A water bath that hasbeen warmed to 33C. Measure the oxygen level in the graduatedcylinder at 1 minute from the time the reaction vessel is turned onits side. What do you think would happen if the potato or liver was boiled before being added to the H2O2? As the dilution factor increased, 1:5 1:10 1:50 so did the amount of time it took for the filter paper to reach the top of the suspension. By adding a catalyst to the H 2O 2, the activation energy needed for the reaction is lowered until the reaction by the catalyst is finished or settles. The group hypothesized that by mixing the Bromelain pills with water and pouring into a gelatin at varying temperatures, the protease within the Bromelain would break down the gelatin causing it to become a liquid. Exercise 2D: An Enzyme-Catalyzed Rate of H2O2 Decomposition The baseline assay was reestablished following the directions of Exercise 2B. Both of these conditions denature the enzyme and cause its active site to change shape.
If you had to rely onthese reactions to take place spontaneously, you would wait a verylong time -- you wouldn't be here! However the addition of more H 20 2 solution to the used liver did produce another visible and fast reaction. At the pH of 9 basic it took the amylase 9 minutes to produce the indication of starch being converted into simpler monosaccharides. Hydrogen peroxide is naturally produced in organisms as a by-product of oxygen metabolism and needs to be broken down because high levels of it are extremely toxic. We then used a transfer pipet to add 1mL of distilled water from the d H 2O cup to the baseline cup used to replace 1mL of catalase from activities A and D. Once the liver was macerated thoroughly, we added 10mL of H 2O 2 with the H 2O 2 syringe and observed the reaction.
Though unlikely, being off by perhaps 10 seconds during your first stagger time would cause the timer to be off by 10 seconds each time they reach the 1 minute interval; after 6 rounds of observations the time would be off by an entire minute. Submerge the graduatedcylinder to fill it with water. After the transfer with the transfer syringe, rinse syringe with water from the wash water cup and discard it to the waste water cup. We first boiled the catalase to see what the boiling would do to an enzyme, our result was that the boiling denatured it, making it not work anymore. Suggested multiple tests are in order to provide any new data. Our data shows a linear trend, however, research shows that the trend is in fact a parabola opening downwards with a maximum at a pH of 5, the acidity of Hydrogen Peroxide alone.
The same steps were repeated for the 30, 60, 120, 180, and 360 second cups, respectively. Also, in discussing your results, in the discussion sectionyou will also need to do some research to explain the effect of thevarious conditions have on catalase activity. As the concentration of enzyme decreased, so did the amount of oxygen that was being produced. Using a clean and unused pipet, we then added 1mL of the boiled catalase after 5 minutes to the unlabeled cup and observed. Mix and wait exactly 5 mintues before making run. Starting ml O2 : 0 ml Part B: Enzyme Concentration.
Lecture 1: secondary structure of proteins. Hypothesis The enzyme catalase, under optimum conditions, effectively speeds the decomposition of hydrogen peroxide. Place a thermometer in the pan and sometime duringPart A, record the temperature of the water. Keep the side with the disks upward andcarefully place the reaction vessel on its side in the pan of water. Materials andMethods: At your lab table, youwill find everything you need to conduct today's experiments. Because of their 3-D shape, enzymes are highly specific for thereactions that they catalyze.
Getting a solid into a graduated cylinder was no easy task and required using water displacement to approximate the measurements. Having a larger piece of liver would have given the liver greater surface area in which to react with the H 20 2 solution and therefore could have created a larger column of oxygen bubbles; having a smaller piece would have resulted in vice versa. A lower concentration of the yeast would take longer to rise to the top; less oxygen bubbles would be provided from catalyzing the decomposition of the H 20 2 solution. The liver at 0C still produced visible results; however the oxygen column they produced was smaller than the one produced when the catalase was at its optimal temperature. Summarize the effect of concentration on enzyme activity.
The catalase found in chicken liver did react. Error: Part 2 Stagger time slightly off This result is entirely human error but is a problem because of the difficultly of properly managing the stagger time for this experiment. Neither you, nor the coeditors you shared it with will be able to recover it again. They allow these complex biochemical reactions to occur at a relatively low temperature and with less energy usage. By understanding a simple catalase reaction with H 2O 2 in activity A, we can determine the baseline amount of H 2O 2 in activity B and use the correct collected data to determine the H 2O 2 decomposition rates in activities C and D.
Keep adding ice to keep temperatureclose to 5C. Summarize the effect of temperature on enzyme activity 1. Inaccurate readings of volume may also have played a critical role in the failure of the experiment. Observing the reaction of catalase on hydrogen peroxide: a. Throughout the three parts of the lab, the ability for enzymes to break down was studied.
Catalase breaks down H202 into water and oxygen. While two students worked on the time trial reactions, the other two will take the time trial cups after the reactions and titrate them by using the titration protocol with potassium permanganate. In this lab forease of calculation, you will measure for one minute. Activity A: Observing the Reaction Hypothesis: If we add catalase to the H 2O 2 solution, a reaction will occur because of the catalase. The rate was determined by taking an accurate reading of the amount of oxygen being produced over a period of 30 seconds. What is the temperature optima foR human enzymes? This disturbes their relative ly weak bonds.