{"title":"Kinetics of Protease Thermal Inactivation.","authors":"Natalija Andrejević, Natalija Polović, Jelica Milošević","doi":"10.1007/978-1-0716-4478-2_21","DOIUrl":null,"url":null,"abstract":"<p><p>Proteolytic enzymes have various applications in biomedicine, biotechnology, pharmaceuticals, and the food industry. Some of these applications require incubation at extreme temperatures, high pressure, different pH of the solution, and the presence of various additives that might destabilize enzyme structure and function. Structural features of proteases define the mechanism of denaturation, which is reflected in different activation energies for the process. Understanding the activation energy gives clues about the kinetic inertness of the enzyme and its resistance to destabilizing factors. Monitoring the kinetics of enzyme thermal inactivation by measuring activity loss upon heating at various temperatures enables the determination of activation energy for the inactivation process using the Arrhenius plot. This way, measuring the activity loss as a result of structural perturbations gives direct information on the enzyme stability in the relevant conditions for applicative processes.</p>","PeriodicalId":18490,"journal":{"name":"Methods in molecular biology","volume":"2917 ","pages":"247-258"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in molecular biology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1007/978-1-0716-4478-2_21","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
Abstract
Proteolytic enzymes have various applications in biomedicine, biotechnology, pharmaceuticals, and the food industry. Some of these applications require incubation at extreme temperatures, high pressure, different pH of the solution, and the presence of various additives that might destabilize enzyme structure and function. Structural features of proteases define the mechanism of denaturation, which is reflected in different activation energies for the process. Understanding the activation energy gives clues about the kinetic inertness of the enzyme and its resistance to destabilizing factors. Monitoring the kinetics of enzyme thermal inactivation by measuring activity loss upon heating at various temperatures enables the determination of activation energy for the inactivation process using the Arrhenius plot. This way, measuring the activity loss as a result of structural perturbations gives direct information on the enzyme stability in the relevant conditions for applicative processes.
期刊介绍:
For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.