The objective of this lab is to test enzyme productivity, whilst evaluating the factors that influence enzyme catalyzation activities and recording the results. The intent was to discover how temperature impacted the enzymes efficiency. Introduction: Enzymes function to catalyze a change in a chemical reaction they are normally made of proteins and can be used repeatedly. The enzyme studied in this experiment is catalase an enzyme known for breaking down hydrogen peroxide. Enzymes are not always able to accomplish what they are designed to accomplish, they can become denatured (lose shape) due to temperatures, pH, inhibition, competitive inhibition, etc. Enzyme efficiency or activity can be affected by external factors, in this experiment …show more content…
what affected the enzymes and to what extent they were affected was looked at in order to answer how the enzymes were affected and why they were affected, this was measured by monitoring O2 levels. Hypothesis: The testing consisted of multiple different parts. The part being focused on in this write up is part A. If Catalase is exposed to varying temperatures, then Catalase will work best at temperatures close to the natural body temperature in humans, because temperatures too low will cause the enzyme to not function/form efficiently; too high and it will become denatured and cease to function. Most likely Catalase will work best at or near the natural human body temp, as it functions in humans and must do so at 98.6 F. This is important to study because it allows one to understand more closely the cycle of products through the human body as well as humans ability to process waste, that otherwise left untreated could cause serious harm. This gives better scientific knowledge of what is currently happening in other organisms as well as catalase is not specific to humans. Methods: Step 1: Part A of the lab is to test the effect of temperature on the enzyme Catalase. Gather 4 clean test tubes, create a mixture of 20mL of H2O2 and 12mL of H2O, distribute 8mL of this mixture to each test tube, label each test tube something different with a wax crayon/pen to easily identify which tube they are. Step 2: Cool one test tube down to 0C using ice and water in a 400mL beaker, keep handy but set aside to chill. Allow one beaker to reach room temp if the mixture isn’t room temp already by letting it sit out keep this tube in a holding rack at table.
Next, place one tube in a warmer that is set to 37C and leave the tube to reach this temp. For the 4th tube you will place this tube in a special warmer that is keeping the water at a constant 55C allow tube contents time to reach temp. Step 3: Take a break until the test tubes have been in temperature baths for 10 minutes. To identify the enzymes activity, take tube 1 out of the ice bath it is in. Add ~10 drops of the catalase solution to the mixture. Now, pour the contents of test tube into a Nalgene bottle capable of holding 250mL and mix contents. Place an O2 sensor in the mouth of the Nalgene bottle thus plugging the bottle ensuring no outside air will contaminate the air the probe/sensor is reading from. One can start data collection after 30 seconds, collect for 3 minutes, retrieve a graph of the O2 data on a time-longitudinal scale disconnect O2 sensor from Nalgene bottle, pour out contents into a safe and disposable waste receptacle/drain. Clean out Nalgene bottle and thoroughly dry it as well as possible. Calculate the slope of the reaction or the % of change of O2 to calculate the reaction rate. Step 5: Using the tube at room temperature (23C) repeat step 3 for this tube and
record. Step 6: Using tube 3 at 37C repeat step 3 and record. Step 7: Using tube 4 at 55C repeat step 3 and record results. Independent variable = Temperature of H2O and H2O2 mixture. Dependent variable = O2 creation rates. Controlled variables = Keep moisture off of O2 probe, thoroughly dry and washed the Nalgene bottle between uses, securely place the O2 sensor in Nalgene bottle neck in order to keep external air from being introduced and contaminating these readings. Control: The readings received in the at room temperature (23C). Results: From conducting the experiment oxygen was observed being created except at the temperature of 55C,
Start with the hot water and first measure the temperature. Record it. 8. Then pour 40 ml into the beaker. You can measure how much water was used by looking at the meniscus.
We then took 1ml of the 0.1% solution from test tube 2 using the glucose pipette and added it to test tube 3, we then used the H2O pipette and added 9ml of H2O into test tube 3 creating 10ml of 0.01% solution.
5.) One at a time, place your test tubes in the water bath and heat the first test tube to 25 , the second to 50 , the third to 75, and the last to 100 degrees c. Remeber to stir with your stirring rod every so often.
When those steps are finished, the temperature is held at 20°C. In step one, the hot start is initiated by incubating the tubes for five minutes at 95°C and adding the water. 0.4 ul Taq DNA polymerase to each tube while disallowing the tubes to cool and without taking. time to mix the reaction solution after adding the Taq polymerase.
The purpose of this experiment was to discover the specificity of the enzyme lactase to a spec...
3. Drop tablet of Alka-Seltzer into the cold beaker of water and time length of reaction with a stopwatch
3. Fill up the 50 milliliter beakers all the way up with deionized water. After, use your scopula to scoop out a small portion of one substance and put in into your beaker. Then put your hot plate onto medium temperature and rest the mixture onto the plate.
The factors that were investigated in the enzyme lab were specificity of substrate, concentration of substrate and enzyme, pH of reaction solution, and temperature of reaction solution.
Analyze each fraction by spotting 10 times with capillary tubes on a TLC plate, which is exposed to iodine vapor for 15 minutes.
2. In the large beaker, put water and boil it completely. After that, remove the beaker from heat. 3. Sample tubes (A-D) should be labeled and capped tightly.
In a 250ml beaker place 100mls of water, measure the temperature of the water and record this initial temperature onto a table. Set the timer and add one teaspoon of Ammonium Nitrate to the water, stir this continuously until the Ammonium Nitrate has dissolved. After 1 minute measure the temperature and record it, do this for a further 2 minutes (3 minutes in total). Repeat this process for a total of 10 teaspoons.
Then add growth media until the sample is submerged beneath the 2-3 ml of liquid, cap the tube and invert several times to mix thoroughly, incubate the tube while shaking vigorously in a shaking incubator at 250 rpm for a couple hours, then allow the sample to sit.
13.Repeat steps 1-12 for the 80℉, 95℉, and 110℉ catalase solutions. Use clean filter paper each time you test. Record the times for the three trials in the appropriate column of the data table.
Then we pulse-spinned the contents for allowing flow to the bottom. Following, we utilized a floating rack to allow incubation for thirty minutes. After incubation, we proceeded to load 2 micro liters of loading dye into each individual test tube. Following, we allowed the samples to rest on ice. Since we utilized agarose gel, at this point we obtained the gel and filled the chamber with the buffer. The buffer was loaded properly, so we had it properly cover the gel just enough, and it covered both negative and positive poles at the proper height. Next, we loaded ten micro liters in each respective well in the gel. Following, we covered the chamber and connected the leads. Then allowed current to flow for 43 minutes is how long it took our group. The handout recommended approx. 30