______________________________________________________________________ UNIVERSITI TEKNOLOGI MARA INTERNATIONAL EDUCATION COLLEGE (INTEC)

DEMONSTRATING OXYGEN UPTAKE

Name ID Group Lecturers Name

: Abdul Afiq bin Abdul Hamid : 2010615814 : 11M11 : ASSOC. Prof. Madya Dr. Noraini binti Mohd. Fadzillah

ABSTRACT Every living organism respires throughout their whole life in order to stay alive. The aim of this experiment is to demonstrate the uptake of oxygen in respiration. In this experiment, yeast is used to represent the organisms and the amount of oxygen uptake for five minutes is measured. A replicate of the experiment is also done by changing the amount of yeast tested and any difference of the oxygen uptake is recorded. From the results obtained, an increase of the amount of yeast seems to increase the amount of oxygen uptake thus the rate of respiration is higher.

INTRODUCTION Yeast Yeast are unicellular fungi. Yeast are characterized by a wide dispersion of natural habitats. Common on plant leaves and flowers, soil and salt water. Yeast are also found on the skin surfaces and in the intestinal tracts of warm-blooded animals, where they may live symbiotically or as parasites. Yeast multiply as single cells that divide by budding (eg Saccharomyces) or direct division (fission, eg. Schizosaccharomyces), or they may grow as simple irregular filaments (mycelium). In sexual reproduction most yeast form asci, which contain up to eight haploid ascospores. These ascospores may fuse with adjoining nuclei and multiply through vegetative division or, as with certain yeast, fuse with other ascospores.1 Yeast species either require oxygen for aerobic cellular respiration (obligate aerobes), or are anaerobic, but also have aerobic methods of energy production (facultative anaerobes). Yeasts grow best in a neutral or slightly acidic pH environment.2 The useful physiological properties of yeast have led to their use in the field of biotechnology. Fermentation of sugars by yeast is the oldest and largest application of this technology. Many types of yeasts are used for making many foods: baker's yeast in bread production; brewer's yeast in beer fermentation; yeast in wine fermentation and for xylitol production. Below is an equation showing yeast fermentation.3 C6H12O6 2(CH3CH2OH) + 2(CO2) + Energy

1 2

http://www.yeastgenome.org/VL-what_are_yeast.shtml http://en.wikipedia.org/wiki/Yeast 3 http://en.wikipedia.org/wiki/Yeast

Figure 1: Bakers yeast, Saccharomyces

cerevisiae

Source: http://www.azurebio.com/img/yeast 1.jpg

Respirometer A respirometer is a device used to measure the rate of respiration of a living organism by measuring its rate of exchange of oxygen and/or carbon dioxide. A simple whole animal respirometer designed to measure oxygen uptake consists of a sealed container with the living specimen together with a substance to absorb the carbon dioxide given off during respiration, such as soda lime pellets or cotton wads soaked with potassium hydroxide. The oxygen uptake is detected by displacement of manometric fluid in a thin glass U-tube connected to the container. When the organism takes in oxygen it gives off an equal volume of carbon dioxide. As this is absorbed by the soda lime, air is sucked in from the U-tube to keep the pressure constant, displacing the liquid.
4

Figure 2: A simple Respirometer Source: http://t3.gstatic.com/images?q=tbn: ANd9GcS0lOMw0fA4DvbNVbn9CX3q uTGL_PQC_PuGdlsVMcjChxOkvdTmA g

http://en.wikipedia.org/wiki/Respirometer

HYPOTHESIS The amount of oxygen uptake is higher with more organisms.

NULL HYPOTHESIS The amount of oxygen uptake is the same despite of the increase in the number of organisms.

VARIABLES Manipulated Responding Fixed : Number of filter paper covered with yeast : The distanced of coloured liquid travelled in U-tube : Temperature of surrounding, volume of potassium hydroxide

MATERIALS Potassium hydroxide, filter paper covered with yeast, coloured liquid

APPARATUS U-tube, boiling tube, stopwatch, micropipette, syringe, three-way tap, gauze basket, measuring cylinder

PROCEDURE 1. The apparatus was set up as shown below without the filter paper rolled and also the seeds are replaced with a filter paper containing yeast. The coloured liquid was placed in the U-tube by using the micropipette.

2. The stopwatch was started and any movement of the coloured liquid are recorded in a five minutes interval. 3. The experiment was repeated by replacing the current yeast with two filter paper containing yeast.

RESULTS Time (min) Yeast 0 1 2 3 4 5 Total 6.30 6.50 6.50 6.60 6.70 6.75 Tube Control 5.60 5.40 5.40 5.30 5.20 5.15 0.20 0.00 0.10 0.10 0.05 0.45 Distance moved by the marker fluid (cm)

Table 1: The amount of oxygen uptake by yeast in 1 filter paper for 5 minutes

Time (min) Yeast 0 1 2 3 4 5 Total 7.90 8.30 8.35 8.40 8.50 8.55

Tube Control 5.60 5.20 5.15 5.10 5.00 4.95

Distance moved by the marker fluid (cm)

