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1. Introduction
2. Cofactor
3. Specificity of Enzyme
4. Naming / Nomenclature
5. Classification of Enzyme
6. Enzyme Kinetic & Michaelis-
Menten equation
7. Factor affect enzyme activity

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Enzyme - Introduction
• Majority biochemical reaction
• can’t take place spontaneously

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Enzyme - Introduction
Biochemical reaction

• Metabolism is usually divided into two

categories:
• Catabolism breaks down organic matter, for
example to get energy in cellular respiration.
• Anabolism uses energy to construct
components of cells such as proteins and
nucleic acids.
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Enzyme - Introduction
• Majority biochemical reaction
• can’t take place spontaneously

Chemical reaction
• Using catalyst
•  rate (acceleration) of chemical
reaction
• Catalyst - remain unchanges

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Enzyme - Introduction
What is enzyme?
• In biochemical reaction
• Catalyst enzymes

Example:
• Oxidation of fatty acid
• Break down of protein

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Enzyme - Introduction
Oxidation of fatty acid
The oxidation of a fatty acid to carbon dioxide and
water is not a gentle process in a test tube -
extremes of pH, high temperatures and corrosive
chemicals are required.

Yet in the body, such a reaction takes place

smoothly and rapidly within a narrow range of pH
and temperature.

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Enzyme - Introduction
Break down of protein

In the laboratory, the average protein must

be boiled for about 24 hours in a 20% HCl
solution to achieve a complete breakdown.

In the body, the breakdown takes place in

four hours or less under conditions of mild
physiological temperature and pH.

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Enzyme – Chemical Nature
Enzymes:
• Protein in nature
• High molecular weight
• ~ 10,000 to 2,000,000
• Chain of amino acid
• Linked by peptide bond
• Denature at high temperature
• Precipitated with salts / solvent…

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Enzyme – Chemical Nature
Enzyme

•posses catalytic activity

• rate of chemical reaction
•No changes in the reaction
•Regenerate afterward

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Cell metabolism required:

1. Extracting matter & energy from

the environment
2. transporting various chemicals
3. synthesizing new biomolecules
4. moving within its environment
5. replicating, new generation

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• Without enzymes
• very slowly or no reaction
• enzymes speed up reactions
• maintain their life
• each reaction within cell
• catalyzed by a specific enzyme

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Enzymes - Protein in nature
• 3o structure

Folding rule
• Polar out / Non-polar In

Bonding
1. Hydrophobic interaction
2. H-bond
3. Ion pair (salt bridges)
4. Disulfide bonds
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What is substrate?

• In enzymatic reaction
• Substrate
• beginning of the process

• Within substrate
• make / break chemical bonds

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Enzyme – active site
• Polypeptide chain folded
• unique 3-D shape
• pocket or cleft on the enzyme
surface
• active site

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• substrate binds to the active site of
the enzyme
• form an Enzyme-Substrate complex
(ES complex)
• further reactions & form product
• then regenerated enzyme

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• enzyme molecule  lock
• Substrate  key,
• active site  keyhole of the lock.
This description is called the lock-
and - key theory of enzyme
structure.

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Some enzymes
-Required NO chemical groups for
activity

Some enzymes
- Require additional chemical
component
- addition component Cofactor

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Three types of cofactors

Nature: organic or inorganic

Cofactor:
**Non Protein chemical component**

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1.Coenzyme
cofactor – enzyme  loosely bound

•Non-protein organic substance

•As carriers for transferring chemical
groups or atoms from one enzyme to
another
•Many coenzymes are derived from
vitamins

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1.Coenzyme

Example:
Nicotinamine adenine dinucledtide (NAD)
acts as a coenzyme to dehydrogenase
(the enzyme) by acting as a hydrogen
acceptor

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2.Prosthetic group
cofactor – enzyme tightly bound
covalent bond

Non-protein organic molecules

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2.Prosthetic group

Example:
Haem 血紅素is a prosthetic group.
It is a ring-shaped organic molecule with
iron at its centre.
Haem is the prosthetic group of the
electron carrier cytochrome 細胞色素
and of the enzyme catalase.

