CHAPTER 5 : METABOLISM 新陈代谢

5.1 Metabolism
Metabolism

 all chemical reactions occur in living organisms.

 process of converting food into energy in the form of ATP.
 process of forming macromolecules (carb, protein, nucleic, lipid) which are essential
to living organisms.
Types :
a. Catabolism / catabolic reaction CUT
o Break down complex substances/ molecules into simpler form/ substances

Substrate A → Product B + Product C

o Release energy
e.g. breakdown of glucose during cellular respiration
Oxygen, 6 O2 + Glucose, C 6 H 12 O 6 → Carbon dioxide, 6 CO 2 + Water, 6 H 2 O + Energy
(ATP)
e.g. Glycolysis in liver cells – breakdown of glucose to produce energy
e.g. Breakdown of protein to amino acids

b. Anabolism / anabolic reaction

o Synthesise complex substances/ molecules from simpler form/ molecules/
substances

Substrate A + Substrate B → Product C

o Use/ absorb energy

e.g. formation of glucose during photosynthesis (growth, flower formation, fruit
development)
Carbon dioxide, 6 CO 2 + Water, 6 H 2 O + Energy (ATP) → Oxygen, 6 O2 + Glucose,
C 6 H 12 O6
e.g. Synthesis of DNA strands form nucleic acid building blocks
e.g. Synthesis of protein form amino acids

5.2 Enzyme
Enzyme

 Organic compound/ molecules found in living organisms

 Specific organic molecules found in biological systems
 Complex protein made up of polypeptide chains that are folded into three-
dimensional shape (3D shape)/ complex structure
  Specialised class of protein
 Macromolecular catalysts
Function:
 Catalyse (speed up) biological reaction/ chemical reaction in living
organisms.
 Allow biological reactions to occur at high and necessary rate to support life/
living processes
Enzyme reaction
o Enzyme and substrate
o Some enzyme reactions may need cofactors

Process/ Reaction :
Substrate will approach and bind with the specific enzyme/ enzyme at its specific active site.
This forms enzyme-substrate complex. The enzyme converts/ break down the substrate into
products. Once the products leave active site, the enzyme is ready to attach / break down
new substrate and repeat the process.

Characteristics of enzyme :
1. It has specific active site which complements to specific substrate molecules. The
binding of substrate molecule on the active site of enzyme is specific like a lock and
key combination.
2. Enzyme remains unchanged after a reaction/ cannot be destroyed after a reaction
but it remains the same. (unless there are certain other factors that might causes
changes to the shape)
3. Enzyme reaction is reversible. An enzyme can synthesise and decompose/ break
down molecules.
e.g. breakdown of polymers into their constituent 构成部分 parts/ simpler form in a
catabolic reaction.
e.g. synthesis of new macromolecules in an anabolic reaction.
4. Enzyme is only in a small quantity in a chemical reaction and reusable.
5. Some enzymes need cofactors to increase its efficiency in a chemical reaction.
Cofactor 辅因子 is non-protein organic compound/ molecule or ions that bind with
 the enzyme. Cofactors is required in biological reaction where enzyme cannot
perform alone.
e.g. Our body cannot synthesise organic compound such as vitamins, hence, these
vitamins are consumed from diet which helps the digestive enzymes in digesting
food more efficiently.
6. Enzyme reaction can be stopped by enzyme inhibitors 缓蚀剂 .
o Competitive inhibitor blocks active site of enzyme in the treatment of Influenza
using neuraminidase inhibitor, Relenza.
o Non-competitive inhibitor changes the shape of active site of enzyme. For
example, the use of cyanide as a poison to prevent aerobic respiration.
o Enzyme inhibitor antibiotic/ antibiotic stops the growth of bacteria. For example,
penicillin inhibit the enzyme which is essential/ important for growth of
bacteria./ stops bacteria from multiplying.

Intracellular enzyme and extracellular enzyme

a. Intracellular enzyme
o Synthesised in a cell for its own use.
o Enzyme that are formed and retained 保留 in a cell for its own use.

e.g. RNA polymerase (allow transcription of DNA to mRNA)

b. Extracellular enzyme
o Synthesised in a cell and then secreted from the cell to catalyse reactions outside
of the cell.
e.g. trypsin enzyme produced by pancreatic cells and secreted to duodenum to
break down proteins.
***amylase,cellulose
Production of extracellular enzymes
Ribosome synthesise protein. After synthesised, the protein enters into Rough Endoplasmic
Reticulum (RER) for further modification and later it is transported through the RER. The
protein comes to the end of the RER. A transport vesicle containing protein is bud off.
The transport vesicles approach/ moves towards/ comes to the Golgi apparatus and then
fuses with its membrane. Then, the protein is released into Golgi apparatus. The proteins
are processed, modified into enzymes. Lastly, the enzyme is released into the cytoplasm as
intracellular enzyme to be used within the cell.
**mark the end of production/ synthesis of intracellular enzyme.

