...responsible for one particular reaction that occurs in the cell. In this lab, you will study an enzyme that is found in the cells of many living tissues. The name of the enzyme is catalase (KAT-uh-LAYSS); it speeds up a reaction which breaks down hydrogen peroxide, a toxic chemical, into 2 harmless substances--water and oxygen. The reaction is: 2 H2O2 ----> 2 H2O + O2 This reaction is important to cells because hydrogen peroxide (H2O2) is produced as a byproduct of many normal cellular reactions. If the cells did not break down the hydrogen peroxide, they would be poisoned and die. In this lab, you will study the catalase found in liver cells. You will be using chicken or beef liver. It might seem strange to use dead cells to study the function of enzymes. This is possible because when a cell dies, the enzymes remain intact and active for several weeks, as long as the tissue is kept refrigerated. MATERIALS: 1molar HCl solution 1molar NaOH solution 6 Test tubes Measuring Pipette 10-ml Graduated cylinder 40 ml 3% Hydrogen peroxide solution (found in stores) Straight-edged razor blade Scissors and Forceps (tweezers) pH paper (optional) Stirring rod Fresh liver, Apple, and Potato Test tube holders Ice bath Warm water bath Boiling water bath PART A - Observe Normal Catalase Reaction 1. Place 2 ml of...
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...affect them. This experiment also seeks to establish the manner in which some enzymes like Catalase affect the rates of reactions (Cohnheim 2009). Methods To establish the factors that affect enzymes, the procedures for the experiments to be carried out had to be almost perfect. For this reason the apparatus to be used had to be cleaned thoroughly just before commencing the experiment. To avoid differentiated results, similar kinds of apparatus were used all through the experiment. In this case glass test tubes were used. Also measuring apparatuses used were of the same size and volume. In this case four experiments were carried out. The first experiment is to establish the manner in which the enzyme Catalase affects reaction rates. The procedure of this experiment is as follows; using a pencil, label tree test tubes as test tube 1, 2 & 3. On these test tubes, label two marks using the pencil. These are at the 1cm mark and at the 5 cm mark. For the first test tube, pour in Catalase enzyme up to the first mark and add Hydrogen Peroxide up to the second mark. Swirl the mixture and leave it for about 20 seconds for it to bubble. Measure the height of the bubble column and record your findings in table. Repeat the same procedure for test tube 2 & 3 but replace the hydrogen peroxide in test tube 2 with water and that in test tube 3 with sucrose solution. In the second test tube add water to the The second experiment is meant to...
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...enzyme: Catalase found in chicken liver. * The name of the organism is chicken (liver). * The substrate is hydrogen peroxide (H2O2)|. * Hypothesis: If the catalase in the chicken liver is exposed to hydrogen peroxide, it will cause a chemical reaction. This reaction will result in the destruction of hydrogen peroxide in the chicken liver, which is harmful to the cells; the enzyme catalase works best in acidic solutions and enzymes are denatured in very high temperatures (boiling). * Method of measuring enzyme activity: observation of normal catalase reaction, the effect of temperature on catalase activity and the effect of pH on catalase activity. * Treatment: test the effect of temperature on catalase activity using different temperature water baths; ice bath, warm bath and boiling water bath. A solution of 1ml of hydrogen peroxide will be placed into 2 separate test tubes and a piece of liver into 2 separate test tubes. Place one test tube of liver and one of H2O2 into each of the following water baths: Ice bath and Warm water bath (not boiling).After 3 minutes, pour each tube of H2O2 into the corresponding tube of liver and observe the reaction. The boiling water bath will be left for 5 minutes, after which the liquid will be poured off and the liver will be placed into a 2ml solution of hydrogen peroxide. This is done to see the effect that boiling has one the enzyme. Record the rate of each reaction from 0(no reaction)-5(very fast). Test the effect...
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... September 24, 2014 Principles of Biology 120.601 Mrs. Annemarie Duncan Abstract: (Burmania) This experiment was performed in order to examine ways in which a potato catalase enzyme reacts to various assays with differing variables. To do so a baseline assay (undiluted extract and room temperature H2O2) was used within the experiment with only one other variable changed in the other assays. These variables included a boiled, frozen and then thawed, and frozen potato extract and dH2O instead of the potato extract. It was noted that the temperature and or way the potato extract was prepared effects how the enzyme with the potato will react. Therefore the results of each assay varied, suggesting there is more than one way that a catalyzed reaction can occur. Introduction: (Burmania) The main purpose for this experiment was to explore how an enzyme catalase caused a reaction to catalyze through doing various assays using potato extracts. Enzymes are catalysts that are crucial in helping to speed up reactions, and catalase is a common enzyme found in almost every living organism that is exposed to oxygen. For this experiment, potatoes were used because they were previously known to have significant catalase activity. The potato catalase was mixed with hydrogen peroxide because the catalase helps to breakdown the hydrogen peroxide which helps determine how long a paper disk takes to float to the surface, which in return provides the time of the complete reaction...
