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Bao kha
Lab Partner: Noah, Harry, Titan.
Effect of Temperature on Enzyme Activity
Honor Code: On my honor, I have neither received nor given any unauthorized aid on this
assignment. Kha

Introduction:

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Enzymes are biological chemicals that help speed up the process of chemical reactions that can
take place within cells. One of the place we can find enzymes in our body is the digestive
system. Enzymes help us break down food and speed up chemical reactions. Quoting NCBI, In
the absence of enzymatic catalysis, most biochemical reactions are so slow that they would not
occur under the mild conditions of temperature and pressure that are compatible with life.
Enzymes accelerate the rates of such reactions by well over a million-fold, so reactions that
would take years in the absence of catalysis can occur in fractions of seconds if catalyzed by the
appropriate enzyme. Also, without enzymes, food in our body will not be able to break up, thus
we would not be able to digest and we would die. There are some factors that affect the enzyme
activity, they are: pH, temperature, enzyme concentration, and substrate concentration. The
purpose of this lab experiment is to know and understand how temperature affect on enzyme, and
how long would it take to affect the enzyme activity. If the temperature is high, it will fasten the
enzyme activity. If the temperature is colder, it will slower the enzyme activity. Quoting Live
Strong, The liver is the largest gland in the body and possesses remarkable regeneration
capabilities. The gland secretes bile, a green-colored digestive enzyme stored in the gall bladder
during fasting. Bile acids aid in digestion, among other functions. Quoting MadSci, Hydrogen
peroxide is made by quite a few enzymes in the body. In particular, some enzymes breaking
down certain amino acids and fatty acids (D-amino acid oxidize and acyl-CoA oxidize) make
significant amounts of hydrogen peroxide. Since hydrogen peroxide can be damaging to normal
Tissue, these enzymes are kept inside specialized organelles inside cells called paroxysms. The
paroxysms also contain large amounts of catalase to break down the hydrogen peroxide before it
can escape. Other enzymes that make significant amounts of hydrogen peroxide are plasma
amine oxidize and xanthenes oxidize. In addition to enzymes that produce hydrogen peroxide
as part of their normal catalytic cycle, many enzymes that undergo oxidation and reduction make
hydrogen peroxide and other reactive oxygen species by autoxidation (a kind of side-reaction
that is not part of their catalytic cycle). This happens quite a bit in the mitochondria. The
chemical equation is H2O2 + Catalase --> H2O + O2 Hypothesis: The hotter the temperature is,
the faster the enzyme activity and it will have more bubbles. The colder the temperature is the
slower the enzyme activity and it will have fewer bubbles. If the temperature is in room
temperature, the enzyme activity will be in a normal rate because it is the controlled variable.
Material:
1) Liver
2) 1% Hydrogen Peroxide
3) Petri Disk
4) Liver Puree
5) 25ml graduated cylinder
6) Five 50ml beakers
7) Filter-paper disks
8) Forceps
9) Glass making pencil
10) Ice Bath
11) Thermometer
12) Warm water bath
13) Clock or watch with second hand
14) Paper towel

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Procedure:
1) Put your safety gear on (gloves, goggles, lab coat)
2) Select the proper equipment and technology to measure activity-either a filter-paper disk
or and oxygen probe. We are going to use the filter disk in this experiment.
3) Measure the activity of catalase by using a graduated cylinder to place 25ml of hydrogen
peroxide solution in a 50ml beaker.
4) Use forceps to dip a filter-paper disk in the liver puree. Place the filter-paper disk on a
paper towel for 4 seconds to remove any excess liquid.
5) Use the forceps to place the filter-paper disk at the bottom of the beaker of hydrogen
peroxide solution. Observe the filter-disk paper disk and record the number of seconds it
takes to float to the top of the liquid.
Data and Result:
Trial
RT 1
RT 2
RT 3
Average

Temp
(oC)

Time
(sec)
26
26
26
26

9
8
5
7.33

Cold 1
Cold 2
Cold 3
Average

9
10
10
9.67

5
12
8
8.33

Hot 1
Hot 2
Hot 3
Average

30
30
30
30

5
7
6
6.00

RESULT (AVERAGE):
Trial
Cold (Ave)
RT (Ave)
Hot (Ave)

Temp (oC)
9.67
26
30

Time (sec)
8.33
7.33
6.00

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Effect of Temperature On Enzyme Activity

9.00
8.00
7.00

Time (sec)

6.00
5.00
4.00

Series1

3.00
2.00
1.00
0.00
0.00

5.00

10.00

15.00

20.00

Temperature

25.00

30.00

35.00

(oC)

