Lab Enzyme (1) (1).docx

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 COMMITME NT MAX. 2

RESEARC H METHOD MAX.6

ANALYS IS

EVALUATI ON

COMMUNICATI ON

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MAX 6.

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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. 7477) 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 coolseason 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:

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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 samples

Original potato

Room temperature potato (piece #1)

Cold temperature potato (piece #2)

Hot temperature potato (piece #3)

High Width Length Weight

3,1 cm 1,3 cm 4 cm 80,9 g

2 cm 1 cm 1,3cm 26,7 g

2,5 cm 1,1cm 1,5 cm 27,4 g

1,7cm 0,8 cm 1,2 cm 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 sample

100°c Hot sample

-18° c Cold 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/wpcontent/uploads/2012/12/CHEM203_Wikipedia_Catalase_12.20.12.pdf

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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