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Evaluating Fruit Enzymes: Effects of Oxidative Enzymes on Fruit Appearance, Taste and Texture and

Effects of Proteolytic Enzymes on Gelatin

NTR 502L
Selina LaVista
October 18, 2016
A wide variety of fruits and vegetables are available in the marketplace in fresh, canned, frozen
and dried forms, (Walter & Beathard, 2015, p. 105). When preparing foods with fruit ingredients it is
important to determine which of these varieties is the most appropriate. This decision can be based on

flavor, texture, appearance and also enzymes. Enzymes are proteins that are able to catalyze a chemical
reaction. When preparing foods with enzymes the reactions they cause may be favorable or unfavorable
depending on the circumstances (McWilliams, 2017). There are two types of fruit enzymes that will be
discussed in this laboratory experiment, oxidative enzymes and proteolytic enzymes.
Oxidative enzymes that are present in fruits and vegetables are polyphenolases, which are
responsible for oxidative browning. This is the oxidation of colorless polyphenolic compounds called
anthoxanthins found in fruits and vegetables (McWilliams, 2017). This reaction often occurs when the
polyphenolic compounds are exposed to oxygen by means of cutting or bruising of the fruit or vegetable
(Walter & Beathard, 2015). During this experiment, different preparation techniques were used in
attempt to preserve the color of the fruit, bananas and apples. The preparations used were as follows:
blanch the fruit, coat in pineapple juice, coat in lemon juice, coat in sucrose and coat in a mixture of
cream of tartar and water. The goal was to determine if each technique prohibited the oxidative enzymes
while preserving the flavor and texture of the fruit. This was evaluated by judging the appearance, texture
and flavor of the fruit. Lowering of the pH, addition of an antioxidant, immersion in a dilute sodium
chloride solution or sugar solution, and blanching are all expected to inhibit oxidative browning (Walter
& Beathard, 2015).
Proteolytic enzymes hydrolyze or breakdown various proteins. Some proteolytic enzymes such
as bromelain in pineapple are denatured when they are cooked while others are activated at higher
temperatures (McWilliams, 2017). For the purpose of this experiment, the goal is to evaluate the effect
of pineapple and kiwi on JELL-O strawberry gelatin dessert. Both fresh and canned pineapple were used
to address the effects of cooking on the enzymes. It is expected that the canned pineapple will not
hydrolyze the gelatin while the fresh pineapple and kiwi will. The purpose of this experiment is to
evaluate both the fruit enzymes discussed as they relate to oxidative browning and proteolytic degradation
of protein.

The enzymatic oxidative browning procedure was completed by following Unit 9, Lab Procedure
A-Enzymatic Oxidative Browning on page 107, from Understanding Food Principles and Preparation,
5th edition (Walter & Beathard, 2015). Exceptions to the procedures by Walter and Beathard (2015) are as
follows: the ascorbic acid solution, step 7 was omitted; the dry ascorbic acid solution, step 8 was omitted;
sucrose water solution, step 10 was omitted; the plastic wrap procedure, step 12 was omitted; the potato
and lettuce were omitted; only one type of apple, tart, was used. The cream of tartar solution made was
one teaspoon of cream of tartar for one cup of water. The fruits used were apple and banana. For each
procedure, the appearance, texture and flavor were evaluated with the exception of the cream of tartar
flavor and texture and the blanched banana flavor and texture.
For the proteolytic enzyme procedure the directions on a three-ounce box of strawberry JELL-O
were followed as per Dr. FitzPatricks instruction. Four bowls of gelatin were prepared: one control, one
with canned pineapple, one with kiwi and one with fresh pineapple. The warning on the box to avoid
fresh or frozen pineapple, kiwi, gingerroot, papaya, figs or guava was disregarded. Once the JELL-O was
prepared the bowls were briefly placed in the freezer to speed up gelatinization. After the JELL-O was
partially gelatinized the fruit was added. The JELL-O with fruit and control were observed shortly after
to determine the appearance of each.

The results of the enzymatic oxidative browning procedures can be found in the tables below.
Table 1-A articulates the results found for the apple. The most notable result would be the change in
texture associated with the blanched, pineapple juice and cream of tartar preparations. The appearance
for the most part remains unchanged except for the apple exposed to air.

