Scribd Logo
Upload

Welcome to Scribd!

  • 6 views

    Catalase Lab

    Uploaded by

    dennymaeperpetua
    This document describes an experiment to test the effect of pH and substrate concentration on the activity of the enzyme catalase. Potato extract is used as the source of catalase enzyme. Filter paper discs soaked in catalase or catalase solutions of varying pH are placed in hydrogen peroxide solutions of different concentrations. The time for discs to rise due to oxygen bubble production is measured and analyzed to understand how pH and substrate concentration affect catalase activity.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd

    Catalase Lab

    Uploaded by

    dennymaeperpetua
    6 views2 pages
    This document describes an experiment to test the effect of pH and substrate concentration on the activity of the enzyme catalase. Potato extract is used as the source of catalase enzyme. Filter paper discs soaked in catalase or catalase solutions of varying pH are placed in hydrogen peroxide solutions of different concentrations. The time for discs to rise due to oxygen bubble production is measured and analyzed to understand how pH and substrate concentration affect catalase activity.

    Original Title

    catalase-lab

    Copyright

    © © All Rights Reserved

    Available Formats

    PDF, TXT or read online from Scribd

    Did you find this document useful?

    Is this content inappropriate?

    This document describes an experiment to test the effect of pH and substrate concentration on the activity of the enzyme catalase. Potato extract is used as the source of catalase enzyme. Filter paper discs soaked in catalase or catalase solutions of varying pH are placed in hydrogen peroxide solutions of different concentrations. The time for discs to rise due to oxygen bubble production is measured and analyzed to understand how pH and substrate concentration affect catalase activity.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    6 views2 pages

    Catalase Lab

    Uploaded by

    dennymaeperpetua
    This document describes an experiment to test the effect of pH and substrate concentration on the activity of the enzyme catalase. Potato extract is used as the source of catalase enzyme. Filter paper discs soaked in catalase or catalase solutions of varying pH are placed in hydrogen peroxide solutions of different concentrations. The time for discs to rise due to oxygen bubble production is measured and analyzed to understand how pH and substrate concentration affect catalase activity.

    Copyright:

    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    of 2

    Catalase and Potatoes

    Introduction: Hydrogen Peroxide (H2O2) is a poisonous byproduct of metabolism that can damage
    cells if not removed. Catalase is an enzyme that speeds up the breakdown of hydrogen peroxide into
    water (H2O) and oxygen gas (O2).

    General Directions: The assay system used in this lab consists of a filter paper disc which is coated with the enzyme and
    then dropped in t a paper cup of substrate. As the enzyme breaks down the hydrogen peroxide into water and oxygen
    gas, the bubbles of oxygen collect underneath the filter paper disc and make it rise to the surface of the hydrogen
    peroxide. The time it takes for the filter paper to rise is an indication of the rate of enzyme activity.

    Catalase soaked paper disc immersed in H2O2 Bubbles of oxygen form beneath the paper disc as catalase
    and placed at the bottom of a cup at time 0. breaks down H2O2 causing the disc to rise.

    Rate of enzyme activity = distance / time where distance is the depth of the hydrogen peroxide in mm and time
    measured in seconds. We will assume that each filter paper disc is coated in the same amount of catalase (except in an
    investigation of enzyme concentration).

    The enzyme has been prepared as follows: 50g of peeled potato was mixed with 50 mL of distilled water and crushed ice
    and homogenized in a blender for 30 seconds. This extract was filtered through cheese cloth and cold distilled water
    was added to a total volume of 100mL. Extract concentration is set at 100 units/mL. ENZYME SHOULD BE KEPT ON ICE
    AT ALL TIMES.

    Materials: Catalase Hydrogen peroxide Forceps


    Filter paper discs Ice Paper cups
    Timer

    Procedure: What is the effect of pH on enzyme activity?


    1. Obtain 1 paper cup of 40mL 3% H2O2. Measure and record the depth of the hydrogen peroxide.
    2. Label 3 small paper cups as follows: pH 1,pH 7, pH 9, and dilute catalase into the appropriate paper cup as
    directed below:
    a. pH 1: 5 mL catalase + 5 mL pH 3 buffer
    b. pH 7: 5 mL catalase + 5 mL pH 7 buffer
    c. pH 9: 5 mL catalase + 5 mL pH 10 buffer
    3. Dip a filter paper disc into the catalase, drain on a paper towel and then drop the filter paper into the 1% H2O2.
    Time how long it takes the filter paper to rise to the top.
    4. Remove the filter paper and repeat this procedure for each of the pH’s.
    5. Record your results.

    Adapted from: The properties of Enzymes: A study of Catalase. Cornell Institute for Biology Teachers
    Procedure: What is the effect of substrate concentration on enzyme activity?
    1. Obtain 1 paper cup of catalase - 15 mL
    2. Dilute the hydrogen peroxide as described below. Each dilution should be made in a separate paper cup.
    Measure and record the depth of the hydrogen peroxide.
    a. 3.0% H2O2 : 50 mL of 3% H2O2
    b. 1.5% H2O2: 25 mL of 3% H2O2 + 25 mL distilled water
    c. 0.90 % H2O2: 15 mL of 3% H2O2 + 35 mL distilled water
    d. 0.60% H2O2 : 10 mL 3% H2O2 + 40 mL distilled water
    e. 0.0% H2O2: 50 mL distilled water
    3. Dip a filter paper disc into the catalase, drain on a paper towel and then drop into the 3% H2O2.
    4. Time how long it takes the filter paper to rise to the top.
    5. Record your results.

    Data Analysis:
    1. Collect all of the class data.
    2. Process the class data.
    3. Create graphs to correctly display the class data.

    Conclusion:
    1. How does pH affect the activity of catalase? Consider both high and low pH, and explain your observations by
    discussing the effect of pH on protein structure.

    2. What is the effect of substrate concentration on enzyme activity? How does enzyme activity change as
    substrate concentration decreases? Explain your observations by discussing this reaction on a molecular level.

    3. Ectothermic organisms have body temperatures that vary with the temperature of their surroundings. Discuss
    the effect this variation might have on the functioning of enzymes in these organisms. Suggest some ways
    ectothermic organisms might cope with this problem.

    Adapted from: The properties of Enzymes: A study of Catalase. Cornell Institute for Biology Teachers

    You might also like