Introduction In the Earth, all living organism obtain the energy necessary to sustain life, from the oxidation of organic substances by molecular oxygen, in the process of respiration. Under anaerobic condition (no oxygen or low oxygen concentration), many organism, including yeast obtain the energy from the process of fermentation. In alcoholic fermentation, characteristic of many yeast species, the fermentation process starts with one molecule of the six carbons sugar- glucose, and terminates with two molecules of the two carbons alcoholic- ethanol, two molecules of CO2: Fermentation is an anaerobic process in which fuel molecules are broken down to create pyruvate and ATP molecules. Both pyruvate and ATP are major energy sources used by the cell to do a variety of things. For example, ATP is used in cell division to divide the chromosomes (Bio 116, Metabolism P 3 of 16). The chemical equation for aerobic respiration: C6H12O6 + 6 O2 ---> 6 CO2 +6 H2O+ enough energy to regenerate 38 ATP.
Material and Method This experiment will working in groups of 3-4 students. Our first week experiment was measure the effect of enzyme inhibitor on alcoholic fermentation, also practice how to do the statistical test. Set up fermentations with the conditions in following (Bio 116, winter 2013): 1. Buffer only ( negative control with no sugar or other additives) 2. 2% glucose 3. 2% glucose + 5 mM sodium fluoride ( NaF) 4. 2% glucose + 25mM sodium fluoride (NaF)
There will be four contents, and each of fermentation mixture will have volume of 24 ml. To find out amount of stock solution needed, we need to label the bottle of each stock, then we find the final concentration of 2% by used the equation following.
Desired finalconcentrationConcentration of stock solution×Final Volume
For each contents will also have 8 ml of yeast suspension, and for the positive contents, we would have 8 ml of glucose and 8 ml of yeast. Add 8 ml of 6% glucose in each tube, except for the negative where we add 8 ml buffer. Then add amount of the buffer and other that record in the table 1 show below. To mixture the fermentation well, we need to place vial over the tube, while holding it on firmly, convert the tube twice and place bath water under condition of 45 F. Mark the line at top of the liquid in the tube before place in bath water. We record the time and measure the tube at 10 minute intervals, using a millimeter (Bio 116, 2013).
Table 1: Delicate of the volume (ml) needed of glucose, glucose + NaF, yeast suspension, and buffer by using the equation above. Tube | Contents | 6% ml glucose | ml buffer | Ml other | ml yeast sus | Total ml | 1 | Negative | 0 | 16 | 0 | 8 | 24 | 2 | Glucose | 8 | 8 | 0 | 8 | 24 | 3 | Glucose+ NaF | 8 | 6.8 | 1.2 | 8 | 24 | 4 | Glucose+ NaF | 8 | 2 | 6 | 8 | 24 |
Second week, was conduct fermentations to test a variable. List below are the possible variable to test. 1. Effect of NaF on cell viability 2. Sodium metabisulfite 3. Sucrose 4. Variable glucose concentration 5. Magnesium 6. EDTA 7. Temperature
My group was choosing Magnesium for the conduct the test variable to be investgated.
Table 2: The final volume (ml) to mixture will have 8 ml o yeast suspension, 8 ml of glucose. For Tube | Contents | ml Buffer | ml glucose | ml other (Mg) | Ml yeast suspension | Total ml | 1 | Glucose | 8
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