Name: De Chavez, Jheslyn M.

Date: September 20, 2021

Year and Section: BSMLS 2E Rating: ______

EXPERIMENT 4
Factors that Affect Enzyme Activities

OVERVIEW (describe enzyme activity)

• Enzyme activity is naturally associated with free energy of transitional states, binding of
the ligand, protonation. It is measured in the units indicating the reaction speed
catalyzed by the enzyme as the micromoles of a changed (or created) substratum per
minute. The only major feature of enzymes is their capacity to boost the reaction rates
in living organisms, known as catalytic activity. Due to the fact that most enzymes are
proteins, their activity is influenced both by protein-disrupting stimuli and by factors
which in general impact catalysts.

OBJECTIVE (state the objective of the experiment as whole)

• Look at how enzymes function
• Test what enzyme factors affect enzyme activity

MATERIALS
• Test tubes (3)
• Vinegar
• Distilled Water
• Starch Solution
• Amylase
• Pipettes

REAGENT
• Benedict’s Solution
PROCEDURE (per test)
Prepare test tube and label it A, B and C.

For Test Tube A:

• Put 2ml of amylase in each test tube.
• Add 2 droppersful of vinegar to test tube A (to see if acid affects the enzyme activity)
• Mix by tapping the test tube.

We now have different volumes in each test tubes, so what we’re going to do is to even the
volume out;
- Get the test tubes
- Using a dropper, add distilled water to test tube B and C until the volumes are the same

For test tube B (amylase with distilled water):

• Get the test tube B and put it under high temperature.
• Let it boil for about 10 minutes until the enzyme breakdown.

After taking the test tube B out of the hot water bath;
- Add 5ml of starch solution in each test tube
- Tap them to mix.
- Let them (starch solution and amylase) sit for about 10 minutes.

✓ At this point, the amylase is breaking down the starch into simple sugar
✓ The benedict’s solution is a test for simple sugars.
- If only starch is present, then benedict’s solution will stay blue.
- If the starch solution converts into simple sugars, the starch solution will turn into
orange to red.

After 10 minutes;
- Add 2 droppesrful of benedict’s solution in each test tube.
- Get the test tubes and mix them.
- Put them into the hot water bath.

Wait and observe if their color change.

Remove the test tubes (from the hot water bath).

For Test Tube C:

• We just added distilled water, starch solution and benedict’s solution to the amylase.
DATA AND RESULT

SAMPLE RESULT

PROCEDURE A (With Acid) Still blue color without orange.

Amylase + Vinegar + Starch Solution + Therefore, vinegar or acid affects enzyme
Benedict’s Solution activity.

PROCEDURE B (Heated) Although it didn’t change from blue, it

Amylase + Distilled Water + Starch Solution + still has an orange
Benedict’s Solution Probably, heat affected the enzyme,
denatured the enzyme and made it non-
functional.

PROCEDURE C The color changed into orange

Amylase + Distilled Water The amylase has turned the starch into
Starch + Benedict’s Solution simple sugars.

GENERALIZATION:
• Finally, I could see how an enzyme functions exactly like how all types of chemical
reactions are catalyzed, and every minor change was observed to determine how many
factors impact the enzyme's activity. Two environmental variables are indicated, as seen
in the data and table of results. First of all, there is a pH, where an enzyme has an ideal
pH of 6.0-7.5, which makes the enzymes at extremes of pH denatured. Furthermore, the
temperature at which enzymes are thermolabile is susceptible to changes in
temperature. The range of optimum enzyme temperature is said to be 25-40 °C, which
indicates that increasing temperatures beyond the optimal temperature would lead to a
change of site form and ultimately loss of activity by the enzyme.