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COLEGIO INTERNACIONAL DE BOGOTÁ

SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

SUBJECT: Biology TEACHER: Ingrid Rocio Marín


DATE: April 22/17 NAME: Carolina Valencia
TITLE: Temperature as a cause of denaturation
GRADE: 11°

PERSONAL RESEARC ANALYS EVALUATI COMMUNICATI TOTA


COMMITME H IS ON ON L
NT METHOD
MAX. 2 MAX.6 MAX 6. MAX. 6 MAX. 4 MAX.
24

INTRODUCTION:

What happens when a potato is combined with hydrogen peroxide?

Enzymes are organic molecules which act as catalysts, more specifically enzymes are
proteins. Thus, enzymes are long chains of amino acids that have a three-dimensional shape.
There is an area in the enzyme that is designed to match a specific molecule known as the
enzymes substrate. This area of the enzyme is called the active site. The active of an enzyme
matches the substrate in a similar way to the way a glove fits over a hand. (Damon et al. 74-
77)

Enzyme catalysis involves molecular motion and the collision of substrates with the active
site (Bioninja, 2.5 Enzymes/ Enzyme catalysis. s.f)
Catalase, is a common enzyme found in nearly all living organisms exposed to oxygen. It
catalyzes the decomposition of hydrogen peroxide to water and oxygen
(Chelikani P, Fita I, Loewen PC (January 2004). "Diversity of structures and properties
among catalases".)
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

Catalase is a tetramer of four polypeptide chains, each over 500 amino acids long. It contains
four porphyrin heme (iron) groups that allow the enzyme to react with the hydrogen peroxide.
Catalase can also catalyze the oxidation, by hydrogen peroxide, of various metabolites and
toxins, including formaldehyde, formic acid, phenols, acetaldehyde and alcohols. It does so
according to the following reaction:
H2 O2 + H2 R → 2H2 O + R
(Boon EM, Downs A, Marcey D. "Catalase: H2 O2 : H2 O2 Oxidoreductase")

This laboratory looks for the acquisition of knowledge of enzymes, how it can be affected
and changed by factors such as the temperature, pH and substrate concentration works and
its importance. The purpose of this laboratory will be to evidence the decomposition of the
hydrogen peroxide by the catalase enzyme, founded in potato at different temperatures.

RESEARCH QUESTION:
¿ How temperature in each sample affects the decomposition of hydrogen peroxide and
why?

HYPOTHESIS: As catalase acts as the catalyzing enzyme in the decomposition of hydrogen


peroxide and temperature is one of the factors that affect the rate of activity of enzymes, it
would be a sure decomposition and a breakdown of molecules, ending into water and oxygen.
But if the sample is at a higher temperature it could denature the protein (the enzyme).
OBJECTIVES:
General aims:
a. Evaluate the mashed potatoes behavior through the decomposition of hydrogen
peroxide.

Specific aims:
a. Describe the changes the samples will have and describe the decomposition process
in each one, describe how temperature affect them.
b. Compare each other and the results they get.
c. Analyze how temperature in each sample could get oxygen and water at the end.

THEORETICAL FRAMEWORK:
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

First let’s start with the enzyme, in this case catalase that is a common enzyme found in
nearly all living organisms exposed to oxygen. It catalyzes the decomposition of hydrogen
peroxide to water and oxygen.
(Chelikani P, Fita I, Loewen PC (January 2004). "Diversity of structures and properties
among catalases".)
In this case the organism that contain catalase is the potato (Solanum tuberosum) is a cool-
season plant originally from the Andes Mountains of South America. The tubers are
underground stems (also known as stolons), not roots. Kaiser, Cheryl, and Matt Ernst
Ernst. Potatoes. 1st ed. KENTUCKY: UNIVERSITY OF KENTUCKY COLLEGE OF
AGRICULTURE, FOOD AND ENVIRONMENT, 2017. Web. 22 Apr. 2017.
Of the 64.46 grams of carbohydrates in a potato, 56.97 grams exist in the form of starch.
Ulmer, Graham, and Graham Ulmer. "Starch And Glucose In
Potatoes". LIVESTRONG.COM. N.p., 2017. Web. 15 May 2011.

“An enzyme is a biological catalyst. Catalysts speed up biochemical reactions, such as


digestion and respiration, but they remain unchanged at the end of the process. If the
three-dimensional shape of an enzyme is destroyed or damaged, it can no longer carry
out its job and is said to be denatured. Extremes of temperature, heavy metals and, in
some cases, pH can cause permanent changes in an enzyme”. (Walpole, Brenda et
al. BIOLOGY for the IB Diploma. 2nd ed. The Edinburgh Building, Cambridge CB2 8RU,
UK: Cambridge University Press, 2011. Fri. 21. Apr. 2017.)

