Download Chapter Nineteen

Fundamentals of General,
Organic, and Biological Chemistry
5th Edition
Chapter Nineteen
Enzymes and Vitamins
James E. Mayhugh
Oklahoma City University
2007 Prentice Hall, Inc.
Outline
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19.1 Catalysis by Enzymes
19.2 Enzyme Cofactors
19.3 Enzyme Classification
19.4 How Enzymes Work
19.5 Effect of Concentration on Enzyme Activity
19.6 Effect of Temperature and pH on Enzyme Activity
19.7 Enzyme Regulation: Feedback and Allosteric Control
19.8 Enzyme Regulation: Inhibition
19.9 Enzyme Regulation: Covalent Modification and Genetic
Control
► 19.10 Vitamins
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19.1 Catalysis by Enzymes
► Enzyme: A protein or other molecule that acts as a
catalyst for a biological reaction.
► Active site: A pocket in an enzyme with the specific
shape and chemical makeup necessary to bind a
substrate.
► Substrate: A reactant in an enzyme-catalyzed
reaction.
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► Specificity (enzyme): The limitation of the activity
of an enzyme to a specific substrate, specific
reaction, or specific type of reaction.
► The enzyme papain from papaya fruit catalyzes the
hydrolysis of peptide bonds in many locations.
► Thrombin is specific for catalyzing hydrolysis of a
peptide bond adjacent to arginine and does so
primarily in a protein essential to blood clotting.
► Catalase is almost completely specific for one
reaction—the decomposition of hydrogen peroxide.
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► The specificity of an enzyme for one
of two enantiomers is a matter of fit. A
chiral reactant fits a chiral reaction
site. The enantiomer at the top fits the
reaction site like a hand in a glove, but
the enantiomer at the bottom does not.
► The enzyme lactate dehydrogenase
catalyzes the removal of hydrogen
from L-lactate but not from D-lactate.
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Catalytic activity is measured by turnover number,
the maximum number of substrate molecules acted
upon per enzyme per unit time. Most enzymes turn
over 10–1000 molecules per second.
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19.2 Enzyme Cofactors
► Cofactor: A nonprotein part of an enzyme that is
essential to the enzyme’s catalytic activity; a metal
ion or a coenzyme.
► Coenzyme: An organic molecule that acts as an
enzyme cofactor.
► Why are cofactors necessary? The functional groups
in proteins are limited to those of the amino acid side
chains. By combining with cofactors, enzymes
acquire chemically reactive groups not available as
side chains.
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Trace minerals and certain vitamins are a dietary
necessity because they function as building blocks
for cofactors and we cannot synthesize them.
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19.3 Enzyme Classification
► Enzymes are divided into six main classes according
to the general kind of reaction they catalyze.
► Oxidoreductases catalyze redox reactions of
substrates, most commonly addition or removal of
oxygen or hydrogen. These enzymes require
coenzymes that are reduced or oxidized as the
substrate is oxidized or reduced.
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Transferases catalyze transfer of a group from one
molecule to another. Kinases transfer a phosphate
group from ATP to give ADP and a phosphorylated
product.
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► Hydrolases catalyze the breaking of bonds with
addition of water. The digestion of carbohydrates
and proteins by hydrolysis requires these enzymes.
► Ligases catalyze the bonding together of two
substrates. Such reactions are generally not favorable
and require energy from ATP hydrolysis.
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Isomerases catalyze the isomerization
(rearrangement of atoms) of a substrate in reactions
that have but one substrate and one product.
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Lyases catalyze the addition of a molecule to a
double bond or the reverse reaction in which a
molecule is eliminated from a double bond.
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► Enzymes have the family-name ending -ase.
► Exceptions occur for enzymes such as papain and
trypsin, which are still referred to by older common
names.
► Modern systematic names always have two parts:
- the first identifies the substrate on which the
enzyme operates
- the second part is an enzyme subclass name like
those shown on the next slide.
► Example: Pyruvate carboxylase is a ligase that acts
on the substrate pyruvate to add a carboxyl group.
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19.4 How Enzymes Work
► Two models are invoked to represent the interaction
between substrates and enzymes.
► Historically, the lock-and-key model came first. The
substrate is described as fitting into the active site as
a key fits into a lock.
► Enzyme molecules are not totally rigid like locks.
