Lab 6 Cellular Respiration

Introduction

Cellular respiration is the release of energy from organic compounds by metabolic chemical oxidation in the mitochondria within each cell. Enzyme mediated reactions are required. The equation for cellular respiration is:

C6H12O6 + 6 O2 6 CO2 + 6 H2O + 686 kilocalories of energy/mole of glucose oxidized

Several different measures can be taken from this equation. The consumption of oxygen, which will tell you how many moles of oxygen are consumed during cellular respiration. That is what was measured in this lab. The production of CO2 can also be measured. And of course the release of energy can be measured. Cellular respiration is a catabolic pathway and the mitochondria houses most of the metabolic equipment for cellular respiration. It will break down glucose in what we call an exergonic reaction. Like previously said, the consumption of oxygen molecules will be measured in a gas form. One must know the physical laws of gases when working with them. The laws are summarized by the following equation.

PV=Nrt

Where:

P stands for the pressure of the gas

V is the volume of the gas

n is the number of molecules of gas

R is the gas constant (fixed value)

T is the temperature of the gas ( in K )

The CO2 produced during cellular respiration will be removed by potassium hydroxide (KOH) and will form a solid potassium carbonate (K2CO3) when the following reaction occurs: CO2 + 2 KOH K2CO3+ H2O

Since the CO2 is removed, the change in the volume of gas in the respirometer will be directly related to the amount of oxygen consumed. If the water temp and volume stay constant then the water will move toward the region of lower pressure. During respiration, oxygen will be consumed and its volume will be reduced because the CO2 is being converted to a solid. The net result is a decrease in gas volume in the tube and a decrease in pressure of the tube. The vial with beads will detect any atmospheric changes.

Hypothesis

Several different things will affect the rate of O2 consumption. The non germinating peas will have a lower rate than the germinating peas and the coldness of the water will slow the rates.

Materials

The materials used for this lab were: a 100 mL graduated cylinder, 6vials,germinating peas, dry peas, glass beads, 2 water baths, absorbent cotton and non-absorbent cotton, weights, KOH, water, stoppers, pipettes, rubber bands, masking tape, glue, thermometer, ice, a pencil, and paper.

Methods

Set up a 25 C and a 10 C water bath. Ice may be used to obtain 10 C.

Respirometer 1:Obtain a 100 mL graduated cylinder and fill it with 50 mL of H2O.

Drop in 25 germinating peas. Determine the amount of water displaced. Pea volume =11 mL. Take peas out and place on paper towel.

Respirometer 2: refill cylinder with 50 mL of H2O. Drop 25 dry peas into the cylinder. Add glass beads to obtain the same volume that you got in respirometer 1. Remove peas and beads to a paper towel.

Respirometer 3: Add 50 mL of water to the cylinder. Put only beads in to get an equivalent volume to the first 2 respirometers. Put on paper towel when finished. Repeat respirometer 1 steps for respirometer 4. And 2 for 5. And 3 for 6. Listen to your teacher on how and where to set up the respirometers. Now fill your vials with the required items shown in the table and in figure 5.1. Seal the vials after your items have been put in to stop any gas or water leaks. Place a weighted collar onto the bottom of your vials so they will stay submerged in the water baths. During equilibration use masking tape attached to each side of the water baths to hold the respirometers out of water for 7 minutes. Vials 1-3 should be in the 25 C water bath and vials 4-6 should be in the 10 C water bath. Finally submerge totally the respirometers and let them equilibrate for 3 more minutes. Read the water line where the oxygen is and record in intervals of 5 minutes all the way up to 25 minutes. Record in table 5.1.

Results

Table 5.1: Measurement of O2 Consumption by Soaked and Dry Pea Seeds at Room Temperature and 10 C Using Volumetric Methods

Beads Alone

Germinating Peas

Dry Peas and Beads

Reading at time X

Diff.

Reading at time X

Diff.

Corrected Diff.

Reading at time X

Diff.

Corrected Diff.

Initial-0

1.35

 

1.62

 

 

1.32

 

 

0-5

1.33

.02

1.20

.42

.4

1.32

.0

.02

5-10

1.33

.02

1.12

.50

.48

1.3

.02

.0

10-15

1.32

.03

1.02

.60

.57

1.29

.03

.0

15-20

1.32

.03

.92

.7

.67

1.3

.02

.01

Initial-0

1.48

 

1.37

 

 

1.46

 

 

0-5

1.48

.0

1.15

.22

.22

1.45

.01

.01

5-10

1.45

.03

.98

.39

.36

1.44

.02

.01

10-15

1.43

.05

.84

.53

.48

1.43

.03

.02

15-20

1.41

.07

.70

.67

.6

1.41

.05

.02

In this activity, you are investigating both the effects of germination versus non-germination and warm temperature versus cold temperature on respiration rate. Identify the hypothesis being tested on this activity.
   
