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THE EFFECT OF TEMPERATURE

TO THE RESPIRATION RATE OF

BEAN SPROUT

NUR RIZQI AKHFIANI (093204028)

INTERNATIONAL CLASS OF BIOLOGY EDUCATION

2009


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CHAPTER 1

PREFACE

1.1BackgroundAlthough we often think of respiration in terms of humans and breathing, it is

important to note that it is one of the 7 characteristic processes shared by all living

organisms, so it must have a universal definition: "processes leading to, and

including the chemical breakdown (oxidation) of food materials to provide energy

for life" (Beth, Asher.2010).

It is important to realise that green plants respire at all times, like all living

organisms. However, they are unique in that they can also make their own food by

photosynthesis . As a by-product of this process, they produce a waste substance,

oxygen, which they release from the cells of their leaves into the air. Of course,

they may re-use some of this oxygen in aerobic respiration, so some people

wrongly assume that plants do not need oxygen, or do not respire in the same way

as other living organisms. What happens in these plants is merely a question of

balance between the processes of photosynthesis and respiration.

Seeds are dormant stages of living organisms, and contain embryo plants,

ready to grow when conditions are right. Most of the time, they appear to be doing

nothing much, but they are respiring only slowly because they do not need much

energy, and need to conserve the food reserves they contain.

However, before they start to grow into plants (and continue the life cycle by

flowering and producing seeds again), seeds must germinate. In order to do this,

seeds absorb water which they need in order to mobilise their food reserves using

enzymes (basically the same process as digestion in animals), then they speed up

their rate of respiration quite dramatically. They then use the energy released in

order to sprout roots to absorb more water and minerals, and grow a shoot which

takes the leaves above ground, so as to make food by photosynthesis. Of course

this usually happens under the ground, but it is not necessarily so.


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The respiration rate of plant is affected by some environmental factors; such

as: temperature, humidity, pH, and so on. For this activity, we will observe about

the effect of temperature to the repiration rate of bean sprout.

1.2ProblemBased on the background above, the problem which we got is How does the

effect of temperature to the respiration rate of bean sprout?

1.3PurposeBased on the problem above, the problem which we got is observing the effect of

temperature to the respiration rate of bean sprout.


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CHAPTER 2

THEORY

Definition and The Process ofPlants Respiration

Respiration is a catabolic reaction that breaks down glucose and other

nutrients. Germinating seeds take in oxygen from the surrounding air. The oxygen is

used to convert nutrients stored in the seeds endosperm into energy that the seed uses

to sprout. Enzymes activated when the seed imbibes (takes in water) make the energy

conversion possible. Carbon dioxide is given off as a waste product during

respiration. Plant respiration is the mechanism by which energy stored in the form of

glucose, a product of photosynthesis, is released for use in plant metabolism.

Photosynthesis uses light energy from the sun along with water and carbon dioxide to

produce glucose, or sugar, and releases oxygen into the atmosphere. Respiration

releases carbon dioxide (Asher,Beth.2010).

Respiration begins when a seed absorbs water. The enzymes inside seeds have

to be in suspension to function. Activated enzymes start respiration and use organic

matter stored in the seed during respiration to make ATP molecules needed for

growth. Respiration continues in the seed after the sprout has emerged until the seeds

endospermic material is used up and the cotyledons have produced the sprout

(Schlesinger, 1997).

Soaking seeds before planting has been a common practice and seeds need

water uptake to begin respiring. Dry seeds do respire, but at an extremely low, nearly

undetectable rate. Stored gases in seeds are released when water is first taken up. This

is not true respiration. True respiration starts when the seeds testa (seed coating) is

softened by water and enough water passes the softened coating to activate seed

enzymes. The seed swells and begins to consume oxygen (Gifford, 1994; Amthor,

1995).

Respiration goes in spurts, like seedling growth. A root and a shoot develop as

the result of the biochemical reactions in the seed. Seed respiration levels off after this

point. When the root and shoot begin the next growth spurt leading up to true leaves,

the respiration rate increases again to fuel the growth (Amthor, 1991).


