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B04-Catalase-Enzyme

Biology

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Studies In Biology: Research (BIOL 493)

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Western Carolina University

Academic year: 2024/2025

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Keirra Battle

Tennessee College of Applied Technology Knoxville

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Name _____________________ Class ______________ Date _________

####### Activity B04: Catalase Enzyme Activity

####### (Pressure Sensor)

Concept DataStudio ScienceWorkshop (Mac) ScienceWorkshop (Win) Biochemistry - enzymes B04 Catalase B05 Catalase Activity B05_CATA Equipment Needed Qty Equipment Needed Qty Pressure Sensor – Abs. (CI-6532) 1 Protective gear PS Balance (SE-8723) 1 Beaker, 600 mL 1 Chemicals and Consumables Qty Connector (640-030) 1 Chicken liver extract 12 mL Flask, 250 mL 1 Glycerin 1 mL Graduated cylinder, 100 mL 1 Hydrochloric acid (HCl), 1 M 10 mL Hot plate 1 hydrogen peroxide, 3% 100 mL Magnetic stirrer & spin bar 1 Ice, crushed 500 mL Stopper, one hole, for flask 1 Sodium fluoride, solid 2 g Test tube 1 Sodium hydroxide (NaOH), 1 M 10 mL Tongs 1 Water 500 mL Tubing (w/ sensor) Water, distilled 500 mL What Do You Think

####### What are some factors that can influence the rate of enzyme activity in an organism?

####### Take time to write an answer to this question in the Lab Report section.

Background

####### Enzymes are very important molecules found in every cell. Enzymes

####### generally act as catalysts that increase the speed or rate at which substances

####### in a cell get converted into other substances. Without enzymes, some

####### reactions would take place too slowly – or might not take place at all.

####### Each enzyme has a different job and many enzymes must work together to

####### keep an organism alive and healthy. In the liver, for example, there are

####### several enzymes that act on certain toxic or poisonous compounds by

####### removing hydrogen atoms from the poisons and transferring them to oxygen

####### molecules. This detoxifies the poison but it creates a new compound, hydrogen peroxide (H2O2)

####### that is very active and can be harmful to the organism. Fortunately there is another enzyme in

####### the liver that helps break down the peroxide into water and oxygen.

####### This enzyme is known as catalase. The catalase enzyme reduces the substrate, peroxide, to water

####### and oxygen by the following decomposition reaction.

2H 2 O 2

####### Catalase

⏐ ⏐ ⏐ ⏐ →2H 2 O + O 2 (gas)

(substrate) (enzyme) (products)

####### Like all enzymes, catalase helps the reaction but does not itself get used up in the reaction. Also

####### like other enzymes, catalase must have a proper environment in which to work. Your body’s

####### enzymes, for example, work best when your temperature is normal (around 37 ̊ C) and when the

####### pH is between 7 to 7. If the environmental conditions are outside the normal range, the

####### catalase will lose its ability to catalyze the peroxide reaction or may even be destroyed.

Biology Labs with Computers Student Workbook B04: Catalase Enzyme Activity 012-06635B

####### Since the breakdown of hydrogen peroxide produces oxygen gas, what is a way to measure the

####### rate of the production of that gas?

####### Take time to write an answer to this question in the Lab Report section.

SAFETY REMINDERS  Wear protective gear while handling chemicals.  Follow directions for using the equipment.  Dispose of all chemicals and solutions properly. For You To Do

####### Use the Pressure Sensor to measure the change in gas pressure inside a flask containing

####### hydrogen peroxide and a source of catalase enzyme. After you measure the rate of activity for

####### catalase and hydrogen peroxide, compare the rate of activity for the mixture under three

####### different conditions: change in pH, change in temperature, and in the presence of an inhibitor

####### (sodium fluoride). Use DataStudio or ScienceWorkshop to record and analyze the data.

PART I: Computer Setup

####### 1. Connect the ScienceWorkshop interface to the computer, turn on the

####### interface, and turn on the computer.

####### 2. Connect the Pressure Sensor DIN plug into Analog Channel A on

####### the interface.

