Place a paper coffee filter in a funnel and then set the funnel in a 100 mL graduated cylinder, as shown in Figure 1, below, to filter the juice from one of the apple preparations. After removing the beakers from the water bath, you may want to use a wooden spoon to gently stir/squeeze the apple pieces in each. The water should come up to the level of the chopped apples, but you do not want so much water that the beakers float and tip over. Your water bath can be as simple as a styrofoam box or insulated cooler, with water at 40☌. Put both beakers into a 40☌ water bath for 25–30 minutes.Stir the chopped apple pieces in each beaker with a separate plastic spoon.Using masking tape and a permanent marker (or sticky notes and a pen), label the beakers ("pectinase" and "water"). Using a different, clean pipet or syringe, add 4 mL of distilled water to the other beaker. Using a pipet or 1 milliliter (mL) syringe, add 4 mL of diluted enzyme to one beaker.Prepare the pectinase enzyme according to the manufacturer's instructions.Use the balance to weigh equal amounts of chopped apple (about 50 grams ) into each beaker.(Use care with the knife!) It is important to chop the apple into very small pieces-added surface area helps the enzyme break down the pectin in the plant cell walls, releasing more juice. Chop the apples into cubes that are roughly 5 millimeters (mm) on a side.The concentration of pectinase used will be much higher than is used in commercial juice production, and the fruit and enzyme have not been handled aseptically. Do not drink or cook with the juice produced in this experiment. Water bath can be as simple as a styrofoam box or insulated chest, big enough to hold the beakers.Disposable plastic spoons for stirring (2).Masking tape and permanent marker or sticky notes and a pen.Balance for weighing out apple pieces, such as the Fast Weigh MS-500-BLK Digital Pocket Scale, 500 by 0.1 G available at.You will also need to gather these items: Alternatively, 1 mL syringes or pipets may be used. You may also be able to find it locally at a store with home winemaking supplies. Alternatively, you could use clean glass jars. These items can be purchased from Carolina Biological Supply Company, a Science Buddies Approved Supplier: "Pectin," Wikipedia, The Free Encyclopedia. "Enzyme," Wikipedia, The Free Encyclopedia. "Cell Wall," Wikipedia, The Free Encyclopedia. "In a Jam and Out of Juice," National Centre for Biotechnology Education, University of Reading, U.K. What are the substrates for the enzymes cellulase and amylase? Do you think that these enzymes would increase juice production?.How does pectinase promote the release of juice from apples?.What is the source of pectinase enzyme?.Some possible conditions to investigate are duration of enzyme treatment, enzyme concentration, and temperature. You can monitor the enzyme activity by measuring how much juice is produced under the various experimental conditions. In this project, you can apply pectinase to fruit under controlled experimental conditions in order to investigate the enzyme activity of pectinase. Pectinase is also used for clarifying the extracted juice. By enzymatically breaking down the cell wall, pectinase releases the juice from within the cells. Pectinase is used commercially to aid in extracting juice from fruit. Pectinase is an enzyme that catalyzes the breakdown of pectin, a component of the cell wall in fruits such as apples and oranges. Enzymes are proteins that catalyze (speed up) specific chemical reactions-increasing reaction rates by factors of at least a million. Enzymes are the workhorses of biochemistry.
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