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Respiration process begins with the arrest of O2 from the environment. Gas

transport processes in plants as a whole takes place in diffusion. Oxygen used in

respiration entry into any plant cell by diffusion through intercellular spaces, cell wall,

cytoplasm and cell membrane. Similarly, the CO2 produced by respiration will

diffuse out of the cell and into the space between cells. This is because the plasma

membrane and the protoplasm of plant cells are highly permeable to both gas. After

taking O2 from the air, O2 and then used in the respiration process with several

stages, among which glycolysis, oxidative decarboxylation, Krebs cycle and electron

transport. The first stage is glycolysis, which stages the conversion of glucose into

two molecules of pyruvic acid (atom C3), this event took place in the cytosol.

Pyruvate acid produced will then be processed in the stage of oxidative

decarboxylation. Besides glycolysis also produces 2 molecules of ATP as energy, and

2 NADH molecules that will be used in electron transport. In anaerobic conditions,

glycolysis results Pyruvate acid is converted into carbon dioxide and ethyl alcohol.

This conversion process is catalyzed by enzymes in the cytoplasm. In anaerobic

respiration the amount of ATP produced only two molecules for every one molecule

of glucose, these results differ greatly with ATP generated from the overall results of

aerobic respiration is 36 ATP. The second stage of respiration is oxidative

decarboxylation, namely the conversion of pyruvic acid (C3 atom) into acetyl CoA

(atom C2) with the release of CO2, this event took place in the cytosol. Acetyl CoA is

produced will be processed in the Krebs cycle. Other results of NADH that will be

used in electron transport. The next stage is the citric acid cycle (Krebs cycle) that

occurs in the matrix and inner mitochondrial membrane, namely the processing stages

of acetyl CoA by the citric acid compound as the compound which was first formed.

Some of the compounds generated in this phase, including the one molecule of ATP

as energy, one molecule of FADH and three molecules of NADH to be used in

electron transfer, as well as two molecules of CO2. Last stage is the transfer of

electrons, ie a series of reactions involving the career system of electrons (electron

carriers). This process occurs in the inner mitochondrial membrane. In this reaction

electrons are transferred in a series of redox reactions and assisted by the enzyme

cytochrome, quinone, pyridoxine, and flavoprotein. This electron transfer reaction

will produce H2O (I Komang Jaya Santika Yasa, 2009).

The process of respiration can be described as follows:


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1.) glycolysis:

Glucose 2 pyruvic acid + 2 NADH + 2 ATP

2. Krebs Cycle:

2 acetyl pyruvate acetyl CoA 2 + 2 CO2 + 2 NADH + 2 ATP

2 acetyl CoA4 CO2 + 6 NADH + 2 FADH2

3. Electron transport chain:

10 NADH + 5O2 10 10 NAD + + H2O + 30 ATP

2 FADH2 + O22 FAD + 2 H2O + 4 ATP

Thus, the total energy generated from the process of respiration is 38 ATP.

(Danang, 2008).

Function of Plants Respiration (Davey et al., 2004).

During respiration, glucose is broken down to become products that will fuel

other processes that the plant will use. Respiration is a multi-stage process that can

take different pathways depending on the presence or absence of oxygen. Glycolysis

breaks glucose into two molecules initially, in the cytoplasm, or the main body of the

cell enclosed within the plant cell wall.

The presence or absence of oxygen determines how the process will progress.

If oxygen is present, products from glycolysis will be used within the mitochondria,

or energy centers, of the cell, to continue respiration. If oxygen is not present,

fermentation, a less efficient use of energy, occurs.

Effects of Plants Respiration (Davey et al., 2004).

During photosynthesis, plants store energy captured during the day, the light

reaction, to fuel the process of creating sugar chiefly at night, during the dark reaction

of photosynthesis. As the name implies, the dark reaction is independent of the

presence of light. Glucose is produced. Glucose will in turn fuel respiration.


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Factors Affecting Respiration Rate

1. Availability of substrate

Respiration dependent on substrate availability. Starch content of plants,

fruktan, or low sugar, do respiration at a low rate. Scarcity of food plants carry on

respiration of sugar often faster when sugar is provided. Even the leaf respiration rate

is often much faster soon after sunset, when the high sugar content as compared to

when the sun rises, while a lower sugar content. In addition, the shaded leaf or leaves

the bottom usually slower respirated than the upper leaves are exposed to more light.

