ENZYMES

dr.Syahrijuita, M.Kes, Sp.THT-KL

Deparment of Biochemistry
Medical Faculty of Hasanuddin University
 Topic
• Definition,composition, structure and
properties of enzyme
• How Enzymes work
• Enzyme activity
• Factors affecting enzyme activity
• Regulation of enzyme activities
• Enzymes in clinical diagnosis
 Definition of enzyme
•Enzymes are biological catalysts.
•A Catalyst is defined as "a substance
that increases the rate of a chemical
reaction without being itself changed in
the process.”
INTRODUCTION
 Enzymes are biological catalysis
- substance of biological origin that accelerate
chemical reactions.
- the vast majority of enzyme are proteins.
- catalytically active ribonucleic acids “ ribozymes”.
- in general, name suffix “-ase”
 The presence and maintenance of a complete and
balanced set of enzymes is essential for
- the breakdown of nutrients to supply energy, and
chemical building block;
- the assembly of those building blocks into proteins,
DNA, membranes, cells, and tissues;
- and the harnessing of energy to power cell motility and
muscle contraction.
Deficiency in quantity and catalytic activity can result
from:
- genetic defects,
- nutritional deficits,
- or, toxins.
Defective enzymes can result from:
- genetic mutation
- infections by viral or bacterial pathogens.
Medical scientists:
- imbalances in enzyme activity
pharmacologic agent to inhibit.
- investigating gene therapy
remedy deficits in enzyme level and function.
 Enzymes as Biological Catalysts
Enzymes are
proteins that
increase the rate of
reaction by lowering
the energy of
activation
They catalyze nearly
all the chemical
reactions taking
place in the cells of
the body
Enzymes have
unique three-
dimensional shapes
that fit the shapes of
reactants
(substrates)
 Properties of enzymes (important!)

• Catalytic efficiency – high efficiency, 103 to

1017 faster than the corresponding
uncatalyzed reactions

• Specificity - high specificity, interacting with

one or a few specific substrates and
catalyzing only one type of chemical reaction.
• Mild reaction conditions- 37℃, physiological
pH, ambient atmospheric pressure
 Chemical composition of enzymes

(1) Simple protein

(2) Conjugated protein

Holoenzyme= Apoenzyme+ Cofactor

Coenzyme : loosely bound to enzyme (non-covalently bound).

Cofactor
Prosthetic group : very tightly or even covalently bound to
enzyme (covalently bound)
 Classification of enzymes

(1). By their composition

1). Monomeric enzyme
2). Oligomeric enzyme
3). Multienzyme complex: such as
Fatty acid synthase
 (2) Nomenclature
• Recommended name
•Enzymes are usually named according to the
reaction they carry out.
•To generate the name of an enzyme, the
suffix -ase is added to the name of its
substrate (e.g., lactase is the enzyme
that cleaves lactose) or the type of
reaction (e.g., DNA polymerase forms
DNA polymers).
•Systematic name (International classification)
• By the reactions they catalyze (Six
classes)
Enzyme Nomenclature
International Union of Biochemist (IUB)
In general:
- Type of reaction catalyzed followed by suffix –ase.
- dehydrogenase, protease, etc.

 IUB:
- Each enzyme has a unique name and code number that reflect the type of
reaction catalyzed and the substrate involved.
- EC 1.2.3.4
- EC, Enzyme catalog
- 1, Class
- 2, subclasses
- 3, subsubclasses
- 4, where the enzyme belongs in the subsubclases.
- EC 2.7.1.1
- class 2, transferase
- subclass 7, transfer of a phosphoryl group.
- subsubclass 1, alcohol is the phosphoryl acceptor.
- 1, hexose-6, alcohol phosphorylated is the of carbon-6 of a
Enzyme Classes (IUB)
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5. How enzymes work (important!)
1) Enzymes lower a
reaction’s activation
energy
All chemical reactions have
an energy barrier, called the
activation energy,
separating the reactants
and the products.
activation energy: amount
of energy needed to disrupt
stable molecule so that
reaction can take place.
Enzymes
Lower a
Reaction’s

Activation
Energy
 What is the difference between an enzyme
and a protein?

Protein Enzymes RNA

•All enzymes are proteins except some RNAs

• not all proteins are enzymes
2) The active site of the enzyme
Enzymes bind substrates to their active site and
stabilize the transition state of the reaction.
The active site of the enzyme is the place where
the substrate binds and at which catalysis occurs.
The active site binds the substrate, forming an
enzyme-substrate(ES) complex.