The active site is the region on the surface of the enzyme which binds to the substrate
molecule. The active site and the substrate complement each other in terms of both shape
and chemical properties. (Bioninja, 2.5 Enzymes/ Enzymes & substrate. s.f)
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

(Bioninja, 2.5 Enzymes/ Enzymes & substrate. s.f)

 An enzyme’s performance depends on how rapidly it can process its substrate. The
rate of an enzyme reaction (V) increases as the substrate concentration increases until
a maximum value (Vmax) is reached.

Stable temperature is important to living things because the range of temperatures in which
biological reactions can occur is quite narrow.
(Alberts, Bruce et al. ESSENTIAL CELL BIOLOGY. 3rd ed. NY: Published by Garland
Science, Taylor & Francis Group, 2017. Fri. 21 Apr. 2017.)
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

“At very low temperatures, enzymes hardly work at all and the rate of reaction is very
low. As the temperature rises, molecular collisions are more frequent and energetic,
and therefore the rate of the enzyme controlled reaction increases.
As the temperature rises above the optimum, the enzyme and substrate molecules move
faster – but atoms within the enzyme molecule itself also move faster, straining the
bonds holding it together. Eventually, these bonds may be stressed or broken to such
an extent that the enzyme loses its three-dimensional shape and the active site can no
longer receive substrate molecules. At these high temperatures, the structure is
permanently destroyed and the enzyme is denatured and can no longer catalyze the
reaction.” (Walpole, Brenda et al. BIOLOGY for the IB Diploma. 2nd ed. The Edinburgh
Building, Cambridge CB2 8RU, UK: Cambridge University Press, 2011. Web. 10 Apr.
2017.)
If there is a set concentration of enzyme present in a reaction mixture, and the concentration
of substrate increases, the rate of production of the products will increase because of the
greater chance of collisions between substrate and enzyme molecules.
(Alberts, Bruce et al. ESSENTIAL CELL BIOLOGY. 3rd ed. NY: Published by Garland
Science, Taylor & Francis Group, 2017. Fri. 21 Apr. 2017.)

METHODOLOGY
Independent variable: Time, type of exercise

Dependent variable: Heart rate/ Target heart-rate zone

Control: Rate of index finger lifts at 20°C, or room temperature.

VARIABLE METHOD OF

CONTROLL

MEASUREMENT
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

TEMPERATURE The environmental

temperature of Bogota city.

16° The environmental

Bogotá temperature

TIME GIVEN 15 minutes per exercise From the chronometer

HEART RATE For 10 seconds then

multiply it by 6

With my hand fingers

Heart rate inside its target

range as you exercise

by subtracting your age

from the number 220


COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

Target heart-rate zone is between 60 and 85

percent of your maximum

rate

Materials:

Calculator

Two fingers

Comfortable exercise clothes (optional)

A jump rope

A bicycle

Notebook

A pen

Chronometer
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

Procedure:

COLEGIO INTERNACIONAL DE BOGOTÁ

SCIENCE DEPARTMENT

LAB REPORT

2016 - 2017

1. Start wearing comfortable exercise clothes.

2. Choose which exercise you want to do first.

3. Before starting it, make sure you have been resting for a few minutes so that your

heart is at its resting heart rate.

4. Put the chronometer ready for 15 minutes.

5. Remember leaving enough time between activities so that your heart rate returns to

its normal resting level.


Materials:
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

 A microscope
 Notebook
 A pen
 A Lab coat
 Gloves
 Lab mask
 One beaker
 3 test tubes
 A marker
 A mask tape
 2 potatoes
 A knife
 Meat hammer
 A spoon
 Hydrogen peroxide (jgb)/ (10 ml)
 Chronometer
 Water
 A dropper
 A graduated pipette
 1 Weight precise balance
 Paper Towel
 A pot
 Ruler
Procedure:
1. Start by performing this procedure in a laboratory with the respective protocol, and the
materials needed there (lab coat, gloves, and lab mask).
2. Measure the weight and the height of your potato and with the knife cut three equal parts
and weight and measure them.
3. Write the data in your notebook with a pen. And take the three test tubes and put a piece
of mask tape in each one and with a marker, write the initials RT: #1Room temperature , H:
#2 Hot and C: #3 Cold.
4. Put the first piece of potato in a little kitchen plate and put into a freezer for 30 min.
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

