Enzyme Action: Testing Catalase Activity 6B
Ava Suda
Cor Jesu Academy
Enzyme Action: Testing Catalase Activity 6B
Introduction
Hydrogen Peroxide, or H2O2, is harmful to most living organisms but can be converted to oxygen and water before the damage is permanent. This is thanks to enzymes, the biological catalysts that increase the rate of reactions. Enzymes can be studied by measuring the rates of enzyme-catalyzed reactions. This can be done in a number of ways, including measuring the pressure of the product as it appears, measuring the rate of disappearance of the substrate, and measuring the rate of appearance of a product.
Purpose
To measure the rate of enzyme activity under different temperatures, enzyme
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Click the green “Collect” button to collect data, which will cease collection after 3 minutes. If the pressure becomes higher than 130 kPa, then the stopper will pop out. If this occurs, then loosen the plastic tubing around the stopper to relieve pressure. Once the data is collected, disconnect the tubing from the stopper, remove the stopper, and discard the contents in a waste beaker or sink. Move the cursor to where the values start increasing, and click. Drag the mouse pointer to the point where the pressure values stop increasing and release the button. Next, click the Linear Fit button to form a linear regression. Record the slope of the line in Table 4. The line regression box can now be closed. Repeat these steps with Test Tubes 2, 3, and 4, adding 2 drops to Test tube 2, 3 drops to Test Tube 3, and 4 drops to Test Tube 4, respectively.
To test the effect of temperature, place 4, clean test tubes in a test tube rack and label them “T 0-5,” “T 20-25,” “T 30-35,” and, “T 50-55.” Add 3 mL of 3% H2O2 and 3 mL of water to each test tube. To measure enzyme activity at 0-5°C, prepare a water bath with a temperature range of 0-5°C in a 600 mL beaker. Place Test Tube T 0-5 in the water bath and record the temperature in Table 4. Add 2 drops of the enzyme solution to the test tube, and connect the free end of the plastic tubing to the connector in the stopper. Click the “Collect” button to collect data, and
The more acidic a substance is the less oxygen it will produce when going through a chemical reaction. During the Lab “How Do Changes in pH Levels Affect Enzymes Activity”, the researcher conducted an experiment to test the effects that an acidic, neutral, and a base substance will have when combine it with hydrogen peroxide. The data table shows that HCL (acidic substance) barley produced any oxygen at all when it was combining with Hydrogen Peroxide. The pH level for HCL was 2.5; this level indicates that the substance was very acidic. When the H2O and NaOH were tested they produced more bubbles than HCL. NaoH produced a little more bubbles than HCL. The pH that NaoH produced was a 9, which is a base. H2O produced more bubbles than both substances;
I put one test tube for each control, substrate, and enzyme in the 4° C (ice bath), 23° C (room temperature), 37° C (body temperature), and 60° C (water bath). Add the inhibitor that was used and what it was used for.
Students will be observing normal catalase reaction, the effect of temperature on enzyme activity, and the effect of pH on enzyme activity in this experiment. The enzymes will all around perform better when exposed in room temperature than when it is exposed to hot and cold temperatures. This is based on the fact that the higher the temperature, the better the enzymes will perform, but as the temperature reaches a certain high degree, the enzymes will start to denature, or lose their function.
These results show how temperature of extreme high, or low affects enzyme activity. The highest rate of enzyme activity occurred at 37 Cº. Anything that was hotter or cold than 37 Cº slowed the reaction rate. As I thought, 100 degrees would denature the enzyme, and that was the case. The data provided shows exactly what temperatures enzymes work best, and worst. The objective was achieved as we discovered the different reaction rates under different temperatures. The results are reliable, as we know enzymes do not work well when under extreme heat or denaturation occurs. What I learned in this experiment was that enzymes don’t work well under cold temperatures because they tend to move slower. My hypothesis did not quite match, because I thought they work best at lower temperatures.
Enzymes are catalysts that function to speed up reactions; for example, the enzyme sucrose speeds up the hydrolysis of sucrose, which breaks down into glucose and fructose. They speed up reactions but are not consumed by the reaction that is taking place. The most important of the enzyme is the shape as it determines which type of reaction the enzyme speeds up. Enzymes work by passing/lowering and energy barrier and in doing so; they need to bind to substrates via the active. Once they do, the reaction speeds up so much more quickly than it would without the enzyme. Coenzymes and cofactors aid the enzyme when it comes to binding with the substrate. They change the shape of the active site so the substrate can bind properly and perform its function.
Hold the IKI spray bottle 25 - 30 cm away from the paper towel, and mist with the IKI solution.
The optimum pH level would be pH 7. This is because this is where the highest amount of enzyme activity is taking place.
Lactose is a sugar that can be put into smaller molecules, glucose and galactose. Lactose is when you are not able to digest milk and dairy meaning that the enzyme lactase that breaks down lactose is not functioning properly. ONPG was used as a substitute for lactase because even though it is colorless it helps show enzyme activity by turning yellow. This experiment measured the absorbance ONPG when exposed to lactase within an environment of different salinity’s. The enzyme, lactase, was obtained by crushing a lactaid pill and then was added into four cuvettes. ONPG and salt solution of different concentrations were added and their levels of absorption was measured by a spectrophotometer. The results showed that higher salt concentrations have a lower level of absorption. There were 4 cuvettes and within those cuvettes that solutions within them were being tested and the results showed the more salt solution added with the lactase the lower the absorbance. The less salt solution there was a higher rate of absorbance. The data supported the hypothesis that with increasing NaCl concentration there would be a decrease in enzyme activity.
