Course Code: BIO 024

Students’ Guide # ENZYMES

Lesson title: ENZYMES Materials:

Lesson Objectives: by the end of this
activity, you should be able to …
1. Examine the action of catalase
on hydrogen peroxide
2. Understand the actions of
catalase under different
conditions.
3. Explain concepts related to
enzyme activity
See materials in each experiment
References:
▪ Bathan, G. I., Crisostomo, A. C., Daya, M. L., De Guia, R. M., Farrow, F. L.,
Gabona, M. G., . . . Ysrael, M. C. (2017). Laboratory Manual in General
Biochemistry. (2nd, Ed.) Quezon: C & E Publishing.
▪ Josue, P. B., & Ocenar, PhD, M. M. (2014). Biochemistry Laboratory Manual
and workbook for Health Sciences . Quezon : C & E Publishing Inc.
▪ Stoker, H. S. (2013). General, organic and biological chemistry (6 ed.).
Retrieved 8 September, 2020
▪ https://www.education.com/science-fair/article/activator/

PRE-LABORATORY CONCEPT

Living organisms are composed of complex but coordinated systems of chemical reactions. Most of these
reactions are catalyzed by enzymes so that they can proceed at the rate necessary to sustain life. Enzymes are proteins
found in all living organisms. As efficient catalysts, they accelerate reactions but are themselves not subject to change.
Thousands of different enzymes can be found within a cell, and each has unique three-dimensional structure that
determines its functions (Bathan, et al., 2017). Thus, many different reactions are catalyzed. (Josue & Ocenar, PhD, 2014).
Enzymes have active sites where molecular chemistry takes place. Substrates are molecules that bind in the active
sites and are acted upon by the enzyme. Enzymes are very specific for substrates. The shape of an enzyme molecules
allows only its substrate molecules to fit to it and be activated for reaction. (Josue & Ocenar, PhD, 2014)

A simple equation for an enzyme-catalyzed reaction can be written as:

E + S ⇆ ES → E + P

Where E is enzyme, S is substrate, E-S is an enzyme-substrate complex, E-P is enzyme-product complex, and P is product.
Enzyme often use metal ions or prosthetic groups like vitamins to assist catalysis. Zn 2+ for example, is the metal
ion in carbonic anhydrase while Fe2+ is present in cytochrome and is needed for biological oxidation.
Measurement of enzyme activities in body fluids is important in diagnosing medical illnesses. Drugs act by
altering the activities of enzymes. The characteristics of enzymes make them practical tools in the laboratory.
In this experiment, we will test the activity characteristics of the enzyme catalase, it’s specificity and actions in
different influences.

STUDENT TASK:

Answer the following activity questions:

1. Biological relationship between hydrogen peroxide and catalase

2. Why aren’t your cells poisoned to death even though they create toxins like hydrogen peroxide?

3. What is enzyme specificity?

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 Course Code: BIO 024
Students’ Guide # ENZYMES

4. Describe the following enzyme specificity.

SPECIFICITY DESCRIPTION
Bond specificity
Group specificity
Substrate specificity
Stereo specificity
Geometrical specificity
Co-factor specificity

5. Describe the relationship of the following concept:

Concept DESCRIPTION
Enzyme influence on reaction
velocity
Effect of temperature on
enzymatic activity
Effect of pH with enzymatic
activity. Include example of
enzymes optimum pH.
Effect of activators on the
activity of enzymes. Include
examples.
Effect of inhibitors on enzyme
activity
Effect of substrate
concentration on enzymatic
activity
Effect of enzyme concentration
on enzymatic activity

6. Describe the following extremophiles extremozymes:

EXTREMOZYMES DESCRIPTION
Piezophiles
Halophiles
Thermophiles
Cryophiles
Acidophiles
Alkaliphiles

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 Course Code: BIO 024
Students’ Guide # ENZYMES

ACTION OF CATALASE ON HYDROGEN PEROXIDE

Material:
▪ 60mL Hydrogen peroxide or agua oxygenada 3% solution
▪ 1 piece of potato (or liver from chicken, pork or beef if available), use only enough
▪ Acid: 20mL Vinegar
▪ Base: 1 tablespoon Baking powder (or chopped to fine powder bath soap, except dove soap or kojic acid soap or
any soap that is not acidic). Note: check pH level of the soap you are using to confirm pH level as basic
▪ 20mL Tap, Cold and Hot water
▪ 5 clear glasses with the same size (preferably taller glass or long neck narrow glass) that will serve as your 100mL
graduated cylinder or 300mL beaker. Hence, please mark the glasses with uniform calibration if possible.
Label the glasses with: Acid, Base, Hot water, Cold water, Tap (neutral)
But make a separate 2 empty glasses for the preliminary experiment. One for water (in room temperature) and one for
hydrogen peroxide.
▪ Kitchen knife for chopping of the potato (you might include peeler for the removal of its skin)
▪ Chopping board
▪ Mortar and pestle (food processor or blender, if available)
▪ Medicine cup, syringe, or calibrated dropper for measuring volumes of hydrogen peroxide and other solutions

PROCEDURE:
Note: Use only an enough portion of potato for the experiment.

2. Peel the potato. Cut, into cubes, equally on your desired size.

Preliminary experiment:
3. On a separate glass, place one (1) cube each of the potato.
4. Add separately 5 drops of water and 5 drops of hydrogen peroxide in the glasses with potato cube.
5. Observe if the results.

Main experiment:
6. Separately mash five (5) cubes to create a fine substance and place it in the assigned glass container.
7. Place the following materials in the assigned glass.
▪ Glass #1: acidic solution (ex. Vinegar)
▪ Glass #2: one (1) tablespoon of the basic powder (ex. Baking powder)
▪ Glass #3: cold water
▪ Glass #4: hot water
▪ Glass #5: tap water (room temp.)
8. Leave for 3 minutes. Drain only the acid, cold, hot and tap water after.
9. Add 10mL of hydrogen peroxide in each of the glasses.
10. Observe the results and compare. You may use calibration as one way to present your observations.

Note: You will have almost the same procedure if you are using chicken, pork, or beef liver.

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 Course Code: BIO 024
Students’ Guide # ENZYMES

RESULTS, OBSERVATIONS, and INTERPRETATION:

Preliminary experiment:

VISUAL OBSERVATION and INTERPRETATION

Potato + Water Potato + Hydrogen peroxide

Explain the rationale or principle behind the results/experiment?

Main experiment:

VISUAL OBSERVATION and INTERPRETATION: Influence of pH

Potato + Acid+ Hydrogen peroxide Potato + Base + Hydrogen peroxide

Explain the rationale or principle behind the results/experiment? (In comparison with potato + hydrogen peroxide alone from the
preliminary experiment conducted.)

VISUAL OBSERVATION and INTERPRETATION: Influence of TEMPERATURE

Potato + Cold water+ Hydrogen Potato + Tap water (room Potato + Hot water+ Hydrogen
peroxide temperature) + Hydrogen peroxide peroxide

Explain the rationale or principle behind the results/experiment?

Post laboratory questions:

1. Using the equation below, supply reaction involved in the experiment

2. How did you account for the physical observation of the action of catalase?
3. Which of the enzyme specificity is represented by the experiment, conducted in the preliminary part? Why?
4. State the optimum pH and temperature of catalase enzyme.

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