浊度法的线性范围较小,为0～10U/mL。3种方法的RSD值分别是2.71%,11.71%,0.65%。结论发酵上清液中的透明质酸酶适合用3,5-二硝基水杨酸法测定,凝胶过滤产物用浊度法测定效果良好。
Objective To detect the activity of recombinant human hyaluronidase by different methods and determine the optimal methods for activity assay of recombinant human hyaluronidase under different conditions. Methods 3,5- dinitrosalicylic acid（DNS）, p-dimethylaminobenzaldehyde（DMAB） and turbidimetry methods were used to measure the activity of hyaluronidase from Pichia pastoris fermentation supematant and gel filtration products, and the results were analyzed. Results The linear ranges of DNS, DMAB and mrbidimetry methods were 50-450 U/mL, 50-350 U/ mL and 0-10 U/mL, respectively. The RSDs were 2.71%, 11.71% and 0.65%, respectively. Conclusion DNS method is suitable for the activity assay of hyaluronidase from the fermentation supernatant and turbidimetry is suitable for that from gel filtration products.
出处
《食品与药品》
CAS
2011年第1期5-8,共4页
Food and Drug
基金
山东省成果转化重大专项(2009ZHZX1C1001)
山东省优秀中青年科学家科研奖励基金(2008BS02011)
济南市企业自主创新计划(200905050)
