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Enzymes: Structure & Function
Lab #3- Enzyme Catalysis w/Catalase
Three Parts to the lab:
• Establish Baseline Amount of H2O2
• Uncatalyzed Decomposition of H2O2
• Time Trials w/Catalase to determine Rxn rate
• Procedure: – 10 ml H2O2 in a beaker
– 1.0 ml (H2O or Catalase)
– 10 ml 1 M H2SO4
– Mix well
– Take a 5 ml sample and titrate in KMnO4
– Read Initial and final measurements on buret
– Record Data
• For a chemical reaction to begin, reactants
must absorb some energy
– This energy is called the energy of activation
(EA)
– This represents the energy barrier that prevents
molecules from breaking down spontaneously
Enzymes speed up the cell’s chemical
reactions by lowering energy barriers
Enzymes
• Catalytic proteins: change
the rate of reactions w/o
being consumed
• Free E of activation
(activation E): the E
required to break bonds
• Substrate: enzyme reactant
• Active site: pocket or
groove on enzyme that
binds to substrate
• Induced fit model
• A protein catalyst called an enzyme can
decrease the energy barrier
EA
barrier
Reactants
1 Products 2
En
zym
e
Figure 5.5A
• Enzymes are selective
– This selectivity determines which chemical
reactions occur in a cell
A Specific Enzyme Catalyzes
each Cellular Reaction
How an Enzyme Works-Sucrase
How Enzymes Work
• http://www.ekcsk12.org/science/aplabrevie
w/lab02.htm
**Description of Enzyme Lab
• Lab Simulation: http://bioweb.wku.edu/courses/Biol114/enzy
me/enzyme1.asp
Effects on Enzyme Activity
• Temperature
• pH
• Substrate conc.
• Enzyme conc.
• Cofactors:
inorganic, nonprotein
helpers; ex.: zinc, iron,
copper
• Coenzymes:
organic helpers
ex. vitamins
Effects on Enzyme Activity
• pH - based on relative number of H+
• Large number of H+ ions can bond with negative charges
on the enzyme or substrate
• Proper charge matching is unable to occur
• Similar scenario occurs with OH- ion
• Numerous extra + and – charges result in the enzyme
losing its shape
Effects on Enzyme Activity
• Temperature – kinetic energy
• Molecules moving faster are more often to
collide with greater energy
• High temperatures can cause intramolecular
bonds to stress
Effects on Enzyme Activity
• Substrate Concentration – activity increases with
concentration
• Increased molecular collisions
• Enzymes have a maximum rate at which they
can work
Effects on Enzyme Activity
• Enzyme Concentration – activity increases with
concentration
• Increased molecular collisions
• Only so much substrate for enzymes to work on
Enzyme Inhibitors
• Irreversible (covalent); reversible (weak bonds)
• Competitive: competes for active site (reversible); mimics
the substrate
• Noncompetitive: bind to another part of enzyme (allosteric
site) altering its conformation (shape); poisons, antibiotics
• Inhibitors interfere with enzymes
– A competitive
inhibitor takes
the place of a
substrate in the
active site
– A noncompetitive
inhibitor alters an
enzyme’s function
by changing its shape
Enzyme inhibitors block enzyme
action
Substrate
Enzyme
Active
site
NORMAL BINDING OF SUBSTRATE
Competitive
inhibitor
Noncompetitive
inhibitor
ENZYME INHIBITION
Figure 5.8
Competitive & Noncompetitive Inhibitors
• Enzyme activity is influenced by
– temperature
– salt concentration
– pH
• Some enzymes require non-protein
cofactors such as: Fe, Zn, Cu, etc.
The Cellular environment affects
enzyme activity
• Certain pesticides are toxic to insects
because they inhibit key enzymes in the
nervous system
• Many antibiotics inhibit enzymes that are
essential to the survival of disease-causing
bacteria
– Penicillin inhibits an enzyme that bacteria use in
making cell walls
Some Pesticides and Antibiotics
inhibit Enzymes
Allosteric Enzymes-How they Work
Feedback Inhibition