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Enzyme

Why Are Enzymes So Important?


Why are we
devoting two whole
lecture topic to a
enzyme?
Nearly all chemical
reactions in
biological cells need
enzymes to make
the reaction occur
fast enough to
support life.
From the Virtual Cell Biology Classroom on ScienceProfOnline.com

Image: Jumping rope, Meagan E. Klein

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

3. Chemical composition of enzymes


(1) Simple protein
(2) Conjugated protein
Holoenzyme= Apoenzyme+ Cofactor

Cofactor

Coenzyme : loosely bound to enzyme (noncovalently bound).


Prosthetic group : very tightly or even
covalently bound to enzyme (covalently bound)

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

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)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.

Substrate approaches active site


Enzyme-substrate complex forms
Substrate transformed into products
Products released
Enzyme recycled

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

Km: Michaelis constant


Km = (k2 + k3)/k1

after

(2) Effectof[E]onvelocity
[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 Bellshapedcurve

(4) Effect of pH value on velocity


Bellshapedcurve

The pH optimum varies for


different enzymes.
Most enzyme: neutral pH (68).

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
common reason for
this phenomenon.

(5) Effect of activator on veloci


Enzyme activators are molecules that bind to
enzymes and increase their activity.

(i). Inorganic ions


Metalions suchasNa+,K+,Mg2+,Ca2+,Cu2+,Zn2+,
Fe2+etal
Anions:suchasCl,Br,I CNetal

(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.
may
be
foreign
Other
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.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