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4.

5 Enzymes
Metabolism
1. Metabolism = biochemical reaction that occurs within a living organism.

2. The biochemical reactions are occurring in an orderly manner (simultaneously) and consists a series of linked
reaction.

3. The reactant in an enzymatic reaction is called substrate and substances formed at the end of the reaction is
called product. Metabolism: Enzyme
Reactant (Substrate) Products

Eg:
Reaction that build up Photosynthesis /
Anabolism
coplex molecules converting glucose to
glycogen
Metabolism
Eg:
Reaction that break
Catabolism Digestion /respiration
down complex molecule

4. Metabolism in cells are catalysed and regulated by enzymes.


Each step in the biochemical reaction (metabolism) is catalysed by different enzymes for its respective
reaction.

Enzymes
1. Enzyme = Biological catalyst that regulates/speeds up biochemical reactions in our body

(a) Are complex globular proteins that made up of one or more polypeptides. (Tertiary structure)

(b) Has 3 dimensional shape which is very precise

(c) The polypeptide chain are folded to form a cleft or a pocket that is called active sites of enzyme.

Active site of enzymes:

(a) Is highly specific


(b) It has distinctive three dimensional
shape and charges that suits its substrate
(c) Only substrate that fits to enzymes
shape will allow reaction to take places

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Naming of Enzymes

1. Earlier on, enzymes were arbitrary named without a system.


Examples are trypsin, pepsin and rennin.

2. Then, it is named according to the name of the substrate.


Is derived by adding the suffix ase at the end of the substrate.

Examples: (a) Sucrose + water Glucose + fructose

(b) Protein + water Amino acid + amino acids / Dipeptide + polypeptide

3. Now enzymes are classified into 6 main groups by the nature of their action:
Class of enzymes Action Eg of Enzymes
1 Hydrolase Hydrolyses (add water) Peptidase, lipase
2 Lyase Break down chemical bonds: Decarboxylase
C-O, C-C or C-N
3 Isomerase Rearrange functional groups Isomerase, mutase
4 Ligase Connects two molecules Synthetase
5 Oxyreductase Include oxidation and reduction (redox) Dehydrogenase, oxidase
6 Transferase Transfer the functional groups from one molecule to Transaminase,
another phosphorylase

Types of Enzymes: Intracellular and Extracellular Enzymes.


1. All enzymes are synthesized inside the cell but they may either be an
(a) Intracellular enzymes
(b) Extracellular enzymes

Intracellular enzymes Extracellular enzymes


Enzymes which are synthesized and retained in Definition Enzymes which are synthesized in the cell
the cell that catalyses reaction. but secreted out from the cell to work
eternally.

These enzymes are found in the cytoplasm, Mainly involved in food digestion
nucleus, mitochondria and chloroplast.

Found on the free ribosome in the cytoplasm Where it On the ribosome attached to the rough
produces endoplasmic reticulum

DNA polymerase, thiokinase, pyruvate Example Amylase, pepsin, trypsin, lipase, maltase
carboxylase, oxydoreductase, ATP synthetase,
ATPase, Carbonic anyhydrase

Function and location of Intracellular enzymes

Name of the intracellular enzymes Type of reaction Location


DNA polymerase Synthesis of DNA Nucleus
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Thiokinase Fatty acids catabolism Outside the endoplasmic reticulum
Pyruvate carboxylase Cellular respiration Mitochondria
Oxydoreductase Cellular respiration Mitochondria
ATP synthetase Synthesis of ATP Mitochondria
ATPase Catabolic release of energy Mitochondria
Carbonic anhydrase CO2 + H2O H2CO3 Red blood cells

Function and location of Extracellular enzymes

Name of the Where it comes Where it works Food acted on Substances


extracellular from produced
enzyme
Amylase Salivary glands Mouth cavity Starch Maltose
Pepsin Stomach wall Stomach Protein Peptides
Trypsin Pancreas Small intestine Starch Maltose
Lipase Pancreas Small intestine Fat Fatty acids
Maltase Walls of small Small intestine Maltose Glucose
intestine

The sites of enzymes,


1. Ribosomes
Are the sites of protein synthesis.
Since, enzymes are protein Ribosomes are also the sites of enzyme synthesis.
Made up of 2 parts, a small subunit and a large subunit

2. The information for the enzyme synthesis is carried by the DNA.


3. The difference sequences of bases in the DNA are codes to make different proteins
4. In the nucleus, the DNA double helix unwinds and exposes its 2 strands to synthesis of
a messenger RNA (mRNA) strands.
5. mRNA then leaves the nucleus and moves towards itself and attached to a ribosome..
6. mRNA translated the codes into a sequence of amino acids.
7. These amino acids are bonded together to form specific enzymes.The
enzymes are released into the cytoplasm.

