ABSTRACT The factors affecting the rate of cellular respiration was determined through the use of controls; one

for the oxygen uptake of multicellular organisms and another for the cellular respiration in yeast. For the oxygen uptake of the organism, three flasks set-ups were made and it was found out that protein was the utilized substrate of the organism. For the cellular respiration of yeast, two methods were used; the Durham tube method and the Smith fermentation method. In the Durham tube method, the second set-up had a faster rate of CO2 production because of the presence of magnesium which activated enzymes. In the Smith fermentation tube, glucose had the fastest rate of CO2 production. Thus, oxygen intake, the co-factors and the type of substrate affects the rate of cellular respiration.

INTRODUCTION

In cellular respiration, energy in food molecules is released and converted to a form that can be used by the cell, which is ATP. The term respiration is often used to mean breathing or simply the process of inhaling and exhaling. For clarity, the term cellular respiration is used to refer specifically to a series of enzymatic reactions that can occur in the presence or absence of oxygen and make energy available to the cell. (Maosa and Talaue, 2007) Organic compounds store energy in their arrangements of atoms. With the help of enzymes, a cell systematically degrades complex organic molecules that are rich in potential energy to simpler waste products that have less energy. Some of the energy taken out of chemical storage can be used to do work, the rest dissipated as heat. Metabolic pathways that release stored energy by breaking down complex molecules are called catabolic pathways. One catabolic process, called fermentation, is a partial degradation of sugars that occurs without the help of oxygen. However, the most

prevalent and efficient catabolic pathway is cellular respiration, in which oxygen is consumed as a reactant along with the organic fuel. (Campbell, 1996)

This study aimed to show the factors affecting the rate of photosynthesis. The specific objectives were: 1. to determine the factors affecting cellular respiration 2. how these factors affect the rate of cellular respiration 3. identify the substrate utilized by the respiring organism

The study took place at room 221, Biology Lab, College of Science and Mathematics, University of the Philippines, Mindanao last August 22, 2012.

MATERIALS AND METHODS To determine the factors that affect the rate of cellular respiration, two control experiments were made; one for the oxygen uptake of organisms and another for the cellular respiration of yeast. In the oxygen uptake of multicellular organisms, three respirometers were obtained. The same number of an experimental animal was placed in two flasks. The two flasks were labelled as flasks 2 and 3. The flask without the experimental animal was labelled as flask 1. Two pleated filter papers were soaked in KOH and were hung inside (between the opening of the flask and the rubber stopper) flasks 1 and 2. The flasks were

covered and were sealed airtight through melted paraffin. The Respiratory Quotient (RQ) for each time interval was computed using the formula: RQ = CO2 output O2 uptake CO2 output and O2 uptake was computed using the formula: vO2 = | v2 v1| and vCO2 = | v3 - vO2 | where: vO2 = volume of O2 consumed vCO2 = volume of CO2 consumed v1, v2, v3 = final volume of each set-up In the cell respiration in yeast, two methods were used; Durham tube method and Smith fermentation method. For the Durham tube method, two test tubes were obtained for each group. 7 ml distilled water and 7 ml glucose were placed in test tube 1. 7 ml glucose and 7ml 0.2M MgSO4 were placed in test tube 2. 7 ml of 10% yeast suspension were poured into each of the test tubes. The mixtures were shaken gently. An inverted Durham tube was slid down into each of the test tubes. To remove air bubbles in the inverted Durham tube, the opening of the bigger test tube was covered tightly with the palm of one hand and let the bubbles escape from the tube through tilting the bigger tube gently from side to side. After ensuring that no more bubbles remain inside the Durham tube, a Pasteur pipette was used to remove excess suspension that covered the tip of the inverted Durham tube. This allowed the measurement of the height of the air space occupied by the CO2 evolved that was trapped at the bottom of the inverted tube. The

bigger tube was plugged with a cotton ball. The set-up prepared by each group was considered as replicates. In the Smith fermentation tube method, six Smith fermentation tubes were obtained. 15 ml of the following solutions (all at 10% concentration) was poured to the respective tubes: 1 starch, 2 lactose, 3 sucrose, 4 glucose, 5 fructose, 6 distilled water. Next, 15 ml of distilled water and 15 ml 10% yeast suspension were added to each tube. The mixtures were shaken gently. Trapped bubbles were removed through covering the opening of the tube with the palm of one hand and tilting the tube horizontally. The openings were plugged with cotton balls. The tubes were tied together at their vertical arms to keep them upright. The tubes were set aside. Figures 1 and 2 shows the set-ups in the Cellular respiration in yeast.