If this does not happen, then the lower leaves will die sooner. Differences due to the

sugar content may not equal of rate of photosynthesis which causes a lower

respiration rate in the shaded leaves. (Salisbury & Ross, 1995)

2. Availability of oxygen

The availability of oxygen will affect the rate of respiration, but the magnitude

of these effects are different for each different species and even between the same

organs in plants. Normal fluctuations in the oxygen content of air is not much affect

the rate of respiration, because the amount of oxygen required for berespirasi plants is

much lower than the available oxygen in the air. (I Komang Jaya Santika Yasa, 2009)

3. Temperature

Effect of temperature factors for respiration rate of plants is strongly

associated with factors Q10, which is generally the reaction rate of respiration will

increase for every increase in temperature of 10 C, but this depends on each species.

For most parts of the plant and plant species, Q10 respiration is usually 2.0 to 2.5 at

temperatures between 5 and 25 C. When the temperature is increased further to 30

or 35 C, respiration rate remained elevated, but more slowly, so Q10 begin to

decline. An explanation of the decline in Q10 at high temperatures is that the rate of

penetration of O2 into the cell through the cuticle or periderma start inhibit respiration

during chemical reactions take place quickly. Diffusion of O2 and CO2 are also

accelerated with increasing temperature, but the Q10 for the physics of this process is

only 1.1, so the temperature does not significantly accelerate the diffusion of solution

through the water. Increasing the temperature to 40 C or more, in fact decreasedrespiration rate, especially when plants are in this state in a long time. It seems that


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the enzymes necessary to quickly start having denaturation at elevated temperatures,

preventing an increase in metabolic proper place. In pea sprouts, increase in

temperature from 25 to 45 C initially rapidly increasing respiration, but after two

hours of its speed began to decrease. Possible explanation is that a period of two hours

is long enough to damage some respiratory enzymes. (Salisbury & Ross, 1995)

4. Type and Age of Plant

Each plant species has metabolsme differences, thus the need for berespirasi

plants will be different in each species. Young plants showed a higher respiration rate

than older plants. Similarly the plant organ that is in its infancy. (I Komang Jaya

Santika Yasa, 2009).


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CHAPTER 3

RESEARCH METHOD

3.1 Kind of Research

The kind of research in this laboratory activity is experimental, because there

is some treatment to solve the problem formula and there are variables used, such as

manipulation variable, control variable, and response variable.

3.2 Variables

Variables which is used in this laboratory activity are:

3.2.1 Manipulation variables : The temperature (incubation temperature: 37C; room

temperature: 32C).

3.2.1 Control variables : Weigh of bean sprout (5gr), Volume of NaOH 0.5M

(30mL), Kind of bean sprout (green bean), Amount

droping of PP solution (two drops), Age of bean sprout

(2 days), Volume of BaCl2 (2.5mL),distance between

hanging bean sprout with NaOH solution in the

Erlenmeyer tube (half of tube), time of saving

(24hours).

3.2.3 Response Variables : The respiration rate of bean sprout (ml/hour)

3.3 Tools and Materials

3.3.1 Tools

Erlenmeyer tube 250mL 6 pieces Kasa silk As enough Rope As enough Pipettes As enough Transparent plastic 6 pieces Rubber 6 pieces Balance 1 piece Burret 1 piece


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Stative and clem 1 piece Cut 1 piece

3.3.2 Materials

Bean sprout age 2 days NaOH solution 0.5M HCl solution 0.5N BaCl2 solution 0.5N Phenolftalin solution

3.4 Procedures

1. Prepare 6 pieces of Erlenmeyer tube, and fill it with 30mL of NaOH 0.5M.2. Measure 5gr of bean sprout for each Erlenmeyer tube. Need 4 erlenmeyer tube

that is contained of bean sprout.

3. Covering the each 5 gr of bean sprout with kasa silk, and tie it using rope.4. Entering the covered bean sprout into Erlenmeyer.5. Hanging the covered bean sprout using rope as half of height of tube. The

Erlenmeyer which is filled by bean sprout is 4 tubes. Two tubes for placed in

room temperature, two others placed in incubator temperature. Covering the

Erlenmeyer tubes hole with transparent plastic and rubber.