Production of Extracellular Enzymes

1. Proteins synthesized by the ribosome are wrapped in vesicles that bud


off from the sides of the rough endoplasmic reticulum.
2. These transport vesicles fuse with the membrane of the Golgi
apparatus and empty their contents into the membranous space.
3. These proteins are then modified during their transport in golgi
apparatus. (Eg: carbohdydrate + protein glycoprotein)
4. Secretory vesicles containing these modified proteins bud off from
the Golgi membrane and travel to the plasma membrane.
5. These vesicles will then fuse with the plasma membrane before
releasing the proteins outside the cell as enzymes.

7 general characteristics of an enzyme:

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(a) All enzymes are COMPLEX PROTEIN (e) Enzyme activities can be slowed down
Are synthesized by ribosome. or stopped by INHIBITOR

Eg: heavy metals (lead and mercury)


(b) SPEED UP chemical reactions rate but REMAIN (f) Most enzyme-catalyzed reactions are
UNCHANGED REVERSIBLE

Activation energy is needed for


reactants/substrate molecules to start a reaction
Enzymes lower the activation energy and this Can catalyze metabolic
helps to speed up the rate of reaction in terms reactions both in forward and
of time reverse directions
(c) Needed in SMALL AMOUNT (g) Require HELPER MOLECULE
Not used up/destroyed but recyclable (COFACTOR) to function
(Is released at the end of the reaction)
Same enzyme molecules can produce large Cofactor= binds to enzymes and
number of substrate molecules weaken the bond in substrate
(d) HIGHLY SPECIFIC molecules.
Can only catalyzed one type of substrate/ a -2 types :
single reaction type (a) Inorganic cofactor = iron and
copper
It has specific active sites which binds to (b) Organic cofactor = coenzymes
specific substrate (water soluble vitamin B complex)

Eg: starch can only fit into the sites of amylase


while protein molecule cannot

The Mechanism Of Enzyme Action (Analogy Used: Lock And Key Hypothesis)

1. Lock and Key Hypothesis: how enzyme binds to or catalyses its substrate.
2. Lock = enzyme.
3. Key = substrate.

4. Mechanism of enzyme action:


(a) Substrate binds to the active site of the enzyme to form enzyme-substrate complex.
(b) Upon the binding, reaction takes place.
(c) The enzyme catalyzes the substrate to form products. (like key fits into the lock).
(d) The product have different shape from the substrate and are therefore repelled from the active site.
(e) The active site of the enzyme is now free to bind another substrate molecule.

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Factors Affecting enzyme activity.

1. The complex three-dimensional structure of an enzyme is sensitive to external factors.


2. The chemical bonds in the enzyme can be altered easily by the chemical and physical changes in the
surroundings.
3. Among the factors are:
(a) Temperature (c) Substrate concentration
(b) Temperature (d) Enzyme concentration

(a) Temperature

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1. At low temperature, an enzyme reaction takes place slowly.

2. Upon heating, rate of most chemical reactions increases because heating:


(a) Increases kinetic energy of enzymes and substrate molecules
(b) Molecules with greater kinetic energy collide with each other more frequently
(c) This will increase the rate of reaction of enzymes and substrate but only up to a peak.

3. In general for every 10C rise in temperature, the rate of reaction is doubled (only up to the optimum C)

**Optimum temperature: Temperature at which an enzyme catalyses a reaction at the


maximum rate.

4. Beyond the optimum temperature, any further increase in the temperature will
cause a sharp decrease in the rate of reaction until it stops completely at
about 60C.

