IB Biology HL 1 Mrs.

Jensen

Miranda Shauger January 17, 2012

Rates of Fermentation: Juiced Natural Fruits vs. Commercialized Fruit Juices

Design:
Question: What is the difference of the rate of fermentation in yeast cells between a juiced fruit and the respective commercialized fruit juice? Hypothesis: This experiment is going to investigate and compare the rate of fermentation in two natural fruits and the two respective commercialized fruit juices. It is predicted that the commercialized fruit juices will have a higher rate of fermentation when compared to their respective natural fruits. This can be predicted because generally, apple juice has a sweeter taste than an apple, as well as orange juice has a sweeter taste than an orange. During cellular respiration when oxygen is not available, known as anaerobic cell respiration, pyruvate can be transformed in the cytoplasm into lactate, or ethanol and carbon dioxide, with no additional yield in ATP. This is known as fermentation. Fermentation has a low energy yield, and few organisms can acquire enough energy for their needs this way. The rate of fermentation of the yeast depends on the relative amount of sugar in the food source. The rate of fermentation is determined by the pressure in a closed test tube. A change in pressure occurs because while the yeast is fermenting the sugars anaerobically, CO2 is being produced and no oxygen is being consumed. CO2 gas pressure sensors can be used to measure the rate of fermentation because CO2 is a by-product of fermentation. The more the CO2 produced in a certain time frame, the faster the rate of fermentation.

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Variables:

Independent Variable Dependent Variable

Variables Juiced fruit or respective commercialized fruit juice Rate of fermentation

Units 2.5 mL kPA/minute

Uncertainties 0.25 mL
Sensors give data to the millionth place data will be rounded to the hundredths place (uncertainty not necessary)

Controlled Variables

Units

Possible effect(s) on results An unstable temperature can cause a fluctuation in fermentation rate. A temperature that is too high can kill the yeast, thus preventing fermentation from occurring for the time measured. More yeast would metabolize more of the sugar in a shorter period of time. This would cause an increase in CO2 gas pressure reading. A higher amount of juice (commercialized or natural) could lead to an increased fermentation rate. A lesser amount could lead to a food shortage, causing a plateau of the CO2 gas pressure reading. The purpose of the vegetable oil was to create an anaerobic environment for the yeast by cutting off its oxygen supply. If an anaerobic environment is not maintained than aerobic fermentation will affect the results. An inconsistent time period in which fermentation is observed would make comparing the data more difficult.

Method for Control Thermometer reading (maintain at 40C, checked every minute) 2.5 mL of yeast was placed in each test tube

Temperature of water bath

40C

Amount of yeast per test tube

mL

Amount of juice per test tube

mL

2.5 mL of juice was placed in each test tube

Amount of vegetable oil per test tube

mL

2 mL of vegetable oil was used to cover the yeast and fruit juice in each test tube Each trial was observed for 10 minutes (600 seconds)

Time observed

seconds

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Photograph of Lab Setup:

Gas Pressure Sensor to measure the rate of fermentation

Thermometer to visualize temperature change

Test tube containing Yeast, Independent Variable, and oil immersed in the hot water bath

Materials: computer 18 150 mm test tube Vernier computer interface 1 L beaker (for water bath) Logger Pro basting bulb or Beral pipette Vernier Gas Pressure Sensor Distilled water l-hole rubber stopper assembly test tube rack plastic tubing with Luer-lock fitting thermometer 5% glucose solution yeast suspension vegetable oil ring stand utility clamp 1mL pipet 10 mL graduated cylinder Commercialized Orange Juice (Tropicana) Juiced Orange (Fruit) Commercialized Apple Juice (Motts) Juiced Apple (Fruit)
3