The nongerminating peas will have a slower rate of respiration than the germinating peas and the coldness of the water will slow down the rate as it gets colder.

This activity uses a number of controls. Identify at least three of the controls, and describe the purpose of each.
    The three controls are the beads in one vial controlling the barometric pressure, the KOH keeps equality in the consumption of CO2, and the time intervals give each vial the same amount of time so the results will not be affected.

Describe and explain the relationship between the amount of oxygen consumed and time.
   
The relationship was pretty constant, there may have been a gradual rising in O2 consumption.

5.

Condition

Calculations

Rate in mL O2/ minute

Germinating Peas/ 10 oC

(1.62-.92)

20

.035

Germinating Peas/ 20 oC

(1.37-.7)

20

.0335

Dry Peas/ 10 oC

(1.32-1.30)

20

.001

Dry Peas/ 20 oC

(1.46-1.41)

20

.0025

Why is it necessary to correct the readings from the peas with the readings from the beads?
    The beads were just a control, experiencing no gas change.

Explain the effects of germination (versus non-germination) on pea seed respiration.
    The germinating seeds had a higher metabolic rate and therefore consumed more oxygen than the nongerminating.

Above is a sample graph of possible data obtained for oxygen consumption by germinating peas up to about 8 oC. Draw in predicted results through 45 oC. Explain your prediction.
    Once the temperature gets above about 30 degrees C, the enzymes will denature and that will be the end of respiration.

What is the purpose of KOH in this experiment?
    The KOH will take the CO2 and turn it to a precipitant at the bottom of the vial and it will have no affect on the O2 readings.

Why did the vial have to be completely sealed under the stopper?
    The vial had to be sealed or gas would leak out and water could leak in and affect the results.

If you used the same experimental design to compare the rates of respiration of a 35g mammal at 10 oC, what results would you expect? Explain your reasoning.
    Respiration would be higher in the mammal because they are warm-blooded.

If respiration in a small mammal were studied at both room temperature (21 oC) and 10 oC, what results would you predict? Explain your reasoning.
   
The rate of respiration would be higher in the 21-degree bath because the mammal would perform better when its body was more comfortable.

Explain why water moved into the respirometer pipettes.
   
The water moved in the vial because it was fully submerged in water but it came to a stop when it met the oxygen coming out of the vial.

14. Design an experiment to examine the rates of cellular respiration in peas that have been germinating for 0, 24, 48, and 72 hours. What results would you expect? Why?
   
You could put peas in vials each from a time interval above. You would have a vial with just started germinating peas, one with 24 hour germinating peas, another with 48 hour peas, and the last with 72 hour peas. Place them in a room temp water bath. Take readings at intervals of 5 min up to 20 min. The 72-hour peas should have more O2 consumption because they will use more oxygen because they have been germinating the longest. The just started germinating peas would use the least O2 because they havent been germinating vary long. The other two will be in the middle of the "just started peas" and the "72 hour peas".

Error Analysis

Many errors could have been made in this lab. There could have been miscalculations when trying to equal the pea volumes. The stoppers might not have been sealed and gas could have been lost from the vials affecting the results with vengeance. The water temperatures had to be maintained precisely or the results would not be what they should be. There was also a lot of math in this lab when figuring results and many numbers could have been affected by this poor math.

Disussion and Conclusion

This lab showed many things about thew rates of cellular respiration. This lab showed that germinating peas consume more O2 than nongerminating peas. The colder temperature also slowed the rate of oxygen consumption. The oxygen could be clearly seen because of the following reaction

CO2+2KOH K2O3 +H2O

This reaction gets rid of the CO2 so that it would not affect the readings of oxygen. It is absorbed by KOH to give you a precipitant K2CO3 + H2O. I conclude that the rate of O2 consumption is directly proportional to the respiration rate in that when the rate increases the gas consumption increases. When the gas consumption is low then the rate is low. Organisms go through cellular respiration more proficiently when the body of the organism is comfortable with its outside temp and environment. This lab showed many things affecting the rate of cellular respiration.

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