Why?
Reason: Substrate can no longer fit into the active sites of enzymes as its 3-dimensional shape of enzymes active site
is altered and destroyed at high temperature. Bonds that hold enzyme molecules together begin to break at high
temperature.

5. The enzymes is said to be denatured when enzymes lose their activities.


Denaturation is irreversible.
It has a permanent change in its molecular structure that cannot be reversed by cooling.

6. Hence, the body needs to maintain its temperature at 37C for the optimal functioning of enzymes.
Most human enzymes have an optimum temperature of around 37C
Most plants optimum temperature is lower that is around 25C.
Some bacteria can live in hot springs between 80C to 100C or higher.

Experiment 4.Studying the effects of temperature on the activity of salivary amylase

Problem statement What are the effects of temperature on the activity of salivary amylase on starch?
Variables
Manipulated: Surrounding temperature
Responding: Rate of enzymatic reaction
Constant Volume of saliva (enzymes), volume and concentration of starch suspension and pH medium
Hypothesis Rate of activity of salivary amylase on starch increases with the increase in temperature until
it reaches the optimum temperature of 37C
Materials 1% starch suspension, saliva solution, iodine solution, ice and distilled water

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Apparatus Beakers, test tubes, syringes, droppers, glass rods, spotting tiles, thermometers, Bunsen
burner. Tripod stand, test tube rack, wire gauze and a stopwatch
Technique Conduct iodine test to determine the presence of starch by measuring and recording the
time taken for the hydrolysis os starch to be completed with a stopwatch

Procedure:

1. The mouth is rinsed with warm water and the saliva is collected.
2. The saliva is diluted with an equal volume of distilled water.
3. The test tubes are labeled as A1, A2, B1,B2, C1, C2, D1,D2,E1 and E2 respectively.
4. 5ml of 1% of starch suspension is put into each test tubes A1,B1,C1,D1 and E1 using a syringe.
5. 2ml of saliva is added into test tubes A2, B2, C2, D2 and E2.
6. All test tube A1 and A2, B1 and B2, C1 and C2, D1 and D2 and E1 and E2 are immersed into 5 different
water bath with temperature kept constant at 0 C , 28 C , 37 C , 45 C and 60
C .respectively
7. The test tube is left for 5 minutes.
8. Meanwhile, a dry piece of white tile with grooves and iodine are drop into each groove.
9. After 5 minutes of immersion,
the starch suspension in test
tube A1 is poured into test
tube A2.
10. The mixture is stirred using a
glass rod. The stopwatch is
started immediately.
11. A dropper is used to remove a drop of mixture from test tube A2
and is placed in the iodine solution in the first groove on the tile. (this first groove is considered as 0 minute)
12. The iodine test is repeated every minute for 10 minutes.
13. The dropper is rinsed in a beaker of water after each sampling.
14. The time taken for the completion of hydrolysis of starch that is when the mixture gives a negative test is
recorded.
15. The test tube is kept with mixture with their respective water baths throughout the experiment.
16. Steps 9 to 15 are repeated for test tubes B1, C1, D1 and E1.
17. The thermometer is used to ensure that the temperatures remain constant throughout the experiment.
18. The results are recorded in a table and a graph is plotted showing the rate of reaction (1/t) against
temperature.

Background: Salivary amylase is secreted by the salivary glands and it begins the carbohydrates digestion. It
functions most efficiently at the body temperature. Iodine is used to test the presence of starch. When amylase is
added to the starch, It will break down the starch and the blue black colour disappear.

Results:

Test tube Temperature (C) Time taken for the hydrolysis of starch Rate of enzymatic reaction 1/t
to be completed (minutes) (minute-1)
A2 0 Not completed after 10 minutes 0.00
B2 28 5 0.20
C2 37 3 0.33
D2 45 10 0.10
E2 60 Not completed after 10 minutes 0

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At 370 C

Above 370 C

370 C

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Discussion:

1. Why do you need to immerse the test tube in their respective water baths for 5 minutes at the beginning of the
experiment?
Expected answer: To allow both solution to reach temperature set.

2. What is the result of the action of salivary amylase on starch?


Expected answer: Starch is hydrolysed by salivary amylase to a reducing sugar.

