DNS比色法测定污泥中淀粉酶活性的研究

Study on Determination of Amylase Activity in Activated Sludge with DNS Spectrophotometry

[目的]研究利用DNS比色法测试污泥中淀粉酶活性,以解决测试条件差异大和测试结果可比性低等问题。[方法]以水解酸化反应器中的污泥为原料,研究工作波长、显色及培养时间、培养pH值、灭菌时长等因素对测试结果产生的影响。[结果]结果表明,最大吸收波长处的背景干扰大且测试不稳定,工作波长应在背景干扰相对较小的长波方向选择。该研究的最适工作波长为510 nm;显色的适宜时间为8 min;在培养阶段添加的缓冲溶液pH值与原污泥pH值不一致,会造成实际酶活力的放大或缩小;灭菌时间过短则灭菌不完全,会造成测试结果偏低,该研究确定的灭菌时间为10min;同时确定了最适的培养反应时间为90 min。[结论]为DNS-淀粉酶测定方法的完善提供依据。

[Objective] The research aimed to study the measurement of amylase activity in activated sludge with DNS spectrophotometry, and solve the large difference of test conditions and the low comparability of test results. [Result] Using the activated sludge of the hydro-acidification reactor as the exper/mental material, effects of the work wavelength, pH value, disinfection and incubation time, the the of color reaction were investigated. [ Result] the resuits showed that the absorbency of the reducing product should be measured at 510 nm to abate the disturbance of DNS.The boiling time for color re.action should be about 10 rain and the pH value of the buffer should be the same as the sample to avoid magnifying or lowering amylase activity. The disinfection time should be at least 10 min. Otherwise, the result of the measurement should be lower because of the survival of bacteria.The best incubation time was about 90 rain. [Conclusion] The study provided the basis for improving the determination method of amylase activity based on DNS spectmphotometry.