The Alcohol Dehydrogenase Kinetics Laboratory: Enhanced Data Analysis and Student-Designed Mini-Projects

Abstract

A highly instructive, wide-ranging laboratory project in which students study the effects of various parameters on the enzymatic activity of alcohol dehydrogenase has been adapted for the upper-division biochemistry and physical biochemistry laboratory. Our two main goals were to provide enhanced data analysis, featuring nonlinear regression, and also to give students experience in experimental design. Students use appropriate kinetic and thermodynamic equations to fit their data from Michaelis–Menten plots, enzyme activity pH profiles, inhibitor and denaturant concentration profiles, and temperature-dependence plots. Experiments at the end of this project are designed and implemented by student pairs, thus preparing them for independent research.

When both Km and Vmax are altered, Is the enzyme inhibited or activated?

“…(3) we can predict that [S] = = 5.7 mM. Using typical laboratory values for YADH in buffer , Fig. shows how the crowder glucose should alter enzyme activity above and below 5.7 mM ethanol.…”

“…To exemplify the effect of a specific crowder on a well‐characterized enzyme, Schneider et al showed in their first figure that glucose (300 g/L) lowered the K m (EtOH) value of yeast alcohol dehydrogenase (YADH) by about fivefold (from 17 mM to 3.4 mM) and V max by about 2.5‐fold; under these conditions, using Eq. (3) we can predict that [S] = = 5.7 mM.…”

“…Predicted Michaelis–Menten plot of YADH (17 nM) in buffer (20 mM Tris, pH 9, with 0.25 mM NAD + ) in the absence (black curve) and presence of the crowder glucose (300 g/L, red curve). Control (buffer‐only) Michaelis–Menten parameters are K m (EtOH) = 17 mM, V max = 1.40 μM/s ; K m '(EtOH) = 3.4 mM, and V max ' = 0.56 μM/s in the presence of glucose . [Color figure can be viewed at wileyonlinelibrary.com]…”

“…To exemplify the effect of a specific crowder on a wellcharacterized enzyme, Schneider et al [12] showed in their [14], Fig. 1 shows how the crowder glucose should alter enzyme activity above and below 5.7 mM ethanol.…”

Kegiatan Laboratorium Kimia Berbasis Mini-Project Terhadap Hasil Belajar dan Aktivitas Siswa pada Praktikum Uji Aldehid

“…Hal ini menunjukkan bahwa penerapan (Rosa et al, 2018). Selain itu terdapat juga penelitian yang menyatakan bahwa kegiatan praktikum berbasis Mini Project dapat memberikan pengalaman eksperimen siswa (Silverstein, 2016).…”