Valls Cristina, Pujadas Gerard, Garcia-Vallve Santi, Mulero Miquel
Nutrigenomics Group, Department of Biochemistry and Biotechnology, Rovira i Virgili University, Tarragona 43007, Spain.
Biochem Mol Biol Educ. 2011 Jul;39(4):280-90. doi: 10.1002/bmb.20488.
Detergent enzymes account for about 30% of the total worldwide production of enzymes and are one of the largest and most successful applications of modern industrial biotechnology. Proteases can improve the wash performance of household, industrial, and institutional laundry detergents used to remove protein-based stains such as blood, grass, body fluids, and food soils. This article describes two easy and cheap laboratory exercises to study the presence, profile, and basic enzymology of detergent proteases. These laboratory practicals are based on the determination of the detergent protease activity of various commercial detergents using the N-succinyl-L-alanyl-L-alanyl-L-prolyl-L-phenylalanine p-nitroanilide method and the bovine serum albumin degradation capacity. Students are also required to elucidate the enzymatic subtype of detergent proteases by studying the inhibitory potential of several types of protease inhibitors revealed by the same experimental methodology. Additionally, the results of the exercises can be used to provide additional insights on elementary enzymology by studying the influence of several important parameters on protease activity such as temperature (in this article) and the influence of pH and effects of surfactants and oxidizers (proposed). Students also develop laboratory skills, problem-solving capacities, and the ability to write a laboratory report. The exercises are mainly designed for an advanced undergraduate project in the biochemistry and biotechnology sciences. Globally, these laboratory practicals show students the biotechnological applications of proteases in the detergent industry and also reinforce important enzymology concepts.
洗涤剂用酶约占全球酶总产量的30%,是现代工业生物技术中规模最大、最成功的应用领域之一。蛋白酶可以提高家用、工业用和公共机构用洗衣洗涤剂的洗涤性能,这些洗涤剂用于去除基于蛋白质的污渍,如血液、草渍、体液和食物污垢。本文介绍了两个简单且成本低廉的实验室实验,用于研究洗涤剂蛋白酶的存在、概况和基本酶学性质。这些实验室实践基于使用N-琥珀酰-L-丙氨酰-L-丙氨酰-L-脯氨酰-L-苯丙氨酸对硝基苯胺法和牛血清白蛋白降解能力来测定各种商用洗涤剂中洗涤剂蛋白酶的活性。学生还需要通过研究用相同实验方法揭示的几种类型蛋白酶抑制剂的抑制潜力,来阐明洗涤剂蛋白酶的酶亚型。此外,通过研究几个重要参数(如本文中的温度)对蛋白酶活性的影响以及pH值、表面活性剂和氧化剂的影响(建议研究),这些实验结果可用于提供关于基础酶学的更多见解。学生还能培养实验室技能、解决问题的能力以及撰写实验室报告的能力。这些实验主要是为生物化学和生物技术专业的高年级本科生项目设计的。在全球范围内,这些实验室实践向学生展示了蛋白酶在洗涤剂行业的生物技术应用,同时也强化了重要的酶学概念。