This document reports on an experiment investigating enzymes in corn seedlings and potato tubers. Amylase and sucrase were found in corn seedling roots through starch and sucrose hydrolysis tests. Dehydrogenase and catalase were detected in mungbean seedlings and potato tubers through color change and gas evolution assays. The effects of enzyme concentration, pH, and temperature on amylase activity were examined. Amylase activity increased with higher concentration and optimal pH and temperature ranges were identified.
This document reports on an experiment investigating enzymes in corn seedlings and potato tubers. Amylase and sucrase were found in corn seedling roots through starch and sucrose hydrolysis tests. Dehydrogenase and catalase were detected in mungbean seedlings and potato tubers through color change and gas evolution assays. The effects of enzyme concentration, pH, and temperature on amylase activity were examined. Amylase activity increased with higher concentration and optimal pH and temperature ranges were identified.
This document reports on an experiment investigating enzymes in corn seedlings and potato tubers. Amylase and sucrase were found in corn seedling roots through starch and sucrose hydrolysis tests. Dehydrogenase and catalase were detected in mungbean seedlings and potato tubers through color change and gas evolution assays. The effects of enzyme concentration, pH, and temperature on amylase activity were examined. Amylase activity increased with higher concentration and optimal pH and temperature ranges were identified.
This document reports on an experiment investigating enzymes in corn seedlings and potato tubers. Amylase and sucrase were found in corn seedling roots through starch and sucrose hydrolysis tests. Dehydrogenase and catalase were detected in mungbean seedlings and potato tubers through color change and gas evolution assays. The effects of enzyme concentration, pH, and temperature on amylase activity were examined. Amylase activity increased with higher concentration and optimal pH and temperature ranges were identified.
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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.
