Frederick A.

Bettelheim
William H. Brown
Mary K. Campbell
Shawn O. Farrell
Omar J. Torres
www.cengage.com/chemistry/bettelheim

Chapter 23
Enzymes
William H. Brown Beloit College

TOC
23.1 What Are Enzymes?
23.2 How Are Enzymes Named and Classified?
23.3 What Is the Terminology Used with Enzymes?
23.4 What Factors Influence Enzyme Activity?
23.5 What Are the Mechanisms of Enzyme Action?
23.6 How Are Enzymes Regulated?
23.7 How Are Enzymes Used in Medicine?
23.8 What Are Transition-State
Analogs and Designer Enzymes?

What Are Enzymes?

Most substances necessary for the function of the human

body are not found in the diet. They are synthesized within the
cells through hundreds of chemical reactions that take place in
your cells every second of your life.
Enzymes: are a biological catalyst
1) Globular proteins: which are large molecules that increase
the rates of chemical reactions without themselves undergoing
any change.
2) Ribozymes (Section 25.4 ): (RNAs that catalyze their own
self-cleavage) are enzymes made of ribonucleic acids. They
catalyze the self-cleavage of certain portions of their own
molecules & involved in generating peptide bonds

What Are Enzymes?

Enzymes: they increase the reaction rate. They cause
reactions to take place faster by lowering the activation
energy.
As catalysts, enzymes are remarkable in two respects:
1. They are extremely effective, can increase the rate of a
reaction by a factor of 109 to 1020 over an uncatalyzed
reaction.
2. Most of them are extremely specific: each of them speeding
up only one particular reaction or class of reactions.
Substrate specificity: The limitation of an enzyme to catalyze
specific reactions with specific substrates
Examples on specificity:

Examples on specificity:
a) Some catalyze the reaction of only one compound. e.g urease
catalyzes only the hydrolysis of urea & not that of other amides, even
closely related ones.

b) Others catalyze reactions of specific types of compounds or bonds.

e.g.
1) Trypsin: cleaves the peptide bonds only on the carboxyl side of Lys
and Arg residues. This enzyme consist of polypeptide chains only.
carboxyl side of
lysine

carboxyl side of
arginine

Examples on specificity:
2) Carboxypeptidase specifically catalyzes the hydrolysis of
only the last amino acid on a protein chain (the one at the
C-terminal end).
3) Lipases are less specific: They catalyze the hydrolysis of
any triglyceride, but they still do not affect carbohydrates
or proteins
c) Some are stereoselective; for example, enzymes that
catalyze the reactions of only L-amino acids. e.g. arginase
hydrolyzes the amino acid L-arginine (the naturally occurring
form) to a compound called L-ornithine and urea (Section
28.8) but has no effect on its mirror image (D-arginine).

Distribution according to bodys need

Location of enzyme

Function

Blood

promote clotting (protein-splitting

enzymes )

Secretions of the stomach &

Pancreas

Digestive enzymes (catalyze

the hydrolysis of proteins)

Intra-cellualr enzymes (enzymes localized according to the

need for specific reactions)

Intracellular location

Specific reaction catalyzed

Mitochondria

oxidation of compounds (part of

the citric acid cycle) (Section 27.4)

Special organelles e.g. lysosomes (lysozyme): catalyzes the

dissolution of bacterial cell walls.

enzymes

Not enzymes

enzyme

Digestive enzymes
classified based on their target substrates:
Name

Function

Proteases and peptidases

Split proteins into small peptides + amino acids.

Lipases:

split fat into three fatty acids + a glycerol molecule.

amylases

split carbohydrates such as starch and sugars into

simple sugars such as glucose.

nucleases

split nucleic acids into nucleotides.

Muscle Relaxants & Enzyme Specificity

Acetylcholine: neurotransmitter. Has a specific receptor in the
muscle end plate. Attachment transmits a signal to the muscle
to contract; then acetylcholinesterase, catalyzes the hydrolysis
of acetylcholine, removing it from the receptor & the muscle
relaxes & prepare for the next signal transmission
(contraction).

