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Enzyme Lab Let’s Juice it Up Academic Biology 2016-17 Learning Target: Enzyme Lab Activity I can… Conduct an experiment to identify the most cost effective enzyme for apple juice production from filtered applesauce. I know I am successful when I… Identify the question being asked or problem that is trying to be solved by this experiment. Conduct an experiment to answer the question. Identify the independent, dependent, and controlled (constant) variables along with the experimental control. Draw a conclusion that answers the original question based on the data and validity of the experimental design. Introduction: A Connecticut company is in the business of making and selling apple juice. To make apple juice, apple sauce is strained through filters to remove the juice. The company would like your help in testing the effect of different enzymes on the production of the apple juice. You will investigate the ability of these enzymes to remove more juice during the process of filtration and decide the most cost effective plan to increase juice production. The following is a list of the enzymes along with their prices: Pectinase: Cellulase: $100.00 per liter $50.00 per liter Background Information: Enzymes are proteins that catalyze chemical reactions (metabolism) in the cell of all living organisms. Enzymes control many vital functions in the cell, including the release of energy during the breakdown of nutrients into smaller molecules (catabolism) and the synthesis of complex cell materials (anabolism) from small molecules. In this lab you will work with two plant enzymes – pectinase and cellulose. The enzyme pectinase breaks down pectin. Pectin is a complex carbohydrate that is part of the plant cell wall. Pectin acts like “glue,” holding plant cell walls together. Pectin is soluble in water, and in a mild acidic environment it becomes sticky. These properties make pectin very useful in the production of jams and jellies. When the enzyme pectinase is added to mashed fruits it breaks down the pectin in the fruit cell walls, thus facilitating the industrial production of fruit juices. The enzyme cellulase breaks down cellulose. Cellulose is a polymer made out of long branching chains of glucose and it is one of the main components of plant cell walls. Cellulose accounts for about 50 percent of all the organic materials on Earth. Unfortunately, humans, like all other mammals, do not synthesize the enzyme cellulase and therefore can’t digest cellulose. Scientists purified the enzyme cellulase and currently it is used in the food industry for the production of wine and juices. The enzyme is also used in the production of plant-based materials such as paper, light basswood, rayon fibers and photographic films. How It’s Made: Apple Juice video https://youtu.be/Vk_ufH6je84 Your Task: You and your two lab partners will conduct an experiment to determine which enzyme or combination of the two enzymes maximizes apple juice production. Once you complete the experiment, you will analyze your data and evaluate the experimental design to determine the most cost effective use of enzymes in apple juice production. Experimental Design Materials: Apple sauce Pectinase enzyme Cellulase enzyme Paper/plastic funnels Filter paper Access to a stopwatch, or clock Stirring rods/spoons Graduated cylinder Access to tap water Paper/plastic cups Droppers Access to a balance Paper towels for cleanup Beaker A Procedure: C B Figure 1. 1. 2. 3. 4. 5. Label 4 separate cups (first cup A, second B, third C and fourth D) (Figure 1) Place cup A on electronic scale and reset to zero (Figure 2). Add applesauce to the cup until measures 50 g. Remove from scale. Add 10 drops of water to cup A (Figure 3). Stir cup A for 10 seconds, then let sit 50 seconds (total of 60 seconds or 1 minute). 6. Transfer cup A applesauce into paper filter and funnel over collection (graduated cylinder) and start timing (Figure 4). 7. Let applesauce filter for 2 minutes collecting juice in the clean graduated Figure 2. cylinder (Figure 4). 8. When 2 minutes are up, discard applesauce, filter and paper funnel in trash receptacle. 9. Measure amount of apple juice in milliliters (ml) produced in graduated cylinder and record in Data Table 1. 10. Rinse out graduated cylinder in sink and thoroughly drain out excess water (to avoid source of error in measuring). 11. Repeat steps 2 and 3 for cup B. 12. Add 10 drops of Pectinase enzyme to cup B. 13. Repeat steps 5 – 10. 14. Repeat steps 2 and 3 for cup C. 15. Add 10 drops of Cellulase enzyme to cup C. 16. Repeat steps 5 – 10. 17. Repeat steps 2 and 3 for cup D. 18. Add 5 drops of Pectinase and 5 drops of Cellulase enzymes to cup D. 19. Repeat steps 5 - 10 20. Clean up lab station according to check-off list. 21. Have Mrs. Kapper inspect your lab station. 22. Record your collected data in entire class data table. D A cup 10 drops A Figure 3. Figure 4. Data Collection Data Table 1: Amount of Apple Juice Produced (ml) 50 g Applesauce Cup and Added Enzyme(s) Amount of Apple Juice Produced (ml) Cup A (10 drops water) Cup B (10 drops pectinase) Cup C (10 drops cellulase) Cup D (5 drops Pectinase + 5 drops cellulase) Graph: Class Averages - Amount of Apple Juice Produced (ml) Class Average Amount of Apple Juice Produced (ml)