1. Concept 8.4: Enzymes speed up metabolic reactions by lowering energy barriers
A catalyst is a chemical agent that speeds up a reaction without being consumed by the reaction
An enzyme is a catalytic protein
Hydrolysis of sucrose by the enzyme sucrase is an example of an enzyme-catalyzed reaction

2. LE 8-13
Sucrose
C12H22O11
Glucose
C6H12O6
Fructose
C6H12O6

3. The Activation Energy Barrier
Every chemical reaction between molecules involves bond breaking and bond forming
The initial energy needed to start a chemical reaction is called the free energy of activation, or activation energy (EA)
Activation energy is often supplied in the form of heat from the surroundings

4. Progress of the reaction
LE 8-14 A B C D
Transition state A B EA
Free energy C D
Reactants A B
DG < O C D
Products
Progress of the reaction

5. How Enzymes Lower the EA Barrier
Enzymes catalyze reactions by lowering the EA barrier
Enzymes do not affect the change in free-energy (∆G); instead, they hasten reactions that would occur eventually
Animation: How Enzymes Work

6. Progress of the reaction
LE 8-15
Course of
reaction
without
enzyme EA
without
enzyme
EA with
enzyme
is lower
Reactants
Free energy
Course of
reaction
with enzyme
DG is unaffected
by enzyme
Products
Progress of the reaction

7. Substrate Specificity of Enzymes
The reactant that an enzyme acts on is called the enzyme’s substrate
The enzyme binds to its substrate, forming an enzyme-substrate complex
The active site is the region on the enzyme where the substrate binds
Induced fit of a substrate brings chemical groups of the active site into positions that enhance their ability to catalyze the reaction

8. LE 8-16
Substrate
Active site
Enzyme
Enzyme-substrate
complex

9. Catalysis in the Enzyme’s Active Site
In an enzymatic reaction, the substrate binds to the active site
The active site can lower an EA barrier by
Orienting substrates correctly
Straining substrate bonds
Providing a favorable microenvironment
Covalently bonding to the substrate

10. LE 8-17 Substrates enter active site; enzyme
changes shape so its active site
embraces the substrates (induced fit).
Substrates held in
active site by weak
interactions, such as
hydrogen bonds and
ionic bonds.
Active site (and R groups of
its amino acids) can lower EA
and speed up a reaction by
acting as a template for
substrate orientation,
stressing the substrates
and stabilizing the
transition state,
providing a favorable
microenvironment,
participating directly in the
catalytic reaction.
Substrates
Enzyme-substrate
complex
Active
site is
available
for two new
substrate
molecules.
Enzyme
Products are
released.
Substrates are
converted into
products.
Products

11. Effects of Local Conditions on Enzyme Activity
An enzyme’s activity can be affected by:
General environmental factors, such as temperature and pH
Chemicals that specifically influence the enzyme

12. Effects of Temperature and pH
Each enzyme has an optimal temperature in which it can function
Each enzyme has an optimal pH in which it can function

13. LE 8-18 Optimal temperature for typical human enzyme
enzyme of thermophilic
(heat-tolerant
bacteria)
Rate of reaction 20 40 60 80
100
Temperature (°C)
Optimal temperature for two enzymes
Optimal pH for pepsin
(stomach enzyme)
Optimal pH
for trypsin
(intestinal
enzyme)
Rate of reaction 1 2 3 4 5 6 7 8 9 10 pH
Optimal pH for two enzymes

14. Cofactors Cofactors are nonprotein enzyme helpers
Coenzymes are organic cofactors

15. Enzyme Inhibitors
Competitive inhibitors bind to the active site of an enzyme, competing with the substrate
Noncompetitive inhibitors bind to another part of an enzyme, causing the enzyme to change shape and making the active site less effective

16. LE 8-19 A substrate can Substrate bind normally to the
active site of an
enzyme.
Substrate
Active site
Enzyme
Normal binding
A competitive
inhibitor mimics the
substrate, competing
for the active site.
Competitive
inhibitor
Competitive inhibition
A noncompetitive
inhibitor binds to the
enzyme away from the
active site, altering the
conformation of the
enzyme so that its
active site no longer
functions.
Noncompetitive inhibitor
Noncompetitive inhibition

17. Concept 8.5: Regulation of enzyme activity helps control metabolism
Chemical chaos would result if a cell’s metabolic pathways were not tightly regulated
To regulate metabolic pathways, the cell switches on or off the genes that encode specific enzymes

18. Allosteric Regulation of Enzymes
Allosteric regulation is the term used to describe cases where a protein’s function at one site is affected by binding of a regulatory molecule at another site
Allosteric regulation may either inhibit or stimulate an enzyme’s activity

19. Allosteric Activation and Inhibition
Most allosterically regulated enzymes are made from polypeptide subunits
Each enzyme has active and inactive forms
The binding of an activator stabilizes the active form of the enzyme
The binding of an inhibitor stabilizes the inactive form of the enzyme

20. LE 8-20a Allosteric activator stabilizes active form.
Allosteric enzyme
with four subunits
Active site
(one of four)
Regulatory
site (one
of four)
Activator
Active form
Stabilized active form
Oscillation
Allosteric inhibitor
stabilizes inactive form.
Non-
functional
active site
Inhibitor
Inactive form
Stabilized inactive
form
Allosteric activators and inhibitors

21. Cooperativity is a form of allosteric regulation that can amplify enzyme activity
In cooperativity, binding by a substrate to one active site stabilizes favorable conformational changes at all other subunits

22. Binding of one substrate molecule to
LE 8-20b
Binding of one substrate molecule to
active site of one subunit locks all
subunits in active conformation.
Substrate
Inactive form
Stabilized active form
Cooperativity another type of allosteric activation

23. Feedback Inhibition
In feedback inhibition, the end product of a metabolic pathway shuts down the pathway
Feedback inhibition prevents a cell from wasting chemical resources by synthesizing more product than is needed

24. LE 8-21 Initial substrate (threonine) Active site available Threonine
in active site
Enzyme 1
(threonine
deaminase)
Isoleucine
used up by
cell
Intermediate A
Feedback
inhibition
Enzyme 2
Active site of
enzyme 1 can’t
bind
theonine
pathway off
Intermediate B
Enzyme 3
Intermediate C
Isoleucine
binds to
allosteric
site
Enzyme 4
Intermediate D
Enzyme 5
End product
(isoleucine)

25. Specific Localization of Enzymes Within the Cell
Structures within the cell help bring order to metabolic pathways
Some enzymes act as structural components of membranes
Some enzymes reside in specific organelles, such as enzymes for cellular respiration being located in mitochondria

26. LE 8-22
Mitochondria,
sites of cellular respiration
1 µm