ENZYMES

Jay Andrea vea D. Israel, RMT, MPH

Learning objectives
 Systematically list down the classes, subclasses, sub-subclasses of
each individual enzyme.
 Differentiate the functions of different enzymes
 Discuss the different properties of enzymes
 Plot and examine the changes in energy for in enzyme kinetics
 Understand the Michaelis – Menten equation
 Synthesize the factors affecting enzymatic action
 Explain the role and mechanism of enzyme inhibitors
BLUFF – ENZYMES game mechanics
 Divide the class into 5 teams.
 Each team will be given a topic on enzyme.
 Each team will strategically provide 15 statements or facts on their
card for their given topic. The first team will put down their cards
and say aloud their statement (Note: only 10 cards are used in the
game, the other 5 will be a reserve card)
 Other teams gets to call out if the player is bluffing or not.
 The other teams who called the bluff must hand down the right
answer to get 5 points
BLUFF – ENZYMES game mechanics
 The player will get a card from the reserve card if
the other teams figure out their bluffs
 if the team does not get the right answer, they will
get a card from the reserve card
 The team who were not opposed on their bluff
question will get additional 10 points per card
 This goes on till a team finishes all their cards.
 Topics
 Properties of Enzymes
 Enzyme Structure
 Cofactors and co-enzyme
 Catalytic power
 Specificity
 Enzyme Classification and Nomenclature
 Enzyme Kinetics
 Factors affecting enzyme activity
 Enzyme Inhibition
ENZYMES
 “biological catalysts” that accelerates biological
reactions.
 Found in all body tissues and appears in serum
after cellular damage or injury.
Properties of an Enzyme
 Catalytic power
 Structure of Enzymes
 Specificity
 Enzyme Naming and Nomenclature
Catalytic Power
 Enzymes have high catalytic activity, speeding up
reactions as much as 1021 over uncatalyzed levels
under optimum temperature and pH conditions.
 Catalytic power is defined as the ratio of the
enzyme-catalyzed Rate of a reaction to the
uncatalyzed rate.
Enzyme Structure

Figure 5.2. Enzyme Structure (taken from

http://www.phschool.com/science/biology_place/labbench/lab2/images/enzyme.gif)
Enzyme Structure
Specificity
 Enzymes are highly selective, catalyzing only specific
substrates (substances where enzyme acts) and specific
reaction
 Absolute: specific to only 1 substrate
 Group: specific to all substrates of a chemical group

 Bond: specific to chemical groups

 Stereoisometric: specific to 1 optical isomer of a compound

©2010 Wolters Kluwer Health I Lippincott Williams & Wilkins
 Figure 5.1. Lock and Key Model and Induce Fit model of Enzyme specificity
(taken from https://alevelbiology.co.uk/wp-content/uploads/2019/02/Enzymes_1.png)
 Figure 5.1. Lock and Key Model and Induce Fit model of Enzyme specificity
(taken from https://alevelbiology.co.uk/wp-content/uploads/2019/02/Enzymes_1.png)
Enzyme classification and Nomenclature
 Enzymes are formally named according to the
reaction they catalyze
 Six classes are recognized:
Oxidoreductases Transferases
Hydrolases Lyases
Isomerases Ligases
Systematic Classification of Enzymes
According to Enzyme Commission
(taken from p384 from Biochemistry 4e by
Garrett & Grisham)
EC. 1 Oxidoreductases
 CATALYZE REDOX REACTIONS
 SOME COMMON TYPES:
 REDUCTASES
 DEHYDROGENASES
 OXIDASES
 OXYGENASES
 LOOK FOR INVOLVEMENT OF COENZYMES
 NAD+ , NADP+, FAD, FMN
EC. 1 Oxidoreductases
E.C. 2 TRANSFERASES
 TRANSFER OF FUNCTIONAL GROUPS
 SOME EXAMPLES:
 TRANSFERASES
 PHOSPHORYLASES
 TRANSFER OF Pi OR PPi
 KINASES
 A PHOSPHATE TRANSFER
 UTILIZE OR GENERATE ATP
E.C. 2 TRANSFERASES
E.C. 3 HYDROLASES
 HYDROLYSIS REACTIONS

 WATER MOLECULES USED TO BREAK BONDS

 EXAMPLES:
 PHOSPHATASES
 PEPTIDASES
 ATPase, GTPase
E.C. 3 HYDROLASES
E.C. 4 LYASES
 CATALYZES REACTIONS THAT
 GENERATE A DOUBLE BOND
 ADDS A SUBSTRATE MOLECULE TO DOUBLE BOND OF
A SECOND SUBSTRATE
 EXAMPLES:
 DECARBOXYLASES
 DEHYDRATASES
 ALDOLASE
E.C. 4 LYASES
E.C. 5 ISOMERASES
 CONVERSION OF ONE ISOMERIC FORM INTO
ANOTHER
 EXAMPLES:
 ISOMERASES
 EPIMERASES

 RACEMASES

 MUTASES
E.C. 5 ISOMERASES
E.C. 6 LIGASES
 TWO MOLECULES ARE JOINED
 ANABOLIC REACTIONS
 REQUIRE NUCLEOTIDES (ATP, GTP) TO DRIVE THEM
 A PYROPHOSPHATE BOND MUST BE BROKEN
 EXAMPLES:
 CARBOXYLASE
 SYNTHETASES
 DON’T CONFUSE SYNTHETASES WITH SYNTHASES
E.C. 6 LIGASES
Enzymatic Activity
 Cofactors and Coenzymes
 Enzyme Kinetics
 Activation Energy and Free Energy
 Factors affecting Enzyme activity
Cofactors and Coenzymes

