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Kami Export - Lab 8 Enzymes Answer Sheet

This document summarizes a biology lab experiment on enzymes. The experiment tested how different factors affect the rate and activity of the enzyme catechol oxidase. The factors tested included substrate concentration, enzyme concentration, temperature, pH, and inhibitors. Increasing substrate or enzyme concentration increased the reaction rate until all enzymes were active. High temperatures above the optimum range caused enzyme denaturation, while very low or high pH impaired enzyme function. Phenylthiourea acted as a competitive inhibitor of catechol oxidase.

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Monica Ochoa
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684 views

Kami Export - Lab 8 Enzymes Answer Sheet

This document summarizes a biology lab experiment on enzymes. The experiment tested how different factors affect the rate and activity of the enzyme catechol oxidase. The factors tested included substrate concentration, enzyme concentration, temperature, pH, and inhibitors. Increasing substrate or enzyme concentration increased the reaction rate until all enzymes were active. High temperatures above the optimum range caused enzyme denaturation, while very low or high pH impaired enzyme function. Phenylthiourea acted as a competitive inhibitor of catechol oxidase.

Uploaded by

Monica Ochoa
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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Download as PDF, TXT or read online on Scribd
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Bio 101 Laboratory Michel Lupian

Name: __________________________________

Laboratory 8
Enzymes: Factors Affecting Their Rate and Activity

Complete these questions (part 1 and part 2) for credit. The answer spaces are all fill-in
or drop-down selections. Once finished with the answer sheet, do not simply “save” the
file. You must “print” the file to “pdf”, generally by selecting a printer named “Adobe
PDF” for windows computers – this does usually involve a choice to “save as pdf”.

QUESTIONS PART 1
1. Name the following for this laboratory:
Catechol oxidase
a. Enzyme............................................................... _______________
Catechol
b. Substrate ............................................................ _______________
Hydroxyquinone
c. Product of the enzymatic reaction ..................... _______________
2. Was a control used in these experiments? If so, what were the contents of the
tube was the control?
The contents of the tube that was the control in these experiments was enzyme.

3. What result do you expect in the control?


The result that I would expect in the control is that altering the amount of substrate available to
an enzyme and the physical environment of the enzyme will affect its function and activity.

4. Briefly describe why scientists use a control.


Scientists use controlled experiments because they allow for precise control of extraneous and
independent variables. This allows a cause and effect relationship to be established. Controlled
experiments also follow a standardized step by step procedure.

5. What would it mean if you got unexpected results in the control tube?
What it would mean if you got unexpected results in the control tube is that the main hypothesis
is not supported by the data collected. It could be that a different tube wasn't used to test the Dl
water and substrate and as a result an unexpected reaction took place.

6. What is the result of increasing substrate concentration?


The result of increasing substrate concentration is that it will increase the rate of reaction but if all
the enzymes are active then the substrate activity will freeze until enzymes have finished and are
available

Adapted from: Nicholson, M., & Newby, S. (Eds.). (2017). General biology. Second Edition. Minneapolis, MN: Bluedoor. (Lab manual)
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Bio 101 Laboratory Name: __________________________________

Table 8.2: Catechol oxidase enzymatic activity after 5 minutes.


Tube Contents Color Intensity
1 Enzyme + Substrate brown 2 or (++)
2 NO Enzyme Choose zero (0)
3 NO Substrate Choose zero (0)

Table 8.4: Degree of color change in each test tube after 5 minutes.
Tube Substrate Concentration Color Intensity
1 0.5x Substrate pink 1 or (+)
2 1.0x Substrate yellow 1 or (+)
3 2.0x Substrate brown 2 or (++)

Table 8.6: The effects of enzyme concentration after 5 minutes.


Tube Enzyme Concentration Color Intensity
1 0.5x Enzyme yellow 1 or (+)
2 1.0x Enzyme brown 2 or (++)
3 2.0x Enzyme brown 2 or (++)

Table 8.8: The effects of temperature on enzyme activity.


Tube Temperature Color Intensity
1 0°C - Ice Bath yellow 1 or (+)
2 22°C - Water Bath yellow-brown 2 or (++)
3 40°C - Water Bath russet 2 or (++)
4 60°C - Water Bath pink zero (0)
5 80°C - Water Bath light-pink zero (0)

Table 8.9: The effect of pH on enzymatic activity.


Tube pH Color Intensity
1 2 Choose zero (0)
2 4 yellow 1 or (+)
3 6 brown 2 or (++)
4 8 brown 2 or (++)
5 10 garnet Choose
6 12 pink zero (0)

Adapted from: Nicholson, M., & Newby, S. (Eds.). (2017). General biology. Second Edition. Minneapolis, MN: Bluedoor. (Lab manual)
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Bio 101 Laboratory Michel Lupian
Name: __________________________________

Table 8.11: The Effects of Enzyme Inhibitors


Tube Contents Color Intensity
1 no PTU, 1x catechol brown 2 or (++)
2 1x PTU, 1x catechol Choose zero (0)
3 0.5x PTU, 1.5x catechol Choose zero (0)
4 0.1x PTU, 2.0x catechol yellow 1 or (+)

QUESTIONS PART 2
1. If the tube incubated at 80 °C was placed back in the optimum temperature,
would a reaction occur? Explain.
If the tube incubated at 80 degrees celsius was placed back in the optimum temperature, a reaction
wouldn't occur because the hotter it is, the more rapidly molecules are vibrating and moving about.
Collisions of water molecules with the enzyme could undo its folded structure that is held in
position by hydrogen bonds; when this happens the molecule is said to have been denatured.

2. Why does the enzyme reaction fail to occur at very low or very high pH?
The enzyme reaction fails to occur at very low or very high pH because the change of pH will
lead to the ionization of amino acids atoms and molecules, change the shape and structure of
proteins, thus damaging the function of proteins. Very low or very high pH will lead to the
complete loss of the activity of most enzymes.

3. Was phenylthiourea a competitive inhibitor or a non-competitive inhibitor?


(Review Figure 8.3 and the preceding text about inhibition.)
The phenylthiourea was a competitive inhibitor.

Adapted from: Nicholson, M., & Newby, S. (Eds.). (2017). General biology. Second Edition. Minneapolis, MN: Bluedoor. (Lab manual)
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