####### 3. Open the file titled as shown;

DataStudio ScienceWorkshop (Mac) ScienceWorkshop (Win) B04 Catalase B05 Catalase Activity B05_CATA

The DataStudio file has a Workbook display. Read the instructions in the Workbook. The

####### file has a Graph of Pressure versus Time and a Digits display of Pressure.

The ScienceWorkshop document opens with a Graph display of Pressure (kPa) versus

####### Time (s).

Data recording is set for 1 measurement per second with a ‘Stop’ condition at 150 s. p. 28 © 1999 PASCO scientific B

Sensor

Pressureport

Quickrelease

connector

Tubing

Biology Labs with Computers Student Workbook B04: Catalase Enzyme Activity 012-06635B PART III: Data Recording  There are six parts to the data recording. Part Description Part Description A Catalase + Hydrogen Peroxide D Catalase + Hydrogen Peroxide + Acid B Catalase + Hydrogen Peroxide + Inhibitor E Chilled Catalase + Hydrogen Peroxide C Catalase + Hydrogen Peroxide + Base F Heated Catalase + Hydrogen Peroxide PART IIIA: Catalase + Hydrogen Peroxide Prepare the Mixture

####### 1. Pour 15 mL of 3% hydrogen peroxide in a 100 mL graduated cylinder.

####### Fill the cylinder to the 100-mL mark with 85 mL of distilled water.

####### 2. Transfer the diluted peroxide solution to the flask.

####### 3. Turn on the stirrer.

####### 4. Add 2 mL of catalase extract to the dilute peroxide solution in the flask.

####### 5. Put the one-hole stopper into the flask.

####### 6. Align the quick-release connector on the end of the

####### plastic tubing with the connector on the pressure

####### port of the Pressure Sensor. Push the connector onto

####### the port, and then turn the connector clockwise until

####### it clicks (about one-eighth turn).

Record the Data

####### 7. Start recording data. (Hint: Click ‘Start’ in DataStudio or click ‘REC’ in

####### ScienceWorkshop.)

Data recording will stop automatically at 150 seconds. Clean Up

####### 8. Disconnect the tubing from the Pressure Sensor. Remove the stopper from the flask.

####### 9. Dispose of the peroxide mixture as directed and clean the flask thoroughly.

Hydrogen

Peroxide

( 15 mL)

Add 85 mLof

DistilledWater

DiluteHydrogen

Peroxide+Catalase

Stopper

Tubingto

sensor

Name _____________________ Class ______________ Date _________ Part IIIB: Catalase + Hydrogen Peroxide + Inhibitor Make a prediction:

####### What effect do you think adding an inhibitor to the hydrogen peroxide will have on the

####### enzymes’s ability to catalyze the breakdown of the peroxide? Put your prediction and a brief

####### explanation in the Lab Report.

Prepare the mixture

####### 1. Repeat the data recording procedure. Put 100 mL of dilute peroxide solution in the flask.

####### Add the spin bar.

####### 2. Add 2 g of sodium fluoride to the peroxide solution. Add 2 mL of catalase extract to the

####### flask and stopper the flask.

####### 3. Re-connect the tubing to the Pressure Sensor.

Record the data

####### 4. Start recording data.

Data recording will stop automatically at 150 seconds. Clean Up

####### 5. Disconnect the tubing from the Pressure Sensor. Remove the stopper from the flask.

####### 6. Dispose of the peroxide mixture as directed and clean the flask thoroughly.

Part IIIC: Catalase + Hydrogen Peroxide + Base Make a prediction:

####### What effect do you think adding a base to the hydrogen peroxide will have on the enzymes’s

####### ability to catalyze the breakdown of the peroxide? Put your prediction and a brief explanation in

####### the Lab Report.

Prepare the mixture

####### 1. Repeat the data recording procedure. Put 100 mL of dilute peroxide solution in the flask.

####### Add the spin bar.

####### 2. Add 10 mL of 1 Molar sodium hydroxide (NaOH) to the peroxide to raise the pH before

####### you add the catalase. Add 2 mL of catalase extract to the flask and stopper the flask.

####### 3. Re-connect the tubing to the Pressure Sensor.

Record the data

####### 4. Start recording data.

Data recording will stop automatically at 150 seconds. Clean Up

####### 5. Disconnect the tubing from the Pressure Sensor. Remove the stopper from the flask.