6. The last two of Erlenmeyer tube is not filled by bean sprout, only filled byNaOH 30mL as the control.

7. Put one of control tube, and two of tube filled bean sprout into incubator (37C). wait until 24 hours.

8. Put one of control tube, and two of tube filled bean sprout into usual room (32C). Wait until 24 hours.

9. After 24 hours. Doing titration. Firstly, fill the burret with HCl solution untilthe scale of burret shown 0 number

10.Take 5mL of NaOH solution in Erlenmeyer. Then, added by 2.5 mL of BaCl211.Drop it with 2 drops of PP till the solution is young red.12.Titration the red solution with HCl in the burret. Titrate it drop by drop of HCl

until the color of solution is changing to be clear (the red color is disappear).

13.Doing the titration until 6 tube of erlenmyer is titrated. Note, the volume ofHCl that used for changing the color of solution.


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3.5 Design Experiment

1. Prepare 6 pieces of Erlenmeyer tube, and fill it with 30mLof NaOH 0.5M.

2. Measure 5gr of bean sprout for each Erlenmeyertube. Need 4 erlenmeyer tube that is contained ofbean sprout.

3. Covering the each 5 gr of bean sprout with kasa silk, and tieit using rope.


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4. Entering the covered bean sprout into Erlenmeyer.

5. Hanging the covered bean sprout using rope as half of heightof tube. The Erlenmeyer which is filled by bean sprout is 4

tubes. Two tubes for placed in room temperature, two others

placed in incubator temperature. Covering the Erlenmeyer

tubes hole with transparent plastic and rubber.

6. The last two of Erlenmeyer tube is not filled by bean sprout,only filled by NaOH 30mL as the control.


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7.

Put one of control tube, and two of tube filled bean sprout intoincubator (37 C). wait until 24 hours.

8. Put one of control tube, and two of tube filled bean sprout intousual room (32 C). Wait until 24 hours.

9. After 24 hours. Doing titration. Firstly, fill the burret with HClsolution until the scale of burret shown 0 number.


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10.Take 5mL of NaOH solution in Erlenmeyer. Then, added by 2.5

mL of BaCl2

11.Drop it with 2 drops of PP till the solution is young red.

13.Titration the red solution with HCl in the burret. Titrate it dropby drop of HCl until the color of solution is changing to be clear

(the red color is disappear).

12.Doing the titration until 6 tube of erlenmyer is titrated. Note, thevolume of HCl that used for changing the color of solution.


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CHAPTER 4

DATA AND ANALYSIS

4.1 Data

After did the experiment, we got the problem as follow:

Table of Data

Table 1.The Effect of Temperature to The Respiration Rate of Bean Sprout

No.

Measurement

Result from

Room Temperature 32C Incubator Temperature 37C

Control 1st bean

sprout

2n

bean

sprout

Control 1st bean

sprout

2n

bean

sprout

1.Volume HCl

(mL)1.5 0.8 1.2 1.8 0.8 0.9

2.

Volume of

NaOH which

isnt binded

by CO2 (mL)

9.0 4.8 7.2 10.8 4.8 5.4

3.

Volume of

NaOH which

is binded by

CO2 (mL)

21 25.2 22.8 19.2 25.2 24.6

4.

Volume of

CO2 that is

respiration

product (mL)

3 5.7

5.

Respiration

Rate

(mL/hour)

0.125 0.2375


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Graphic1.The Relationship of Temperature to The Respiration Rate of

Bean Sprout

4.2 Analysis of Data

Based on the data above, we can see the relationship of temperature in room to the

respiration rate of bean sprout. In incubation room with temperature 37 C has

respiration rate 0.2375 mL/hour. In room temperature with 32 C has respiration rate

0.125 mL/hour. So, the higher of temperature, the faster of respiration rate but just

until the optimum temperature to do respiration.

