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Enzymes

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by

Stephanie Cerone

on 1 May 2015

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Transcript of Enzymes

How does temperature influence the rates of enzymatic reactions?
Primary Question
Enzymes
Background Information
If turnips are located in soil that has a temperature of 5 degrees Celsius (41 degrees F), then the enzyme, turnip peroxidase, will have an optimal rate at the temperature closest to 5 and will degrade at a higher temperature.
Hypothesis
Experimental Group:
Temperatures (15, 37, 50, 65, and 100 degrees C)
Control Group:
Room temp turnips (19 degrees C)
Independent Variable:
Temperature
Dependent Variable:
Rate of enzyme activity
Constants:
Amount of enzyme/substrate, time, sunlight, volume, pH

Experimental Design:
1. Using clean test tubes make six sets of pairs of original substrate and enzyme tubes. Prepare the test tubes as follows and be sure to label them. We are doing two trials.
For each substrate in a pair (6), add 7 mL of distilled water, 0.3 mL of hydrogen peroxide, and 0.2 mL of guaiacol for a total volume of 7.5 mL. Cover each test tube with a piece of Parafilm and gently mix.
For each enzyme tube in a pair (6) add 6.0 mL of distilled water and 1.5 mL of peroxidase.
2. Obtain the room temp using a thermometer, this is your constant.
3.Set up an ice bath at 15 C, place substrate in the ice bath for 5 minutes, then test temp using the distiller water constant. Additional time may be needed.
4. Once the substrate is cooled, take out the enzyme and pour into Enzyme 1. Cover the test tube with a price of Parafilm and gently mix.
5. Record the observed color for the test tube at 0 minutes and continue recording at 30 second intervals for 2 minutes.
7. Repeat steps 4 &5.
Procedure
Turnip Peroxidase Color
Graphs
Our results showed that the optimal condition for turnip peroxidase would be around 37 degrees Celsius.
15 C
At 15 C there was minimal activity, since the enzyme was working at a slower rate. The trials average from starting at color 1.5 to reaching color 9.
19 C
At room temperature, the first trial did not have any activity, due to some form of error. However, the second trial showed minimal activity again, but more than at 15 degrees C. This vial's color increased at a steady rate until reaching 10 as its final color chance.
37 C
At the temperature inside humans, there was the highest amount of activity in peroxidase, allowing the reaction to run much quicker than at the colder temperatures. From the initial recording, both trials were are color level 9 and exceed ti 10 after 30 seconds.
50 C
At 50 degrees C there was still enzyme activity. However, it had slowed down compared to 37 degrees C. Both tubes initially started a color 4 but increased to only color 7 within the 2 minute interval.
65 C
At this temperature, there was no enzyme activity due to the high heat denaturing the enzyme, rendering it ineffective and unable to perform its function. Both trials had 0 color change overall.
100 C
We did not bother to test this, since the enzyme had already denatured at 65 C and would do the so at any higher temps.

Analysis
Lab Partners: Stephanie, Liz, Nicole, and Allie
Materials
Turnip peroxidase
0.1% hydrogen peroxide
Guaiacol
Ice/ice bucket
Incubator