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...DATA The Unknown #5 had the following morphology after Gram staining and observed under a brightfiled microscope: purple color, spherical shape and clustered like grape. After determining that it was a Gram positive staphylococcus, it was inoculated on a MSA plate, Blood Agar, DNA agar and Catalase test was also performed to help figure out the staphylococcus type. The Table below lists all of the biochemical tests, their purpose and results. TEST PURPOSE REAGENTS OBSERVATIONS RESULTS Gram Stain To determine the Gram reaction of the bacterium Crystal violet, Iodine, Alcohol, Safranin Purple grape cluster arrangement cocci Gram Positive coccus Mannitol Salt Agar To determine the if the bacterium can tolerate high saline levels and grow If the bacterium can ferment mannitol None There was a full growth and the mannitol turned to yellow (acidic) Positive Blood Agar To determine if the bacterium can hemolysis blood None Metallic gray, Creamy – golden color ß-hemolysis Positive DNA Agar To determine if bacterium has DNase and can hydrolyze DNA None Clearing around the bacterial growth Positive Catalase Test To determine the presence of the enzyme catalase (converts H2O2 to H2O and O2) Hydrogen peroxide Immediate bubbles Positive DISCUSSION/CONCLUSION After...
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...Ultimately, the data somewhat supported the hypothesis because it was justified as the amount of hydrogen peroxide (mL) increases, then the displaced volume of oxygen (mL) that is produced will increase and eventually level out. This is because the hydrogen peroxide is substrate specific to catalase which is found in the liver solution tab. The two products react to create an active site which produces water and oxygen. Therefore, as the amount of hydrogen peroxide increased then the displaced oxygen will increase but eventually it should level out due to the effect of substrate concentration on enzyme activity (Allot, 99-100, 2014). Catalase is an enzyme that is found in the liver of animals and it aids to catalyze the breakdown of hydrogen peroxide into water and oxygen (Allot, 99-100, 2014)....
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...Effect of Temperature on Rate of Reaction of Catalase Abstract Properties of Enzymes focused on the variations of reaction rates amongst enzymes subjected to various circumstances such as temperature, pH levels, different concentrations of substrate, salt concentrations, Metal Copper Sulfate and lastly, the presence of an Enzyme Inhibitor. The assigned section of this laboratory for our efforts was the effect of temperature variations on enzyme reactions. To perform the experiment, we used a spectrophotometer to monitor the baseline catalase activity when they are placed in these two temperatures. In this way, absorbance can be measured over time to monitor catalase activity of the main baseline reaction. Our results showed that temperatures at higher degrees led to being inactive, whereas those at lower degrees lowered the reaction time. This comes to show that each enzyme can have a different optimal temperature and this experiment helped us to understand how reaction rate can be affected by temperature change. Introduction Thousands of complex biological processes are constantly taking place within our bodies. We require material transport, energy synthesis, and the manufacturing of various proteins, hormones, and other molecules (Source 1). Almost all of these everyday processes rely on the function of enzymes to take place. Enzymes are specifically grouped according to their function, and this information can often provide us with clues regarding what type of reaction...
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...LAB #3: An Enzyme in Plant and Animal Tissues BACKGROUND INFORMATION Liver and other living tissues contain the enzyme catalase. This enzyme breaks down hydrogen peroxide (H2O2), which is a harmful by-product of cellular respiration if it builds up in cells. In this lab, you will perform reactions with the enzyme catalase. OBJECTIVES: 1. Investigate the enzyme catalase in various tissues. 2. Measure the effect of changes in temperature on reaction rates of an enzyme-catalyzed reaction in a controlled experiment. INTRODUCTION: What would happen to your cells if they made a poisonous chemical? You might think that they would die. In fact, your cells are always making poisonous chemicals. They do not die because your cells use enzymes to break down these poisonous chemicals into harmless substances. Enzymes are proteins that speed up the rate of reactions that would otherwise happen more slowly. The enzyme is not altered by the reaction. You have hundreds of different enzymes in each of your cells. Each of these enzymes is responsible for one particular reaction that occurs in the cell. In this lab, you will study an enzyme that is found in the cells of many living tissues. The name of the enzyme is catalase (KAT-uh-LAYSS); it speed up a reaction that breaks down hydrogen peroxide, a toxic chemical, into 2 harmless substances--water and oxygen. The reaction is as follows: 2H2O2 à 2H2O + O2 This reaction is important to cells because hydrogen peroxide...
