Bettelheim / Brown / Campbell / Farrell / Torres

Introduction to General, Organic, and Biochemistry 11e

Chapter 23
Enzymes

William H. Brown Beloit College www.cengage.com/chemistry/bettelheim

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Enzymes
Ribbon diagram of cytochrome c oxidase, the enzyme that
directly uses oxygen during respiration.

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Enzyme Catalysis
Enzyme: A biological catalyst.
• With the exception of some RNAs that catalyze their own
self-cleavage, all enzymes are proteins.
• Enzymes can increase the rate of a reaction by a factor of 109 to
1020 over an uncatalyzed reaction.
• Some catalyze the reaction of only one compound.

• Others are stereoselective; for example, enzymes that catalyze the

reactions of only L-amino acids.
• Others catalyze reactions of specific types of compounds or
bonds; for example, trypsin catalyzes hydrolysis of peptide bonds
formed by the carboxyl groups of Lys and Arg.

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Enzyme Catalysis
Figure 23-1 Trypsin catalyzes the hydrolysis of peptide
bonds formed by the carboxyl group of lysine and
arginine.

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Classification of Enzymes
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 two groups to a C-C double bond,
or removal of two groups to create a C-C double bond.
• Isomerases: Isomerization reactions.
• Ligases: The joining to two molecules.

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Classification of Enzymes
1. Oxidoreductase: Oxidation-reduction reactions

2. Transferase: Group transfer reactions

3. Hydrolase: Hydrolysis reactions(breaking of bonds with

the help of water to form two products)

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Classification of Enzymes
4. Lyase:Addition of two groups to a C-C double bond, or
removal of two groups to create a C-C double bond
COO - COO-
CH2 CH2
C-COO- Aconitase C-COO-
+ H2O
CH HO C-H
COO - COO-
cis-Aconitate Isocitrate

5. Isomerase: Isomerization reactions.

CH2 OPO3 2- CH2 OPO3 2-

O Phosphohexose O CH2 OH
isomerase H HO
OH
HO OH H OH
OH HO H
a-D- Glucose-6-phosphate a-D-Fructose-6-phosphate

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Classification of Enzymes
6. Ligase: The joining to two molecules
Tyrosine-tRNA
synthetase L-tyrosyl-tRNA + AMP + PPi
ATP + L-tyrosine + t-RNA

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Enzyme Terminology
Apoenzyme: The protein part of an enzyme.
Cofactor: A nonprotein portion of an enzyme that is
necessary for catalytic function; examples are metallic
ions such as Zn2+ and Mg2+.
Coenzyme: A nonprotein organic molecule, frequently
a B vitamin, that acts as a cofactor.
Substrate: The compound or compounds whose
reaction an enzyme catalyzes.
Active site: The specific portion of the enzyme to which
a substrate binds during reaction.

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Schematic of an Active Site
Figure 23.2 Schematic
diagram of the active site
of an enzyme and the
participating components.

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Mechanism of Enzyme Action
A. Lock and Key Model
Mechanism of Enzyme Action
B. Induced Fit Model
Terms in Enzyme Chemistry
Activation: Any process that initiates or increases the
activity of an enzyme.
Inhibition: Any process that makes an active enzyme less
active or inactive.
Competitive inhibitor: A substance that binds to the
active site of an enzyme thereby preventing binding of
substrate.
Noncompetitive inhibitor: Any substance that binds to a
portion of the enzyme other than the active site and
thereby inhibits the activity of the enzyme.

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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.

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Factors Affecting Enzyme Action
1. Enzyme activity and Substrate Concentration
Enzyme Activity
Figure 23-3 The effect of enzyme concentration on the rate
of an enzyme-catalyzed reaction. Substrate concentration,
temperature, and pH are constant.

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Enzyme Activity
Figure 23-4 The effect of substrate concentration on the
rate of an enzyme-catalyzed reaction. Enzyme
concentration, temperature, and pH are constant.

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Factors Affecting Enzyme Action
1. Enzyme activity and Substrate Concentration
Enzyme Activity
Figure 23-5 The effect of temperature on the rate of an
enzyme-catalyzed reaction. Substrate and enzyme
concentrations and pH are constant.

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Factors Affecting Enzyme Action
2. Temperature
Enzyme Activity
Figure 23-6 The effect of pH on the rate of an enzyme-
catalyzed reaction. Substrate and enzyme concentrations
and temperature are constant.

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Factors Affecting Enzyme Action
4. Presence of Inhibitor
A. Competitive
B. Non-competive/
Uncompetitve
Mechanism of Action
Figure 23-7 Lock-and-key model of enzyme mechanism.
• The enzyme is a rigid three-dimensional body.
• The enzyme surface contains the active site.