It is packaged and secreted in secretory vesicles 分泌小泡 .The secretory vesicles is released
from the Golgi apparatus to cytoplasm before moving towards the plasma membrane. It
fuses with the membrane and then protein is released and secreted out of the cell as
extracellular enzyme.

Activation energy
Activation energy is the energy which must be available/ the energy needed to break the
bond in the substrate before a reaction occur/ to produce a reaction/ to catalyse a reaction.
Most reaction in a cell requires high amount of activation energy. However, with the
existence of enzyme in our cells, all the chemical reaction is catalysed/ accelerated by
lowering the activation energy.
The Mechanism of Enzyme Action and Factor Changes
Factors affecting the rate of enzymatic reaction/ chemical reaction controlled by enzymes.
o Temperature, pH, substrate concentration, enzyme concentration.

1. Temperature

Optimum
temperature
最佳反应

10 20 30 37 40 50 60

o Lower temperature – rate of reaction catalysed by enzyme/ rate of enzymatic

reaction is low
 o High temperature – rate of reaction is faster/ higher. At high temperature/ when
the temperature increases, the substrate and enzyme molecules gain more
kinetic energy. This increases collision between substrate and enzyme molecules.
This means that the rate of reaction between substrate and enzyme increases.
o For every rise of 10°C in temperature, the rate of reaction doubled until it
reaches the optimum temperature of 37°C.
o At the optimum temperature, the rate of reaction is the maximum. After which/
after reaching the optimum temperature, the rate of reaction starts to decrease
with the further rise in temperature and stops at 60°C.
o At 60°C, the enzyme is denatured/ becomes denatured because the chemical
bond in the enzyme breaks at high temperature. This changes the shape and
active site of the enzyme. Hence, the substrate can no longer complement to its
active site.
e.g. Fever increases our body temperature and may denature the enzymes which are
crucial for our metabolism/ enzymatic reaction. High fever alter/ change normal cell
metabolism and which may leads to cell injury or death.

2. pH
Enzyme activity/ reaction is affected by the surrounding pH. The enzymes can react
with substrate/ work properly and effectively at optimum pH.
o pH can affect the ionization of enzyme which can change the chemical bond that
maintain the enzyme shape.
o pH can change the charge on the active site of enzyme.
o At optimum pH, enzyme can function well by forming bond and allow substrate
to bind to its active site.
o At extreme pH, the chemical bond that maintain the shape and active site of
enzyme will change/ will break, leading to denaturation of enzyme. This causes
enzyme and substrate cannot form bond and bind to each other.
o When pH returns to optimum level, the charge on active site of enzyme is
restored. This allows enzyme to function normally again. (provided enzyme is not
denatured)
 e.g. pepsin works in acidic condition in our stomach. It has the optimum pH of 1-3.
Trypsin works in alkaline solution in duodenum. It has the optimum pH 7. 5-8.
Amylase works in neutral situation. It has the optimum pH of 6.8-7.
3. Substrate Concentration
With the fixed concentration of enzyme, when the substrate concentration
increases, the rate of enzymatic reaction will increase with the fixed concentration of
enzyme. This leads to more product produced/ synthesised.
o The increase in substrate concentration increases the effective collision
between the enzyme and substrate. This increases the rate of reaction
between enzyme and substrate until it reaches the maximum level.
o At the maximum level, the reaction becomes constant because the enzyme
concentration becomes the limiting factor. All the active site of enzyme are
occupied/ saturated with substrates and all are involved in respective
chemical reaction.

e.g. binging on large meal can cause indigestion, nausea or even diarrhea. This is due to the
insufficient enzyme concentration in our body to digest the great amount of food in time.
4. Enzyme Concentration
With the fixed concentration of substrate, when the enzyme concentration
increases, the rate of enzymatic reaction will increase until it reaches the maximum
level.
o The increase in enzyme concentration means there are more active sites for
substrates to bind to/ attach to for catalytic reaction/ chemical reaction. This
 increases the rate of reaction between enzyme and substrate until it reaches
the maximum level.
o At the maximum level, the reaction becomes constant because the substrate
concentration becomes the limiting factor. The available enzyme is more
than it is needed to react with the limited amount of substrate.

o With the excessive amount of substrate, if the enzyme concentration is

doubled, the product concentration is also doubled.