####### 6. Dispose of the peroxide mixture as directed and clean the flask thoroughly.

Name _____________________ Class ______________ Date _________ PART IIIF: Heated Catalase + Hydrogen Peroxide Make a prediction:

####### What effect do you think boiling the catalase will have on the enzymes’s ability to catalyze the

####### breakdown of the peroxide? Put your prediction and a brief explanation in the Lab Report.

Prepare the mixture

####### 1. Repeat the data recording procedure. Put 100 mL of dilute peroxide solution in the flask.

####### Add the spin bar.

####### 2. Put 2 mL of catalase extract into a test tube. Use tongs to hold

####### the test tube in a beaker of boiling water. Heat the test tube in

####### boiling water for 5 minutes.

####### 3. Add the heated catalase extract to the flask and stopper the

####### flask.

####### 4. Re-connect the tubing to the Pressure Sensor.

Record the data

####### 5. Start recording data.

Data recording will stop automatically at 150 seconds. Clean Up

####### 6. Disconnect the tubing from the Pressure Sensor. Remove the stopper from the flask.

####### 7. Dispose of the peroxide mixture as directed and clean the flask thoroughly.

Analyzing the Data

####### 1. Set up the Graph display to show all your data

Hint: DataStudio automatically shows all six runs of data. In ScienceWorkshop, do the

####### following to put two runs of data in a top plot, two runs in a middle plot, and two runs of

####### data in a bottom plot.

Click the ‘Add Plot Menu’ button ( ) to add a second plot to the Graph window.

####### Select Analog Channel A, Pressure from the Add Plot

####### Menu.

Click the ‘Add Plot Menu’ button again to add a third

####### plot to the Graph window. Select Analog Channel A,

####### Pressure from the Add Plot Menu.

Click the ‘DATA Menu’ button ( ) in the top plot. Select Run #1 from

####### the DATA Menu. Repeat to select Run #2 from the DATA menu.

Click the ‘DATA Menu’ button in the middle plot. Select No Data first.
Use the Data Menu in the second plot to select Run #3 and then Run #4.
Click the ‘DATA Menu’ button in the bottom plot. Use the Data Menu to add

####### Run #5 to the plot.

Biology Labs with Computers Student Workbook B04: Catalase Enzyme Activity 012-06635B

####### 2. Use the built-in statistics for the Graph display to find the minimum pressure and the

####### maximum pressure for each run of data.

Hint: In DataStudio, click the ‘Statistics Menu’ button ( ). The default statistics are

####### ‘Minimum’ and ‘Maximum’. The values appear in the legend in the display area.

####### In ScienceWorkshop, click the ‘Statistics’ button ( ) to open the statistics area. Click

####### the ‘Statistics Menu’ button ( ). Select ‘Maximum’ from the menu. Repeat and select

####### ‘Minimum’ from the menu.

####### 3. Record the minimum pressure as the starting pressure. Record the maximum pressure as

####### the ending pressure.

####### 4. Calculate the difference in pressure.

####### 5. Calculate the enzyme activity. Divide the difference in pressure by the amount of time (in

####### minutes).

(measuredby therateofoxygen production)

CatalaseActivity -

-

EndingPressureStartingPressure

= 2

####### Record your results in the Lab Report section.

Biology Labs with Computers Student Workbook B04: Catalase Enzyme Activity 012-06635B Data Table Item Part IIIA Part IIIB NaF Part IIIC NaOH Part IIID HCl Part IIIE Chilled Part IIIF Heated Starting Pressure kPa kPa kPa kPa kPa kPa Ending Pressure kPa kPa kPa kPa kPa kPa Pressure Difference kPa kPa kPa kPa kPa kPa Activity (kPa/min) Questions

####### 1. What does the graph of the reaction between hydrogen peroxide and catalase tell you about

####### enzyme activity?

####### 2. Describe the effect of adding the inhibitor (sodium fluoride) to the peroxide before you

####### add the catalase to the solution of peroxide? What explanation can you give for the results?

####### 3. Describe the effect of adding the base (sodium hydroxide) to the solution of peroxide?

####### What did the sodium hydroxide do to the pH of the solution in the flask? What does this

####### tell you about the range of conditions in which catalase may be effective?