3. What is the function of iodine solution?


Expected answer:
(a) To test the presence of starch.
(b) If the iodine solution changes from brownish-yellow to blue-black, this indicates that starch is still present in
test tube.
(c) If iodine solution remains brown (unchanged), this indicates all starch has been hydrolsed by salivary
amylase.

4. Based on the graph plotted, explain the effects of temperature on enzyme activity.

Expected answer:

(a) Graph shows that rate of enzyme activity against temperature is a bell shaped curve.
(b) At low temperature, for every 10 C, increase in temperature, the rate of enzymatic reaction is doubled.
(c) The maximum rate of reaction occurs at 37C which is the optimum temperature for salivary amylase.
(d) The average human body temperature is also 37C.

5. Discuss the results of experiment when the temperature is set at


(a) O C (b) 37C (c) 60C

Expected answer:

(a) At O C, the enzyme is not active. Therefore the salivary amylase cannot hydrolyse the starch. As the
temperature increases, the rate of enzymatic reaction increases until it reaches an optimum temperature of
37 C.
(b) 37C is the optimum temperature for the activity of salicary amulase because the hydrolysis of starch is
completed in the shortest period.
(c) Beyond the optimum temperature, the rate of enzymatic reaction decreases and cease altogether at 60C.
there is no enzyme activity at 60C. this is because the enzyme has denatured due to high temperature.

Conclusion:

Changes in temperature affect the activity of salivary amylase on starch. Salivary amylase is inactive at 0C and the
highest rate of reaction catalysed by salivary amylase is 37C denoting the optimum temperature. the hypothesis is
accepted.

(b) pH

1. Each enzymes has an optimum pH at which its rate of reaction is fastest.


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The optimum pH is the pH at which the rate of reaction is at maximum.

2. Below is the optimum pH of different enzymes.

Pepsin Acidic condition (pH2) in stomach


Trypsin alkaline condition (pH 8.5) in duodenum.
Most cells pH range from 6 to 8

3. Acidity and alkalinity affect the active sites of an enzyme.


Free hydrogen ions (H+) and hydroxyl ions (OH-) can affect the charges on the amino acids of the enzymes
active site
This reduces the ability of both molecules to bind each other.
For example, if the enzymes and substrate both have the same charges, they repel each other and the
enzyme-substrate complex cannot be formed

4. However, the effects of pH on enzymes are normally reversible.


(a) When the pH reverts to the optimal level, the ionic charges on the active sites are restored.
(b) Enzymes resume their normal function.

Experiment 4.2 Studying the effects of pH on the pepsin activity.

Problem What are the effects of pH on the activity of pepsin?


statement

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Variables
Manipulated pH of medium
: clarity of solution
Responding: volume and concentration of albumen suspension, volume and concentration of pepsin solution
Constant and medium temperature
Hypothesis The optimum pH for the activity of pepsin is an acidic medium for pH 3
Materials Albumen suspension, 1% pepsin solution, 0.1M hydrochloric acid, 0.1 M sodium hydroxide solution
and distilled water
Apparatus Beakers. Droppers, thermometers, test tubes, 5ml syringes, pH paper, wire gauze, stopwatch, test
tube rack
Technique Observe and record the condition of mixture before and after 20 minutes
Procedure:

1. An albumen suspension is prepared by mixing egg white with 500ml of distilled wate
2. The suspension is boiled and left it to cool
3. The large particles are discarded using glass wool.
4. The test tubes are labeled as P, Q and R.
5. 5ml of albumen is put into each test tube using a syringe
6. The following solution is added into each test tube:
(a) P: 1ml of 0.1M hydrochloric acid + 1ml of 1% pepsin solution
(b) Q: 1 ml of distilled water + 1 ml of 1% pepsin solution
(c) R: 1ml of 0.1M sodium hydroxide + 1 ml of 1% pepsin solution.
7. A piece of pH paper is dipped into each test tube and the pH value is recoded.
8. All the test tubes are immersed in the water bath wherer the temperature is set at 37 C for 20 minutes.
9. The conditions of the mixture are observed at the beginning og the experiment and again after 20 minutes.
10. The results is recorded in a table.

