活性污泥总蛋白提取方法的比较

Comparison of Total Protein Extraction Methods for Activated Sludge

有效提取总蛋白是活性污泥蛋白质组学研究的第一步,由于活性污泥中含有腐殖质以及其他复杂的有机物质,因此常规的提取方法均难以获得高质量的蛋白质。文章考察了SDS-酚-TCA法、SDS-酚法、柠檬酸钠-酚法、SDS提取法和SDS震荡提取法抽提活性污泥总蛋白的效果,比较了蛋白得率,并通过SDS-PAGE对蛋白的质量进行了检测。结果表明,5种方法均可从活性污泥中提取蛋白质,但5种方法在提取的蛋白的得率和质量上存在差异,其中采用SDS-酚法抽提的蛋白质效果最好,以该方法提取的蛋白质得率最高并且蛋白条带最多。该文为活性污泥总蛋白的提取提供了一种有效的方法。

Isolation of high-quality protein from activated sludge is the first step to study activated sludge metaproteomics. Activated sludge is rich in humic substances and other organic substances which may cause low yield of protein during protein extraction with the conventional method. Five protein extraction methods such as SDS-phenol-TCA extract, SDS- phenol extract, citrate-phenol extract, SDS extract and SDS-shaking-extract were compared and investigated. Yield of the extracted protein and protein quality were tested by SDS-PAGE. The results showed that all these methods can extract certain protein from activated sludge but with different yield and quality, SDS-phenol extract obtained the highest total protein yield and bands. This study provides an efficient method for extracting total protein from activated sludge.