Evaluation of the Effect of Fruit Enzymes on Browning

and Protein Degradation

Katherine Kern
Fruit Lab
NTR402L
October 14, 2016

INTRODUCTION
Fruit enzymes are responsible for both enzymatic oxidative browning and protein
degradation. Enzymatic browning occurs when an enzyme acts on a phenolic compound in the
presence of oxygen. As the pigments oxidize, the color changes from white to brown. These
enzymes are responsible for the browning that occurs in cut fruit. Different methods may be used
to reduce this enzymatic browning by interfering with the oxidation reaction. This reaction may
be inhibited by lowering the pH, preventing exposure to oxygen in air, and via application of
heat to denature the enzymes. Reducing enzymatic browning may help reduce food waste and
may be important in presentation of dishes. However, different treatments may have effect the
taste and texture of the product, other important considerations.
Fruits such as papaya, pineapple, kiwi, and figs contain proteolytic enzymes that degrade
protein. Papain is a proteolytic enzyme found in papaya, and is an ingredient in commercial meat
tenderizer (McWilliams, 2017). The enzyme destroys the sarcolemma of muscle fibers and
hydrolyzes various proteins contained in the fiber, resulting in tenderness (McWilliams, 2017).
Other proteolytic enzymes include bromelain found in pineapple, and ficin found in figs
(McWilliams, 2017). The degradation of protein caused by these enzymes may be desirable for
meat tenderization. However, it may be undesirable if the meat becomes mushy or if protein
structure must be maintained in dishes containing gelatin.
During the first part of the experiment, enzymatic oxidative browning will be evaluated
in samples of apples and bananas. Various treatments will be compared to a control for each fruit
to observe the treatments effect on browning. Treatments to be used are exposure to air,
blanching, pineapple juice, lemon juice, cream of tarter solution, and dry sucrose. Appearance,
taste, and texture will be evaluated for each sample. The goal of the experiment is to explain the

mechanism by which the treatments inhibit browning, and determine the treatment that both
inhibits browning and has the smallest negative effect on taste and texture. It is expected that
treatments that lower pH, prevent exposure to oxygen, or denature enzymes will reduce
browning but may negatively affect taste or texture.
For the second part of the experiment, the effect of proteolytic fruit enzymes on proteins
in gelatin will be examined. Samples of fresh pineapple, canned pineapple, and kiwi will be
placed in gelatin to determine whether enzymes in these fruits degrade the gelatin protein
structure. The goal of this experiment is to determine which fruits degrade the protein structure
of the gelatin. It is expected that both fresh pineapple and kiwi will degrade protein structure, but
that canned pineapple will not due to denaturation of enzymes during the canning process.
The purpose of these laboratory experiments was to evaluate the effects of fruit enzymes
and demonstrate these effects via observation of browning of cut fruit and degradation of gelatin.
MATERIALS AND METHODS
Enzymatic Oxidative Browning
This experiment was completed by following the lab procedures in Unit 9, Part A of the
Lab Manual for Browns Understanding Food Principles and Preparation, 5th edition (Walter &
Beathard, 2015). Treatment solutions were prepared before cutting fruit. Samples of apples and
bananas were used. The following ingredients were omitted: Fruit-Fresh, ascorbic acid solution,
and sucrose solution. Modifications to the procedure included omission of steps 7, 8, 10, and 12.
The only treatments used were exposure to air, blanched, pineapple juice, lemon juice, cream of
tarter solution, and dry sucrose. Each sample was evaluated based on appearance, taste, and
texture and results were recorded in a table.
Degradation of Protein

For this experiment, four packages of Jell-O were prepared in separate according to
directions on box and cooled until partially set. While Jell-O was cooling, fresh pineapple and
kiwi were cut and a sample of canned pineapple was obtained. After Jello was partially set, fresh
pineapple was added to the first bowl, canned pineapple to the second, fresh kiwi to the third,
and no fruit was added to the last bowl to serve as a control. The bowls were returned to the
cooler to finish setting. After setting, the four samples were evaluated for appearance and results
recorded in a table.
RESULTS
Enzymatic Oxidative Browning
For the samples of apple, all treatments reduced the appearance of browning to various
degrees when compared to the apple exposed to air only. Lemon juice treatment was observed to
have the least amount of browning. When compared to the apple exposed to air, all treatments
had some effect on either taste or texture except for the cream of tarter solution. Results of the
evaluation of appearance, taste, and texture for the apple samples are summarized in Table 1.
Table 1. Results of Evaluation of Apple Samples
Treatment
pH
Appearance
Exposed to Air
n/a
Very brown
Blanched
n/a
No browning
Pineapple Juice
4
Minimal
browning