5. Put the second piece of potato in a plate and leave it in your working place, this case the
laboratory.
6. Put the third piece of potato in the pot and start boiling it for 5 minutes.
7. When all the samples are complete, then with the knife cut into smaller pieces each one
and with a spoon and a meat hammer mash them.
8. Then each sample put it into the test tubes.
9. When all the samples are into the test tubes, with a graduated pipette take from the
hydrogen peroxide 10 ml for each sample and put into the test tubes to observe the changes.
10. Take into account the time where this changes happens, and write it in your notebook,
indispensable to take pictures.
11. Compare each sample and its changes (Bubbles or not) (Color taken) (rate of reaction).
12. After that for understanding more the decomposition of the substrate that exists between
the catalase enzyme and the substrate (hydrogen peroxide), take the other potato in a small
cup and put a drops of hydrogen peroxide and observe in a microscope.
13. Watch the changes, take pictures and take notes (a register).
14. Qualitative changes are registered.

RESULTS AND ANALYSIS.


The results that get this laboratory are going to be explained in four sections.
The first one is the moment the lab starts with all the materials prepared, including the three
test tubes marked with the initials of RT: Room temperature, H: Hot and C: Cold, the
hydrogen peroxide, the beaker, the ruler and your notebook/this is where the measures and
the weight are taken from the potato in first instance and then from the three equal parts that
were cut from the potato and putting them in three different plates.
The results they have were:
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

Potatoes Original potato Room Cold Hot


samples temperature temperature temperature
potato (piece potato (piece potato (piece
#1) #2) #3)

High 3,1 cm 2 cm 2,5 cm 1,7cm


Width 1,3 cm 1 cm 1,1cm 0,8 cm
Length 4 cm 1,3cm 1,5 cm 1,2 cm
Weight 80,9 g 26,7 g 27,4 g 26.8 g

The second part consists in putting one plate in the freezing for 30 minutes, boiling other one
in a pot for 5 minutes and letting the third one outside.
The third part is based on smashing the potato helped by a knife and a meat hammer.
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

16° c Room temperature 100°c Hot sample -18° c Cold sample


sample
 After 5 minutes the sample #1: 16°c Room
temperature sample took a reddish color. This
happens because when a potato tissues are broken,
an enzyme called polyphenol oxidase (PFO) is
released this enzyme has the ability to oxidize the
polyphenols and trigger a chain of reactions that
lead to dark compounds (melanoidines). (“¿Por
Qué Se Oscurecen Las Patatas Cuando Se Pelan O
Se Cortan? - Clickmica". Clickmica. N.p., 2017.
Web. 28 Apr. 2017.)
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

Step number 4, Getting ready with the 10 ml of hydrogen


peroxide in the beaker, the 3 smashed potatoes into the
respective test tube, and the 10 ml of hydrogen peroxide in
each one at the same time so they threw the results of
enzyme activity and the catalase. It catalyzes the
decomposition of hydrogen peroxide to water and oxygen
in three different temperatures: RT: 16°c, H: 100°c and C:
-18°c test tubes respectively.

After 4.03 seconds, the enzyme reaction took effects, the effects produced were at a very fast
rate. The presence of bubbles that means the decomposition and the breakdown of molecules,
ending into water and oxygen was evidenced in sample all the samples but mostly and with
a big difference , the presence was founded in sample #1 and # 3. RT has the fastest enzyme
activity and decomposition. C having so many bubbles in the bottom and on top has also a
fast enzyme activity and catalysis.
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

With the pass of milliseconds, the decomposition and catalysis activity raises and bubbles
increases. In the first sample the bubbles get out from the test tubes in questions of 6.02
seconds. The decomposition of the hydrogen peroxide by the catalase enzyme, founded in
each potato, show how temperature affects its reaction rate.
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

DISCUSSION.
The hypothesis that was planted was totally proved, the decomposition process, enzyme
activity and rate of reaction were evaluated in a small potato of 80,9 grams divided in three
relative equal parts and putted into totally different temperatures for a period of time, finally
being mashed with a meat hammer, (in first instance it was tried with a spoon but it was so
hard to do it so then it was with a meat hammer). The results expected were totally achieved,
at the same time each of the objectives this laboratory have, helping us to understand how
the temperature factor could affect the decomposition of hydrogen peroxide. Potatoes
behavior during the experiment was being watched all the time and give us at the same time
the proof of the effects of temperature. All the samples has a specific process, each one with
different characteristics and different times, each one with a different aspect and each one
representing a respective phenomenon. The experiment catalase activity in potatoes show us
curious chemical and physical changes in the test tubes, being in contact with the hydrogen
peroxide. The presence of bubbles was a factor that show us more proofs. This laboratory let
us know how catalase found in potatoes decompose hydrogen peroxide, and how temperature
affect it. The different effects in each one taking into account the time, the presence of
catalase enzyme in potato (Solanum tuberosum), which of the 64.46 grams of carbohydrates
it has, 56.97 grams exist in the form of starch, so the carbohydrate macro molecule presence
too, the hydrogen peroxide from jgb Colombian brand, the importance of temperature and
the proportions.
CONCLUSION
For this lab the effect of temperature in the decomposition of the hydrogen peroxide by the
catalase enzyme, founded in potato was examined and analyzed. All objectives were
achieved. Three different parts from an original potatoes were exposed to different
temperatures from each other: 16°C at the room temperature, -18°C at the freezing
temperature and 100°C at a boiling temperature (This sample was into water). This solutions
produce and got different effects in each one of the three sections of the potato , the first over
the span of 33 minutes of exposure to the room temperature(waiting for the others samples
for being experimented), the second over the span of 5 minutes into boiling water, and then
28 minutes waiting for the last sample for then being experimented and the last one that has
the span of 30 minutes of exposure in the school kitchen freezer and 3 more minutes for
being mashed and putted into the test tubes, the following reaction occurs:

H2 O2 + H2 R → 2H2 O + R
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
LAB REPORT
2016 - 2017

“Enzyme catalysis involves molecular motion and the collision of substrates” in this lab the
substrate were 10 ml of Hydrogen peroxide from jgb in each test tube matching and
interacting with mashed potatoes that act at the same time as the active site of the catalase
enzyme, “Catalase is a common enzyme found in nearly all living organisms exposed to
oxygen”. It catalyzes the decomposition of hydrogen peroxide to water and oxygen. The
oxygen getting out was noticed in the bubbles in sample #1 and #3, but in sample #2, the hot
one there was not an evidence of a rate of reaction. An enzyme’s performance depends on
how rapidly it can process its substrate. “The rate of an enzyme reaction (V) increases as the
substrate concentration increases until a maximum value (Vmax) is reached.”, so that means
in sample #2 the enzyme denatures, for being at the presence of higher boiling temperatures,
so it doesn’t has any process with the substrate.

Every objective planned in this lab was achieved and the results let us the evidence of a
decomposition and a separation of water and oxygen process on the three test tubes of mashed
potatoes, by the enzyme catalase and the substrate used in a product that is common in the
industry, the hydrogen peroxide. The behavior and the physical changes as the chemical ones
that the samples have through the experiment where evaluated and observed. All the changes
the potatoes represented where taken onto account for describing them and the process they
experimented with the specific substrate used, samples #1 and #3 act so fast, in questions of
milliseconds they get a rapid rate of reaction. But #1 being the fastest, during this shortly
period of 6.2 seconds, the changes were drastic and there was an evident decomposition
process, also proved by the comparison with the other samples ones and their aspect. The
results were tangent and evident, that help us a lot to analyses them.

All of this contributed to reach the greatest objective that consist in the evaluation and the
analysis of decomposition process and temperatures effects in potatoes. The experiment
could prove successfully the functioning of catalase enzyme in the decomposition of the
hydrogen peroxide.

BIBLIOGRAPHY:
 Damon, Alan et al. BIOLOGY SL. 1st ed. England: Pearson Education Inc, 2017.
Print.
 https://www.saylor.org/site/wp-
content/uploads/2012/12/CHEM203_Wikipedia_Catalase_12.20.12.pdf
COLEGIO INTERNACIONAL DE BOGOTÁ
SCIENCE DEPARTMENT
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2016 - 2017

 Chelikani P, Fita I, Loewen PC (January 2004). "Diversity of structures and


properties among catalases". Cell. Mol. Life Sci. 61 (2): 192–208.
doi:10.1007/s00018-003-3206-5. PMID 14745498.
 tfssbio.pbworks.com/w/file/fetch/54980708/Biology_for_the_IB_Diploma.pdf
 http://ftp.dsma.dp.ua/211/ENG/Other/Alberts%20-
%20Essential%20Cell%20Biology%203rd%20ed.pdf
 Boon EM, Downs A, Marcey D. "Catalase: H2 O2 : H2 O2 Oxidoreductase"
(http://biology.kenyon.edu/BMB/Chime/catalase/frames/ cattx.htm). Catalase
Structural Tutorial Text. . Retrieved 2007-02-11.

 Kaiser, Cheryl, and Matt Ernst Ernst. Potatoes. 1st ed. KENTUCKY:
UNIVERSITY OF KENTUCKY COLLEGE OF AGRICULTURE, FOOD AND
ENVIRONMENT, 2017. Web. 22 Apr. 2017.
 http://www.livestrong.com/article/440617-can-you-live-off-potatoes/
 https://clickmica.fundaciondescubre.es