0.40 0.05 0.05 0.10 0.05 0.65

Table 2: The amount of oxygen uptake by yeast in 2 filter papers for 5 minutes

DISCUSSION When carrying out this experiment, certain factors need to be taken note to enable the experiment can be done without any problems. First of all, the usage of potassium hydroxide is to absorb carbon dioxide in the boiling tube. Since respiration uses oxygen and produce carbon dioxide, potassium hydroxide reacts with the carbon dioxide thus allowing measurement the oxygen uptake to be done without measuring the carbon dioxide as well. The reaction shows how carbon dioxide reacts with potassium hydroxide which produce potassium hydrogen carbonate, KOH + CO2 KHCO3. Besides that, the usage of coloured liquid in the U-tube allow easier reading of the scale since the U-tube is very small and the readings could be deceiving. In this experiment, the aim is to demonstrate the respiration in living organisms and from the results obtained in this experiment, it is clear that living organisms do carry out respiration in order to stay alive. It can be seen in both table (table 1 and 2), the coloured liquid seems to move at a certain distance. These shows that there are some reduction in the volume of the boiling tubes containing yeast, thus allowing the coloured liquid in the U-tube to move towards it. As a result, it can be concluded that the yeast used up the oxygen in the boiling tube for respiration leading to a lower volume. Besides than demonstrating respiration in this experiment, another aim of the experiment which is to determine either different number of organisms have different total uptake of oxygen or not is also demonstrated. In table 1, it can be seen that with only one filter paper, the distance moved by the coloured liquid in the U-tube is only 0.45 cm. In contrast, in table 2 which contained two filter papers covered with yeast, the colour liquid moved by 0.65 cm during the five minutes. This means that more oxygen is taken up in the boiling tube with two filter papers than with one filter paper. Consequently, the movement of coloured liquid is further in the boiling tube with two filter papers. Therefore, the numbers of organisms do affect the total oxygen uptake. respiration. As for ethical issue, the usage of yeast does not rise any ethical issue since yeast are used widely in many things besides in experiment such as in bakery. Therefore, this experiment is quite ethical and also humane. More organisms are present causing more oxygen being used up for

SAFETY MEASURES There are a few safety measures that must be taken when carrying out this experiment. Firstly, as usual, all the laboratory safety measure must be followed. This is to ensure that experiment can be conducted without problems. Lab coats must be worn at all times in the laboratory as a standard protocol when conducting the experiment. Besides that, in this

experiment potassium hydroxide is used to absorb the carbon dioxide. Potassium hydroxide is a corrosive substance therefore can be quiet dangerous. If any potassium hydroxide is split on our hand, quickly wash it with water to dilute it. Eye goggles should also be worn to prevent any splash of the potassium hydroxide from entering the eyes.

LIMITATION AND SOURCE OF ERRORS There are some errors that might contribute to inaccuracy of the results of the experiment. One of it is the setting up of apparatus. The apparatus set up may be not well done and may causes air to leak out from the boiling tube. This may cause the reading to move only a little or not at all. Besides that, unequal pressure on both boiling tube when connecting the tubes to the U-tube may also cause the reading to be imbalanced at the start of the experiment. Parallax error might also in this experiment, since the reading of the scale on the U-tube is very small, some reading may be taken wrongly if the eye is not perpendicular at the right level. As for the limitations in this experiment, the amount of oxygen in the boiling tubes may be limited once it is sealed and closed using the three-way tap, this may causes the yeast to carry out anaerobic respiration if the oxygen content is depleted. These anaerobe yeast does not need oxygen to respire but will produce carbon dioxide and ethanol instead. The production of carbon dioxide is not a problem because it will react with potassium hydroxide but ethanol may causes an inaccuracy in the results. The production of ethanol may cause the volume of the boiling tube to increase thus moving the coloured liquid in an opposite way.

SUGGESTION FOR IMPROVEMENT Some improvements can be done in this experiment to allow it to give more accurate results. First of all, the set up of the apparatus must be completely tight to prevent any air from leaking out of the tube or entering the tube. As to prevent the imbalance pressure, make sure that both three-way tap on each tubes are not close when setting up the experiment. Having someone with a good eye sight or use a magnifying glass to read the scale of the U-tube is also a good alternative to avoid any parallax error. Besides that, since in this experiment

temperature of surrounding is set to constant, immersing the boiling tubes in a water bath could be done to avoid any unnecessary error. At the same time, limitations could also be reduced in this experiment during the set up of the experiment. Making sure that the three-way tap is not close at all before starting the experiment is a good way to avoid the oxygen content from being used before the experiment is even started. Only close the three-way tap when the experiment start could minimise the

probability of oxygen being depleted during the five minutes. This in turn would also prevent the yeast from carrying out fermentation and therefore avoiding the production of ethanol.

FURTHER WORK A further work that can be done using the same set of apparatus is by demonstrating respiration in different temperature. Besides that, the experiment could also be done to study the uptake of oxygen at different temperature. This experiment can be done by immersing the boiling tubes in water baths of different temperature allows the temperature to be manipulated.

Figure 4: A Respirometer immersed in a water bath Source: http://www.practicalbiology.org/data /images/originals/measuring-rate-ofmetabolism-respirometer-in-use-500346.jpg

CONCLUSION From the results obtained in this experiment, it can be seen that the increase in the number of yeast increases the amount of oxygen uptake. Therefore, the amount of oxygen uptake is higher with more organisms. Hypothesis is accepted.

REFRENCE 1. Edexcel A2 Biology Students Book, Ann Fullick Pearson Education 2. http://www.practicalbiology.org/areas/advanced/energy/gas-balance-in-respiration-andphotosynthesis/measuring-the-rate-of-metabolism,117,EXP.html 3. http://en.wikipedia.org/wiki/Potassium_hydroxide 4. http://en.wikipedia.org/wiki/Yeast 5. http://www.yeastgenome.org/VL-what_are_yeast.shtml 6. http://en.wikipedia.org/wiki/Respirometer