Table 1-A: Enzymatic Oxidation Browning of Apple

Exposed to air
White, unchanged
Pineapple juice, pH= 4

Not browned, very slightly yellow

Sweeter, a bit of
pineapple flavor

Soft, mushy

Lemon juice, pH=2.5

Cream of tartar solution,

White, unchanged

Very sour

Slightly softer
Crunchy, slightly

White, unchanged
White, unchanged

Slightly sweeter

Slightly grainy

The results for the banana oxidation are presented in Table 1-B. The texture changes associated
with the banana were more drastic than the apple. The pineapple juice, lemon juice, cream of tartar and
dry sucrose preparations all resulted in a slimy texture. It is also important to note the browning that
occurred in the blanched, cream of tartar and dry sucrose preparations that did not occur for the apple.
Table 1-B: Enzymatic Oxidation Browning of Banana
Exposed to air
Pineapple juice, pH= 4
Lemon juice, pH=2.5
Cream of tartar solution,



Mostly white, a little browned

in the center
Not browned, very slightly
White, unchanged

Sweeter, a bit of pineapple
Very sour


Slightly browned, rigid edges

Slightly browned

Slightly sweeter


Slightly slimey

Table 2 describes the results of the proteolytic enzyme effects on JELL-O. The canned pineapple
did not change the appearance of the JELL-O while the fresh pineapple and kiwi both broke down the
gelatin and exposed liquid. The fresh pineapple had already begun to fall into the gelatin while the kiwi
just made some breaks in the surface and exposed liquid.
Table 2: Protelytic Enxyme Affect on Jello

Control, no fruit
Canned pinapple added
Fresh pineapple added
Fresh kiwi added

Solid gel
Very firm surface, with slight indentations from fruit
Surface breaking down, fruit sinking in, some liquid exposed
Surface breaking down, some liquid exposed

The relationship between an enzyme and its specific substrate in food catalyzes a chemical reaction
that causes a change to occur in the food (McWilliams, 2017). For the purpose of this experiment, those
reactions were evaluated relating to oxidative browning and proteolytic degradation of gelatin.


predicted many of the preparations used in the experiment prevented the oxidative browning to occur
especially in the apples, as can be seen in Table 1-A and Table 1-B. Blanching is used to denature the
proteins, which inactivate the enzymes however, this process did not work for the bananas but did for the
apples (McWilliams, 2017). It is possible that the boiling water bruised the tender banana as the apple is
more firm and would hold up better. The cream of tartar solution and dry sucrose also resulted in a slight
browning of the banana, Table 1-B. The pH of the cream of tartar was the same as the pineapple juice so
it can be concluded that the pH of four would be acidic enough to inhibit oxidative browning. It is possible
that human error occurred when the cream of tartar solution was made which could have caused the
browning. The same goes for the dry sucrose solution. It is possible that the banana was not thoroughly
coated with sucrose and still exposed to oxygen.
It can be determined that the predictions made for the proteolytic experiment were accurate.
Canned pineapple has undergone cooking proving that the bromelain, proteolytic enzyme, is in fact
denatured at a heat of between 77 and 82 degrees Celsius (McWilliams, 2017). Therefore, the canned
pineapple had little to no effect on the JELL-O as per Table 2. The fresh pineapple and kiwi both hydrolyzed
the gelatin releasing some liquid, Table 2. It appeared that the fresh pineapple was falling into the gelatin
more so than the kiwi but that could be due to the weight of the pineapple.

The findings of this experiment can be applied often in the foodservice, food science and nutrition
industries. When recommending or making a fruit salad, being able to prevent oxidative enzymatic
browning is crucial for the salad to have a vibrant and desirable appearance. Many would avoid a fruit
salad that looks browned. Knowledge of the changes in texture related to oxidative inhibition techniques
would help to determine which preparation would be best to use. Creating a crunchy fruit salad that is not
too tart would be ideal. Tables 1-A and 1-B could help determine which preparations would preserve the
texture and flavor of the fruits. The proteolytic enzymes are not only important for gelatin preparation but
also for meat tenderizing. Proteolytic enzymes are often used to increase tenderness in tougher cuts of
meat. Some of these tenderizers include using fresh pineapple in a marinade, papaya and salt as a rub and
ficin, an enzyme found in figs (McWilliams, 2017).
In conclusion, some fruits contain oxidative enzymes that cause browning while others may contain
proteolytic enzymes that cause degradation of proteins. It is important to be aware of these enzymes in
fruits and vegetables when preparing them or including them in recipes. The results of these enzymes can
be desirable or not depending on the product but the enzyme activity should always be considered when
preparing foods (McWilliams, 2017).


Walter, J.M. & Beathard, K. (2015). Understanding Food Principles and Preparation (5th ed.). Stanford,
CT: Cengage Learning.
McWilliams, M. (2017). Foods: Experimental Perspectives (8th ed.). Upper Saddle River, NJ: Pearson
Education, Inc.