2.1.

4 a the role of enzymes in catalysing reactions that affect metabolism at a cellular and
whole organism level
Enzyme= biological catalysts-> they speed up the rate of chemical reactions without being
used up or undergoing permanent change. They’re globular proteins. Metabolic pathways
are controlled by enzymes.
Intracellular Enzymes= produced + function inside the cell.
Extracellular Enzymes= secreted by cells + catalyse reactions outside cells.
2.1.4 b the role of enzymes in catalysing both intracellular and extracellular reactions
Amylase starch into maltose, other
extracellular digestive enzyme that
is secreted by pancreas + enters
small intestine is trypsin (breaks
proteins down into amino acids)
The macromolecules or large
molecules are broken down or
hydrolysed by the enzyme called
hydrolases. Hydrolases are enzymes
which help in splitting up of a
molecule by adding water and the
process is hydrolysis e.g. protease
The enzyme catalase converts one
such ROS hydrogen peroxide to
water and oxygen according to the
reaction: 2 H 2 O2 →2 H 2 0+ O2

2.1.4 c the mechanism of enzyme action

Lock and Key:
Enzymes have an active site where specific substrates bind
forming an enzyme-substrate complex. Extremes of heat or
pH can change the shape of the active site, preventing
substrate binding – this is called denaturation. Substrates collide with the enzymes active
site at the correct orientation + speed in order for a reaction to occur
Enzyme specificity: result of complementary active site + substrate. So shape of active site
is based on the complex tertiary structure of protein that makes up enzyme. enzyme-
substrate complex is only formed temporarily before the enzyme catalyses the reaction
and the product(s) are released.
Induced Fit Hypothesis:
The enzyme and its active site (and sometimes the substrate) can change shape slightly as
the substrate molecule enters the enzyme. These changes in shape are known as
conformational changes. This maximises the ability of the enzyme to catalyse the reaction.
Initial interaction between active site + substrate= weak but then these weak interactions
induce changes in the enzyme’s tertiary structure which strengthens binding.
Enzymes in lowering Activation Energy:
Activation energy is the amount of energy needed by the
substrate to become just unstable enough for a reaction to
occur and for products to be formed. Enzymes speed up
chemical reactions because they reduce the stability of bonds
in the reactants-> destabilisation of bonds in the substrate
makes it more reactive.
Enzymes provide an alternative energy pathway with a lower
activation energy.
2.1.4 d (i) the effects of pH, temperature, enzyme concentration and substrate
concentration on enzyme activity
pH-> enzymes need optimum pH bc if pH is too high or too low, the ionic +
hydrogen bonds in the tertiary structure will break. This means that enzyme-
substrate complexes can’t form.
Temperature-> similar w pH, optimum temp is needed for
complexes to form. If temp is increased, the kinetic energy inc=>
forms more complexes. If it’s too high then bonds in tertiary
break (disulphide, ionic, hydrogen etc). changes shape of active
site= denatured. Temperature coefficient (Q) for a biological
reaction= ratio between rates of that reaction at two diff temps.
Tempe coefficient= (rate of reaction at (x + 10) °C) ÷ (ROR at x °C).
Enzyme concentration-> As enzyme concentration increases so will
the rate: As there are more enzyme-substrate complexes forming.
At very high concentrations of enzyme the rate remains constant as
substrate becomes the limiting factor Add more substrate will
cause rate to increase again.
Substrate Concentration-> As the substrate concentration inc, the
rate inc bc more substrate molecules can collide with enzyme
molecules, so more reactions will take place. At higher conc the
enzyme molecules become saturated with substrate, so there are
few free enzyme molecules, so adding more substrate doesn't make
much difference.
Vmax=> maximum rate of reaction.
(ii) practical investigations into the effects of pH, temperature, enzyme concentration and
substrate concentration on enzyme activity
Equipment: hydrogen peroxide, Distilled water, 50 cm3 measuring cylinder, 250 cm3
beakers (x5), Marker pen, Trough to hold water, conical flask, Delivery tube and bung to
fit conical flask, Cork borer, Knife, White tile, Ruler, Potato.
Aim: To investigate the effect of changing the substrate concentration on the rate of
hydrogen peroxide breakdown by catalase.