Succinylcholine is sufficiently similar to acetylcholine binds to

the receptor of the muscle end plate. However,
acetylcholinesterase can hydrolyze succinylcholine very slowly
keeping the muscle relaxed especially when performing
bronchoscopy.

How Are Enzymes Named & Classified?

Enzymes are commonly named after the reaction or
reactions they catalyze.
Example: lactate dehydrogenase, acid phosphatase.
Enzymes are classified into six major groups according to
the type of reaction catalyzed:

Oxidoreductases: Oxidation-reduction reactions.

Transferases: Group transfer reactions.
Hydrolases: Hydrolysis reactions.
Lyases: Addition of 2 groups to a double bond, or
removal of 2 groups to create a double bond.
Isomerases: Isomerization reactions.
Ligases: The joining to 2 molecules.

How Are Enzymes Named &Classified?

Oxidation-reduction
reactions.

Group transfer
reactions

Hydrolysis
reactions

Classification of Enzymes
1. Oxidoreductase:

2. Transferase:

3. Hydrolase:

How Are Enzymes Named &Classified?

Addition of 2
groups to a
double bond, or
removal of 2
groups to create
a double bond.

Isomerization
reactions.

transfer RNA

The joining to
2 molecules.

Classification of Enzymes
4. Lyase:

COOCH2
C-COOCH

+ H2O

Aconitase

COOcis-Aconitate

COOCH2
C-COOHO C-H
COOIsocitrate

5. Isomerase:
CH2OPO32CH2OPO32Phosphohexose
CH2OH
O
O
isomerase
H HO
OH
OH
H
HO
OH
H
OH
HO
-D- Glucose-6-phosphate
-D-Fructose-6-phosphate

6. Ligase:
ATP + L-tyrosine +t-RNA
transfer
RNA

Tyrosine-tRNA
synthetase

L-tyrosyl-tRNA +AMP +PPi

Enzyme Terminology
Cofactor: A nonprotein portion of an enzyme that is
necessary for catalytic function.
Coenzyme: A nonprotein organic molecule.

Inorganic
cofactors

Zn2+ & Mg2+.

Organic
cofactors

B vitamin/ Heme

Schematic of an Active Site

Figure 23.2 Schematic diagram of the active site of an enzyme
& the participating components.
Apoenzyme: The protein part of
an enzyme.
Substrate: The compound or
compounds whose reaction an
enzyme catalyzes.
Active site:
site A 3-D cavity of the
enzyme with specific chemical
properties that the substrate
binds during reaction. The
coenzyme will be located there
too.

Terms in Enzyme Chemistry

Activation: Any process that initiates or increases the
activity of an enzyme. e.g. addition of a cofactor to an
apoenzyme or the cleavage of a polypeptide chain of a
proenzyme (Section 23.6B).
Inhibition: Any process that makes an active enzyme less
active or inactive. (Section 23.5)
Inhibitors: are compounds that accomplish inhibition,
there are many types of enzyme inhibition
Competitive inhibitor: bind to the active site of the

enzyme surface, thereby preventing binding of substrate.

Terms in Enzyme Chemistry

Noncompetitive inhibitor: Any substance that binds to a

portion of the enzyme other than the active site &

sufficiently alter the tertiary structure of the enzyme so that its
catalytic effectiveness is eliminated.& thereby inhibits the
activity of the enzyme while the inhibitor is bound.
Both competitive & noncompetitive inhibition are

reversible, but some compounds alter the structure of the

enzyme permanently & thus make it irreversibly inactive.

Factors that Influence Enzyme Activity?

Enzyme activity: A measure of how much a reaction rate
is increased.
We examine how the rate of an enzyme-catalyzed reaction
is affected by:
Enzyme concentration.
Substrate concentration.
Temperature.
pH.

A. Enzyme and Substrate Concentration

Figure 23.3 Substrate concentration, temperature, and pH
are constant.

Linear curve

Only change (increase) the concentration of enzyme, the rate

increases linearly. i.e. if the enzyme concentration doubles, the rate
doubles as well and so on.