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Procedure: 1. A gas pressure sensor was connected to the computer interface. 2. Plastic tubing was connected to the valve on the Gas Pressure Sensor. 3. A water bath was prepared for the yeast by filling a large 1-L beaker with around 700 mL of water and this beaker was placed on a hot plate. Temperature was kept at 40C and monitored using a thermometer. 4. The yeast was activated by adding 100mL of distilled water to 7 grams of yeast in a flask. The flask with the yeast was placed in a water bath heated to nearly 40C for at least 10 minutes. 5. 5 Test tubes were obtained and labeled with Natural Apple. 6. 2.5 mL of the Natural Apple, which was previously juiced, was placed into 1 test tube. 7. The yeast suspension was obtained and gently swirled. 2.5 mL of yeast was measured and added to the Natural Apple Juice already in the tube. This mixture was mixed gently. 8. Using a 1 mL pipet, 2 mL of vegetable oil was added to the top of the yeast/Natural Apple Juice mixture. 9. Steps 5-7 was repeated 4 more times. 10. 3 of the 5 test tubes were placed in the hot water bath. 11. 3 single-holed rubber-stoppers were inserted into the 3 test tubes, which were then secured with a utility clamp and ring stand. 12. Most of the test tube was covered by the water in the hot water bath. Test tubes were incubated for 10 minutes and the temperature was checked and accordingly adjusted every minute. 13. The rate of fermentation, measured in kPA, was collected for 10 minutes using the LoggerPro program and Vernier Gas Pressure Sensor. 14. Steps 10-13 were repeated for the remaining 2 trials. 15. Steps 5-14 were repeated for each Commercialized Apple, Natural Orange, and Commercialized Orange. 16. 1 test tube was obtained and labeled Negative Control.
4

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

17. 2.5 mL of Distilled water was placed into the test tube. 18. The yeast suspension was obtained and gently swirled. 2.5 mL of yeast was measured and added to the Distilled water already in the tube. This mixture was mixed gently. 19. Using a 1 mL pipet, 2 mL of vegetable oil was added to the top of the yeast/Distilled water mixture. 20. The test tube was placed in the hot water bath. 21. A single-holed rubber-stopper was inserted into the test tube, which was then secured with a utility clamp and ring stand. 22. Steps 12 and 13 were repeated. 23. 1 test tube was obtained and labeled Positive Control 24. 2.5 mL of 5% glucose solution was placed into the test tube. 25. Steps 18-22 were repeated for the positive control. 26. As soon as the data collection was complete, the plastic tubing connector was disconnected from the rubber stopper, and the rubber stopper was removed from the test tube. Contents of the test tubes were discarded.

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Data Collection
Raw Data Tables:
Commercialized Orange Juice Pressure Values (kPa) Time (Minute) 0 1 2 3 4 5 6 7 8 9 10 Trial 1 103.65 104.62 105.81 107 108.41 109.83 111.3 112.66 114.13 115.66 117.19 Trial 2 104.42 105.5 106.58 107.6 108.85 110.05 111.3 112.72 114.03 115.56 117.04 Trial 3 104.05 105.12 106.43 107.79 109.09 110.34 111.7 113.12 113.29 114.48 115.78 Trial 4 105.44 106.41 107.37 108.28 109.36 110.5 111.58 112.72 113.97 115.33 116.7 Trial 5 105.35 106.31 107.28 108.13 108.98 109.94 110.91 112.1 113.18 114.31 115.61

*Data was collected by Julia Vanzelli

*Uncertainties not necessary, data logger measures up to the millionth decimal place
Natural Orange Fruit Pressure Values (kPA) Time (Minute) 0 1 2 3 4 5 6 7 8 9 10 Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 107.17 104.87 103.99 105.16 103.51 107.74 105.38 104.73 105.74 104.64 108.32 105.781 105.29 106.48 106.01 108.95 106.24 105.97 107.34 107.43 109.92 106.86 106.94 108.26 108.91 111.07 107.71 108.07 109.18 110.39 112.39 108.68 109.26 110.09 111.81 113.76 109.59 110.57 111.07 113.11 115.25 110.78 112.04 111.98 114.48 116.8 112.04 113.46 112.73 115.73 118.52 113.29 115.1 113.42 116.81

*Data was collected by Evelyn Abramson

*Uncertainties not necessary, data logger measures up to the millionth decimal place

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Commercialized Apple Juice Pressure Values (kPa) Time (Minute) 0 1 2 3 4 5 6 7 8 9 10 Trial 1 108.21 108.61 109.24 110.03 110.83 111.73 112.64 113.78 114.80 116.11 117.24 Trial 2 106.83 107.35 107.93 108.63 109.20 109.90 110.53 111.29 112.04 112.85 114.00 Trial 3 105.49 106.11 106.79 107.65 108.16 108.67 109.24 110.03 110.71 111.73 112.81 Trial 4 104.00 104.46 105.22 106.03 106.78 107.41 108.11 108.92 109.78 110.94 112.04 Trial 5 108.16 110.09 112.02 114.06 116.11 117.98 120.19 122.41 124.68 127.01 129.68 Trial 6 104.90 105.18 105.57 105.97 106.36 106.76 106.98 101.45 103.47 103.09 103.82

