3 Enzymes
3.1 Mode of action of enzymes The course of a reaction
• enzymes are globular proteins that catalyse • when the enzyme and substrate are first mixed,
metabolic reactions there’s a large number of substrate molecules
therefore almost every enzyme has a substrate in its
• function as biological catalysts
active site
• specific in nature
• this makes the rate of enzyme-controlled reaction
• precise 3D shape with hydrophilic R-groups on the fastest at the beginning
outside ensuring they’re soluble
• possess active sites which are clefts/depressions to
which a substrate can bind
ENZYMES
INTRACELLULAR EXTRACELLULAR
functions inside of cells functions outside of cells
synthesised and retained synthesised in cell but Image: [Link]
in cell secreted out
Lock and key
3.2 Factors that affect enzyme
• idea that enzymes have particular shapes into which action
their substrate fits into exactly 1) Temperature
• enzyme is said to be specific for a substrate
• rate of reaction is slow at lower temperatures as
molecules are moving slowly which makes collisions
Induced fit hypothesis happen less frequently
• substrate is partially complementary to the active • as temperature rises, enzymes and substrates move
site faster, and collisions happen more frequently
• the active site changes shape slightly to ensure a • when they collide, they do so with more energy
better fit and stronger binding of substrate which makes it easier for bonds to be formed and
• this makes catalysis even more efficient broken
• if temperature keeps increasing, bonds holding
enzyme in shape (ionic, hydrogen bonds) break and
the enzyme is said to be denatured
• the temperature at which enzymes catalyse a
reaction at maximum rate is the ‘optimum
temperature’
• in humans, this is around 40°C
2) pH
• pH is a measure of the H+ ions in a solution
• H+ ions can affect the R-groups of amino acids
Image: [Link] which affects the ionic bonding between groups
which in turn affects the 3D structure of the enzyme
Enzymes reduce activation energy (Ea)
• Active site may also be changed, reducing chances
• in many chemical reactions, the substrate will not be of a substrate fitting in
converted to a product unless it’s temporarily given
extra energy
• this extra energy is activation energy (Ea)
3) Enzyme concentration
• the more enzymes present, the more active sites are
• enzymes do this by holding their substrates in a way
available for substrates to fit in
that bonds can be broken more easily hence
reducing Ea • as long as there’s plenty of substrate available, initial
rate of reaction increasing linearly with enzyme
• or the shape is slightly changed, making it easier to
change the substrate to a product (induced fit concentration
theory)
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4) Substrate concentration • disrupts the three-dimensional shape of enzyme
• as substrate concentration increases, initial rate of preventing the substrate from fitting into the active
reaction also increases site as its distorted
• the more substrate molecules there are around, the • increasing the substrate concentration has no
more often an enzyme’s active site can bind with change on the rate of reaction here
one
• saturation point – enzymes working at max (Vmax)
• all active sites are filled up
• enzyme moves to find substrates as it gets less,
collision forces start to decrease
STARCH → MALTOSE → GLUCOSE
too much product so
difficult to find substrate acting as inhibiting agent
5) Inhibitor concentration
Decreases enzyme activity, slowing down the reaction. • End product inhibition – as enzyme converts
substrate into product, rate is slowed down at the
end as the product binds to another part of the
enzyme and prevents more substrate binding
Enzyme affinities
• affinity – enzyme willingness to bind to a substrate
• at Vmax, all enzyme molecules are bound to substrate
molecules; the enzyme is saturated with substrate
As substrate concentration is increased, reaction rate
a) Competitive inhibition rises until the max rate i.e., Vmax
• compete with the substrate for the active site
• molecule similar in shape to the enzyme’s substrate
binds with the active site inhibiting the function
Image: [Link]
Km (Michaelis-Menten constant)
• the substrate concentration at which enzyme works
at half its maximum rate
• if the concentration of inhibitor rises or substrate • half the active sites of enzymes are occupied by
falls, it becomes less likely that the substrate will substrate
collide with an active site
• can be reversed by increasing the concentration of
substrate
b) Non-competitive inhibitor
Molecule fits into the allosteric site of the enzyme
rather than the active site.
Image: [Link]
• An enzyme with a lower value of Km has a high
affinity to its substrate
Image: [Link]
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Image: [Link]
uncompetitive-inhibitors/
Immobilising enzymes
• enzyme is mixed with a solution of sodium alginate
• droplets of this mixture are added to calcium
chloride solution
• a reaction occurs forming jelly/beads
• enzyme is immobilised in the bead
Advantages of immobilising enzymes
1) enzyme is reused
2) enzyme is easily recovered
3) product isn’t contaminated with enzymes
4) reduces product inhibition
5) enzyme is more stable/less likely to denature
6) longer shelf-line of enzyme
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