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Project Summary

Difficulty	4
Time required	Very Short (a day or less)
Prerequisites	None
Material Availability	Readily Available
Cost	Very Low (under $20)
Safety	Adult supervision recommended for making Jell-O (uses boiling water).

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Objective

The purpose of this project is to test the hypothesis that food items containing a protein-digesting enzyme will prevent gelatin from solidifying.

Introduction

If you like making jello for dessert, you may have noticed that the box recommends against adding certain kinds of fresh or frozen fruit, e.g., pineapple, kiwi, mango, ginger root, papaya, figs, or guava. This is because these fruits contain the protein-digesting enzymes papain or bromelain that can prevent the gelatin from setting.

So why would a protein-digesting enzyme interfere with the setting of gelatin? To find out, you should do some background research on the ingredients that go into gelatin, and how it normally sets (see Howstuffworks, 2006, for a good start).

Although papain and bromelain can digest proteins, these enzymes themselves are proteins. Most proteins can be inactivated by denaturation. Denaturation changes the structure of the protein, without breaking the chemical bonds between the amino acids that make up the protein backbone. Exposure to heat is one method of denaturing proteins. A good example is cooking an egg. When the egg is raw, the egg white—which has lots of proteins called albumins—is transparent and liquid, but after cooking it becomes opaque and solid. In the case of eggs (and most enzymes) denaturation of the protein causes an irreversible change. Most enzymes are inactivated by denaturation.

In this experiment, you will perform two tests.

• First, you will choose one or more of the fresh fruits from the list above to see if they prevent gelatin from setting.
• Second, in order to test if enzyme activity is responsible for the effect, you will also test the fruit(s) after cooking (to denature any enzymes present in the fruit).

With this experiment, you can have some of your results for dessert!

Terms, Concepts and Questions to Start Background Research

To do this project, you should do research that enables you to understand the following terms and concepts:

• gelatin,
• protein,
• enzyme,
• protease,
• papain,
• bromelain.

More advanced students should also study:

• proteolysis,
• protein denaturation,
• protein structure (primary, secondary, tertiary, quaternary).

Questions

• How do papain and bromelain digest proteins?

Bibliography

• Here's a good article on the ingredients in gelatin and how it sets:
  Howstuffworks, 2006. "What Exactly Is Jell-O Made From?" Howstuffworks.com [accessed July 11, 2006] http://home.howstuffworks.com/question557.htm.
• Wikipedia contributors, 2006a. "Papain," Wikipedia, The Free Encyclopedia [accessed July 11, 2006] http://en.wikipedia.org/w/index.php?title=Papain&oldid=63703978.
• Wikipedia contributors, 2006b. "Denaturation (Biochemistry)," Wikipedia, The Free Encyclopedia [accessed July 11, 2006] http://en.wikipedia.org/w/index.php?title=Denaturation_%28biochemistry%29&oldid=61308564.

Materials and Equipment

To do this experiment you will need the following materials and equipment:

• dessert gelatin mix (e.g., Jell-O),
• clear plastic cups,
• fresh fruit such as pineapple, kiwi, mango, ginger root, papaya, figs or guava.
• Optional: you can try papain tablets, available at vitamin/health food stores, as a positive control.

Experimental Procedure

Safety Note. Preparing Jell-O involves pouring and stirring boiling hot water. Adult supervision or assistance is highly recommended.

1. Determine how many different fruits you will be testing, then calculate the number of cups of gelatin to make. For each fruit tested, you should make at least four cups of gelatin: 2 for uncooked fruit and 2 for cooked fruit. You will also need 2 extra cups for plain gelatin (as a negative control, to make sure that gelatin alone sets properly).
2. Note: if you want to try different amounts of each fruit, you will need 4 cups for each amount tested (again, 2 cups for cooked fruit, and 2 cups for uncooked fruit). You will also need extra cups for ground-up papain tablets, if you are using them.
3. Label the cups: "fresh" or "cooked", type of fruit, amount of fruit.
4. Make the Jello-O according to the instructions on the box.
5. For each type and amount of fruit to be tested, add fresh (uncooked) fruit to two cups.
6. For each type and amount of fruit to be tested, add fruit that you have cooked (boil or steam for 5 minutes) to two cups. Use the same amount of cooked fruit as you used for the fresh fruit.
7. Optional: grind up two papain tablets and dissolve in a small amount of water. Add equal parts of this solution to two cups of gelatin as a positive control. Mix well.
8. Finally, add gelatin only to two cups. This is your negative control (to make sure that gelatin alone sets properly).
9. Refrigerate all of the cups, noting the time.
10. Check the consistency of the gelatin in each cup at regular intervals (once or twice an hour). Examine the gelatin carefully and record your observations in your lab notebook. In which conditions does the gelatin set? In which conditions does the gelatin remain liquid? Are there any in-between cases?

Variations

• Does freezing fruit denature protein-digesting enzymes? Design an experiment to compare the enzyme activity of fresh and frozen pineapple.
• Do other methods of fruit processing denature enzymes? How about drying or canning? Design an experiment to find out.
• Design an experiment to test which type of fruit has the most enzyme activity per unit weight.
• Papain and bromelain are also the active ingredients in many meat tenderizers. Design an experiment to see if meat tenderizer can prevent gelatin from setting. Does the effect depend on how much tenderizer is added? Can meat tenderizer be denatured to neutralize the effect?
• Do research on alternative methods of protein denaturation, or other methods for inactivating enzymes. Choose one or more of these methods and see if it works on protein-digesting enzymes in fruit. (Don't eat the results of this experiment!)
• For other Science Buddies projects involving enzymes, see: Liver Stinks!, A Juicy Project: Extracting Apple Juice with Pectinase and Enzyme-Catalyzed Reactions -- What Affects Their Rates?.

• For more science project ideas in this area of science, see Cooking & Food Science Project Ideas.

Credits

Andrew Olson, Ph.D., Science Buddies

Sources

• Shields, R.K., 2003. "Jell-o or Gel-no: Which Fruits Contain a Protein-Digesting Enzyme that Prevents Gelatin from Solidifying?" California State Science Fair Abstract [accessed July 11, 2006] http://www.usc.edu/CSSF/History/2003/Projects/J0411.pdf.

Last edit date: 2006-07-17 23:00:00

Career Focus

If you like this project, you might want to think about career opportunities in Cooking & Food Science.

Good taste, texture, quality, and safety are all very important in the food industry. Food science technicians test and catalog the physical and chemical properties of food to help ensure these aspects. Learn more about this career: Food Science Technician.