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...Motility Positive control - P.vulgaris A. non motile is negative test B. motile is a positive test 5. Carbohydrates fermentation (test for gram -) ( glucose , manitol, lactose) Positive control – ecoli (yellow) Red- no color change – negative test Yellow – color change – positive test 6. Mae Conkey’s Agar Ecoli – positive control Selective for negative gram stain Differential for organism that could ferment lactase Growth pink – positive No growth – negative 7. Gelatin Hydrolysis ( Gelatinase) Positive control – P.aeruginosa Liquid –positive test Solid - Negative test 8. Blood agar Positive control – S. aureus A. Betahemolysis B. alphahemolysis C. gammahemolysis 9. FTM *broth – O2 relationship with 10. MRVP (mix acid fermentation) MR- methyl red (PH lower than 4.4) Positive control – e.coli red color- positive test no red – negative test 11. Voges Proskauer ( acetone) –precursor for those who fermented butane Positive control- E. aerogenosa Red color – positive test No color change – negative test 12. Nitrate Reduction Enzyme – nitrate redactase Positive control- E.coli Pink color- positive No pink – negative 13. Starch hydrolysis (iodine), test for enzyme laminase Positive control- B. subtillus Zone of clearing- positive test No zone of clearing – negative test 14. Fat hydrolysis, test for enzyme lipase Positive control –...
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...Biology HL 20/9/2013 How Substrate Concentration affects a Catalase Enzyme Reaction RQ How does changing the substrate concentration affect the rate of a catalase reaction in an enzyme? Hydrogen peroxide was used as the substrate and the rate was measured by oxygen production. Hypothesis If the substrate concentration is increased then the rate of a catalase reaction will also increase until it reaches the optimal concentration or saturation point and will remain constant. This is because there will be more substrate molecules in a higher concentration therefore a higher frequency of collisions. This increases enzyme activity and more product will be formed. However at a certain concentration the enzymes will become saturated (all active sites are full), so an increase in substrate molecules will stop affecting the rate. Variables Independent Variable - Hydrogen peroxide concentration Dependent Variable – rate of catalase reaction by measuring the volume of oxygen produced after five minutes Controlled Variables – performed at room temperature (21 Celsius), catalase used (potato), weight of potato (1g) in each trial, time given for reaction to take place (5 minutes), method & apparatus Apparatus 1. Safety goggles & apron 2. Measuring cylinder, 100 cm3 (+/- 0.5cm3) 3. Large plastic tub 4. Access to sink/water 5. Clamp stand, boss and clamp 6. Rubber bung and delivery tube 7. 9 large test tubes 8. Stopwatch (+/- 0.01s) 9. Hydrogen Peroxide, range of concentration:...
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...Matriculation No.: A0094403 Name: FOO KAI JIE MARCUS Group 1. Describe your observation in the following table. If we set the gas produced during the first two minutes in the first reaction tube as a reference, describe the relative amount of gas produced during the same time in the rest of reaction tubes. (2 marks) |No of the reaction tube |Solution |Material |Relative amount of gas produced during the first two| | |(substrate + buffer) |(enzyme source) |minutes after the reaction started | |1 |3ml H2O2 + 3ml DDW |Fresh potato |Potato remained at bottom of test tube, | | | | |effervescence observed, potato floated to the top | | | | |after awhile. | |2 |// |Fresh chicken liver |Liver floated to the top immediately, thick layer of| | | | |bubbles formed. Much more gas produced as compared | | | | |to tube 1. ...