Table 5.2 Cofactors and Coenzymes (from pp 385 of your reference by Garett &
Grisham)
Enzyme Kinetics
 General Principle
 Study of the rate of catalyzed reactions by enzymes
 Provides a mechanism of enzyme activity and how it is
regulated
 The reaction catalysed by an enzyme uses exactly
the same reactants and produces exactly the same
products as the uncatalysed reaction.
Enzyme Kinetics
 Like other catalysts, enzymes do not alter the
position of equilibrium between substrates and
products.
 However, unlike normal chemical reactions,
enzymes are saturable.
 This means as more substrate is added, the
reaction rate will increase, because more active
sites become occupied.
 This can continue until all the enzyme becomes saturated
with substrate and the rate reaches a maximum.
 The two most important kinetic properties of an enzyme are
how quickly the enzyme becomes saturated with a
particular substrate, and the maximum rate it can achieve.
 Knowing these properties suggests what an enzyme might
do in the environment of the cell and can show how the
enzyme will respond to changes in these conditions
Enzyme kinetics
E+S ES EP E+P
Where:
E represents enzyme
S represents the substrate
P represents the product
ES represents enzyme-substrate complex
EP represents enzyme – product complex
 Activation Energy
The beginning of reaction starts with
the reactants’ certain level of energy
and reach a transition state or
activation energy (ΔG*) where an
additional energy is absorbed and
lastly energy is released to form a
product.
The change in free energy (ΔG°) is the
difference in the free energy of
reactants and products.
Enzyme Kinetics
 Michaelis-Menten Equation
 Basis for most single-substrate enzyme kinetics
 Vmax – the maximum rate of reaction when all enzyme
active sites are saturated with substrate
 Km – the substrate concentration that gives half
maximal velocity. Km is a measure of the affinity an
enzyme has for its substrate, as a lower Km means that
less of the substrate is required to reach half of Vmax.
Enzyme Kinetics

Michaelis-Menten Curve
Factors affecting Enzyme activity
 Temperature
 pH
 Enzyme concentration
 Substrate concentration
 Product concentration
 Activators
Temperature
 Rate of enzyme-
catalyzed
reactions generally
increases with
temperature
 At temperature
50-60°C, activity
of enzyme declines
Effect of Temperature on Enzyme activity
(taken from p397 from Biochemistry 4e by Garrett & Grisham)
Temperature
 Enzymes are most active at optimum temperature
(37C)
 Two effects
 Increaseof reaction rate with temperature
 Denaturation of protein structure due to high
temperature
 Effect of pH on Enzyme activity

pH (taken from p397 from Biochemistry 4e by Garrett & Grisham)

 Enzymes in
general are
active only over
a limited pH
range, and most
have a
particular pH at
which their
catalytic activity
is optimal
pH

(taken from p397 from Biochemistry

4e by Garrett & Grisham)
 Enzyme Concentration
• Increasing enzyme concentration
will speed up the reaction, as long
as there is substrate available to
bind to.
• Once all of the substrate is bound,
the reaction will no longer speed
up, since there will be nothing for
additional enzymes to bind to.

Image taken from http://science.halleyhosting.com/sci/ibbio/chem/notes/chpt8/substraterxnrate.gif

 Substrate concentration
• Increasing substrate
concentration also increases the
rate of reaction to a certain
point.
• Once all of the enzymes have
bound, any substrate increase
will have no effect on the rate of
reaction, as the available
enzymes will be saturated and
working at their maximum rate.
https://cdn.kastatic.org/ka-perseus-
images/e3a3422f253ec134f28a7a7aca27d9e08623b755.png
Enzyme Inhibition
 Enzyme inhibitors cause a decrease in enzyme
activity by binding to the enzymes.
 Can be reversible or irreversible
 Types of Enzyme Inhibition
 Competitive Inhibition
 Non-competitive Inhibition
 Uncompetitive Inhibition
Competitive Inhibition

Figure 5.4. Competitive inhibition

(taken from https://ib.bioninja.com.au/_Media/competitive-inhibition_med.jpeg)
Non-Competitive Inhibition

Figure 5.5. Non-competitive Inhibition

(taken from https://ib.bioninja.com.au/_Media/noncompetitive-inhibition_med.jpeg)
Uncompetitive Inhibition

Taken from https://alchetron.com/cdn/uncompetitive-inhibitor-f6ae6cfa-a4d2-40fc-9093-ae60816b69f-resize-750.jpeg

Irreversible Inhibition
 Causes enzyme to lose all activity
 Often toxic substance that destroys enzymes
 Usually forms covalent bond with an amino acid
side chain preventing catalytic activity
 May be nerve gas, an insecticide, or an antibiotic
Differences in the types of Enzyme Inhibition
Competitive Non-Competitive Uncompetitive

Inhibition Binds with Binds at the Cause inhibitor

active site alternative site / to bind at the
mechanism allosteric site active site

Km Increase Unchanged Increase

Vm Unchanged Decrease Unchanged

 taken from https://cdn.kastatic.org/ka-perseus-
images/63518c984a09ce17b78b60831e2d6eae8bbacd56.png