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To test your saliva’s activity, use a disposable plastic pipet to add 1 mL of your diluted
saliva solution to a test tube (you will reuse this pipet for the whole experiment). In a
second test tube, mix 1 mL (20 drops) of pH 7 buffer and 4 mL of 1% starch. Put both test
tubes in a 37 °C water bath for a few minutes to allow them to warm up to physiological
temperature. When you are ready, get a clean plastic pipet ready and pour the buffered
starch solution into your saliva solution and mix well. Start a stopwatch and take a sample
of the mixture right away. Add 5 drops of the reaction mixture to the well labeled “0
minutes,” and squirt any extra mixture back into the reaction (reuse this pipet for the
whole trial). Each minute the reaction goes, take another sample of the mixture and add 5
drops to the next well. Keep the reaction in the water bath during this process so that it
stays at the expected temperature.
At the end of the trial, swirl the well plate to make sure the solutions are mixed well, and
decide whether the reaction finished within the 5‐7 minute window. If the reaction is not
done within this window, set up the next two columns (8 wells) in your plate with iodine
solution. Repeat the procedure with a different amount of diluted saliva until you find an
amount that results in complete reaction within 5‐7 minutes. Once you have worked out
how much diluted saliva to use, record your data: the color of each solution, and your
interpretation of whether the mixture is still starch, or has turned to dextrins or plain
glucose. If you can, take a picture of this run and save it (do not dump it out) so you can
compare it to your other trials. Make sure your plate is well‐labeled in your picture so you
don’t have to guess what you are looking at.
Part II. Enzyme activity and pH
Now that you have established a set of conditions that causes hydrolysis of your added
starch in 5 to 7 minutes at pH 7, you will test the effect of changing the pH.
sucrose
fructose
glucose
7. All of these sugar molecules have the same nutritional value of about 4 Calories/g, and
each is eventually converted to glucose during metabolism. What is the possible
significance of their different flavor profiles?
Substance Type of R group Flavor notes
glycine
proline
arginine
glutamine
glutamic
acid
8. Which of the R groups above would change form in acidic or basic solutions? Does this
give insight into why enzymes are sensitive to changes in pH?
9. Use your textbook or another reference to find and draw the structures of three of the
amino acids you tasted. Label each one with its name.