A. Enzyme and Substrate Concentration

Figure 23.4. Enzyme concentration, temperature, and pH are
constant.
saturation
point

saturation curve

Reaction rate increases up to a point then stays the same even if we

increase the substrate concentration further. Because at the
saturation point, substrate molecules are bound to all available
active sites of the enzymes. No more active sites for substrate to
bind to .

B. Temperature
Figure 23.5 Substrate & enzyme concentrations & pH are constant.
optimal
temperature
changes in conformation
are reversible

protein
denatures
irreversibly

Alteration of the
enzyme
conformation:
substrate may
then not fit
properly.

Temperature changes the conformation of the enzyme.

Enzyme Inactivation: When the enzyme is irreversibly denatured: the
polypeptide chain cannot refold to its native conformation. Enzyme
activity cannot be restored by changing back to the optimal temperature.

Applications of temperature and effect on

enzymes in real life

Enzymes (bacteria and higher organisms have an optimal

temperature around 37C).
PCR enzymes: hyperthermophile polymerases that catalyze
the polymerization of DNA (Section 25.6) are obtained from
organisms that live in ocean vents under extreme conditions,
their enzymes optimal conditions 90 -105C, also need
pressures up to 100 atm, and some of them have an optimal
pH in the range of 1 to 4.

C. pH
Figure 23.6 Substrate, enzyme concentrations & temperature
are constant.
However,

optimal pH

Alteration of the enzyme

conformation at either too
acidic or too basic :
substrate may then not fit
properly.

within a narrow pH range,

changes in enzyme activity are
reversible.

pH changes the conformation of a protein (resemble

observation when the temperature changes). Each enzyme
operates best at a certain pH (Figure 23.6).
When enzymes are denatured irreversibly, enzyme activity
cannot be restored by changing back to the optimal pH.

Mechanisms of Enzyme Action

A. Lock-and-Key Model
B. Induced-Fit Model

A. Lock-and-Key Model
A model that explains the
specificity of enzyme action by
comparing the active site to a
lock and the substrate to a key
Assumptions:
The enzyme is a rigid threedimensional body.
The surface containing the
active site has a restricted
opening into which only one
kind of substrate can fit, just as
only the proper key can fit
exactly into a lock and turn it
open.

A. Lock-and-Key Model
An enzyme molecule is very large (typically consisting of
100 to 200 amino acid residues)
The active site is usually composed of only two or a few
amino acid residues, and can be located at different
places in the chain.
The other amino acids (not part of the active site) are
located in a sequence that causes the molecule as a
whole to fold up in exactly the required way.
The shape and the functional groups on the surface of the
active site are of utmost importance in recognizing a
substrate

B. Induced-Fit Model
A model that explains the
specificity of enzyme
action by comparing the
active site to a glove and
the substrate to a hand
Assumption:

the enzyme modifies the

shape of the active site to
accommodate the
substrate..

Competitive/non competitive Inhibitors

Figure 23.9. when inhibitor enters
the active site, the substrate
cannot enter.

Figure 23.10. Inhibitor binds itself to

a site other than the active site
(allosterism), changing the
conformation of the active site.
substrate still binds but there is no
catalysis.

No binding

competitive inhibitor

Noncompetitive inhibitor

Enzyme kinetics in the presence &

absence of inhibitors.

Figure 23.11

Competitive inhibitor

Noncompetitive inhibitor

There is structural similarity between

the inhibitor and substrate.

No structural similarity between

the inhibitor and the substrate

Inhibitor & substrate compete with

each other to bind to the same catalytic
site of the enzyme.

The inhibitor does not bind to

the active site, it binds to
another site

The inhibition is reversible. usually

temporary, and the Inhibitor eventually
leaves the enzyme.

The inhibition is irreversible.

The maximum reaction rate is not

affected (Vmax)

Inhibitor lowers the maximum

rate of the reaction (as if
enzyme available). It decreases
Vmax.

The maximum reaction rate achieved at

high substrate concentration when an
inhibitor is present.

The maximum reaction rate is

not achieved even by increasing
substrate concentration.

How: If the substrate is sufficiently

inhibitor will be displaced from active
site allowing binding of the substrate

Why: inhibitor cannot be

displaced from its binding site
by addition of excess substrate.

