Enzymes

LEARNING OUTCOMES
• Understand enzyme reaction
• Identify factors affecting enzyme reaction
• Distinguish between lock-and-key model and induced-fitmodel
• Describe enzyme kinetics
• Compare allosteric enzyme with non-allosteric enzyme
• Relate Michaelis-Menten and Lineweaver-Burk plots to enzyme
reaction
WHAT IS ENZYME?
• Globular proteins mostly, exception for ribosomal RNA (ribozyme)
• Catalyst for most biological reactions
• Molecules at the beginning of the chemical reaction are called
substrates, and the enzyme converts them into different molecules,
called the products.
• Change the rate of reaction without being consumed by the reaction
(reusable)
• The active site in an enzyme’s catalytic centre, highly specific for
what they will catalyze
• End in–ase (sucrase, lactase, maltase)
ACTIVE SITE
• Region within an enzyme that fits the shape of molecules called
substrate
• Contain amino acid R group that align and bind the substrate
• Substrate are held in the active site by weakin teraction
(Handionicbonds)
• R group of a few amino acid on the active site catalyse the conversion
of substrate to product
• Release product when there action is complete
Classes of enzymes
ENZYME NOMENCLATURE
• Enzyme Commission number (EC number) is a numerical classification
scheme for enzymes, based on the chemical reactions they catalyze
• For example, the tripeptide amino peptidase have the code "EC3.4.11.4"
whose components indicate the following groups of enzymes
• EC3 enzymes are hydrolases (enzymes that use water to break up some
other molecule)
• EC3.4 are hydrolases that act on peptide bonds
• EC3.4.11 are those hydrolases that cleave off the amino/N-terminal amino
acid from a polypeptide
• EC3.4.11.4 are those that cleave off the N-terminal end from a tripeptide
How?
• Enzyme works by weakening bond
which lowers activation energy
• Activation energy is the amount of
energy necessary to push the
reactant over an energy barrier
• At the summit, the molecule are at
an unstable point, the transition
state
• The different between free energy
of the product and the free energy
of the reactant is the delta G
ENZYME ARE SUBSTRATE SPECIFIC
• A substrate is a reactant which bind to an enzyme
• When a substrate bind to an enzyme, the enzyme will catalyse the
conversion of substrate to the product
• Two model have been developed to describe formation of enzyme-
substrate complex:
• Lock and key model
• Induced fit model
LOCK AND KEY MODEL
• Substrate bind to that portion of the enzyme with a complementary
shape
• Active site have a rigid shape
• Only substrate with the matching
shape can fit
• The substrate is a key that fits the
lock of the active site
INDUCED FIT MODEL

• Binding of the substrate induced a change in the conformation of the

enzyme that result in the complementary fit
• Enzyme is flexible, not rigid greater range of substrate specificity
• The shapes of the enzyme, active site and substrate adjust to the
maximum fit which improves catalysis
Induce fit model (2)

• Enzyme change shape leading to a tighter induce fit. Bringing

chemical group in position to catalyse reaction.
• Involved change of Hand ionic bonds
• Once the reaction complete, attraction between substrate/molecule
and enzyme cease, freeing the enzyme from product enabling to
catalysed the second reaction of the same type
ENZYME REACTION
cofactor vs coenzyme
...cofactors
Function of coenzyme

A coenzyme prepares the active site for catalytic activity

Isozymes (isoenzymes)
• Enzymes that differ in amino acid sequence but catalyse the same
chemical reaction
• The existence of isozymes permits the fine-tuning of metabolism to
meet the particular needs of a given tissue or develop mental stage
(for example lactate dehydrogenase (LDH)
ALLOSTERIC REGULATION
• Allosteric regulation is the regulation of an enzyme or other protein
by binding an effector molecule at the protein's allosteric site (that is,
a site other than the protein's active site)
• Effectors that enhance the enzyme's activity are referred to as
allosteric activators
• Whereas those that decrease the enzyme's activity are called
allosteric inhibitors.
ALLOSTERIC ENZYME
• Most allosterically regulated enzyme are constructed of two or more
polypeptide chains
• Each subunit has its own active site and allosteric sites (located where
subunit joins)
ALLOSTERIC ENZYMES
• Some allosteric regulators activators, stabilize the conformation that
has a function active site
• Other regulator inhibitor, stabilize the conformation that lacks an
active site
• R group of amino
acids are properly
charged at optimum
pH
• Activity low at higher
or lower pH than the
optimum is because
of a distrupted
tertiary structer
Effect of substrait concentration on enzyme
activity
Substrate concentration on enzyme activity
Enzyme concentration and enzyme action
• When enzyme concentration increases, the rate of reaction also
increases
• More abundant enzymes means more substrates can be catalysed in a
given amount of time  more substrates can bind with the active site
of enzymes
• Therefore, given the unlimited amount of substrates, rate of reaction
keeps on increasing as enzyme concentration increases
ENZYME INHIBITION
Enzyme kinetics
Kinetics of enzyme reaction is described with
Km and Vmax
The end!