Lipase and protease extraction from activated sludge
Lipase and protease extraction from activated sludge
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DOI：
10.1016/s0043-1354(03)00241-0
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发表时间：
2003-09-01
期刊：
影响因子：
12.8
通讯作者：
Nielsen, PH
中科院分区：
文献类型：
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作者：
Gessesse, A;Dueholm, T;Nielsen, PH
在废水处理过程中，聚合物的水解是第一步，也是限速步骤。更深入地研究催化这些反应的酶对于更好地了解废水处理过程中的微生物活动是必不可少的。因此，开发温和高效的环境样品酶提取方法具有重要意义。本研究提出了一种利用非离子清洗剂Triton X-100、EDTA和阳离子交换树脂(CER)单独或联合提取活性污泥中的脂肪酶和蛋白酶的方法。在单独存在缓冲液的情况下或在洗涤剂和/或螯合剂存在的情况下，连续搅拌污泥。在所有情况下，连续搅拌后，可以观察到絮体尺寸显著减小。然而，在单独的缓冲液存在下，没有提取出脂肪酶活性和可以忽略的蛋白酶活性，这表明酶的提取不是单独的剪切力所致。0.1%Triton X-100提取的脂肪酶活力最高，高于0.1%时酶活力逐渐下降。对于蛋白水解酶，当Triton X-100的浓度为0.5%时，活性最高，且活性没有下降。观察到两种酶提取效率的差异，表明需要优化从活性污泥中提取不同酶或胞外聚合物的工艺。(C)2003爱思唯尔科学有限公司。保留所有权利。
In the process of wastewater treatment hydrolysis of polymeric substances is the first and rate-limiting step. A closer study of the enzymes catalysing these reactions is essential for a better understanding of the microbial activity in the wastewater treatment process. Therefore, development of gentle and efficient enzyme extraction methods from environmental samples is very important. In this study we present a method for the extraction of lipases and proteases from activated sludge using the non-ionic detergent Triton X-100, EDTA, and cation exchange resin (CER), alone or in combination for the extraction of lipases and proteases from activated sludge. The sludge was continuously stirred in the presence of either buffer alone or in the presence of detergent and/or chelating agents. In all cases, a marked reduction in floc size was observed upon continuous stirring. However, no lipase activity and negligible protease activity was extracted in the presence of buffer alone, indicating that enzyme extraction was not due to shear force alone. The highest lipase activity was extracted using 0.1% Triton X-100 above which the activity was gradually decreasing. For proteases, the highest activity was obtained in the presence of 0.5% Triton X-100 and no decrease in activity was observed. Differences observed in the extraction efficiency of the two enzymes indicate the need for optimisation of the extraction process for the different enzymes or the extracellular polymeric substances from activated sludge. (C) 2003 Elsevier Science Ltd. All rights reserved.