RESULTS AND DISCUSSIONS Table 1 shows the height of water in the three set-ups in oxygen uptake of mulitcellular organisms. The first set-up contained a filter paper soaked in KOH; the second set-up contained mongo sprout and filter paper soaked in KOH; the third set-up contained a mouse. Table 1. Height of water in the three set-ups in oxygen uptake of multicellular organisms. Time interval 5 10 15 20 25 30 Height of water E-flask with mongo sprout and KOH (mm) 130 15 10 20 10 0

E-flask with only KOH (mm) 0 4 4 4 4 4

E-flask with mouse only (mm) 3 2 4 6 7 7

As seen in Table 1, the first set-up had a constant height of water since it is the controlled variable. In the second set-up, the height of water lowered down as time passed. In set-up three, the height of water, increased and decreased as the time went on. The Respiratory Quotient was computed as: vO2 = | 0 4| = 4 vCO2 = | 7 - 4 | = 3

RQ = 3 4 = 0.75 = 0.8 The RQ value is 0.8, this means that protein is the substrate. However, this may not be the substrate used because two different living organisms were used in the experiment. This may have affected the RQ value. In Table 2.1, the height of the area occupied by the gas in the Durham tube set-up is shown. Two test tube set-ups were prepared by each group; the first test tube with distilled water, glucose, and yeast; the second test tube with glucose, magnesium sulfate and yeast. In Table 2.2, the volume and rate of CO2 evolved in the Durham tube method is shown. Table 2.1. Height of the area occupied by the gas in the Durham tube set-up. Time (min) 3 6 9 12 15 18 21 24 27 30 Height of the Area Occupied by the gas (mm) Group 1 Group 2 Group 3 Group 4
Test tube1 Test tube2 Test tube1 Test tube2 Test tube1 Test tube2 Test tube1 Test tube2

3 3 3 4 4 3 4 4 4 4

3 3 4 4 5 4 5 5 5 5

1 1 2 3 3 3 4 4 3 4

7.5 1 1 4 4 4 5 5 4 5

2 1 1 1 1 2 2 3 3 3

3 2 3 4 2 3 3 4 4 4

2 3 3 4 3 3 4 4 4 4

3 3 4 4 3 4 4 4 5 5

As seen in Table 2.1, the rate of height increase is faster in the second test tube. The second test tube had magnesium sulfate which sped up the rate of height increase. Table 2.2. Volume and rate of CO2 evolved in the Durham tube method. Time Group 1 Volume of CO2 evolved V=r2h (mm3) Group 2 Group 3 Group 4

Testtube1 (min) 3 84.82 6 84.82 9 84.82 12 113.10 15 113.10 18 84.82 21 113.10 24 113.10 27 113.10 30 113.10 RateofCO2 3.77

