Exercise 6C (Oxidases and Peroxidases)

OXIDASES AND
PEROXIDASES:
SOME ENZYMES INVOLVED
IN METABOLISM
Lazaro, Miciano, Pobre, Villanueva

OBJECTIVES
 To determine what oxidases and peroxidases are
 To explain why browning in fruits/vegetables occur
 To show that browning can be prevented by some
solutions or conditions
 To determine the enzymes involved in plant
oxidation
METHODOLOGY
• Peel the fruit/vegetables (apple,

cucumber, potato)
• Cut 12 strips of each fruit/

vegetable
• Put 3 strips of each type in the

following conditions
CONDITIONS
A. open petri B. 1% ascorbic C. boiled in D. immersed in

plate acid solution distilled water distilled water
for 3 min.
Set aside after Set aside

boiling
• Observe after
30 min.
• Record results
SETUP
RESULTS
A. Open Petri Plate
Observation: specimen had a discoloration
B. 1% ascorbic acid solution
Observation: no discoloration of specimen
C. Boiled in distilled water
D. Immersed in distilled water

Summary of Results:
OPEN PETRI 1% BOILED IN IMMERSED IN
PLATE ASCORBIC DISTILLED DISTILLED
ACID WATER WATER
SOLUTION
Apple
Potato
Cucumber
DISCUSSION
Why did the fruits and vegetables turn
brown?
 The cells of some fruits and vegetables
contain an enzyme called polyphenol
oxidase or tryosinase that reacts with
oxygen in the air.
 When in contact with oxygen, the

enzyme catalyzes one step of the
biochemical conversion of plant
phenolic compounds to brown pigments
known as melanins.
Why did ascorbic acid prevent browning
and discoloration of fruits and vegetables?
 The enzyme activity is pH dependent.

Lowering of the pH to 4.0 by the
addition of citric, ascorbic or other acids
inhibits the enzyme activity.
 Ascorbic acid acts as an antioxidant
(inhibit oxidation)
 It makes fruits or vegetable more acidic
which prevents enzymatic browning
Why did boiling prevent browning and
discoloration of fruits and vegetables?
 Heat destroys enzymes that promote

oxidation
 At temperatures around boiling, the
chemical bonds that hold together the
structure of enzymes begin to break
down.
 The resulting loss of three-dimensional
structure causes enzymes to no longer
fit their target substrate molecules,
and enzymes entirely stop functioning.
Why did boiling affect the texture of
fruits and vegetables?
 Heating in boiling water also affect the
product’s texture
 It is softer after boiling because heat breaks

down cell wall
Why did immersing in water prevent browning
or discoloration of fruits and vegetables?
 Immersing them in water blocks out

atmospheric oxygen. There's
oxygen in the water, but it's mostly
tied up with hydrogen and not
available to react with the enzymes
and phenolic compounds.
DISCUSSION
 Two Types of Browning Reaction:
(i) Non enzymatic browning in which no

enzymes are involved to bring about the color
changes. The two main forms include
caramelization and the Maillard reaction.
DISCUSSION
(ii) Enzymatic browning in which enzymes are involved
 Polyphenols- also called phenolic compounds, are

group of chemical substances present in plants (fruits,
vegetables) which play an important role during
enzymatic browning, because they are substrates for
the browning-enzymes.
 Polyphenol oxidase (PPO)- a substrate from a family

of chemicals called polyphenols
DISCUSSION
 Oxidation Catalysts
Enzymes that promote oxidation or browning in fruits
and vegetables:
 Oxidases- acting on oxygen
 Peroxidases- acting on naturally occurring

phenols
How does enzymatic browning work?
The products of this chemical reaction cause the brown color. The
PPO and polyphenols are present in the plant cells and are
released when the cells are cut or mashed. The oxygen comes
from the air.
GUIDE QUESTIONS
1. Why do some fruits and vegetables become
brown when exposed to air?
Answer:
Apples and other fruit turn brown once the cut surface is
exposed to air due to a reaction with oxygen in the
surrounding air, a chemical reaction called oxidation .
GUIDE QUESTIONS
2. How can you prevent the browning of some fruits
and vegetables?
Answer:
The reaction can be slowed or prevented by inactivating
the enzyme with:
 Heat (cooking)
 Reducing pH on the surface of the fruit
Brushing the surface of the fruit Soak in Acidulated Water

with juice from citrus fruits
 Putting fruit underwater
vacuum packaging
putting fruit underwater
 Reducing the reaction rate
Refrigeration can
slow enzymatic
browning. At temp.
below 7˚C, the PPO
enzyme activity is
inhibited. Home
refrigerator are
usually between 3˚C
and 5˚C
By storing in the refrigerator
 By adding certain preservative chemicals

GUIDE QUESTIONS
3. What organelle is responsible for respiration in
plants?
Answer: MITOCHONDRIA
Mitochondria is
the organelle responsible
for cellular respiration by
converting the energy
stored in glucose into ATP.
GUIDE QUESTIONS
4. If oxygen is not available, will cellular respiration take place
in plants? Explain your answer.
Answer: Yes, cellular respiration will still take place. The oxygen supply
to plants can also run out, this happens for example if the soil gets
waterlogged. In this case they have to obtain their energy via
anaerobic respiration. Below is the word and chemical equation
for anaerobic respiration in plants:
 Glucose Ethanol + Carbon dioxide + Energy
 C6H12O6 2C2H5OH + 2CO2 + Energy
When the above reaction occurs in yeast cells it is referred to

as fermentation. Fermentation is the process used for baking bread and brewing
alcohol.
SOURCES
 http://www.food-
info.net/uk/colour/enzymaticbrowning.htm
 http://classroom.synonym.com/effects-boiling-
freezing-enzyme-activity-23207.html
 http://nieonline.com/downloads/fighting_with_food
/keeping_it_fresh.pdf
 seplessons.ucsf.edu
 www.education.com

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OXIDASES AND

Lazaro, Miciano, Pobre, Villanueva

• Peel the fruit/vegetables (apple,

• Cut 12 strips of each fruit/

• Put 3 strips of each type in the

A. open petri B. 1% ascorbic C. boiled in D. immersed in

Set aside after Set aside

Observation: no discoloration of specimen

 When in contact with oxygen, the

 The enzyme activity is pH dependent.

 Heat destroys enzymes that promote

 It is softer after boiling because heat breaks

 Immersing them in water blocks out

(i) Non enzymatic browning in which no

(ii) Enzymatic browning in which enzymes are involved

 Polyphenols- also called phenolic compounds, are

 Polyphenol oxidase (PPO)- a substrate from a family

 Oxidases- acting on oxygen

 Peroxidases- acting on naturally occurring

Brushing the surface of the fruit Soak in Acidulated Water

 By adding certain preservative chemicals

 C6H12O6 2C2H5OH + 2CO2 + Energy

When the above reaction occurs in yeast cells it is referred to

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