EXERCISE 6:

ENZYMES

GROUP 3
ENZYMES
● Special proteins
● Catalysts for reactions
● absolute/relative
● reversible/irreversible
● competitive/uncompetitive/
noncompetitive
 HYDROLASES:
● Hydrolysis reaction
● Insoluble food storage materials
to soluble
TRANSFERASES:
● Donor to acceptor molecule
OXIDOREDUCTASES:
● Oxidation-reduction reactions
 LYASES:
● Removes or add groups to double
bonds
 ISOMERASES:
● Rearrangement of compounds
 LIGASES:
● Formation of bonds coupled with
breakdown of pyrophosphate
bond in ATP
A. Hydrolases
AMYLASE SUCRASE
A.a.

● Corn seedling root was source of amylase

● Hydrolyzed starch solution
● Iodine test for presence of starch
● Test tube 1 glucose monomers
 A.b.
● Corn seedling root was source of sucrase
● Hydrolyzed sucrose solution
● Benedict’s test as test for reducing sugars
● Brick red precipitate indicated positive
presence of monomer sugars (glucose/fructose)
from degraded sucrose
B. Oxidoreductases
 B. Oxidoreductases
Dehydrogenase Put
stopper Catalase Observe for any
gas evolution
Stand
overnight
Methylene Methylene 5 ml 3 %
blue 5 ml 3 %
blue H202 H202
Record
any
change

Aerate the
solution

Record
any
change

3 3
Fresh Boiled
Mungbe potato potato
an strips strips
sprouts
Dehydrogenase
Anaerobic Aerobic

Colorless With
Mungbean Blue

Without
Mungbean
Blue Blue
 Dehydrogenase
• The mung bean served as a source of dehydrogenase, an enzyme that
catalyzes reduction reaction
• Methylene blue:
*colorless, when reduced

*blue, when oxidized

• Upon exposure to Oxygen, the solution turned blue, indicating that an
oxidation reaction occurred
Catalase
Fresh potato
strips plus 3% Effervescence
H2O2

Boiled potato
strips plus 3% No effervescence
H2 O 2
 ● Fresh potato contains active catalase enzyme
● Catalase enzyme catalyzes the decomposition of hydrogen peroxide
to water and oxygen.
● Effervescence (bubble formation) indicates that Oxygen was released.
● Boiling (high temperature) denatures enzymes
● In the case of the boiled potato, the catalase enzyme was denatured.
Thus, no reaction occurred.
C. Factors Affecting Enzyme
Activity
a. Enzyme concentration
 C.a Enzyme concentration

● Higher concentration, higher frequency

of collision, higher rates of reaction between
enzyme and substrate
● Amylase catalyzes breakdown of starch
to glucose, thus lesser starch molecules in
higher amylase concentration,
less intense color in I2KI test.
b. Hydrogen in concentration (pH)
 C.b Hydrogen-ion
concentration (pH)

● Enzymes have an optimum

pH wherein they are most
active
● Enzymes denature at high
and low pH
● Amylase optimum pH is 7
c. Temperature
 C.c Temperature

● Enzymes have an optimum temperature

in which they are most active
● At low temperatures - enzyme inactivation
● At high temperatures - enzyme denaturation
● Amylase optimum temperature is 28-30 ⁰C
 Answers to guide questions
1. Give (i) the name of the enzymes catalyzing the ff. chemical reactions, (ii)
their cellular localization, and (iii) the plant physiological process they are
involved
● Pyruvate + NAD+ + CoA acetyl-coA + NADH + H+ + CO2

○ Catalyzed by: pyruvate dehydrogenase

○ Found in: mitochondrial matrix

○ Involved in: cellular respiration (pyruvate oxidation)

● Ribulose-1,5 bisphosphate + CO2 2 (3-phosphoglyceric acid)

○ Catalyzed by: RuBisCO

○ Found in: stroma

○ Involved in: Calvin cycle

 Answers to guide questions
● 1. (continuation)
● Fructose-6-phosphate + ATP fructose-1,6-bisphosphate

○ Catalyzed by: phosphofructokinase-1

○ Found in: cytosol

○ Involved in: glycolysis

 Answers to guide questions
2. Describe the rate of enzyme-catalyzed reaction with
increasing substrate concentration at constant enzyme
concentration.
-The rate of reaction will plateau when the enzymes are
saturated with the substrate
- increasing the concentration of
substrate won’t affect the rate of
reaction at point x
 Answers to guide questions
3. In what ways does hydrogen ion concentration affect
enzyme activity?
● Hydrogen ions are positively charged particles that interact with the
negatively charged parts of protein macromolecules

● It can change the shape of the enzyme which will then affect the active site
and disable the ability of the enzyme to interact with the substrate
 Sources

● http://www.namrata.co/factors-affecting-enzyme-activity-2/