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Mechanism of Action
Figure 23-8 The Induced-fit model of an enzyme mechanism.
• The active site becomes modified to accommodate the
substrate.

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Type of Inhibitors
1. Competitive Inhibitor
Mechanism of Action
Figure 23-9 The mechanism of competitive inhibition.
When a competitive inhibitor enters the active site, the
substrate cannot enter.

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Type of Inhibitors
2. Non-competitive Inhibitor
Mechanism of Action
Figure 23-10 Mechanism of noncompetitive inhibition. The
inhibitor binds itself to a site other than the active site
(allosterism), thereby changing the conformation of the
active site. The substrate still binds but there is no
catalysis.

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Type of Inhibitors
2. Non-competitive Inhibitor

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Mechanism of Action

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Summary

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Models of Enzyme Kinetics

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Mechanism of Action
Figure 23-11 Enzyme kinetics in the presence and the
absence of inhibitors.

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Lineweaver-Burk Plots for Enzyme Inhibition

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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.
• Just five amino acids participate in the active site in
more than 65% of the enzymes studied to date.
• These five are His > Cys > Asp > Arg > Glu.
• Four of these amino acids have either acidic or basic
side chains; the fifth has a sulfhydryl group (-SH).

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Catalytic Power
• 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.

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Enzyme Regulation
Feedback control: An enzyme-regulation process where
the product of a series of enzyme-catalyzed reactions
inhibits an earlier reaction in the sequence.

• The inhibition may be competitive or noncompetitive.

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Enzyme Regulation
• Proenzyme (zymogen): An inactive form of an enzyme that
must have part of its polypeptide chain hydrolyzed and
removed before it becomes active.
• An example is trypsin, a digestive enzyme.
• It is synthesized and stored as trypsinogen, which has no enzyme
activity.
• It becomes active only after a six-amino acid fragment is
hydrolyzed and removed from the N-terminal end of its chain.
• Removal of this small fragment changes not only the primary
structure but also the tertiary structure, allowing the molecule to
achieve its active form.

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Enzyme Regulation
Allosterism: Enzyme regulation based on an event
occurring at a place other than the active site but that
creates a change in the active site.
• An enzyme regulated by this mechanism is called an
allosteric enzyme.
• Allosteric enzymes often have multiple polypeptide
chains.
• Negative modulation: Inhibition of an allosteric
enzyme.
• Positive modulation: Stimulation of an allosteric
enzyme.
• Regulator: A substance that binds to an allosteric
enzyme.

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Enzyme Regulation
• 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.

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

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Enzyme Regulation
Protein modification: The process of affecting enzyme
activity by covalently modifying it.
• The best known examples of protein modification
involve phosphorylation/dephosphorylation.
• Example: Pyruvate kinase (PK) is the active form of the
enzyme; it is inactivated by phosphorylation to
pyruvate kinase phosphate (PKP).

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Enzyme Regulation
Isoenzyme (Isozymes): An enzyme that occurs in multiple
forms; each catalyzes the same reaction. Formation of
different isoforms of enzyme to different tissues of the
body
• Example: lactate dehydrogenase (LDH) catalyzes the oxidation of lactate
to pyruvate.
 The enzyme is a tetramer of H and M chains.
 H4 is present predominately in heart muscle.
 M4 is present predominantly in the liver and in skeletal muscle.
 See Figure 23-16 for the relative distribution of these isoenzymes.
• Example: Glucose Transporters
 brain – highest affinity to glucose
 liver – low affinity
 blood – low affinity
 muscles – moderate affinity

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Enzyme Regulation
6. Genetic Expression Induction – some proteins are only
synthesized if they are needed

Examples
non-constitutive enzymes
antibodies
Enzyme Regulation
Figure 23-16 The isozymes of lactate dehydrogenase
(LDH). The electrophoresis gel depicts the relative
isozyme types found in different tissues.

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Enzymes Used in Medicine

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Transition-State Analogs
• Transition state analog: A molecule whose shape mimics
the transition state of a substrate.
• Have the same structure as the substrate in the ES
complex
• Have lower energy requirement compared to substrate
• Preferred by enzyme compared to substrate
• Figure 23.17 The proline racemase reaction. Pyrrole-2-
carboxylate mimics the planar transition state of the
reaction (next screen).

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Transition-State Analog
Figure 23-17 The proline racemase reaction and a mimic for
the transition state.

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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.

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Transition-State Analogs

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Chapter 23 Enzymes

End
Chapter 23

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