Dynamic Metabolism of Yeast 3/11/12

Common yeast, Saccharomyces cerevisiae was used to test the effects of the MgSO4 and NaF on yeast cellular metabolism. Two sets of test tubes were used to test the production of CO2. A third set of test tubes was used as a control. Each test tube was placed in a water bath for one hour. The conclusion of our test reveal that there is not a significant difference between the variables. Common day bakers yeast, Saccharomyces cerevisiae , is a eukaryotic fungi. Very much like our own cells, the yeast cells require energy, which is derived from the catabolic breakdown of organic molecules to obtain. This catabolic pathway is referred to as cellular respiration. Cellular respiration occurs in the presence of oxygen, which is known as aerobic respiration; or it can occur without oxygen, which is known as anaerobic respiration. However, unlike our bodys cells, yeast cells go through fermentation, instead of glycolysis( Faller, 1997). The net products of anaerobic respiration differ compared to the net products of aerobic respiration differ greatly, as do the differences between glycolysis and fermentation. Furthermore, fermentation yields less energy than glycolysis. However, additional products are formed from fermentation, which includes ethanol, known as ethyl alcohol (DOMBEK & INGRAM, 1986). Since yeast is easily available to obtain, and its a simple one celled fungus that respires either aerobically or anaerobically, it is the center of many studies. The purpose of this study is to test the idea that Magnesium can be used to affect the rate of fermentation in yeast, and that it can be added to yeast as a soluble salt, or removed by precipitating it as fluoride (Gamboa, 2007). In addition, if the NaF is added to a solution containing Mg ions, the Mg will be removed from the solution as it forms the precipitate, MgF2. Our experiment hypothesizes that we can obtain significantly different results in our experimental groups, when compared to the control. Studies for our experiment began . We obtained 9 carefully cleaned test tubes and vials, each of which was washed with water and rinsed with distilled water. We then used tape to label each of the nine test tubes, 1-9. In each of the nine test tubes, 8 ml of yeast suspension was added, then 8 mL of 1.0M glucose was added to each of the nine test tubes. In three of the control test tubes, 8 ml of deionized water was added. Then, 8 ml of 0.01M MgSO4 was added to the next three test tubes. Finally, 8 ml of 0.1M NaF was added to the last of the three test tubes. We then placed inverted vials over each of the test tubs, and inverted each of the test tubes and vials. Using a grease pen, we visibly marked the fluids meniscus level on each of the tubes. Then the test tubes were placed in a ~37 water bath. After an hour, we recorded the distance from the grease markings, to the new meniscus level using a ruler. Gathering data from the rest of the class, we averaged our data, and analyzed the combined data using the Kruskal-Wallis test.

Table 1

MgSO4 NaF H2O 1 4.7 6.0 2.7 2 4.0 4.3 2.3 3 4.3 4.3 2.3 4 2.8 2.5 3.3 5 3.0 5.3 4.7 6 5.0 3.0 4.6 7 6.3 2.0 2.5 In table 1, the raw data has been given in regards to the millimeters difference after the test tubes were placed in the water baths. Each column dictates which test tubes were placed and is organized by the respective differences, MgSO4, NaF, and H2O. Table 2 MgSO4 NaF H2O

1 2.8(7) 2.0(1) 2.3(2.5) 2 3.0(8.5) 2.5(4.5) 2.3(2.5) 3 4.0(11) 3(8.5) 2.5(4.5) 4 4.3(13) 4.3(13) 2.7(6) 5 4.7(16.5) 4.3(13) 3.3(10) 6 5.0(18) 5.3(19) 4.6(15) 7 6.3(21) 6.0(20) 4.7(16.5) Sum/average 95-13.6 79-11.3 57-8.14 In table 2, the raw data has been ordered by each compound from lowest to highest in regards to millimeters difference. The numbers in parenthesis have been given to show their ranking. In the last row, the numbers are the sum of all the ranks, and then the average is given. Table 3 MgSO4 NaF H2O

Counts 7 7 7 21 Sum 95 79 57 231 Average 13.6 11.3 8.14 11 Table 3 shows the total counts and sums of data and averages from the raw data after it has been organized.