Binding site
Active site
Catalytic site
 S
E
E
E

Enzyme- Enzyme may

substrate be used again
complex P

Reaction coordinate
© 2007 Paul Billiet ODWS
The Lock and Key Hypothesis
This explains enzyme specificity
This explains the loss of activity when enzymes
denature

© 2007 Paul Billiet ODWS

Induced Fit Model
Enzymes can form to the shape of its substrate.

http://en.wikipedia.org/wiki/File:Induced_fit_diagram.svg

Life Sciences-HHMI Outreach. Copyright 2009 President and Fellows of Harvard College
The Induced Fit Hypothesis

Hexokinase (a) without (b) with glucose substrate

http://www.biochem.arizona.edu/classes/bioc462/462a/NOTES/ENZYMES/enzyme_mechanism
.html

This explains the enzymes that can react with a

range of substrates of similar types
© 2007 Paul Billiet ODWS
1. Substrate approaches active site
2. Enzyme-substrate complex forms
3. Substrate transformed into products
4. Products released
5. Enzyme recycled
6. Enzyme activity
Enzymes are never expressed in terms of their
concentration (as mg or μg etc.), but are expressed only
as activities.
Enzyme activity = moles of substrate converted to
product per unit time.
The rate of appearance of product or the rate of
disappearance of substrate
Test the absorbance: spectrophotometer
7. Factors affecting enzyme activity
Concentration of substrate
Concentration of enzyme
Temperature
pH
Activators
Inhibitors
 Enzyme velocity
Enzyme activity is commonly expressed by the intial
rate (V0) of the reaction being catalyzed. (why?)

• Enzyme activity = moles of

substrate converted to product
per unit time.
 1. Michaelis-Menten equation describes how
reaction velocity (V) varies with substrate
concentration [S].
• The following equation is obtained after
suitable algebraic manipulation.
[S]
V = Vmax Note: V means V0
[S] + KM

Km: Michaelis constant

Km = (k2 + k3)/k1
Persamaan Michaelis-Menten

Vomax [S]
Vo= ----------
Km + [S]

Apabila
[S] <<< Km
[S] >>> Km

[S] = Km

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(2) Effect of [E] on velocity

[S]>>[E] V∝[E]
• The initial rate of an
enzyme-catalyzed
reaction is always
proportionate to the
concentration of enzyme.
• This property of enzyme
is made use in determining
the serum enzyme for the
diagnosis of diseases.
(3) Effect of temperature on velocity
Bell-shaped curve
 (4) Effect of pH value on velocity
Bell-shaped curve
• Each enzyme has
an optimal pH or pH
range (where the
enzyme has maximal
activity).
• Requirements for
the catalytic groups
in the active site in
appropriate
ionization state is a
•The pH optimum varies for different
enzymes.
common reason for
•Most enzyme: neutral pH (6-8). this phenomenon.
 Optimum pH values

Enzyme
activity Trypsin

Pepsin

1 3 5 7 9 11

© 2007 Paul Billiet ODWS

pH
(5) Effect of activator on velocity
•Enzyme activators are molecules that bind to enzymes and increase
their activity.

(i). Inorganic ions

• Metal ions ， such as Na+, K+, Mg2+, Ca2+, Cu2+, Zn2+, Fe2+ et al

• Anions: such as Cl-, Br-, I- 、 CN- et al

(ii). Organic
• Reducing agents, such as Cys 、 GSH

(iii). Proteins
 (6) Inhibition of enzyme activities
(very important!)
• Inhibitor: any molecule which acts
directly on an enzyme to lower its
catalytic rate is called an inhibitor.
(not denaturation)
• Some enzyme inhibitors are normal
body metabolites.
• Other may be foreign
substances,such as drugs or toxins.
8. REGULATION OF ENZYME ACTIVITY
1. Allosteric binding sites: Allosteric enzymes are
regulated by molecules called effectors
(modifiers) that binds nonconvalently at a site
other than the active site.
2. By Covalent Modification: Many enzymes are
regulated by covalent modification, most
frequently by the addition or removal of
‘phosphate’ group to serine, threonine or
tyrosine residue of the enzyme by kinases.
(enzyme)
3. Induction and repression of enzyme
sysnthesis: Cells can also regulate the amount
of enzymes present by altering the rate of
enzyme synthesis.
REGULATION CONT….
4. Zymogen Cleavage: Some enzyme are
synthesized as inactive precursor, called
zymogens, that are activated by proteolysis (e.g.,
digestive enzyme, pepsinogen is inactive and
cleaved to pepsin which is active chymotrypsin)
5.Location within the cell: Many enzymes are
localized in specific organelles within the cell.
This, compartmentation helps in the regulation of
the metabolic pathway.
9. Enzymes in clinical diagnosis
An enzyme test is a blood test or urine test that
measures levels of certain enzymes to assess how well
the body’s systems are functioning and whether there
has been any tissue damage. (why?)
Principal Serum Enzymes Used in Clinical Diagnosis.

Note: Many of the enzymes are not specific for the disease listed.
Some diseases caused by enzyme malfunctions

diseases Enzyme malfunction

Albino 3-monooksigenase tyrosine

Alkaptonuria 1,2-dioksigenase homogentisat
Galactosemia Uridilil transferase galactosa 1-fosfat
Homosistinuria -sintase sintationine
phenylketonuria 4-monooksigenase phenylalanine
Tay-sachs Heksosaminidase A
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http://www.chem.qmul.ac.uk/iubmb/enzyme/

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