六种活性污泥酶活测定及条件优化

The Optimization Parameters of Activity Measurement for 6 Kinds of Enzymes from Activated Sludge

活性污泥的生物降解能力与其酶活性相关。在各种涉及活性污泥的环境工程和生物降解分析测试过程中,快速测定活性污泥的酶活性有助于各种活性污泥的生物降解能力分析和实时监测,对于监控降解过程有着重要意义。文章选择6种与生物降解直接相关的活性污泥酶进行测定方法的研究,从反应体系的pH、反应温度、反应时间、反应终止方法和污泥浓度等4个方面对L-亮氨酸氨基肽酶(L-AP)、棕榈酸脂酶(LIP)、磷酸酯酶(PPA)、α-葡萄糖苷酶(α-GLC)、磷酸酶(APA)和脱氢酶(DHA)的酶活测定条件进行优化,结果显示,L-AP最适反应pH为7.4,PPA、APA和DHA反应最适pH为7.9,LIP和a-GLC最适pH为8.4。6种酶活的最适反应温度均为37℃。a-GLC适合反应时间为90 min,其余酶活为60 min。L-AP、LIP、PPA、a-GLC和APA反应终止方法为高温加热,DHA反应的产物萃取剂为乙醇:二甲基甲酰胺=1:1。在优化测定条件的基础上提出一种快速测定6种与生物降解相关的酶活方法,用该方法测定采自4个不同污水处理厂的活性污泥的酶活,除了LIP外,发现工业污水处理厂的活性污泥酶活普遍要高。优化后的测定方法方便快捷,对仪器设备要求低,更能直接、真实反映活性污泥的生物降解活性。

The biodegradability of activated sludge is related to its enzyme activity. In the process of environmental engineering and biological degradation test, it is of importance to monitor the degradation process when the enzyme activity of activated sludge could be rapidly measure in time. This paper aimed to study the assay method of 6 kinds of emzymes in activated sludge which is directly related to the biodegradation. These enzymes are L-leucine aminopeptidase （L - AP）, palmitic acid lipase （LIP） , phosphatase （PPA） , ~t-glycosidase （a-GLC） , phosphatase （APA） and dehydrogenase （DHA） .From 4 aspects , including the reaction system of pH, reaction temperature, reaction time, reaction termination methods, reaction conditions of these enzymes are optimized. 4 kinds of activated sludge were collected from different wastewater treatment plants and their enzyme activities were measured with this method. Results show that the best reaction pH is 7.4 （ L-AP）, 7.9 （PPA, APA and DHA）, 8.4 （LIP, α-GLC）. The optimum reaction temperature is 37 ℃. The most suitable reaction time of a-GLC is 90 min, the rest of enzymes is 60 min. The reaction termination method of hydrolase is high temperature heating and DHA reaction termination agent is extraction agent （ethanol：dimethyl formamide - 1：1 ）. A rapid method was suggested on the base of conditions optimization. The emzyme activities of activated sludge from induslrial wastewater treatment plant are generally higher than others except LIP. This assay method is not only more conveniently, quickly and low requirements for equipment, but also can be more directly reflect the biodegradation activity of activated sludge.