####### 4. Describe the effect of adding the acid (hydrochloric acid) to the solution of peroxide?

####### What did the hydrochloric acid do to the pH of the solution in the flask? What does this

####### tell you about the range of conditions in which catalase may be effective?

####### 5. Describe the effect of cooling the catalase before adding it to the solution of peroxide?

Name _____________________ Class ______________ Date _________

####### 6. Describe the effect of heating the catalase to boiling before adding it to the solution of

####### peroxide? How did the effect of cooling compare to the effect of boiling the catalase? How

####### can you explain the difference between these two trials?

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B04-Catalase-Enzyme

Course: Studies In Biology: Research (BIOL 493)

3 Documents

Students shared 3 documents in this course

University: Western Carolina University

Was this document helpful?

Name _____________________ Class ______________ Date _________

Activity B04: Catalase Enzyme Activity

(Pressure Sensor)

Concept DataStudio ScienceWorkshop (Mac) ScienceWorkshop (Win)

Biochemistry - enzymes B04 Catalase.DS B05 Catalase Activity B05_CATA.SWS

Equipment Needed Qty Equipment Needed Qty

Pressure Sensor – Abs. (CI-6532) 1 Protective gear PS

Balance (SE-8723) 1

Beaker, 600 mL 1 Chemicals and Consumables Qty

Connector (640-030) 1 Chicken liver extract 12 mL

Flask, 250 mL 1 Glycerin 1 mL

Graduated cylinder, 100 mL 1 Hydrochloric acid (HCl), 1 M 10 mL

Hot plate 1 hydrogen peroxide, 3% 100 mL

Magnetic stirrer & spin bar 1 Ice, crushed 500 mL

Stopper, one hole, for flask 1 Sodium fluoride, solid 2.0 g

Test tube 1 Sodium hydroxide (NaOH), 1 M 10 mL

Tongs 1 Water 500 mL

Tubing (w/ sensor) Water, distilled 500 mL

What Do You Think

What are some factors that can influence the rate of enzyme activity in an organism?

Take time to write an answer to this question in the Lab Report section.

Background

Enzymes are very important molecules found in every cell. Enzymes

generally act as catalysts that increase the speed or rate at which substances

in a cell get converted into other substances. Without enzymes, some

reactions would take place too slowly – or might not take place at all.

Each enzyme has a different job and many enzymes must work together to

keep an organism alive and healthy. In the liver, for example, there are

several enzymes that act on certain toxic or poisonous compounds by

removing hydrogen atoms from the poisons and transferring them to oxygen

molecules. This detoxifies the poison but it creates a new compound, hydrogen peroxide (H2O2)

that is very active and can be harmful to the organism. Fortunately there is another enzyme in

the liver that helps break down the peroxide into water and oxygen.

This enzyme is known as catalase. The catalase enzyme reduces the substrate, peroxide, to water

and oxygen by the following decomposition reaction.

2H 2O2

Catalase

⏐ → ⏐ ⏐ ⏐ 2H 2O+O2(gas)

(substrate) (products)(enzyme)

Like all enzymes, catalase helps the reaction but does not itself get used up in the reaction. Also

like other enzymes, catalase must have a proper environment in which to work. Your body’s

enzymes, for example, work best when your temperature is normal (around 37˚ C) and when the

pH is between 7.3 to 7.4. If the environmental conditions are outside the normal range, the

catalase will lose its ability to catalyze the peroxide reaction or may even be destroyed.

B04-Catalase-Enzyme

Studies In Biology: Research (BIOL 493)

Western Carolina University

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Students also viewed

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Preview text

2H 2 O 2

⏐ ⏐ ⏐ ⏐ →2H 2 O + O 2 (gas)

Sensor

Pressureport

Quickrelease

connector

Tubing

Hydrogen

Peroxide

( 15 mL)

Add 85 mLof

DistilledWater

DiluteHydrogen

Peroxide+Catalase

Stopper

Tubingto

sensor

(measuredby therateofoxygen production)

CatalaseActivity -

-

EndingPressureStartingPressure

= 2

B04-Catalase-Enzyme

Course: Studies In Biology: Research (BIOL 493)

University: Western Carolina University

Students also viewed

Related documents