School of Math and Natural Sciences, University of Arkansas at Monticello, Monticello, Arkansas, USA.
Department of Chemistry and Biochemistry, University of Arkansas at Fayetteville, Fayetteville, Arkansas, USA.
Biochem Mol Biol Educ. 2024 Sep-Oct;52(5):588-598. doi: 10.1002/bmb.21845. Epub 2024 Jun 15.
Enzyme kinetics and inhibition studies are crucial in biochemistry education and research. Conventional methods often require expensive equipment and reagents, potentially limiting their accessibility in limited resource settings. Our approach sought to develop a cost-effective experimental design for studying enzyme kinetics and inhibition. Lactase was chosen as a protein model and its activity was investigated by measuring glucose production from lactose hydrolysis. In the study, commercially available lactase pills were used as an enzyme source, while milk was used as a substrate. Instead of scientific equipment, glucometers were used to measure lactase activity. Enzyme kinetics were evaluated using Michaelis-Menten and Lineweaver-Burk plots. In the study, the effects of temperature, pH, and inhibitors were also investigated. The results of our study aligned with established enzyme kinetics theories and previous studies. Lactase showed temperature and pH-dependent activity, with decreased activity observed at both low and high extremes. Results also showed that galactose acts as a competitive inhibitor of lactase. The approach presented here offers a cost-effective procedure for studying enzyme kinetics and inhibition. It can act as a valuable tool for educational purposes and for preliminary research in settings with limited resources.
酶动力学和抑制研究在生物化学教育和研究中至关重要。传统方法通常需要昂贵的设备和试剂，这可能限制了它们在资源有限的环境中的可及性。我们的方法旨在开发一种经济有效的实验设计，用于研究酶动力学和抑制。乳糖酶被选为蛋白质模型，并通过测量乳糖水解产生的葡萄糖来研究其活性。在这项研究中，市售的乳糖酶片被用作酶源，而牛奶则被用作底物。我们使用血糖仪而非科学仪器来测量乳糖酶的活性。使用米氏-门坦和林维伯克图来评估酶动力学。在研究中，还研究了温度、pH 和抑制剂的影响。我们的研究结果与已建立的酶动力学理论和先前的研究一致。乳糖酶表现出温度和 pH 依赖性活性，在低温和高温极端条件下活性均降低。结果还表明半乳糖是乳糖酶的竞争性抑制剂。本文提出的方法为研究酶动力学和抑制提供了一种经济有效的方法。它可以作为教育目的和资源有限环境中初步研究的有价值的工具。