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...as the ideal temperature, the right pH, and a higher enzyme or substrate concentration, the activity of the catalase enzyme would increase. HYPOTHESIS: Exercise 5.1 Catalase Activity-Test tube one, which contains catalase buffered at pH 7 and hydrogen peroxide, will produce the highest bubble height because it contains the proper substrate/enzyme combo. Exercise 5.2 Effect of temperature on Enzyme Activity-The test tube that is place in the incubator will produce the highest bubble height, because warm temperatures speed chemical reactions. Exercise 5.3 Effect of Concentration on Enzyme Activity-Test tube three will produce the greatest bubble height, because it has highest enzyme concentration out of all of the tubes. Exercise 5.4 Effect of pH on Enzyme Activity-The test tube with a pH 7, will produce the highest bubble column, since pH 7 is the optimum pH for catalase. EQUIPMENT: Exercise 5.1 Catalase Activity- Materials used were three clean test tubes, a black sharpie, and a ruler. Additional materials included 10mm of catalase buffered at pH 7.0, and 40mm of hydrogen peroxide for test tube. Ten mm of water, and 40mm of hydrogen peroxide were used for test tube 2. And, 10 mm of catalase, and 40mm of sucrose solution were used for test tube 3. Exercise 5.2 Effect of Temperature- Materials used included three clean test tubes, a black sharpie, 10mm catalase buffered at pH 7.0, refrigerator, incubator, warm water bath, and 40 mm hydrogen peroxide, and a ruler. Exercise...
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...Data: See attached sheet Discussion: The catalase enzyme does not work at high temperatures around or above 50 degrees celsius. We proved this in our lab when the slope changed from 0.021 kPa/sec at 34 degrees celsius to negative 0.009 kPa/sec at 54 degrees celsius. With just a change in 20 degrees the slope, or how well the enzyme is working went from really good to really bad. The catalase enzyme works best around a pH of 10. At a pH of 4 and 7 the slopes were 0.007 and 0.009 kPa/sec. At the pH of 10 it went way up to 0.028 kPa/sec. We can tell the enzyme needs a fairly basic pH for it to work, and not a neutral or acidic one. The more catalase enzyme there is, the better it will work. The rate and slope increased greatly from 0.002 kPa/sec...
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...of the enzymes catalase in a cell is it breaks down the substrate hydrogen peroxide. Hydrogen peroxide is produced during normal metallic activities in the cell, but it is poisonous to the cell if it accumulates. Eventually, hydrogen peroxide will break down into water and oxygen. The accumulation of the poison in the cell would damage the cell long before it breaks down naturally. The cell naturally produces the enzyme catalase to react with the hydrogen peroxide and break it down as rapidly as it is produced to avoid destruction of the cell. The purpose of this lab was to analyze the enzymes in living tissues, represented by the pieces of liver. Also, hydrogen peroxide was used to demonstrate these effects. Procedure In this lab was tested five different substances. In part A the purpose was to observe untreated normal catalase reaction by using small tweezers to obtain two pieces of liver and one piece of each of the other tissues. Using a small beaker 30ml of hydrogen peroxide was obtained. A graduated cylinder was used to measure 3ml of hydrogen peroxide into 5 different test tubes. Then used an expo marker to mark the top of the hydrogen peroxide. A piece of liver was put into the first test tube containing hydrogen peroxide. In part B the purpose was to figure out which tissues contained catalase. 3 ml of hydrogen peroxide was places in 4 additional test tubes. Then one piece of each of the 4 tissues were put into the hydrogen peroxide in separate test tubes. A second...
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...Effect of Catalase on Hydrogen Peroxide Introduction Catalase is an oxidative catalyst enzyme that is found in all organisms that require oxygen or can survive in its presence. Catalase is an enzyme which catalyzes the breakdown of hydrogen peroxide into oxygen and water (Everything Bio, 2007). Catalase is found in large amounts in the liver and serves as the liver’s cleansing function. Hydrogen peroxide is a colorless, dense liquid that is often used as bleach or is diluted with water for use as an antiseptic (Science, 2010). Hydrogen peroxide causes processes similar to rusting. Metals rust as they react with oxygen and such molecules that cause rusting. Oxidative reactions can occur in plant or animal tissues if oxidative molecules are present. These anti-oxidants prevent tissue from oxidizing by aborting oxidizing molecules like hydrogen peroxide. Catalase is one of these anti-oxidant molecules. When the catalase comes in contact with hydrogen peroxide, it turns the hydrogen peroxide (H2O2) into water (H2O) and oxygen gas (O2). Catalase does this extremely efficiently, up to 200,000 reactions per second (How Stuff Works, 2013). The purpose of this experiment is to test the effect of catalase on hydrogen peroxide at different temperatures. My hypothesis is that the beaker with room temperature water will produce a higher level of bubbles showing that catalase is in high frequency at this temperature. Methods Performing the lab was as fun as it was...
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