taking place. pH may also be a variable that could affect enzyme activity and so Iwill control pH using a buffer solution. It is important to control these variables asthey can affect reaction rate so will affect my results. If these variables are notcontrolled it cannot identified clearly how phosphate concentration inhibits theactivity of beansprout phosphatase as other factors will influence the results.Alternative Hypothesis: The higher the concentration of phosphate present, theless the coloration of the end solution, indicating more enzyme inhibition. This isa one-tailed hypothesis.Null Hypothesis: Phosphate concentration will have no significant effect onenzyme activity so coloration of end solution will not vary significantly.
1 Measuring cylinder 5 Teat pipettes7 Test tubes1 250ml beaker Muslin Filter 1 Colorimeter 1 Cuvette30 C Water bath1 Mortar and PestleDistilled water 150 ml Phenolphthalein phosphateAbout 150 Beansprouts
Sodium carbonate is an alkali and therefore is an irritant to the skin and eyes.Contact should be avoided by wearing a lab coat and safety goggles. Freephenolphthalein is a suspected carcinogen and therefore contact should beavoided, especially ingested. However the small amounts present do notsurmount a large risk.
For my test experiment I will use a large range (0-100%) of 0.3 phosphatesolution concentrations so as to get a sense of scale for the main experiment. Iwill prepare solutions of 0% 20% 33.3% 40% 66.6% 80% and 100% phosphatesolutions by using appropriate parts 0.3M phosphate solution and distilled water.I will extract phosphatase enzyme from the bean sprouts. Using a mortar andpestle and water and filter through muslin filter. I will then add phenolphthaleinphosphate to 7 test tubes and the phosphatase solution prepared. The test tubesmust then be placed in a 30 C water bath controlled and left to allow the reactionto take place. After 20 minutes I will take the test tubes out of the water bath,add 5cm of sodium carbonate solution to each test tube and mix. This solution