Testtube2

Testtube1

Testtube2

Testtube1

Testtube2

Testtube1

Testtube2

84.82 84.82 113.10 113.10 141.37 113.10 141.37 141.37 141.37 141.37 35.34

28.27 28.27 56.55 84.82 84.82 84.82 113.10 113.10 84.82 113.10 3.77

212.06 28.27 28.27 113.10 113.10 113.10 141.37 141.37 113.10 141.37 35.34

56.55 28.27 28.27 28.27 28.27 56.55 56.55 84.82 84.82 84.82 35.34

84.82 56.55 84.82 113.10 56.55 84.82 84.82 113.10 113.10 113.10 3.77

56.55 84.82 84.82 113.10 84.82 84.82 113.10 113.10 113.10 113.10 3.77

84.82 84.82 113.10 113.10 84.82 113.10 113.10 113.10 141.37 141.37 35.34

As seen in Table 2.2, the rate of CO2 evolved in the 2nd test tubes of groups 1, 2, and 4 are greater than the 1st test tube. This is also because of the magnesium present in the second test tube. It activated enzymes that sped up the reactions. However, the results may not be accurate because of not plugging the set-ups with cotton balls. In Table 3.1, the height of the area occupied by CO2, in the Smith fermentation tube method is shown. In Table 3.2 the volume of CO2 evolved in the Smith fermentation tube method is shown. Table 3.1. Height of the area occupied by CO2, in the Smith fermentation tube method.
Time interval (min)

5 10 15 20 25 30

Water 5 9 11 12 12 13

Height of the area occupied by CO2 (mm) Glucose Fructose Sucrose Starch 10 6 9 4 10 9 9 4 14 11 13 6 14 11 13 11 15 13 14 11 15 13 15 12

Lactose 6 8 9 10 12 12

As seen in Table 3.1, the height of the area occupied by carbon dioxide increased in all the Smith fermentation set-ups.

Table 3.2. Volume of CO2 evolved in the Smith fermentation tube method.
Time interval (min)

5 10 15 20 25 30 RateofCO2

Water 769.69 1385.44 1693.32 1847.26 1847.26 2001.19 66.71

Volume of CO2 evolved V=r2h (mm3) Glucose Fructose Sucrose Starch 2269.80 1361.88 1385.44 804.24 2269.80 2042.82 1385.44 804.24 3177.72 2496.78 2001.19 1206.37 3177.72 2496.78 2001.19 2211.68 3404.70 2950.74 2155.13 2211.68 3404.70 2950.74 2309.07 2412.74 113.49 98.36 76.97 80.42

Lactose 1206.37 1608.50 1809.56 2010.62 2412.74 2412.74 80.42

As seen in Table 3.2, the volume evolved by the carbon dioxide in the Smith fermentation increased. The rate of CO2 in the set-ups is also shown. The rank of substrates readily utilized from greatest to least, glucose, fructose, starch & lactose, sucrose, water. Simpler carbohydrates (glucose) can speed up the rate of cellular respiration more that the complex carbohydrates (starch, etc.).

SUMMARY AND CONCLUSION

This experiment was conducted to determine the factors affecting cellular respiration. Two control experiments were made; one for the oxygen uptake of multicellular organisms and another for the cellular respiration of yeast. RQ, CO2 and O2 were measured and computed through the oxygen uptake experiment wherein respirometer set-ups were used. The RQ value was 0.8, so the substrate used by the organisms was proteins. Durham tube method and Smith Fermentation tube method were used to determine the cellular respiration in yeast. In the Durham tube set-up, the rate of CO2 production was faster in the test tube with the Magnesium Sulfate, which is a cofactor that activated the enzymes. In the Smith fermentation set-up, glucose had the fastest rate of CO2 production. The rate of carbon dioxide production depends on the complexity of the substrate. In yeast, carbohydrates are primarily used. The simpler the carbohydrate, the faster the rate of CO2 production.

The rate of cellular respiration depends on the oxygen uptake, the co-factors which activate enzymes and the type of substrate utilized by an organism.

LITERATURE CITED Susan Maosa, Frederick Talaue. 2007. Breaking through Biology. South Triangle, Quezon City. C&E Publishing, Inc. pp. 114-125 Duka, Diaz, Villa. 2009. Biology 1 Laboratory Manual: An Investigative Approach. 9th edition. Laguna, Philippines. Genetics and Molecular Division, Institute of Biological Sciences, College of Arts and Sciences, University of the Philippines Los Baos College, Laguna, Philippines. pp. 50-55 Neil A. Campbell. 1996. Biology. 4th Edition. The Benjamin/Cummings Company, Inc.Menlo, Park, California pp 159-179 Discospinster. 2012. Cellular Respiration. < http://en.wikipedia.org/wiki/ Cellular_respiration >. Accessed August 27, 2012.