Taste
sweet
sweet
Sweet,
noticable
pineapple
flavor
very sour

Texture
crisp
soft, mushy
firm

Lemon Juice

No browning

Cream of tartar solution

Minimal
browning

sweet

slightly
crisp
firm

Dry Sucrose

n/a

Some browning
in a few areas

very sweet

not crisp

For the samples of banana, all treatments reduced the appearance of browning to various
degrees when compared to the banana exposed to air only. Lemon juice and dry sucrose
treatments were observed to have the least amount of browning. When compared to the banana
exposed to air, all treatments had some effect on taste. All had some effect on texture except for
the pineapple juice and cream of tarter solution treatments. Results of the evaluation of
appearance, taste, and texture for the banana samples are summarized in Table 2.
Table 2. Results of Evaluation of Banana Samples
Treatment
pH
Appearance
Exposed to Air
n/a
Very brown
Blanched
n/a
Some browning
Pineapple Juice
4
Minimal
browning
Lemon Juice

No browning

Taste
sweet
less sweet
sweet,
noticeable
pineapple
flavor
sour

Cream of tarter solution

Minimal
browning

sweet, slight
bitter aftertaste

Dry Sucrose

n/a

No browning

more sweet

Texture
soft
very mushy
soft

slightly
mushy
soft
slightly
mushy

Degradation of Protein
The four bowls of Jell-O were evaluated for appearance. The three bowls with fruit added
were compared to the fourth bowl of plain Jell-O, which served as a control. The plain Jell-O and
the Jell-O with canned pineapple both remained gels. No liquid was observed on the surface or
around the pieces of canned pineapple. The Jell-O with fresh pineapple and the Jell-O with fresh
kiwi were both observed to be watery on the surface around the pieces of fruit.
DISCUSSION
As illustrated in Table 1 and Table 2, the cut apple and banana became brown when
exposed to air. This is explained by the action of polyphenoloxidases, a group of enzymes that

oxidize anthoxanthins (McWilliams, 2017). Anthoxanthins, which are white pigments, become
brown when oxidized (McWilliams, 2017). The treatments inhibit browning by inhibiting the
reaction. Through the application of heat, blanching denatures enzymes. The pineapple juice,
lemon juice, and cream of tartar solution had a pH of 4, 2, and 4, respectively (Table 1, Table 2).
Acid inhibits the enzyme action, thus reducing browning (McWilliams, 2017). The dry sucrose
draws water to the cut surface via osmosis, and the water provides a barrier to the oxygen in air.
Eliminating oxygen also inhibits the reaction and reduces browning (McWilliams, 2017).
Although effective at reducing enzymatic browning, some undesirable changes in taste
and texture were observed as expected. Notably, the pineapple juice imparted a fairly strong
pineapple flavor, the lemon juice a very sour taste, and the dry sucrose a sweeter taste to both the
apple (Table 1) and banana (Table 2). The cream of tarter solution had an undesirable bitter
aftertaste for the banana only (Table 2). Many of the treatments caused the texture of both the
apple and banana to become softer, even mushy at times. These effects on taste and texture
should not be ignored as they may make a food product less appealing or unacceptable.
As stated in the results of the degradation of protein in Jell-O, the Jell-O with fresh
pineapple and fresh kiwi became watery. As expected, the protein structure of the gelatin was
broken down. This is explained by the action of proteolytic enzymes in these fruits. These
enzymes hydrolyze the protein structure in the gelatin, and as the structure is broken down the
gel cannot be maintained. The liquid observed around the fruit is evidence of this enzyme action.
As expected, there was no protein degradation observed in the Jell-O with canned pineapple.
This indicates that the proteolytic enzymes have been denatured in the canned pineapple and so
the hydrolysis reaction cannot occur, leaving the gelatin structure intact.

In conclusion, enzymes present in fruit can have a significant impact on food products.
Enzymatic browning that occurs may be unappealing to people, and treatments used to prevent it
may also have unappealing affects. However, this oxidation reaction is sometimes desirable, as it
is during the production of black tea (McWilliams, 2017). Proteolytic enzymes in fruits may
have undesirable effects on protein in some foods such as gelatin, but they are also useful for
tenderizing meat. An understanding of these enzymes and their reactions is important to the
application of food science principles so a desirable product can be produced. In addition,
improper application of these principles could increase food waste due to the production of
undesirable food.

References
Walter, J. M., & Beathard, K. (2015). Lab Manual for Browns Understanding Food Principles
and Preparation (5th ed.). Stamford, CT: Cengage Learning.
McWilliams, M. (2016) Foods: Experimental Perspectives (8th ed.). Upper Saddle River, NJ:
Pearson.