Investigating Alcohol Dehydrogenase Kinetics Using Spectrophotometry

This report delves into the kinetics of the enzyme alcohol dehydrogenase (ADH) under varying temperature conditions, employing spectrophotometry to determine reaction rates using absorbance measurements and Beer's law. The experiment involved analyzing the reaction's progress with and without the enzyme, using different substrate concentrations, and assessing enzyme activity at temperatures of 4°C and 80°C. The results, illustrated through Lineweaver-Burk, Eadie-Hofstee, and Michaelis-Menten plots, demonstrate an optimal temperature range for enzyme activity before denaturation occurs. The experiment successfully quantified the enzyme's maximum rate (Vmax) and Michaelis-Menten constant (Km) at different temperatures, providing insights into the enzyme's catalytic efficiency. Data collected from the spectrophotometer at different time intervals was used to determine initial reaction rates, which were then converted to molar concentrations using Beer's law. The study contributes to a better understanding of enzyme behavior and the factors influencing its activity.