In usual room temperature 32 C, the volume of HCl used for titration of the 5mL

of NaOH (until the pink color is being colorless/white) is 1.5mL in control

Erlenmeyer, 0.8mL in 1st Erlenmeyer of bean sprout, and 1.2 mL in 2nd Erlenmeyer of

bean sprout. The volume of NaOH (from 30mL of NaOH volume) which isnt binded

by CO2 is 9.0mL in control Erlenmeyer, 4.8mL in 1st Erlenmeyer of bean sprout, and

7.2 mL in 2nd

Erlenmeyer of bean sprout.The volume of NaOH (from 30mL of NaOHvolume) which is binded by CO2 is 21mL in control Erlenmeyer, 25.2mL in 1

st


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Erlenmeyer of bean sprout, and 22.8 mL in 2nd

Erlenmeyer of bean sprout. The

volume of CO2 that as respiration product is 3mL. And the respiration rate is the

comparison between volume CO2 as the respiration product with the time of saving

(24 hours), as much 0.125mL/hour.

In incubation temperature 37 C, the volume of HCl used for titration of the 5mL

of NaOH (until the pink color is being colorless/white) is 1.8mL in control

Erlenmeyer, 0.8mL in 1st Erlenmeyer of bean sprout, and 0.9 mL in 2nd Erlenmeyer of

bean sprout. The volume of NaOH (from 30mL of NaOH volume) which isnt binded

by CO2 is 10.8mL in control Erlenmeyer, 4.8mL in 1st Erlenmeyer of bean sprout, and

5.4 mL in 2nd Erlenmeyer of bean sprout. The volume of NaOH (from 30mL of NaOH

volume) which is binded by CO2 is 19.2mL in control Erlenmeyer, 25.2mL in 1st

Erlenmeyer of bean sprout, and 24.6 mL in 2nd Erlenmeyer of bean sprout. The

volume of CO2 that as respiration product is 5.7 mL. And the respiration rate is the

comparison between volume CO2 as the respiration product with the time of saving

(24 hours), as much 0.2375mL/hour

4.3 Explanation

Based on the data analysis above we can see that, the higher of temperature, the

faster of respiration rate but just until the optimum temperature to do reaction rate. If

the optimum temperature has reached, so the reaction rate will be decreased. It

because respiration is the enzymatic reaction which is involving a lot of enzymes

work inside. So, if the temperature is so high and in long time in that condition, the

enzyme which is needed for doing respiration rate is denaturated.

In this experiment we used bean sprout of green bean with age 2 days,

because the bean sprout is very active doing metabolism for growth, and there is still

no photosynthesize process in these bean sprout because they have not chloroplast

organ yet in their body, so the energy from metabolism is concerning for growing

activity of these bean. In other hand, the bean sprout in age 2 days starts to have

cotiledone as the storage tissue (food storage) which contains of a lot of carbohydrate.

The carbohydrate is the substrate in repiration process to get energy by carbohydrate

combine with oxygen. So, the carbohydrate contain is decreasing for bean sprout

growth. The reaction of respiration in bean sprout, as follow:

C6H12O6 (carbohydrate)+ 6 O2 6 CO2 + 6H2O + Energi


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Bean sprouts do breathing to get the energy carried by involving gaseous oxygen

(O2) as the material is absorbed / required and produce the gas carbon dioxide (CO2),

water (H2O) and the amount of energy. Basically, the process of respiration has

functions to get the energy used in metabolism and growth and developmental

processes to become a mature plant. The larger a plant, then the greater needed for

energy, so in respiration process is requiring a lot of oxygen too.

The tested bean sprout us placed in the Erlenmeyer tube which is filled by NaOH

solution. These NaOH has function to catch the carbon dioxide which is resulted by

these bean sprout respiration. The respiration process that we tested to sprout is 24

hours, then the NaOH will be reacted with BaCl2 solution, and droped by PP solution

as indicator. After it, these NaOH solution will be titrated using HCl to produce salt.

The whole reaction of our practice activity to know the respiration rate, as follow:

- When NaOH catch the carbon dioxide as the result of respiration process:NaOH + CO2 NaHCO3

- When 5mL NaOH is reacted with 2.5 mL of BaCl2 :2NaOH + BaCl2 2NaCl+ Ba(OH)2 Base properties because when it

reacted again with indicator PP, the color is being redish purple.

- When doing titration with HCl:2Ba(OH)2 + 2HCl 2BaCl + H2O

Base + Acid Salt + water

The amount of HCl needed to change the color of Ba(OH)2 reddish purple to

the recent disappear (become white or transparent) is the amount of NaOH which is

not binding the CO2 , or can be written like this: HCl needed = NaOH which is

not binding CO2. So, from the volume of HCl needed, we can know how many NaOH

which isnt binding the carbon dioxide.

