Exercise 6

ENZYMES
Results and Discussion
Name: Reymound S. Desdechado Section: HJK

Date Performed: Dec. 4-5, 2009

Date Submitted: Dec. 22, 2009

A. Hydrolyses

a.

Amylase
Iodine test of 0.1% starch solution
Set-up

Color

Test tube 1 (with corn seedling root)

Dark violet (positive for amylase)

Test tube 2 (without corn seedling root)

control

Darker violet

b. Sucrase
Benedicts test of 2% sucrose solution
Set-up

Color

Test tube 1 (with fresh corn seedling root)

Yellow

Test tube 2 (without corn seedling root)

control

Clear

B. Oxireductases
a.

Dehydrogenase
Color

Set-up

After incubation

Test tube 1 (with fresh

mungbean seedlings)
Test tube 2 (without
mungbean seedlings)
control

After aeration

White

White

Clear blue

Clear blue

b. Catalase
Set-up

Observation

Test tube 1 (potato tuber strips in 3%

H2O2)
Test tube 2 (boiled potato tuber strips in
3% H2O2)

Gas evolution observed

No gas evolution observed

C. Factors affecting enzyme activity

a.

Enzyme concentration
Set-up

Color intensity of starch-iodine

solutions
Yellow
lightest yellow / white
pale yellow

Test tube 1 (with undiluted amylase)

Test tube 2 (with half diluted amylase)
Test tube 3 (with diluted amylase)

b. Hydrogen-ion concentration (pH)

Color Intensity of Starch-Iodine Solutions + Amylase
(scale of 1 to 7, dark to light)
pH 4 buffer

(purplish black)

pH 5 buffer

pH 6 buffer

pH 7 buffer
pH 8 buffer

(bluish green)

pH 9 buffer
pH 10 buffer

(light blue)

c. Temperature
Color intensity of starch-iodine
solutions + amylase

Set-up
Test tube 1 (at 5-100C)

violet (darker)

Test tube 2 (at 28-300C)

violet (light)

Test tube 3 (at 98-1000C)

violet (darkest)

Study Questions
1.

Give (i) the name of the enzymes catalyzing the following chemical reactions, (ii)

their cellular localization, and (iii) the plant physiological process they are involved

a) Pyruvate + NAD+ + CoA ----------- acetyl- CoA + NADH + H+ + CO

(i)

Pyruvate dehydrogenase

(ii)

Mitochondrion

(iii)

Citric acid cycle

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

(i)

Ribulose-1,5- bisphosphate carboxylase

(ii)

Chloroplast

(ii)

Carbon fixation

c)

2.

Fructose-6-phosphate + ATP ---------------- Fructose-1,6-biphosphate

(i)

Phosphofructokinase

(ii)

Chloroplast

(iii)

Glycolysis

Describe the rate of enzyme-catalyzed reaction with increasing substrate

concentration.
According to the rate law and the explanation stated in the
collision theory, reaction rate increases with respect to concentration. Thus,as the
concentration of reactants increases the tendency is that, the frequency of collision
also increases.
3. In what ways does hydrogen concentration affect enzyme activity?
Enzyme systems operate at an optimal hydrogen ion
concentration ([H+]), and variation from this optimal can markedly affect enzyme
activity.
Moreover, because [H+] is so critical to enzyme function yet the absolute
concentration is small and difficult to manipulate, the concept of pH was
developed and is now universally used to represent [H+].
Besides, a little variation in [H+] is already very detrimental to an organism.
This explains the existence of buffer systems, which prevent such damage.