The induced-fit model accounts for changes in the
shape of the enzyme active site that accommodate
the substrate and facilitate the reaction. As an
enzyme and substrate come together, their
interaction induces exactly the right fit for catalysis
of the reaction.
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The induced-fit model model of enzyme action has a
flexible active site that changes shape to best fit the
substrate and catalyze the reaction.
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Hydrolysis of a peptide bond by chymotrypsin.
(a) The polypeptide enters the active site (b)
Hydrogen transfer allows formation of a strained
intermediate (c) The peptide bond is broken.
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► Enzymes act as catalysts because of their ability to:
► Bring substrate(s) and catalytic sites together
(proximity effect).
► Hold substrate(s) at the exact distance and in the
exact orientation necessary for reaction (orientation
effect).
► Provide acidic, basic, or other types of groups
required for catalysis (catalytic effect).
► Lower the energy barrier by inducing strain in bonds
in the substrate molecule (energy effect).
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19.5 Effect of Concentration on Enzyme
Activity
► Under most conditions the rate of an enzyme
catalyzed reaction is controlled by the amount of
substrate and the overall efficiency of the enzyme.
► If the enzyme–substrate complex is rapidly
converted to product, the rate at which enzyme and
substrate combine to form the complex becomes the
limiting factor.
► Enzyme and substrate molecules moving at random
in solution can collide with each other no more often
than about 108 collisions per (mole/liter) per second.
► One of the most efficient enzymes is catalase; this
enzyme can break down H2O2 at a rate of up to 107
catalytic events per second.
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► At low substrate
concentration, the
reaction rate is directly
proportional to the
substrate concentration.
► With increasing
substrate concentration,
the rate drops off as
more of the active sites
are occupied.
► With all active sites
occupied, the rate
reaches a maximum.
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► In the presence of
excess substrate, the
concentration of an
enzyme can vary
according to our
metabolic needs.
► If the concentration of
substrate does not
become a limitation, the
reaction rate varies
directly with the
enzyme concentration.
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19.6 Effect of Temperature and pH
on Enzyme Activity
► Enzyme catalytic activity is highly dependent on pH
and temperature.
► Optimum conditions vary slightly for each enzyme but
are generally near normal body temperature and the pH
of the body fluid in which the enzyme functions.
► Pepsin, which initiates protein digestion in the highly
acidic environment of the stomach, has its optimum
activity at pH 2.
► Trypsin, which acts in the alkaline environment of the
small intestine, has optimum activity at pH 8.
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(a) The rate increases with increasing temperature
until the protein begins to denature; then the rate
decreases rapidly. (b) The optimum activity for an
enzyme occurs at the pH where it acts.
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19.7 Enzyme Regulation: Feedback and
Allosteric Control
► A variety of strategies are utilized to adjust the rates
of enzyme-catalyzed reactions.
► Any process that starts or increases the action of an
enzyme is an activation.
► Any process that slows or stops the action of an
enzyme is an inhibition.
► Feedback and allosteric control are two strategies for
enzyme regulation.
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► Feedback control: Regulation of an enzyme’s
activity by the product of a reaction later in a
pathway.
► If a product near the end of a metabolic pathway
inhibits an enzyme that functions near the beginning
of that pathway, then no energy is wasted making the
ingredients for a plentiful substance.
► Allosteric control: An interaction in which the
binding of a regulator at one site on a protein affects
the protein’s ability to bind another molecule at a
different site.
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► Allosteric control can be either positive or negative.
► Binding a positive regulator changes the active sites
so that the enzyme becomes a better catalyst and the
rate accelerates.
► Binding a negative regulator changes the active sites
so that the enzyme is a less effective catalyst and the
rate slows down.
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19.8 Enzyme Regulation: Inhibition
► Enzyme inhibition can be reversible or irreversible.
► In reversible inhibition, the inhibitor can leave,
restoring the enzyme to its uninhibited level of
activity.
► In irreversible inhibition, the inhibitor remains
permanently bound and the enzyme is permanently
inhibited.
► The inhibition can also be competitive or
noncompetitive, depending on whether the inhibitor
binds to the active site or elsewhere.
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A competitive inhibitor can eventually be overcome
by higher substrate concentrations. With a
noncompetitive inhibitor the maximum rate is
lowered for all substrate concentrations.
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► Hg+2 and Pb+2 ions are irreversible inhibitors that
bond to the S atoms in cysteine residues.
