Catalase Enzyme Lab

When placing the large test tube in boiling water, be sure to keep the face of the large test tube away from you or any lab partners with safety tongs.

After 30 seconds the same enzyme volume was added to the second flask. This was continued to the third, fourth and fifth flasks with interval of 30 seconds until the final addition occurred at 2.5 minutes.

Students will place a potato cube into a test tube and add 3 ml of H202 into each tube. Then students will wait one minute and record the height in cm of the bubbles and rate how rapidly the solution bubbles on a scale of 0-5. (0=no reaction, 1=slow,……. 5= very fast.)

12.Stir then pour 2.5ml of the enzyme mixture into one of the test tubes not allowing any water from the saucepan into the test tube.

* The beakers labelled; Solutions of lipase respectively at concentrations of 1%, 2%, 3%, and 4% were made using lipase at 5% and distilled water.

How could you test to see if an enzyme was completely depleted during an experiment?

1. How many “chainobeads” was your enzyme able to make per minute in the 0 – 15 second interval?

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,

The volume of a small test tube and a thin-stemmed pipet were determined in this section of the lab. Water was poured into a small test tube until the water reached the very top edge of the test tube. The test tube was then emptied into a plastic 25 mL graduated cylinder and volume was measured and recorded into data table 3. A think-stemmed pipet was completely filled with water. Drops were carefully counted and emptied into the empty plastic 25 mL graduated cylinder until the water level reached 1 mL. The number of drops in 1 mL was recorded into data table 3. The thin-stemmed pipet had a total volume of 4 mL and that was also recorded into data table 3.

In part II of the lab six small glass tubes were obtained in a test tube rack. Ten drops of distilled water were then added to test tube 1, five drops to tubes 2-4, and no drops in tubes 5 and 6. Five drops of 0.1M HCl were added to test tube 5 and five drops of 0.1M NaOH to test tube 6. Five drops of enzyme were then added to all tubes except tube 1. Tube 3 was then placed in the ice bucket and tube 4 was placed in the hot bucket at 80-900C for five minutes, the remaining tubes were left in the test tube rack. After the five minutes five drops of 1% starch was added to every tube and left to sit for ten minutes. After ten minutes five drops of DNSA were then added to all the tubes. All the tubes were then taken and placed in the

Discussion: The purpose of this experiment was to observe what the normal enzyme1 (proteins that catalyze chemical reactions) activity of beef liver catalase was, and to measure the effects of extreme changes in temperature and pH on the beef liver catalase function. We also observed what the bromelain1 (proteins in pineapple) activity was in a fresh pineapple compared to canned pineapple. Enzymes are present in all living things.1 These specialized proteins speed up chemical reactions fast enough to sustain life.1 This lab was driven by the question; if there is an extreme change in temperature or pH on an enzyme catalase, or a bromelain enzyme, then that catalase or bromelain will be affected or denatured.

Also, the enzyme was treated with different urea concentrations before it was added to the assay mixture. The amount of the second and third enzymes was essential. Likewise, this would have had an effect on the rate of the measured aldolase activity. Hence, the concentrations for both enzymes was measured and calculated prior to the experiment whilst preparing the assay mixture. Furthermore, in order to measure thiol group reactivity the base line was adjusted to zero for each cuvette. This was done to make sure that the results were

Using Logger Pro, the pressure increase of oxygen gas, O2, released for each of the three solutions was collected over a three minute time period. From this data, three graphs were created. After data collection was finished, a linear regression was performed on the graphs to find the slope, m, of each of the graph, which the the rate of enzyme activity. For the hydrogen peroxide solution with the pH 4 buffer, the slope was -0.000501 kPa/s. For the solution with the pH 7 buffer, the slope was 0.00153 kPa/s and for the solution with the pH10 buffer, the slope was 0.000619 kPa/s.

The test tube labelled I was subjected to an ice bath for a minimum of 20 minutes. The test tube was positioned in a way that it won’t touch the ice, and the experiment test was conducted on top of the ice. The test tube labelled 37 was subjected to a 37°C water bath for a minimum of 10 minutes. The experiment test was performed while it was still in the water bath. Finally, the test tube labelled R was left in room temperature because it was the control for the experiment.

This question is important to understand because Catalase plays a vital role in the sustenance of cells and tissues