Mechanism of Action
Both the lock-and-key model and the induced-fit model
emphasize the shape of the active site.
However, the chemistry of the active site is the most
important factor..
Only five amino acids participate in the active site in
more than 65% of the enzymes studied to date. They
are, in order of their dominance:
His > Cys > Asp > Arg > Glu.
Four of these amino acids have either acidic or basic
side chains; the fifth has a sulfhydryl group (-SH).

Catalytic Power of Enzymes

Figure 23.12 Enzymes provide an alternative pathway for reaction.
(a) The activation energy profile for a typical reaction. (b) A
comparison of the activation energy profiles for a catalyzed
and uncatalyzed reactions.

Ex Catalytic Power of Enzymes

Papain is a protease (cleaves peptide
bonds). 2 critical amino acids are present
in the active site of papain (Figure 23.13)

Cysteine side chain

contains sulfur &
performs a reaction
called a nucleophilic
attack
The nucleophilic attack on the carbonyl
carbon of C-N bond & breaks it.
This nucleophilic attack is possible
because of the precise arrangement of
the amino acid side chains that can
participate in this reaction
Nucleophilic attack a reaction where an
electron-rich atom, such as oxygen, bonds
to an electron-deficient atom, such as a
carbonyl carbon.

Histidine (blue) helps

attract the peptide &
hold it in the correct
orientation via
hydrogen bonding
(red dashes).

Medical Uses of Inhibitors

A- HIV protease: It catalyzes the processing and production of
viral proteins (particles) in an infected cell new virus in infected
cells. Without these proteins, viable virus particles cannot be
released to cause further infection.
Solution: Designed & synthesized competitive HIV protease
inhibitors to bind to the active site of this enzyme
Results: Lowering of levels of the virus in the bloodstream (viral
load) are obtained when HIV protease inhibitors are part of drug
regimens for AIDS
Products in the market:
1. Saquinavir from Hoffmann-La Roche,
2.Ritonavir from Abbott,
3.Indinavir from Merck,
4.Viracept from Agouron Pharmaceuticals,
5.Amprenavir from Vertex Pharmaceuticals.

Medical Uses of Inhibitors

b. alleviating side effects from cancer chemotherapy.
Drug: camptothecin (CPT-11). Used to treat colon cancer
Problem: it causes severe diarrhea.
Why: CPT-11 processed in the body by the enzyme
-glucoronidase found in intestinal bacteria. The product of
the reaction is ultimately responsible for the diarrhea.
Solution: researchers synthesized compounds that were
likely to bind to the active site of -glucoronidase.

How Are Enzymes Regulated

A. Feedback control: an
enzyme regulation process in
which formation of a product
inhibits an earlier reaction in
the sequence. i.e. final product
in the chain may inhibit the
activity of the first enzyme (by
competitive, noncompetitive, or
some other type of inhibition).

How Are Enzymes Regulated

B. Proenzyme (zymogen): An inactive form of an enzyme that
must have part of its polypeptide chain hydrolyzed and
removed (a chemical change ) for it to become active.
Trypsin: a digestive enzyme.
Synthesized by pancreas & stored as trypsinogen, which has
no enzyme activity.
Becomes active when a 6-amino acid fragment is hydrolyzed &
removed from the N-terminal end of its chain. Changes the
primary & tertiary structure, allowing the molecule to achieve
its active form.
Why make an inactive form and activate it:
Trypsin: hydrolysis of peptide bonds (digestion of the proteins we
eat). And Also can cleave the proteins of which our own bodies
are made. When trypsin enters the digestive tract does it
become active.

How Are Enzymes Regulated

C. Allosterism: Enzyme regulation based on an event occurring at regulatory site other than the active site but creates a change in the active site.

Allosteric enzyme: the binding of a regulator on one site on the enzyme modifies the enzymes ability to bind the substrate in the active site.