*Data was collected by Miranda Shauger

*Uncertainties not necessary, data logger measures up to the millionth decimal place

Natural Apple Fruit Pressure Values (kPa) Time (Minute) 0 1 2 3 4 5 6 7 8 9 10 Trial 1 103.70 104.49 105.29 106.37 107.39 108.41 109.38 110.63 111.70 112.61 113.46 Trial 2 104.77 105.17 105.74 106.43 107.12 107.86 108.72 109.63 110.55 111.57 112.66 Trial 3 104.48 105.16 106.24 107.26 108.17 108.85 109.59 110.39 111.24 111.98 112.66 Trial 4 107.40 107.80 108.66 109.80 110.89 111.98 113.06 114.32 115.58 116.83 118.03 Trial 5 107.71 108.62 109.53 110.50 111.47 112.38 113.46 114.60 115.62 116.70 117.89

*Data was collected by Kunal Manocha

*Uncertainties not necessary, data logger measures up to the millionth decimal place

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Control Values (kPA) Time (Minute) Positive Negative 0 106.54 102.92 1 107.00 103.09 2 107.92 103.26 3 108.78 103.48 4 109.81 103.6 5 6 7 8 9 10 110.78 111.70 112.56 113.42 114.28 115.25 103.65 103.65 103.82 103.88 104.05 104.11

*Positive control data was collected by Julia Vanzelli; Negative control data was collected by Miranda Shauger

*Uncertainties not necessary, data logger measures up to the millionth decimal place

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Data Processing & Presentation

Overview: The positive and negative control have the same average as the original data point and a standard deviation of 0, as there was only one trial of each performed. The pressures of the trials were averaged by minute. Standard deviation was done in order to show how close the data points were at during that minute. The change of pressure was found, and then divided by the number of minutes which the data was observed. This produced the rate of fermentation in kPA/minute. A scatter plot for each independent variable was done next. The scatter plots show the individual pressure readings and how they changed as the time advanced. The last scatter plot has data representing the average rate of fermentation for each of the independent variables. This was done in order to compare the slopes of the lines. Sample Calculation: Pressure Change for Commercialized Apple Juice Trial 1 Maximum Value Minimum Value 117.24108.21 = 9.03 kPA Fermentation Rate for Commercialized Apple Juice Trial 1 Difference of fermentation rate / 10 (amount of minutes tested for) 9.03 / 10 = .903 kPA/minute Average Rate of Fermentation for Commercialized Apple Juice .903+ .717 + .733 + .803 + 2.15 / 5 = 1.06 kPA/minute Standard Deviation for Commercialized Apple Juice Pressure Value minute 0
(108.21-106.538)2 +(106.83-106.538)2 +(105.49-106.538)2 +(104.00-106.538)2 +(108.61-106.538)2 =

(1.672)2+(.292)2+(-1.048)2+(-2.538)2+(2.072)2= (2.795584)+(.085264)+(1.098304)+(5.4414444)+(4.293184)= 13.2215 13.2215/5=2.62443 2.624431/2=1.62

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Presentation:

Commercialized Apple Juice Pressure Value Averages (kPa) Time Average Standard (Minute) between Deviation trials 0 106.538 1.62 1 107.324 1.95 2 108.24 2.31 3 109.28 2.72 4 110.216 3.23 5 111.138 3.71 6 112.142 4.30 7 113.286 4.84 8 114.402 5.41 9 115.728 5.91 10 117.154 6.51

Natural Apple Fruit Pressure Value Averages (kPa) Time Average Standard (Minute) between Deviation trials 0 106.81 1.10 1 106.90 0.95 2 107.16 0.99 3 107.76 1.19 4 108.44 1.38 5 109.17 1.53 6 110.01 1.62 7 111.00 1.76 8 112.08 1.84 9 113.25 1.91 10 114.41 1.95

Commercialized Orange Juice Pressure Value Averages (kPa) Time Average Standard (Minute) between Deviation trials 0 104.58 0.71 1 105.59 0.69 2 106.69 0.58 3 107.76 0.45 4 108.94 0.31 5 110.13 0.25 6 111.36 0.27 7 112.66 0.33 8 113.72 0.40 9 115.07 0.56 10 116.46 0.65