Results:

Test tube pH Mixture


At the beginning of the experiment After 20 minutes
P 3 Cloudy Clear
Q 7 Cloudy Cloudy
R 8 Cloudy Cloudy

Discussion:

1. Why do the test tubes need to be immersed in the water bath with temperature maintained at 37C?
Expected answer:
This is the optimum temperature for the action of pepsin

2. What are the effects of pH on the action of pepsin on albumen?


Expected answer:
(a) Pepsin hydrolyses albumen (protein) into polypeptides in an acidic medium.
(b) The solution turns clear because polypeptised are soluble in water.

3. Discuss the results you obtained for test tubes P, Q and R?


Expected answer:
(c) pH condition for test tube P (pH 3) is optimum for the function of pepsin. This is because contents of test
tube P become clear at the end of the experiment, showing that albumen has been completely digested or
hydrolsed by pepsin

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(d) The contents of test Q and R are still cloudy at the end of the experiment. This shows that a neutral (test
tube Q) and alkaline pH (test tube R) are not suitable for the activity of pepsin.

Conclusion:

Activity of pepsin is affected by the pH of its medium. Acidic medium (pH 3) is the most suitable medium for pepsin
to function efficiently. The hypothesis is accepted.

(c) Substrate concentration.

Graph below shows the how the rate of reaction changes at different substrate concentration with the fixed
amount of enzymes molecules, pH and temperature.

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Graph description: the rate of reaction is directly proportional to the substrate concentration until the
reaction reaches a maximum rate.

From C1 to C3,
1. As substrate concentration increases, the collisions between the substrate and enzymes molecules increase.
2. Hence more substrate molecules bind to the active sites of the enzymes for catalytic reaction to occur.
3. This increases the rate of reaction increases, provided they are enough of enzymes molecules to catalyse the
additional substrate.
4. Hence the rate of reaction increase from V 1 to Vmax as the substrate concentration increases from C 1 to C3.

From C3 to C4, (limiting factor)


1. All active sites of the enzyme molecules are filled and engaged in catalysis.
2. The enzyme is said to be saturated and the concentration of enzyme becomes a limiting factor.
3. An excess in the substrate molecules does not increase the rate of reaction.
4. To increase the rate of reaction, concentration of the enzyme need to be increased.

(d) Enzyme concentration.

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Graph Description: the rate of an enzyme-catalysed reaction is directly proportional to the concentration of
enzyme present until a maximum rate is achieved.

1. As the enzyme concentration increases, more enzyme molecules are available. This will also make more active
sites of enzyme available for the catalytic reaction.
2. Therefore, rate of reaction will increase, only if there is
(a) An abundant supply of substrate molecule
(b) Other factors such as pH, temperature and pressure are constant (no other limiting factors)

3. Thus, the rate of reaction is directly proportional to the concentration of enzyme present until a maximum rate
is achieved.
4. After the maximum rate, the concentration of substrate becomes a limiting factor.
5. If the concentration of enzyme is doubled, the amount of substrate molecule that is converted to products per
unit time is also doubled. This means when the enzyme concentration is double, the rate of reaction is
doubled, provided that substrate concentration are in excess.

Experiment 4.3 Studying the effect of substrate concentration on the activity of salivary amylase

Problem What are the effects of substrate concentration on the activity of salivary amylase?
statement
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Variables
Manipulated Concentration of starch suspension
: Time taken for the hydrolysis of starch to be completed
Responding: Enzymes concentration, temperature and pH medium
Constant
Hypothesis The rate of enzymatic reaction increases with increase substrate concentration until it
reaches a maximum rate
Materials Starch suspension at various concentrations (0.1%, 0.2%, 0.3%, 0.4%, 0.5% and 0.6%),
0.1% amylase or saliva suspension, iodine solution and distilled water
Apparatus 5 ml of syringes, 1 ml sytinges, test tubes, glass rods, stopwatch, white spotting tiles,
droppers and measuring cyclinders
Technique Conduct iodine test to determine the presence of starch by measuring and recording
the time taken for the hydrolysis of starch to be completed with a stopwatch

Procedure:

1. 10ml of 0.1% salivary amylase is prepared.