The purpose of this experiment was to record catalase enzyme activity with different temperatures and substrate concentrations. It was hypothesized that, until all active sites were bound, as the substrate concentration increased, the reaction rate would increase. The first experiment consisted of five different substrate concentrations, 0.8%, 0.4%, 0.2%, 0.1%, and 0% H2O2. The second experiment was completed using 0.8% substrate concentration and four different temperatures of enzymes ranging from cold to boiled. It was hypothesized that as the temperature increased, the reaction rate would increase. This would occur until the enzyme was denatured. The results from the two experiments show that the more substrate concentration,
An Enzyme is a protein, which is capable of starting a chemical reaction, which involves the formation or breakage of chemical bonds. A substrate is the surface or material on or from which an organism lives, grows, or obtains its nourishment. In this case it is hydrogen peroxide. This lab report will be explaining the experiment held to understand the effects of the changes in the amount of substrate on the enzyme’s reaction.
“Enzymes are proteins that have catalytic functions” [1], “that speed up or slow down reactions”[2], “indispensable to maintenance and activity of life”[1]. They are each very specific, and will only work when a particular substrate fits in their active site. An active site is “a region on the surface of an enzyme where the substrate binds, and where the reaction occurs”[2].
Organisms cannot depend solely on spontaneous reactions for the production of materials because they occur slowly and are not responsive to the organism's needs (Martineau, Dean, et al, Laboratory Manual, 43). In order to speed up the reaction process, cells use enzymes as biological catalysts. Enzymes are able to speed up the reaction through lowering activation energy. Additionally, enzymes facilitate reactions without being consumed (manual,43). Each enzyme acts on a specific molecule or set of molecules referred to as the enzyme's substrate and the results of this reaction are called products (manual 43). As a result, enzymes promote a reaction so that substrates are converted into products on a faster pace (manual 43). Most enzymes are proteins whose structure is determined by its sequence of its amino acids. Enzymes are designed to function the best under physiological conditions of PH and temperature. Any change of these variables that change the conformation of the enzyme will destroy or enhance enzyme activity(manual, 43).
One of the methods I could use to improve the quality and precision of my data is to utilize a micropipette to precisely transfer my substrate solution to the the enzyme. This would help me prevent molecules remaining in contact with the sides of the test tube and so that all the substrate molecules are in direct contact to react with all the enzymes in a more efficient and precise manner. In addition, one method I would follow to maintain the accuracy of my results would be to utilize an oxygen sensor for the measurement of the rate of enzyme activity more precisely and avoid any initial escape of oxygen from the test tube. This would also help me compare my data in a more accurate manner, which would give me a broader spectrum of factors that I could incorporate to the analysis of my experiment. Moreover, in regards to time management, by collecting my data through the course of a single day will serve a significant role within the consistency of enzyme activity and help minimize inaccurate results. Furthermore, it is also important to ensure that equipment such as temperature probes, pipettes or micropipettes, and gas pressure sensors, are all calibrated prior to the course of
To find the effect of temperature on the activity of an enzyme, the experiment deals with the steps as follows. First, 3 mL if pH 7 phosphate buffer was used to fill three different test tubes that were labeled 10, 24, and 50. These three test tubes were set in three different temperature settings. The first test tube was placed in an ice-water bath for ten minutes until it reached a temperature of 2° C or less. The second tube’s temperature setting was at room temperature until a temperature of 21°C was reached. The third tube was placed in a beaker of warm-water until the contents of the beaker reached a temperature setting of 60° C. There were four more test tubes that were included in the procedure. Two of the test tubes contained potato juice were one was put in ice and the other was placed in warm-water. The other two test tubes contained catechol. One test tube was put in ice and the other in warm water. After
Hydrogen peroxide is a toxic byproduct of cellular functions. To maintain hydrogen peroxide levels the catalase enzyme deconstructs hydrogen peroxide and reconstructs the reactants into oxygen gas and water. The catalase enzyme is found inside cells of most plants and animals. Regulating the levels of hydrogen peroxide is crucial in homeostasis and analyzing it’s optimal conditions for performance is just as important. To understand the optimal environment for this enzyme, they are put into different environments based off protein activity (enzymes are proteins). Catalase samples will be put into different hydrogen peroxide environments based off pH and temperature. The more active the enzyme, the more oxygen and water it will produce. Enzyme activity can be seen through the release of oxygen in the hydrogen peroxide. Since oxygen cannot be accurately measured, the data will consist of the longevity of the reaction in different environments. If the pH is higher than 7, then the reaction rate will increase due to the ample amount of hydrogen ions in the hydrogen peroxide. However the pH level cannot be higher than 10 or else there will be too many hydrogen atoms in the peroxide for the enzyme to be able to deconstruct them. If the temperature is increased, then the reaction rate will increase due to the ample amount of energy and movement in the hydrogen peroxide and enzyme.