STUDYING THE EFFECT OF TEMPERATURE ON THE AMYLASE ENZYME ACTIVITY

Problem statement :
What is the effect of temperature on the amylase enzyme activity/ amylase enzymatic
reaction?
Hypothesis :
An increase in temperature will increase the amylase activity/ amylase enzymatic reaction
until it reaches the optimum temperature. After which, the rate of enzymatic reaction drops
to zero.
Variables :
Manipulated : Temperature
Responding : Amylase enzyme activity/ Rate of enzymatic reaction
Fixed : pH, concentration of amylase enzyme, concentration of starch
Materials and apparatus :
1% starch suspension, 0.5% amylase enzyme solution, iodine solution, beaker, test tube,
syringe, dropper, glass rod, white groove tile, thermometer, Bunsen burner, tripod stand,
measuring cylinder, stopwatch, wire gauze
Procedure :
1. Prepare water bath at these temperature : 20, 40, 50, 60 and 80
2. Add 5ml of 1% starch suspension using a syringe to 5 respective test tubes and mark
the test tubes as A, B, C, D and E.
3. Using another syringe, add 5ml of amylase enzyme solution to 5 respective test
tubes and mark the test tubes as a, b, c, d and e.
4. Place the test tubes containing starch and amylase into each water bath of different
temperature of 20, 40, 50, 60 and 80 as follows : A and a, B and b, C and c, D and d,
and E and e.
5. Incubate/ leave the test tubes in the water bath for 5 minutes.
6. At the same time, prepare white groove tile/ palette and put a single drop of iodine
solution onto it.
7. After incubating for 5 minutes, mix the amylase and the starch suspension and stir it
with a glass rod. Then, start the stopwatch immediately.
8. For every 30 seconds, add a drop of the mixture into the iodine solution until the
iodine solution no longer turns into dark blue.
9. Record the time taken for the iodine solution to remain yellowish brown when mixed
with the mixture.
10. Repeat step 7 to 9 for each pair of test tube at different temperature.
Results : https://www.youtube.com/watch?v=1Fa2sSlt4_l
Temperature Time taken for starch to break down/ starch
hydrolysis to complete (minute)
20
40
50
60
80
STUDYING THE EFFECT OF pH ON PEPSINE ENZYME ACTIVITY
Problem statement :
What is the optimum pH doe pepsin enzymatic reaction?
Hypothesis :
pH 2 is the optimum pH for pepsin enzymatic reaction.
Variables :
Manipulated : pH of solution/ pH of reaction medium/ pH
Responding : Pepsin enzymatic reaction
Fixed : Temperature, Pepsin concentration, albumen suspension concentration
Procedure :
1. Prepare 3 test tubes and label them as P, Q and R.
2. Add 5ml albumen suspension in each test tube.
3. Add the following solution into the test tubes as follows :
a. P : 1ml of 0.1M hydrochloric acid (HCl) + 1ml of 1% pepsin solution
b. Q : 1ml distilled water + 1ml of 1% pepsin solution
c. R : 1ml of 0.1M sodium hydroxide + 1ml of 1% pepsin solution
 4. Test the pH of the mixture in each test tube and observe the initial mixture state and
record.
5. Put all the test tubes into a water bath of 37‫ﹾ‬C for 20 minutes.
6. After 2 minutes, observe the mixture and record.

Results :
Test tubes pH Clarity 清晰度 of the mixture

0 min 20 mins
P 2 Cloudy Clear
Q 7 Cloudy Cloudy
R 12 Cloudy Cloudy

n = chromosome number
c = dna content number
Graph 1 illustrates the change of number of DNA content in each daughter cells during cell
division, mitosis.
The cell undergoes mitosis and each daughter cell now contain 2n and 2c because each
daughter cell receives half of the DNA but the number of chromosomes remain 2n which is
similar parent cell.
In contrast, the cell after undergoing meiosis I and II, each daughter cell will end up with
haploid number of chromosomes (n = 23) and 1c.
After G1 phase, the DNA content doubled
in S phase. As a result, 30 pg DNA becomes
60 pg of DNA. After meiosis I, the DNA
content is halved to 30 pg of DNA content.
After meiosis II, the DNA content further
decrease by half and becomes 15 pg of
DNA content. The daughter cells have the
same number of chromosomes from
Meiosis I and Meiosis II but the DNA
content is reduced to half.

**pg - picogram