Thermometer
Hot Plate
Beaker
Tongs
Heat Gloves
Null Hypothesis:
The rate of the enzyme will not be
affected by the different temperatures of the soil.
15 degrees C
Turnip Peroxidase Color
Turnip Peroxidase Color
Turnips have the ability to grow in a range of temperatures, even when the temp is not optimal
Turnips grow best in low temperatures (4-6 degrees Celsius)
Turnips contain an enzyme- turnip peroxidase
Enzymes have the ability to increase the rate of reaction by lowering the activation energy
Turnip Peroxidase Color
Time: 0 sec.
Trial 1: 1
Trial 2:2
Time: 90 sec.
Trial 1: 5
Trial 2: 9
Room temp: 19 degrees C
37 degrees C
50 degrees C
Turnip Peroxidase Color
65 degrees C
Time: 120 sec.
Trial 1: 8
Trial 2: 10
Time: 30 sec.
Trial 1: 4
Trial 2: 6
Time: 60 sec.
Trial 1: 5
Trial 2: 8
Being unable to invert the tube successfully when heated.
Human Error- mixing an enzyme or substrate tube incorrectly.
Temperature fluctuations after taken off hot plate
Time: 0 sec.
Trial 1: 0
Trial 2: 2
Time: 30 sec.
Trial 1: 0
Trial 2: 4
Time: 90 sec.
Trial 1: 0
Trial 2: 8
Time: 120 sec.
Trial 1: 0
Trial 2: 10
Conclusion
Time: 0 sec
Trial 1: 7
Trial 2: 9
Time: 30 sec
Trial 1: 9
Trial 2: 10
Time: 60 sec
Trial 1: 9
Trial 2: 10
Further and improve experiment by:
Multiple trials (more than 2)
Focusing the temperature range closer to the optimal rate (at 37 degrees C)
Focusing on the temperature when the enzyme denatures (roughly 65 degrees C)
Extend the time period
Research if there are other factors that help the turnip as a whole grow best in colder temperatures
Time: 90 sec
Trial 1: 9
Trial 2: 10
We learned that turnip peroxidase best functions at about 37 degrees C and denatures somewhere around 65 degrees C.
Time: 120 sec
Trial 1: 9
Trial 2: 10
The results of our conclusion do not support out hypothesis. We believed that the optimal temperature would be closer to 15 degrees C. Instead we found that the optimal temperature for turnip peroxidase was at 37 degrees C.
Time: 0 sec.
Trial 1: 4
Trial 2: 4
Introduction
Purpose:
Determine the influence temperature has on the enzyme turnip peroxidase
Compare the different rates of reaction at the different temperatures.
Find the optimal temperature that produces the highest peroxidase activity.
Scientific Principles:
The function of an enzyme depends on its shape. At high temperatures, enzymes can denature, results in a deformity in their shape so they cannot perform their function.
If the reaction runs for an extended period of time, it will eventually reach equilibrium, meaning that the amount of substrate is either at zero or almost zero.
A current issue is global warming; how will that affect natural processes if the temperature keeps increasing and certain enzymes denautre?
Time: 30 sec
Trial: 1: 5
Trial 2: 5
Time: 60 sec
Trial 1: 6
Trial 2: 6
Time: 90 sec
Trial 1: 7
Trial 2: 6
Time: 120 sec
Trial 1: 7
Trial 2: 7
Time: 0 sec
Trial 1: 0
Trial 2: 0
Time: 30 sec
Trial 1: 0
Trial 2: 0
Time: 60 sec
Trial 1: 0
Trial 2: 0
Time: 90 sec
Trial 1: 0
Trial 2: 0
Time: 120 sec
Trial 1: 0
Trial 2: 0
In this experiment we made six sets of pairs of test tubes, one containing the enzyme and the other containing the substrate. The enzyme consisted of 6.0 mL distilled water and 1.5 mL of peroxidase, while the substrate was made up of 7.0 mL of distilled water, 0.3 L of hydrogen peroxide, and 0.2 of guaiacol. Our constant of the experiment was at room temperature, which at the times was 19 degrees Celsius. Other temperatures that we tested were varied by either the ice bath or the hot plates. After being heated or cooled, the mixtures then were gently inverted to start the reaction.
Sources of error:
Summary:
Statistical Data
Trial 1:
Rate: .0583
Trial 2:
Rate: .0667
Statistical Data
Trial 1:
Rate: 0
Trial 2:
Rate: .0571
Statistical Data
Trial 1:
Rate: .0167
Trial 2:
Rate: .0083
Statistical Data
Trial 1:
Rate: .025
Trial 2:
Rate: .025
Statistical Data
Trial 1:
Rate: 0
Trial 2:
Rate: 0
Average: .0625
Standard Deviation: .0042
Average: .0286
Standard Deviation: .0286
Average: .0125
Standard Deviation: .0042
Average: 0
Standard Deviation: 0
Average: .025
Standard Deviation: 0
Full transcript