2. 6 test tubes lablled A to F are prepared.
3. 4ml of starch suspension at various concentrations are poured inot the following test tubes using different
syringes.
4. The test tubes are immersed in a water bath at 37 C
5. 5 drops of iodine solution are added separately onto the grooves of the white tile.
6. 1 ml of 0.1% amylase is added to test tube A using a syringe.
7. The stopwatch is activated immediately (0 minutes). The contents are stirred with a glass rod.
8. A drop of mixture is tested with the iodine solution on the white tile
9. This step is repeated at 30 seconds interval until the mixture stops turning blue-black in colour when tested
with iodine solution.
10. The time taken for the hydrolysis of starch to be completed is recorded.
11. Steps 6 to 10 are repeated with test tubes B, C, D, E, and F.
12. At every sampling, the dropper must be rinsed with clean distilled water.
13. The results are recorded in the table below and a graph of rate of reaction against concentration of starch
suspension (%) is plotted.

Results:
Test Concentration Time taken for the hydrolysis of Rate of reaction
tube of starch starch to be completed substrate concentration
= (%
suspension (%) (seconds) (Minutes) time
minutes-1)
A 0.1 240 4.0 0.025
B 0.2 240 4.0 0.050
C 0.3 240 4.0 0.075
D 0.4 240 4.0 0.100
E 0.5 300 5.0 0.100
F 0.6 300 6.0 0.100

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Discussion:

1.

How do you determine the rate of reaction?


Expected answer: divide the total substrate concentration that has been catalysed with unit time (minutes)

2. Describe the trend of the graph that you obtain if the rate of reaction is plotted against the concentration of
starch suspension (%).
Expected answer:
(a) The graph shows that rate of reaction increases with the concentration of starch suspension until a
maximum point.
(b) After this point, a further increase in starch concentration does not increase the rate of reaction.

3. What is the substrate concentration when the enzymes reaches its saturation point?
Expected answer:
(a) 0.4% is substrate concentration is the saturation point of enzymes

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4. At a certain substrate concentration, the rate of reaction does not increase even though the substrate
concentration continues to increase. Explain why.
Expected answer:
(a) Rate of reaction does not increase eventhough the substrate concentration is increased because the
concentration of the enzyme has become a limiting factor
(b) The time taken to hydrolyse the starch compeletely at low substrate concentration is a constant since the
enzymes are not yet saturated.
(c) When the starch suspension concentration increases, the time taken for the hydrolysis of starch to be
competed also increases as the enzymes molecules become saturated.

Conclusion:

The rate of enzymatic reaction is increase with the increase in substrate concentration until it reaches a maximum
rate. The hypothesis is accepted

Experiment 4.4 Studying the effect of enzyme concentration on the activity of salivary amylase

Problem What are the effects of enzymes concentration on the activity of salivary amylase?
statement
Variables
Manipulated: Concentration of enzymes
Responding: Time taken for the starch hydrolysis to be completed
Constant Substrate concentration, temperature and pH medium
Hypothesis The rate of enzymatic reaction increases with the increase in enzymes concentration as long as
there are no other factors limiting the rate of reaction
Materials 1% starch suspension, 0.8% amylase or saliva suspension, iodine solution and distilled water

Apparatus 5ml of syringes, 1ml of syringes, test tubes, glass rods, stopwatch, spotting tile, measuring cylinder
and droppers

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Technique Conduct iodine test to determine the presence of starch by measuring and recording the time taken
for the hydrolysis of starch to be completed with a stopwatch
Procedure:
1. 6 test tunes are labeled A to F.
2. The tes tubes contain the following mixtures:
A: 0.5ml of 0.8% amylase + 2.5ml distilled water D: 2.0ml of 0.8% amylase+ 1.0ml distilled water
B: 1.0ml of 0.8% amylase + 2.0ml distilled water E: 2.5ml of 0.8% amylase + 0.5ml distilled water
C: 1.5ml of 0.8% amylase + 1.5ml distilled water F: 3.0ml of 0.8% amylase
3. Test tubes A to F are immersed in a water bath set at 37C
4. Meanwhile, drops of iodine are added separately onto the groove of the whitetile.
5. 4ml of 1% starch suspension is added to test tube A using syringe.
6. The stopwatch is started immeadiaetely and the time recorded is 0 minute.
7. The mixture in the test tube is stirred using a glass rod. A small amount of the misture is removed and
tested with iodine solution on the tile.
8. The iodine test is repeated every 30 seconds intervals untl the mixture does not turn blue black when
tested with iodine solution.
9. The time taken for hydrolysis of starch to be completed is recorded.
10. Steps 4 to 9 are repeated with the contents of test tubes B, C, D, E and F.
11. All the results are recorded and tabulated in a table
12. The rate of enzymatic reaction ( 1/time) is then calculated. A graph pf the rate of enzymatic reaction (1/t)
against enzymatic reaction is plotted.