Regulator: A substance that binds to an allosteric enzyme. the site to which it attaches is called a regulatory site.
Allosteric enzymes contain more than one polypeptide chain (subunits); regulatory site is on one polypeptide chain, & the active site is on another.

substance binds
noncovalently & reversibly
to a site other than the
active site

Negative modulation: inhibit

allosteric enzyme action.
Positive modulation: stimulate
allosteric enzyme action

How Are Enzymes Regulated

Figure 23.14 The allosteric effect.
Binding of the regulator to a site
other than the active site changes
the shape of the active site.
Specific regulators can bind reversibly
to the regulatory sites. This enzyme has
only one polypeptide chain, so it carries
both active site & regulatory site at
different points in this chain.
As long as the regulator remains bound
to the regulatory site, the total enzyme
regulator complex will be inactive.

How Are Enzymes Regulated

Figure 23.15 Effects of binding activators & inhibitors to allosteric
enzymes. The enzyme has an equilibrium between the T form and the R
form.

R form: The more active form (R stands for relaxed).

T form : The less active form (T stands for taut): (tense; not relaxed)
There is an equilibrium between the T & R form. When the enzyme is in
the R form, it will bind substrate well and catalyze the reaction.

How Are Enzymes Regulated

D. Protein modification: usually a change in the primary
structure, typically by addition of a functional group
covalently bound to the apoenzyme (protein part of the
enzyme)
The best known examples of protein modification
involve phosphorylation/dephosphorylation.
Example:
1. glycogen phosphorylase: (Section 29.1), the
phosphorylated form is the active form of the enzyme.
Without it, the enzyme is less active or inactive.

How Are Enzymes Regulated

2. Pyruvate kinase (PK) is the active form of the enzyme; it is
inactivated by phosphorylation to pyruvate kinase
phosphate (PKP).

How Are Enzymes Regulated

E. Isoenzymes (Isozymes): Enzymes that perform the same
function but have different combinations of subunits and thus
different quaternary structures. in different forms in different
tissues.
Example: lactate dehydrogenase (LDH) catalyzes the oxidation of
lactate to pyruvate.
The enzyme has four subunits (tetramer) of H and M chains.
H4 is present predominately in heart muscle.
M4 is present predominantly in liver & skeletal muscle.
H3M, H2M2, and HM3 also exist.
H4 is allosterically inhibited by high levels of pyruvate (its product)
& has a higher affinity for lactate (its substrate) than does the M4
enzyme, which is optimized for the opposite reaction.
H4 in serum correlates with the severity of heart attack.

How Are Enzymes Regulated

Figure 23.16 The isozymes of lactate dehydrogenase (LDH).

Electrophoresis: samples are separated in a gel using an electric field. the

two subunits of LDH carry different charges. Therefore, each combination
of subunits travels in the electric field at a different rate.

Enzymes Used in Medicine

Insert Table 23.2, page 648

Transition-State Analog
Transition state analog:
analog A molecule whose shape mimics

the transition state of a substrate. Is used to inhibit an

enzyme selectively.
Figure 23.17 The proline racemase reaction. Pyrrole-2-

carboxylate mimics the planar transition state of the

reaction (next screen).

Transition-State Analog

Orientation
reversed

During this reaction, the -carbon must change from a tetrahedral

arrangement to a planar form & then back to a tetrahedral form, but
with the orientation of 2 bonds reversed (pick box)
Pyrrole-2-carboxylate: a chemical that is
structurally similar to what proline would
look like at its transition state because it is
always planar at the equivalent carbon. This
inhibitor binds to proline racemase 160
times more strongly than proline does.

Transition-State Analogs
Abzyme: An antibody that has catalytic activity because it was
created using a transition state analog as an immunogen.
(a) The molecule below is a transition analog for the reaction of an
amino acid with pyridoxal-5-phosphate.
(b) The abzyme is then used to catalyze the reaction on the next
screen.

Transition-State Analogs
This is a very important
reaction in amino acid
metabolism:
The reaction of pyridoxal
phosphate & an amino
acid to form the corresponding
-ketoacid & pyridoxamine
phosphate. The molecule
N -(5 phosphopyridoxyl)-Llysine serves as a transitionstate analog for
this reaction.
designer enzymes to catalyze a
wide variety of reactions

Chapter 23 Enzymes

End
Chapter 23