NaOH has ability to bind the Carbon dioxide as the product of respiration, but

not all of CO2 can be binded by NaOH. So, the NaOH which is not binded the carbon

dioxide is not all of them can be reacted with BaCl2 and produce Ba(OH)2 that has

white color. Ba(OH)2 is tested with indicator PP, the color is changed to be redish

purple that indicates the properties of that solution is base. When we titrates the 5 mL

of Ba(OH)2 with HCl, it results the salt of BaCl2. We can know the solution has

changed to be salt BaCl2 from the changing of color, if the redish purple color is


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changed to be disappear recently of color being transparent, so the solution has been

the salt.

In the temperature range of 0C to 45C, a temperature increase will be

followed by an increase in respiration rate. In incubation room with temperature 37 C

has respiration rate 0.2375 mL/hour. In room temperature with 32 C has respiration

rate 0.125 mL/hour. It means that in incubator temperature, the respiration rate is

faster than in room temperature. Its because in incubator, the temperature is stable or

constant. The enzyme work is more optimal if it is placed in stable temperature

without the denaturation of enzyme. As we know, all the respiration reaction is

involving the enzymes. If the enzyme is not denaturate, so the work of enzyme will be

optimum in respiration. It means that the enzyme will make faster the glucosechanging to be carbon dioxide, and the number of carbon dioxide produced in

respiration of bean sprout is more number. If the temperature is higher, the volume of

carbon dioxide is big binded by NaOH, so that the concentration of carbon dioxide

which is produced in respiration is bigger too.

In room temperature the volume of carbon dioxide as the product of

respiration is fewer than in incubator room. It is because if the temperature is lower,

the enzyme work is not optimal. It means that the enzyme will make slower theglucose changing to be carbon dioxide, and the number of carbon dioxide produced in

respiration of bean sprout is fewer. If the temperature is lower, the volume of carbon

dioxide is few binded by NaOH, so that the concentration of carbon dioxide which is

produced in respiration is fewer too.

In control Erlenmeyer which is not filled by bean sprout is also showing the

respiration process. Because there is another microorganism which is did respiration

except the bean sprout. We can know it because there is NaOH which bind the carbon

dioxide, it means there is carbon dioxide production in the Erlenmeyer. Maybe from

microorganism which live in the Erlenmeyer because we didnt sterilize the

Erlenmeyer firstly before doing those practice.

The respiration rate of Erlenmeyer with bean sprout is faster than without bean

sporut and the carbon dioxide product is more than in without bean sprout, it because

of the oxidizing substrate in respiratory metabolism. And generally substrates for

respiration are substances that accumulate in number and relatively many metabolic


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processes involves a series of enzymatic reactions that involve enzymes, the

respiration rate on the existing Erlenmeyer sprouts are also influenced by the enzymes

contained in the sprouts and enzymes will increase when the temperature also high

when the temperature is too high but will also damage the enzyme. While the

erlenmeyer containing only NaOH, the process of respiration is slower and less CO2

emitted. This is because it is not influenced by enzyme


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CHAPTER 5

CONCLUSION

From the experiment we did and the explanation above, we conclude that

temperature is affecting the respiration rate of bean sprout. The higher of temperature,

the faster of respiration rate happen, but just until the optimum temperature to do

reaction rate. In the temperature range of 0C to 45C, a temperature increase will be

followed by an increase in respiration rate. More of CO2 is produced by the bean

sprout, the faster of respiration rate.


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Asimov, Isaac (1968). Respiration. New York, London: Basic Books, Inc.

Campbell, Neil A., Lawrence G. Mitchell, Jane B. Reece. 1999. Biology, 5th Ed.

Benjamin/Cummings Publ. Co., Inc. Menlo Park, CA. (plus earlier editions)

Affourtit C, Krab K, Moore AL. 2001. Controlof plant mitochondrial respiration.

Biochimica Biophysica ActaBioenergetics 1504: 5869

Amthor JS. 1991. Respiration in a future, higher CO2 world. Plant, Cell and

Environment 14: 1320.

H,Lambers.2008.Plants respiration. Online at:

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