ENZYMES

DR. WAN HAFIZAH W. JUSOF

27 March 2018
 Lecture Outlines
• Introduction of enzymes
• Mechanism of enzyme actions
• Enzyme Cofactors
• Enzymes classification
• Factors affecting of reaction velocity of
enzyme
• Enzyme Kinetics
 Learning Outcomes
• Describe enzymes and their mechanism of
actions
• Give examples of cofactors and their function in
enzymes activities
• Identify enzymes according to their classification
• Explain factors affecting velocity reactions of
enzymes
 What are ENZYMES?
Definition:
•Enzymes are biological catalyst that can speed up the
chemical reactions without being consumed in the process.
•Most enzymes are Proteins (tertiary/quaternary structures)
•Not permanently changed in the process
•Catalyze nearly all the chemical reactions in the cells
•Have unique 3 dimensional shapes that fit the shapes of
reactants
 Enzymes
• They are very specific for their substrates
 Enzyme vocabulary
• Substrate
– molecule that enzymes work on
• Products
– what the enzyme helps produce from the reaction
• Active site
– part of enzyme that substrate molecule fits into
Enzymes
 HOW DO ENZYMES WORK
Energy Changes Occur During the Reaction
•All chemical reactions have an energy barrier (activation
energy) – the minimum energy required to start a chemical
reaction
 HOW DO ENZYMES WORK
Energy Changes Occur During the Reaction
•Enzyme lowers the activation energy
•Without enzyme, the reaction will occur only if enough heat
energy is added
•With an enzyme as a catalyst, the reaction may easily
proceed at the normal physiological temperature
 MECHANISM OF ENZYME ACTION
• Enzyme attracts substrates to its active site 🡪 catalyzes
the chemical reaction by which products are formed
🡪allows the products to dissociate (separate from the
enzyme surface)
• The combination formed by an enzyme and its
substrates is called the enzyme–substrate complex. 
 Enzyme Active Site
• Amino acid side chains interact, metal ions,
• Various types of polar, non-polar, ionic interactions
• TWO models for substrate binding to the
active site of the enzyme:
1) Lock and key model
2) Induced fit model (Hand-in glove model)
LOCK AND KEY MODEL
• Proposed by Emil Fisher in 1890
• Lock = enzyme, key = substrate
 LOCK AND KEY MODEL
• The active site has a rigid shape
• Only substrates with the matching shape can fit
• The substrate is a key that fits the lock of the active site
LOCK AND KEY MODEL
LOCK AND KEY MODEL
LOCK AND KEY MODEL
Basic Enzyme Diagram
 INDUCED FIT THEORY
Proposed by Daniel E Koshland in 1958
•The active site is flexible, not rigid
•Shape of the active site can be modified by the binding of
the substrate.
•Placing a hand (substrate) into a glove (enzyme) induces
changes in the glove’s shape
 COFACTORS
• Additional nonprotein component required by some enzymes for
its optimum activity
• Bound to the protein portion of the enzyme
• These cofactors may be:
– Organic compounds – coenzymes
– Inorganic ions - activators
• Apoenzyme = enzymes without its cofactor
• Holoenzyme = Apoenzyme + cofactor
• Many vitamins function as coenzymes
COFACTORS (ACTIVATORS)
Enzymes requiring inorganic elements as cofactors
 Enzyme Cofactors
• Coenzyme - non protein organic, maybe a vitamin.
ENZYME CLASSIFICATION
• Six major classes:
 FACTORS AFFECTING
REACTION VELOCITY OF
ENZYMES
- Substrate concentration
- Temperature
- pH
(Vmax)
• The optimum temperature for most human enzymes is between 35
and 40oC. Enzymes start to denature at temperatures above 40oC
• The optimum pH differs from enzyme to enzyme; Pepsin
=1.2; Trypsin = 8.0
• Enzyme activity is related to the ionic state of active site of
the enzyme.
Michaelis-Menten
Kinetics Dr. Wan Hafizah Binti W.
Jusof
2017
 MICHAELIS – MENTEN KINETICS
MICHAELIS- MENTEN EQUATION

k1 k2
[E] + [S] [ES] [P] + [E]
It is based on a model ofkenzyme
-1 action in which the enzyme (E) binds to
a single substrate (S) to form an enzyme-substrate complex (ES), which
breaks down to form products (P) and to liberate free enzyme (E)
FIGURE A
The relationship between reaction rate and substrate
concentration shown in FIGURE A is described as
follows:
 SIGNIFICANCE OF KM
(MICHAELIS CONSTANT)
1. Km is characteristic of an enzyme and its particular substrate &
reflects the affinity of the enzyme for that substrate.
• Low Km indicates strong binding with its substrate because a
low concentration of substrate is needed to half-saturate the
enzyme
• High Km indicates weak binding with its substrate because a
high concentration of substrate is needed to half-saturate the
enzyme

1. The Michaelis constant. Km has two significances:

a) Km provides a measure of the substrate concentration
required for significant catalysis to occur
b) It is a measure of the affinity of the enzyme for its substrate
• Low Km indicates strong
binding with its substrate
because a low
concentration of substrate
is needed to half-saturate
the enzyme

• High Km indicates weak

binding with its substrate
because a high
concentration of substrate
is needed to half-saturate
the enzyme