► Organophosphorus insecticides such as parathion
and malathion, and nerve gases like Sarin are
irreversible inhibitors of the enzyme
acetylcholinesterase.
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19.9 Enzyme Regulation: Covalent
Modification and Genetic Control
► There are two general modes of enzyme regulation
by covalent modification, removal of a covalently
bonded portion of an enzyme, or addition of a group.
► Some enzymes are synthesized in inactive forms
known as zymogens or proenzymes, activation
requires a chemical reaction that splits off part of the
molecule.
► Genetic (enzyme) control: Regulation of enzyme
activity by hormonal control of the synthesis of
enzymes is especially useful for enzymes needed
only at certain times.
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Enzymes that cause blood clotting or digest proteins
are examples of enzymes that must not be active at
the time and place of their synthesis.
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Glycogen phosphorylase, the enzyme that initiates
glycogen breakdown, is more active when
phosphorylated. When glycogen stored in muscles
must be hydrolyzed to glucose for quick energy, two
serine residues are phosphorylated. The groups are
removed once the need to break down glycogen for
quick energy has passed.
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19.10 Vitamins
► Vitamin: An organic molecule, essential in trace
amounts that must be obtained in the diet because it
is not synthesized in the body.
► Scurvy, pellagra, and rickets are caused by
deficiencies of vitamins.
► Vitamins are grouped by solubility into two classes:
water-soluble and fat-soluble.
► Some vitamins are valuable as antioxidants.
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Water soluble vitamin C is biologically active without
any change in structure, biotin is connected to enzymes
by an amide bond at its carboxyl group but otherwise
undergoes no structural change from dietary biotin.
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Other water-soluble vitamins incorporate into coenzymes.
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► The fat-soluble vitamins A, D, E, and K are stored in
the body’s fat deposits.
► Although the clinical effects of deficiencies of these
vitamins are well documented, the molecular
mechanisms by which they act are not nearly as well
understood as those of the water-soluble vitamins.
None has been identified as a coenzyme.
► The hazards of overdosing on fat-soluble vitamins
are greater than those of the water-soluble vitamins
because of their ability to accumulate in body fats.
► Excesses of the water-soluble vitamins are more
likely to be excreted in the urine.
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► An antioxidant is a substance that prevents
oxidation.
► In the body, we need protection against active
oxidizing agents that are by-products of normal
metabolism.
► Our principal dietary antioxidants are vitamin C,
vitamin E, b-carotene, and the mineral selenium.
► They work together to defuse the potentially harmful
action of free radicals, highly reactive molecular
fragments with unpaired electrons.
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Chapter Summary
► Enzymes are mostly water-soluble, globular proteins,
and many incorporate cofactors, which are either metal
ions or organic molecules known as coenzymes.
► Six major classes and many subclasses of reactions are
catalyzed by enzymes.
► As the substrate enters the active site, the enzyme
shape adjusts to best accommodate the substrate and
catalyze the reaction (the induced fit). Within the
enzyme–substrate complex, the substrate is held in the
best orientation for reaction and in a strained condition
that allows the activation energy to be lowered. When
the reaction is complete, the product is released and the
enzyme returns to its original condition.
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Chapter Summary Cont.
► Enzyme reaction rate is maximal at a temperature
and pH that reflects the conditions at the site of
action in the body.
► In the presence of excess substrate, reaction rate is
directly proportional to enzyme concentration. With
fixed enzyme concentration, reaction rate first
increases with increasing substrate concentration and
then approaches a fixed maximum.
► A product of a later reaction can exercise feedback
control over an enzyme for an earlier reaction in a
pathway. Binding a regulator induces a change of
shape in the active site, increasing or decreasing the
efficiency of the enzyme.
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Chapter Summary Cont.
► Competitive inhibitors typically resemble the
substrate and reversibly block the active site; they
slow the reaction rate but do not change the
maximum rate. Irreversible inhibitors form bonds to
an enzyme that permanently inactivate it.
► Enzyme activity is regulated by covalent
modification and genetic control.
► Water-soluble vitamins are coenzymes or parts of
coenzymes. Excesses of water-soluble vitamins are
excreted and excesses of fat-soluble vitamins are
stored in body fat. Vitamin C, b-carotene, vitamin E,
and selenium work together as antioxidants to
protect biomolecules from damage by free radicals.
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End of Chapter 19
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