Variables: IV-> concentration of hydrogen peroxide. DV-> rate of gas produced from each
hydrogen peroxide conc. CV-> temperature, size of potato and volume of water in beaker.
1. Serial dilution + label beakers
2. Cut 20-cylinder potatoes using cork borer tile + knife. Remove skin
3. Set up conical flask, delivery tube, measuring cylinder full of water inverted in
water trough
4. Place four cylinders in flask + add 2vol hydrogen peroxide and secure bung
5. Record vol of gas given off every 30sec for 3minutes
6. Dispose of contents and rinse flask + refill measuring cylinder. Repeat with other
concentrations of hydrogen peroxide.
7. Exchange data to get three values for each concentration.
8. Calculate mean + standard deviation and record in a table
2.1.4 e the need for coenzymes, cofactors, and prosthetic groups in some enzyme-
controlled reactions
Cofactor-> Some enzymes require inorganic ions to function properly. Particular inorganic
ions may help to stabilise the structure of the enzyme or may actually take part in the
reaction at the active site. Obtained via the diet as minerals. E.g. Chloride ions act as a
cofactor for amylase. Meaning that in order for amylase to digest starch into maltose,
chloride ions need to be present.
Coenzymes-> organic cofactors. Obtained via vitamins(e.g. vitamin B3 -has nicotinic acid-
used to synthesise NAD, a coenzyme that transfers hydrogen atoms between molecules in
respiration)
Prosthetic groups-> they are cofactors bc they are required by certain enzymes. Some
cofactors are loosely/ temporarily bound to enzymes, but prosthetic groups are tightly
bound to enzymes + they’re permanent. E.g. Z n2 +¿¿ is bound to carbonic anhydrase for
metabolism of carbon dioxide.
Precursors-> enzymes produced in an inactive form. They are enzymes which can cause
damage within cells producing them/ enzymes whose action needs to be controlled and
only activated under certain conditions.
Precursor enzymes need to undergo a change in tertiary shape so they can be activated.
This is achieved by adding a cofactor. Before the cofactor the precursor protein is called an
apoenzyme. After cofactor is added, it’s called a holoenzyme.
2.1.4 f the effects of inhibitors on the rate of enzyme-controlled reactions.
Enzymes can be inactivated using inhibitors. There are two types of inhibition->
competitive or non-competitive.
Competitive-> a molecule which has a similar shape to the substrate fits into the active
site of an enzyme. This inhibits the enzyme and prevents it from catalysing a reaction. This
reduces the number of substrate molecules binding to active sites which slows down rate
of reaction.
Non-Competitive-> inhibitor binds to enzyme in a location other than active site
(alternative site= allosteric site). This inhibitor alters the tertiary structure of the enzyme
so active site shape changes. Means enzyme can’t carry out it’s function so it’s inhibited.
Effect of the rate of reaction (ROR):-
Competitive= reduces ROR but it doesn’t change Vmax of enzyme it inhibits. If substrate
conc is increased, then there will be more substrate than inhibitor an Vmax is reached.
Non-competitive= increasing conc of enzyme/ substrate is not going to change anything so
the ROR will keep reducing if the inhibitor keeps increasing.

Reversible inhibitor= slow down/ stop enzyme activity, decreasing ROR. They act as
regulators in metabolic pathways.
Non-reversible inhibitor= form covalent bonds with enzymes, inhibiting them
permanently
End-product inhibition:
Term used for enzyme inhibition that occurs when the product of a reaction acts as an
inhibitor. This shows Negative- Feedback mechanism.
Metabolic reactions can be controlled by using the end-product of a particular sequence
of metabolic reactions as a non-competitive, reversible inhibitor:
- Enzyme converts substrate into product. Process is slowed down as end-product
binds to an alternative site on original enzyme-> changes active site. No more
enzyme-substrate complexes.
- End-product can then detach from enzyme + used elsewhere, this allows the active
site to return to its ‘active state’.

Non-reversible inhibitors: cause the complete inactivation of an enzyme-> dangerous

because the reaction that is being catalysed is completely stopped. Only way to avoid this,
is to produce more of the enzymes being inhibited-> this is a relatively slow process.
This is why some non-reversible inhibitors are considered to be metabolic poisons, e.g.
cyanide which is an irreversible inhibitor of cytochrome oxidase (mitochondrial enzyme)
that catalyses one of key reactions in aerobic respiration.