Natural Orange Fruit Pressure Value Averages (kPa) Time Average Standard (Minute) between Deviation trials 0 104.94 1.26 1 105.65 1.13 2 106.38 1.04 3 107.19 1.05 4 108.18 1.17 5 109.28 1.29 6 110.45 1.43 7 111.62 1.57 8 112.91 1.68 9 114.15 1.82 10 115.43 2.01

10

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Commercialized Orange Juice Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average Pressure Change (kPa) 13.54 12.62 11.73 11.26 10.26 --Fermentation Rate (kPa/minute) 1.35 1.26 1.17 1.13 1.03 1.19

Natural Orange Fruit Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average Pressure Change (kPa) 11.35 8.42 11.11 8.26 13.3 --Fermentation Rate (kPa/minute) 1.14 0.84 1.11 0.83 1.33 1.05 Commercialized Apple Juice Trial 1 Trial 2 Trial 3 Trial4 Trial 5 Average Pressure Change (kPa) 9.03 7.17 7.33 8.03 21.51 --Fermentation Rate (kPa/minute) 0.9 0.72 0.73 0.8 2.15 1.06

Natural Apple Fruit Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Average Pressure Change (kPa) 9.77 7.89 8.17 10.63 10.18 --Fermentation Rate (kPa/minute) 0.98 0.79 0.82 1.06 1.02 0.93 Controls Positive 8.71 .87

Pressure Change (kPa) Fermentation Rate (kPa/minute)

Negative 1.19 .12

11

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

12

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

13

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

Conclusion & Evaluation

Conclusion: The results showed that the commercialized orange juice had the highest average rate of fermentation, the commercialized apple juice and the second highest average rate of fermentation, the natural orange fruit had the third highest average rate of fermentation, while the natural apple fruit had the lowest average fermentation rate. The results support the hypothesis which predicted that the commercialized fruit juices will have a higher rate of fermentation when compared to their respective natural fruits. This indicates that, on average, the commercialized fruit juice samples contained more sugar than the natural fruit samples. This is indicated because the fermentation rates are higher in the commercialized fruit juices than the natural fruits in this investigation. One anomaly which occurred during this experiment was trial 5 of the commercialized apple juice. This trial had a fermentation rate which was nearly double the other rates of fermentation. This may have been caused by the sugar in the juice settling to the bottom of the container and this sample being from an area which the sugar was more concentrated. This

14

IB Biology HL 1 Mrs. Jensen

Miranda Shauger January 17, 2012

may also have happened with the yeast, which would mean more yeast in this trial than the others. Further investigations of this experiment could include the same experiment with more trials therefore a t-test may be performed. Additionally, the rate of fermentation of fruits with varying pHs would be a good way to further investigate this topic, since oranges have a lower pH than apples. Evaluation: Evaluating the Procedure: One anomoly which occurred during this investigation, raw data trial 6 of the commercialized apple juice, was caused by the rubber stopper not staying in the test tube. This was caused by excessive vegetable oil near the rim of the test tube on the inside. The vegetable oil provided lubrication, so as the pressure increased, the rubber stopper was pushed further out of the test tube, not maintaining the pressure once it popped out of the test tube. This data point was removed from the processed data. Improving the Investigation: To alter the procedure to avoid this from happening in future trials, it is suggested that after the vegetable oil is added, the inner part of the test tube which the rubber stopper will be should be whiped clean with a tissue/paper towel/cloth of some sort to remove the oil. Additionally, more trials would improve the investigation. Bibliography FDA. (2008). Raw fruits poster. U S Food and Drug Administration. Retrieved January 15, 2012, from: http://www.fda.gov/Food/LabelingNutrition/FoodLabelingGuidanceRegulatoryInformati on/InformationforRestaurantsRetailEstablishments/ucm063482.htm Masterman, D., & Redding, K. (2006). Biology with computers. (3 ed.). Beaverton, OR: Vernier Software & Technology. Motts. (2011). Motts products. Retrieved January 15, 2012, from: http://www.motts.com/Products/FamilyHealthyFavorites/MottsOriginal100AppleJuice. aspx Tropicana. (2011). Tropicana products. Retrieved January 15, 2012, from: http://www.tropicana.com/#/trop_products/productsLanding.swf?TropicanaPurePremi um/21
15