Andrejević Natalija, Polović Natalija, Milošević Jelica
Department of Biochemistry, University of Belgrade - Faculty of Chemistry, Belgrade, Serbia.
Methods Mol Biol. 2025;2917:247-258. doi: 10.1007/978-1-0716-4478-2_21.
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.
蛋白水解酶在生物医学、生物技术、制药和食品工业中有多种应用。其中一些应用需要在极端温度、高压、溶液的不同pH值以及可能使酶结构和功能不稳定的各种添加剂存在的条件下进行孵育。蛋白酶的结构特征决定了变性机制，这反映在该过程的不同活化能上。了解活化能可以提供有关酶的动力学惰性及其对不稳定因素的抗性的线索。通过测量在不同温度下加热时的活性损失来监测酶热失活的动力学，能够使用阿伦尼乌斯图确定失活过程的活化能。这样，测量由于结构扰动导致的活性损失可以直接提供有关酶在应用过程相关条件下稳定性的信息。