Observations: As the hotter the temperature is, the faster the enzyme activity and it will have
more bubbles. The colder the temperature is the slower the enzyme activity and it will have
fewer bubbles. If the temperature is in room temperature, the enzyme activity will be in a normal
rate because it is the controlled variable.
Discussion:
The trend of the graph shows the relationship between how the temperature affect the enzyme
and the time it took to affect the enzyme, like when the temperature is at 10(oC), the time seem to
be slower. When the temperature is 30(oC), the time it took to make the filter-disk float up is
much faster. While in the room temperature, we have an average time (not too fast or not too
slow). We would need to graph the data to understand more about the relationship between the
time it took for the enzyme to react and the temperature that makes the enzyme react. Without
the graph, it is much harder to differentiate between which temperature affect the enzyme faster
and which one is slower. The trend and the graph would also help us observe and understand

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better about the data because it is much clearer, with just the data (no graph), we would need a
longer time to describe the trend. When we look at the graph, we can see clearly that the trend
drop when the temperature is higher, without the graph, we would have to look at the
temperature, then the time to figure out which temperature the enzyme reacts faster (which
would, indeed, take a longer time to describe). The graph did support my hypothesis because
when the temperature is at 10(oC), the time it took for the enzyme to float up is about 8.5
seconds, while if the temperature is 30(oC), the time it took for the enzyme to float up was about
6 seconds which is faster than the cold temperature one. Some of the experimental errors that can
affect my result is that a few times we push the filter-disk in a place where it was locked in the
beaker and sometimes we forget to let go of the forceps, thus the filter-disk could stay in the
beaker longer than we anticipated. The result for the timing could be different every time we try,
thus we have to do it three times so we need to repeat the experiment three times to find out the
average and to be more precise. We need the average because the three trials have different
results and we needed the average of these three outcomes so our results could be more precise.
Research Questions:
Quoting Live Science, In the lock-and-key model, the active site of an enzyme is precisely
shaped to hold specific substrates. In the induced-fit model, the active site and substrate don't fit
perfectly together; instead, they both alter their shape to connect. Whatever the case, the
reactions that occur accelerate greatly over a million fold once the substrates bind to the
active site of the enzyme. The chemical reactions result in a new product or molecule that then
separates from the enzyme, which goes on to catalyze other reactions. Quoting Brooklyn,
Changes in pH may not only affect the shape of an enzyme but it may also change the shape or

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charge properties of the substrate so that either the substrate cannot bind to the active site or it
cannot undergo catalysis.
Reflection:
After I did the lab, I learned that you should be careful with thermometers because they contain
mercury which would kill you on contact. I also learn that liver puree stinks. One more thing I
learn is you have to be precise on every data or your result will be wrong. I like about this
experiment is that it is much quicker than most labs, and we can see the thermometer rise or drop
as we put the thermometer in the liver puree. The part I dislike about this lab is that it requires
the scientist or the one doing the lab to be incredibly precise, also I did not like the smell of the
liver puree. The challenging part in this lab is being precise on how high or low is the
temperature of the liver puree and how long the filter-disk float up. To me, I think my group
work collaboratively because I was the person who did the timing, Noah and Titan took the
samples, while Harry write the data in the notebook. We could improve our team work if me and
Harry also took the samples rather sitting on our chairs doing nothing, just waiting for Noah and
Titan to take the samples back.

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Works Cited
Cooper, Geoffrey M. "The Central Role of Enzymes as Biological Catalysts - The Cell - NCBI
Bookshelf." National Center for Biotechnology Information. U.S. National Library of Medicine,
01 Jan. 1970. Web. 28 Oct. 2016. https://www.ncbi.nlm.nih.gov/books/NBK9921/

"What Digestive Enzyme Is Produced by the Liver?" LIVESTRONG.COM.

LIVESTRONG.COM, 16 Aug. 2013. Web. 28 Oct. 2016.
http://www.livestrong.com/article/17785-digestive-enzyme-produced-liver/

"Re: Where Does Hydrogen Peroxide Come from in the Body?" Re: Where Does Hydrogen
Peroxide Come from in the Body? N.p., n.d. Web. 28 Oct. 2016.
http://www.madsci.org/posts/archives/1999-09/938519528.Bc.r.html

"How Do Enzymes Work?" LiveScience. TechMedia Network, n.d. Web. 28 Oct. 2016.
http://www.livescience.com/45145-how-do-enzymes-work.html

"PH and Enzymes." PH and Enzymes. N.p., n.d. Web. 28 Oct.

2016.http://academic.brooklyn.cuny.edu/biology/bio4fv/page/ph_and_.htm