Results:
Test tube Concentration of Time taken for the hydrolysis of Rate of reaction
starch suspension starch to be completed substrate concentration
= (% minutes-1)
(%) time
(seconds) (Minutes)
A 0.17 330 5.5 0.18
B 0.33 150 2.5 0.40
C 0.50 90 1.5 0.67
D 0.67 60 1.0 1.0
E 0.83 60 1.0 1.0
F 1.00 60 1.0 1.0

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Discussion:

1. What is the relationship between the time taken for amylase to catalyse starch and the enzymes
concentration?
Expected answer: The time taken for amylase to catalyse starch decreases as the enzymes concentration
increases.

2. Why does it take less time to complete hydrolyse the starch when a more concentrated enzyme solution is
used?
Expected answerv:
A higher concentration of enzyme concentration contains more the enzyme molecules to hydrolyses the
starch molecules and hence, less time for hydrolysis of starch to be completed.

Conclusion:

The rate of reaction increases with the increase in enzyme concentration until a certain concentration of enzyme is
reached. The hypothesis is accepted.

The uses of enzymes.

1. Enzymes are widely used in our daily life as well as in various industries such as food, leather and textile
industries and in the manufacturing of detergents. It is known as enzyme technology.
2. They can be obtained from
(a) Plants
(b) Animals
(c) Microorganisms: bacteria and fungi (main source). Hence, microorganisms are grown on a large scale in
industrial fermenters to produce large amounts of enzymes.
Types of industry Enzyme used Uses
1. Food processing
industry

(a) Meat products Protease Tenderizes meat


(b) Starch products Amlylase and Making syrup: Starch sugar
amyloglucoxidase

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Glucose isomerase Produce high fructose syrup: Glucose fructose (sweeter).
- In sliming products (only needed in small amount)
(d) Fish products Protease Removes skin of fish
(e) Dairy products Lipase Ripening of cheese
lactase Making ice-cream: Lactose glucose + galactose
Rennin Solidifies milk protein
(f) Beer/wine industry zymase Wine/Beer: Sugars ethanol
EG: Alcoholic drinks

(g) Baking industry -amylase Making bread or dough: Starch flour sugar
EG: Bread and baking products

(h) Cereal grain Cellulase Extracts agar from seaweeds


products
(i) Fruit juice making Pectinase Breakdown the pectin in fruits into simple sugars
industry ** pectin makes the fruit juice cloudy.
Pectinase is added to crushed fruits like apples to obtain more
juice and more colour from the skin
2. Leather products Tyrpsin Removal of hair from animal hides
3. Textiles products Amylase Remove starch that is used as stiffeners from fabrics
4. Medical/ - Pancreatic - Treats inflammation
pharmaceutical trypsin - Dissolves blood clots
products - Microbial trypsin

5. Biological washing Protease,Amylase, Protease: Dissolve protein present in blood, grass and egg
powder/ Lipase and stains
detergents cellulase
Amylase: remove starch stains in clothes

Lipase: digest fats, oil and grease stain in clothes

Cellulose: digest free cellulose microfibrils on damaged


clothes(clothes becomes softer and brighter)

3.6 The importance of Chemical Composition in Cell.

1. Enzymes which are made of proteins will not be synthesized by cells.


2. Without enzymes, all biochemical reactions will proceed too slowly to sustain life
3. Thus, the chemical substances are very important to the cell.
4. Even trace element such as iodine, manganese and selenium are important.
5. Deficiency in iodine leads to enlargement of the thyroid gland (goitre).
6. Deficiency in selenium can cause muscle pains and possibly leads to deterioration of heart muscles.
7. All minerals or trace elements are harmful to body when consumed in excess.

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