生物膜活性测定中TTC-脱氢酶活性测定法的改进

Improvement of Determination on TTC-Dehydrogenase Activity in Biofilms

对生物膜活性测定中氯化三苯基四氮唑(TTC)-脱氢酶活性测定法进行了改进,解决了标准曲线制作稳定性差的问题,对测定中的诸多影响因素进行比较分析,确定了改进后的脱氢酶活性测定的最佳条件.结果表明,以甲苯作为萃取剂的液-液分层明显,提取效果好,操作简便.以硫化钠代替连二亚硫酸钠作还原剂,效果较好,显色稳定不褪色.反应的适宜pH值为8.6,适宜温度为38℃.同时确定了生物膜的最佳培养反应时间为6h.

The method of 3,5,5-triphenyltetrazolium (TTC) determining biofilms activity was improved,and the problem that the standard curve of determination on TTC-dehydrogenase activity can’t be made conveniently and stably was resolved.The factors which influence the determination were studied.The results showed that sodium sulfide as reducer,toluene as TF organic extractant,Tris-HCl as buffer solution,pH 8.6 and 38 ℃ were the best condition on the developed method,and the optimum reaction time to cultivate biofilms is 6 h.