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Outline
Composition, structure and properties
of enzyme
How Enzymes work
Enzyme activity
Factors affecting enzyme activity
Regulation of enzyme activities
Enzymes in clinical diagnosis
1. 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.
EnzymesasBiologicalCatalysts
Enzymesareproteins
thatincreasetherate
ofreactionby
loweringtheenergy
ofactivation
Theycatalyzenearly
allthechemical
reactionstakingplace
inthecellsofthe
body
Enzymeshaveunique
threedimensional
shapesthatfitthe
shapesofreactants
(substrates)
2.Propertiesofenzymes(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
Cofactor
4. 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)
5.Howenzymeswork(important!)
1)Enzymeslowerareactions
activationenergy
Allchemicalreactions
haveanenergybarrier,
calledtheactivation
energy,separatingthe
reactantsandthe
products.
activationenergy:
amountofenergyneeded
todisruptstablemolecule
sothatreactioncantake
place.
Enzymes
Lower a
Reactions
Activation
Energy
2)Theactivesiteoftheenzyme
Enzymes bind substrates to their active site and
stabilizethetransitionstateofthereaction.
The active site of the enzyme is theplacewhere the
substratebindsandatwhichcatalysisoccurs.
The active site binds the substrate, forming an
enzymesubstrate(ES)complex.
Binding site
Active site
Catalytic site
Enzymaticreactionsteps
1.
2.
3.
4.
5.
6.Enzymeactivity
Enzymes are never expressed in terms of their
concentration (as mg or g etc.), but are
expressedonlyasactivities.
Enzyme activity = moles of substrate converted
toproductperunittime.
The rate of appearance of product or the rate of
disappearanceofsubstrate
Testtheabsorbance:spectrophotometer
7.Factorsaffectingenzymeactivity
Concentrationofsubstrate
Concentrationofenzyme
Temperature
pH
Activators
Inhibitors
Enzymevelocity
Enzyme activity is commonly expressed by the
intial rate (V0) of the reaction being catalyzed.
(why?)
Enzymeactivity=moles
ofsubstrateconvertedto
productperunittime.
MichaelisMentenequation(veryimportant!)
1. Michaelis-Menten equation describes how
reaction velocity (V) varies with substrate
concentration [S].
The following equation is obtained
suitable algebraic manipulation.
V=Vmax
[S]
[S]+KM
Note: V means V0
after
(2) Effectof[E]onvelocity
[S]>>[E] V[E]
(ii). Organic
(iii). Proteins
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.Enzymesinclinicaldiagnosis
Anenzymetestisabloodtestorurinetest
that measures levels of certain enzymes to
assess how well the bodys systems are
functioningandwhethertherehasbeenany
tissuedamage.(why?)
Commonenzymesusedforclinicaldiagnosis
include:
alanineaminotransferase(ALT,alsocalled
glutamatepyruvatetransaminase,GPT)
alkalinephosphatase
amylase
aspartateaminotransferase
creatinekinase
lactatedehydrogenase