Flurkey William H, Inlow Jennifer K
Department of Chemistry and Physics, Indiana State University, Terre Haute, Indiana, 47809.
Biochem Mol Biol Educ. 2017 May;45(3):270-276. doi: 10.1002/bmb.21029. Epub 2016 Nov 7.
An inexpensive enzyme kinetics laboratory exercise for undergraduate biochemistry students is described utilizing tyrosinase from white button mushrooms. The exercise can be completed in one or two three-hour lab sessions. The optimal amounts of enzyme, substrate (catechol), and inhibitor (kojic acid) are first determined, and then kinetic data is collected in the absence and presence of the inhibitor. A Microsoft Excel template is used to plot the data and to fit the Michaelis-Menten equation to the data to determine the kinetic parameters V and K . The exercise is designed to clarify and reinforce concepts covered in an accompanying biochemistry lecture course. It has been used with positive results in an upper-level biochemistry laboratory course for junior/senior students majoring in chemistry or biology. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(3):270-276, 2017.
本文描述了一种为本科生物化学专业学生设计的、成本低廉的酶动力学实验室练习,该练习使用白蘑菇中的酪氨酸酶。该练习可在一或两个三小时的实验课中完成。首先确定酶、底物(儿茶酚)和抑制剂(曲酸)的最佳用量,然后在不存在和存在抑制剂的情况下收集动力学数据。使用Microsoft Excel模板绘制数据,并将米氏方程拟合到数据中以确定动力学参数V和K。该练习旨在阐明和强化配套生物化学讲座课程中涵盖的概念。它已在面向化学或生物学专业大三/大四学生的高级生物化学实验室课程中使用,并取得了积极成果。© 2016年,国际生物化学与分子生物学联盟,45(3):270 - 276,2017。