Dmitri N. Piquero
Biological Discovery I Laboratory
Section I Abstract
Amylase is an enzyme that speeds up the conversion rate from starch to glucose. Four test tubes were filled with a starch solution and a different concentration of amylase solution ranging from 0 μL to 1000 μL. The test tubes were then left to soak in a 37°C water bath before having 2 mL of 1 M hydrochloric acid added to a test tube of pure amylase solution. Lugol’s iodine was used to test for the existence of starch in the samples, while Benedict’s reagent detected any broken-down glucose. The same procedures were then performed waiting 5, 10, and 15 minutes before deactivating the amylase. Then again repeating the procedures with 4°C and 95°C water baths. Five test tubes were filled with pH buffers ranging from 2 to 10 in each tube before 2 mL of starch solution was added and the tubes were left to incubate at 37°C for 5 minutes. 500 μL of enzyme was added to each tube, then immediately quenched by adding 2 mL of 1 M hydrochloric acid. Lugol’s iodine was again used to detect the presence of starch. The remaining mixtures were left in the tubes with the addition of 3 mL of Benedict’s …show more content…
Increasing amounts of amylase were added to 1% starch solution incubating at 37⁰C, 4⁰C, and 95⁰C respectively, and then the reactions were subsequently quenched immediately, at 5 minutes, 10 minutes, and 15 minutes. Activity of amylase was recorded by adding 10 μL of Lugol’s Iodine to 100 μL of the reaction mixture. At 4⁰C, the enzyme reacted slightly, turning the mixture a light and transparent yellow color. At 37⁰C, the mixture reacted the most aggressively, turning several wells to an orange a few shades darker than the 4⁰C sample. Lastly, at 95⁰C, the mixture failed to substantially react, indicated by almost every well turning a dark black, meaning a high concentration of starch still existed in the