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Three types of cofactors
Nature: organic or organic

3.Metal ions / inorganic ions

various metal ion
e.g. Mg+2, Mn+2, Zn+2, Cu+2
enzyme – metalloenzymes

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3.Metal ions / inorganic ions

Example:
Salivary amylase requires the
presence of chloride (Cl-) ions before
it will efficiently change starch into
maltose

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Apoenzyme
• an enzyme requires cofactor
• but does not have bound
• Inactive enzyme

Holoenzyme
• an apoenzyme + cofactor
• catalytically active

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Active site (key hole)

•Specific 3 dimensional shape

•Only one type of substrate

• “Specificity”

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Few enzymes
•absolute specificity
•Catalyze only one particular reaction

Other enzymes
•Specificity for particular chemical
bond / functional group

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4 main classes of specificity :

1.Absolute specificity

2.Group specificity

3.Linkage specificity

4.Stereospecificity

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1. Absolute specificity
Enzyme only catalyse on one
reaction

Example
1. succinate dehydrogenase, that is specific for
succinate (In citric acid cycle / Krebs cycle)

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• enzyme act on substrate
• which substrate with
• specific functional groups

Example
Phosphatases –
phosphate functional group

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• enzyme only act on substrate
• which substrate having
• particular type of chemical bond /
linkage

Example
Esterases - hydrolysis of esters

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• enzyme only act on substrate
• which substrate
• having steric / optical isomer

Example:
• Enzyme can discriminate between D-
stereoisomers and L-stereoisomers
• Catalyse the reaction on one of them

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Example: (a) & (b) are stereoisomer

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How polypeptide chain folds &
make the active site

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1. -ve charged amino acid
• enzyme  –ve charged
• substrate  +ve charged
• Electrostatic interaction
e.g. aspartic acid & glutamic acid

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2. +ve charged amino acid
• Enzyme  +ve charged
• Substrate -ve charged
• Electrostatic interaction
• E.g. histidine, lysine and arginine

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3. Hydrophobic amino acid
• Enzyme  hydrophobic
• Substrate  hydrophobic
• Hydrophobic/hydrophobic interaction
• E.g. phenylalanine

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4. Hydrophilic amino acid
• Enzyme hydrophilic
• Substrate hydrophilic
• Hydrogen bonding

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Simple naming system

1. According to substrate that the

enzyme acted on
• Added suffix “-ase”

For example,
• Enzyme acted on substrate - arginine
called “arginase”

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Simple naming system

2. According to the type of reaction

catalyzed
• Added suffix “-ase”

For example,
• Enzyme “glucose oxidase” catalyzes
oxidation of glucose

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•Systematize naming enzyme

•International Enzyme Commission

•IEC

•based on type of catalytic reaction

•6 major categories

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1. Oxidoreductases catalyze oxidation-
reduction reactions
Example:

Oxidoreductases - require a cofactor

NAD+, which accepts the hydrogens released during oxidation.

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1. Oxidoreductases catalyze oxidation-
reduction reactions
Example:

Oxidases - oxygen is used as an acceptor

glucose oxidase, glucose  gluconic acid

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2. Transferases catalyze - transfer
functional groups of substrate
Example:
methyltransferase, transfers a methyl group

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2. Transferases catalyze - transfer
functional groups of substrate
Example:
transaminase, transferring an amino functional group from one
molecule to another. This allows for the interconversion of certain
amino acids, and also allows amino acids to enter into glucose
metabolism.

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3.Hydrolases catalyze
•addition of a water molecule
•process called hydration
•cleaves the bond

Example:
Lipases, phosphatases,
acetylcholinesterase and proteases

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4. Lyases catalyze
• removal of various groups
• from substrates
• forming double bonds
• cleavage of C-C, C-O and C-N bonds
• not by means of hydrolysis

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5. Isomerases catalyze
• intramolecular rearrangements
• E.g. interconversion of D &L-isomeric

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6. Ligases catalyze the formation of
chemical bonds.
• E.g. C-C, C-S, C-O and C-N bond
• need input of chemical energy, ATP
to form new bond

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