ENZIMAS I

Curso Química Biológica

Licenciatura en Química
Química Ambiental

2017
 Enzymes
Enzymes are proteins that serve as catalysts for
biochemical reactions.

Enzymes are characterized by:

1. Catalysis for rate enhancement

2. Specificity for substrate

3. Regulation of reaction or pathway

Some are regulatory, some are not
4. Cofactors
Some use cofactors, some do not
 Enzyme Classification
Enzyme Commission System:
Lactate dehydrogenase = EC 1.1.1.27
Know the six main classes and the EC class no.
 Rate Enhancement
Formation of carbonic acid from CO2 and H2O.

Uncatalyzed rate = 1.3 x 10-1 molecules/sec

Catalyzed rate = 1.0 x 106 molecules/sec
Rate enhancement = 7.7 x 106 times

Carbonic
Anhydrase
Lock & Key – Fischer (1894)
A proposal for ES
Induced Fit – Koshland (1963)
A proposal for ES
Examples of Enzymatic Activity.
Proteolytic Activity
Catalyzes cleavage of a peptide bond

A protease
Cleavage of a Peptide Bond
Enzymatic cleavage occurs on the
carboxyl side of the recognized sidechain.
Trypsin
Cofactors
Some
enzymes
require
cofactors
for activity

(Apoenzyme
+
Cofactor =

Holoenzyme)
Enzyme Substrate Complex, ES
(noncovalent)
 Active Site Residues

Need not be
adjacent in
the sequence.
3o structure
must have them
all positioned
around the
active site.
 Types of Catalysis

1. Covalent
2. Acid-Base
1. General acid-base (Bronstead
acid or base, HA or A-)
2. Specific acid-base (solvent,
e.g. water, H+ or OH-)
3. Metal ion
4. Binding Effects
1. Approximation (proximity)
2. Transition state stabilization
 Chymotrypsin
Chymotrypsin is an intestinal protease that
Recognizes and binds non-polar sidechains,
primarily aromatic sidechains: Phe, Tyr, Trp

Cleaves
slower

Cleaves
 Mechanism
Covalent catalysis – two steps

Fast Slow

A covalent
intermediate
 Catalytic Triad in Chymotrypsin
The cataytic triad makes Ser-195 the only acidic
Ser in chymotrypsin.
The developing alkoxide is an excellent nucleophile.
Mechanism

Attack at the
Peptide bond
Stabilization of the tetrahedral
intermediate
The O- forms ion-dipoles with two peptide N-H
hydrogens in the oxyanion hole.
Reform carbonyl and release
N-terminus
 New
substrate
(water)
enters
Stabilization of the tetrahedral
intermediate
Again, the O- forms ion-dipoles with two
peptide N-H hydrogens in the oxyanion hole.
Stabilization of the tetrahedral
intermediate
Again, the O- forms ion-dipoles with two
peptide N-H hydrogens in the oxyanion hole.
Dissociation
of the
C-Terminus
Binding in other seryl enzymes
Carbonic Anhydrase, a Zn++ enzyme
Carbonic Anhydrase reaction
 Carbonic Anhydrase Mechanism
Proton release
His64 assists
in H+ removal

HCO3-
CO2
released.
enters
H2O
enters

HO- attacks CO2

His64 Participation
 ATP:Mg++ Complexes
Most enzymes that require ATP, actually require
ATP:Mg++ as substrate and will not use ATP
alone.
Kinases are of this type.

Isomeric forms
Additional conformational
change occurs after NMP
binds.