Table 4

MgSO4

NaF

2.8(3) 2.0(1) 3.0(4.5) 2.5(2) 4.0(6) 3(4.5) 4.3(8) 4.3(8) 4.7(10) 4.3(8) 5.0(11) 5.3(12) 6.3(14) 6.0(12) 56.5-8.1 48.5-6.9 Table 4 shows the rankings between just magnesium sulfate and sodium fluoride. The bottom shows the sum and average of each column, respectively. Table 5 MgSO4 NaF

Sum 56.7 48.5 n 7 7 Table 5 indicates the sum of both MgSO4 and NaF and the N- value. Upon performing the Mann-Whitney test, the value of Magnesium sulfate was at 20.5.This indicates that Magnesium was higher than the critical value test, 8. Table 6 MgSO4 H2O

2.8(5) 2.3(1.5) 3.0(6) 2.3(1.5) 4.0(8) 2.5(3) 4.3(9) 2.7(4) 4.7(11.5) 3.3(7) 5.0(13) 4.6(10) 6.3(14) 4.7(11.5) 66.5-9.5 38.5-5.5 Table 6 shows the rankings between just magnesium sulfate and water. The bottom shows the sum and average of each column, respectively.

Table 7

MgSO4

H2O

Sum 66.5 38.5 n 7 7 Table 7 indicates the sum of both MgSO4 and H2O. Upon performing the Mann-Whitney test, the value of Magnesium sulfate was at 10.5. Magnesium was higher than the critical value test, 8. Table 8 NaF H2O

2.0(1) 2.3(2.5) 2.5(4.5) 2.3(2.5) 3(7) 2.5(4.5) 4.3(9.5) 2.7(6) 4.3(9.5) 3.3(8) 5.3(13) 4.6(11) 6.0(14) 4.7(12) 58.5-8.4 46.5-6.64 Table 8 shows the rankings between just sodium fluoride and water. The bottom shows the sum and average of each column, respectively. Table 9 NaF H2O

Sum 58.5 46.5 n 7 7 Table 9 indicates the sum of both NaF and H2O. Upon performing the Mann-Whitney test, the value of Magnesium sulfate was at 18.5. Magnesium was higher than the critical value test, 8.

After performing the Kruskal-Wallis test, the H value has been computed at 2.7.

H0 was set to equal test tubes 1,2,3 not equaling 4,5,6, and 4,5,6 not equaling 7,8,9; which would indicate that either MgSO4 or NaF would have a significant effect on the metabolic processes of yeast. At the conclusion of the Kruskal-Wallis test, we determined that there was no

difference at the conclusion of our test, as 2.7<5.99 at a p-value of .005. Thus, our null hypothesis, H0 was invalid. A type II error couldve been made. Errors could have been made during the preparation stage of the experiment, grease markings could have been lower than the meniscus was actually at, meaning a lower difference level in millimeters; indicating a higher HValue. This higher value couldve exceeded the critical value of 5.99, making the difference significant. However, there was a difference between MgSO4 and NaF; with the value of MgSO4 at 20.5> the critical value equaling 8. This indicates that magnesium sulfate has a positive influence on the metabolic processes of yeast, compared to sodium fluoride. A lower level of carbon dioxide in the metabolic functions of yeast indicates that an absence of magnesium in the presence of NaF will result in lower metabolic functions. Magnesium is required for fermentation to operate.

Literature Cited Dombek, K. M., & Ingram, L. O. (1986). Magnesium limitation and its role in apparent toxicity of ethanol during yeast fermentation. Manuscript submitted for publication, Deapartment of Microbiology and Cell Science, University of Florida, Gainesville, Florida. Gamboa, G. J. (2007). Experience biology laboratory manual. (3 ed.). Dubusque, Iowa: Kendall/Hunt Publishing Company. Larry D. Faller, Bahige M. Baroudy, Alan M. Johnson, and Ralph X. Ewall Biochemistry 1977 16 (17), 3864-3869