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Introduction:
The purpose of this experiment is to monitor the level of enzymatic browning of apple
slices. Apples contain an enzyme called polyphenol oxidase (phenolase). In the presence of
oxygen from the air, this enzyme catalyses the formation of brown pigments called melanins.
Treatment of the apple slices with ascorbic acid, citric acid, or acetic acid will reduce the
level of browning. The reduction in browning is dependent on the type of substance and its
concentration. Soaking in water alone will temporarily reduce the level of browning by
restricting the amount of oxygen in contact with the apple slice. When fruits or vegetables are
peeled or cut, enzymes contained in the plant cells are released. In the presence of oxygen
from the air, the enzyme phenolase catalyses one step in the biochemical conversion of plant
phenolic compounds to form brown pigments known as melanins. This reaction, called
enzymatic browning, occurs readily at warm temperatures when the pH is between 5.0 and
7.0. The presence of iron or copper can increase the rate of reaction. This can be easily
observed when fruit is cut with a rusty knife or mixed in a copper bowl. Bruising or other
injury to the plant tissue disrupts the arrangement of chemicals within the cells and allows
these chemicals to make contact with oxygen. This may lead to browning of uncooked fruits.
Enzymatic browning can be a significant problem, limiting the shelf life of many fruits and
vegetables which have had little heat applied during processing. However, enzymatic
browning is not always a defect. The browning reaction contributes to the desirable colour
and flavour of raisins, prunes, coffee, tea, and cocoa.
Objective:
The monitor the level of enzymatic browning of apple slices.
Materials/Apparatus:
Fresh fruit/Vegetable, (Apple)
1% ascorbic acid
1% citric acid
1% acetic acid
2% acetic acid
Beaker or cup
Paper towel
Tong
Glove
Knife
Stopwatch or timer
Procedure:
1. The fruit (apple) was slice into 6 pieces with same size.
2. The unheated fruit slice was place on a paper towel. The towel was label as control.
3. Another apple slice was dip into one of the test solution (1% ascorbic acid) using tong
for 10 minutes.
4. The tong was rinse and repeat the same procedure for the other three solutions (1%
citric acid, 1% acetic acid, 2% acetic acid).
5. The one apple slice was soak in water for 10 minutes, the apple slice was place it on a
paper towel and label the towel water soak.
6. The time and level of browning in table was recorded. The slices every 10 minutes for
2 hours and record your observations.
Results:
Level of Browning
Time (min)
Untreated
(control)
Ascorbic
acid (1%)
Citric acid
(1%)
10
20
30
40
50
60
70
80
90
100
110
120
Key
5 = Completely dark brown
4 = Fully covered light brown
3 = Half-covered light brown
2 = Slight or scant brown patches
1 = No browning present
Discussion:
Water
soaked
Conclusion:
In conclusion, recent researches have shown that the browning of apples can be
prevented or inhibited using a combination of combinations, either natural products or their
derivatives that are food additives. Various type combinations of enzymatic inhibitors,
reducing agents and anti-microbial compounds containing Ca2+ were tested. They all slowed
down microbial decay and changes in organic acids and sugars during storage. It was also
found that irradiating gamma rays could reduce rate of browning of apple juice and increase
anti-oxidant activity. This is found to have importance especially in improving the shelf-life
of food. More evidently, fresh-cut fruits could now be preserved longer without the need of
special packaging.
Questions:
1.
What causes browning when fresh fruits and some vegetables are peeled or cut?
The browning is caused by the conversion of phenolic compounds to melanins
catalysed by the enzyme polyphenol oxidase.
2.
3.
How do food additives or treatment processes in use today prevent or retard browning
in fruits and vegetables?
Sulfites will inhibit melanin formation. Ascorbic acid (vitamin C) will prevent
oxygen from reacting with the polyphenol oxidase. Citric acid and acetic acid will
lower the pH, which decreases the enzyme activity. Citric acid also ties up copper
ions and prevents them from participating in the reaction. Heating process inactivates
the polyphenol oxidase.
4.
References:
1. http://www.sciencedaily.com/releases/1999/01/990113074812.htm.com
2. http://www.nps.ars.usda.gov/publications/publications.htm.com.?
SEQ_NO_115=122938
3. Polyphenol Oxidase". Worthington Enzyme Manual. Retrieved 13 September 2011.
4. Mayer, AM (November 2006). "Polyphenol oxidases in plants and fungi: Going
places? A review". Phytochemistry 67 (21):
23182331. doi:10.1016/j.phytochem.2006.08.006.PMID 16973188.
5. Questions and Answers: Okanagan Specialty Fruits Non-Browning Apple (Events
GD743 and GS784)" (PDF). APHIS. Retrieved 18 March 2014.
6. Milkovich, M. "Non-browning apples cause controversy" Fruit Growers News. April
29, 2011.
References:
1. http://www.sciencedaily.com/releases/1999/01/990113074812.htm
2. http://www.nps.ars.usda.gov/publications/publications.htm?SEQ_NO_115=122938