Chem 203

Biochemistry Laboratory

Name Course & Year

Group no./Section Date

EXPERIMENT NO. 11
ENZYMES

Objective(s): To prepare enzymes and be familiar with its physical and chemical
properties.

Apparatus: Test tubes, mortar and pestle, beaker, bamboo splint

Materials: Potatoes, apples¸ guava, and chicos or other available fruits,

10 % hydrogen peroxide, liver

Theory:
Enzymes are complex organic compounds with definite chemical
structure, secreted by living things. They have the property of initiating and
hastening chemical reactions without themselves being affected in the process.
Enzyme activity is influenced by concentration of the enzyme, concentration of the
substrate, and concentration of the products of the reaction, temperature, pH,
inorganic salts and the presence of inhibitors and activators. All these factors
therefore should be considered while performing the following experiment.

Procedure:

A. Oxidase from fruits: (kindly do this one at your homes)

Cut thin slices of either apple, guava, or chico and expose them to the air.
Observe the darkening of the exposed sliced sides. Explain the color change.
Observations?
B. Catalase:

Catalase from potato:

PREPARATION:

1. Prepare a small potato and grate it into fine pulp.

2. Mix this pulp with 50 ml of water and let it stand for 15 minutes and
strain this thru a piece of cheese cloth.
3. Filter the extract.

ENZYME ACTION: (peroxidase)

1. Divide the filtrate into four parts and boil one part for 1 minute.

2. Place 5 ml of the boiled filtrate in one test tube and 5 ml of the

unboiled filtrate in another test tube.

3. Add to each a few drops of 3% hydrogen peroxide. Note what

happens.
Observation:

NOTE: The action of peroxidases depends to some extend upon phenol oxides which
decomposes the hydrogen peroxide. Potato has both peroxides and phenol oxides. This
accounts for the rapidity of actions in the presence of hydrogen peroxide, H2O2.

C. Amylase from saliva:

1. Take two 5 mL of starch solution and transfer them in each test tube.

2. Heat one of the starch solution in a water bath until it boils.

3. To both unboiled and boiled starch solutions, add 1 mL of the saliva.

4. Heat both test tubes in controlled temperature at 38 degrees Celsius in the water
bath

5. Remove the solutions from the water bath then add Fehling’s reagent. Then
continue heating them in the water bath

6. Remove the solutions and observe.

Observations:

Questions:

1. What does a catalase do? Explain.

2. What is a substrate?

3. What are the different kinds of catalyst and define each type.

4. How does an enzyme work?

5. Give the biological importance of enzyme.