2.

Biological Molecules
Biological Molecules
Carbohydrates:
1. Contain the elements Carbon, Hydrogen and Oxygen.

2. Are split into groups polysaccharides (e.g. starch, glycogen), disaccharide (e.g. sucrose)
and monosaccharides (e.g. glucose).

Protein:
1. Contain the elements Carbon, Hydrogen, Oxygen and nitogen.

2. Made of subunits called amino acids.

3. The test for protein involves the use of a biuret solution (Sodium Hydroxide and
Copper Sulphate). If the solution goes from blue to pink-purple then protein is present.

Lipids:
1. Contain the elements Carbon, Hydrogen and Oxygen.

2. Made of a molecule of Glycerol and three fatty acids.

Test for glucose

1) Add the sample solution into a test tube
2) Add drops of Benedict’s solution into the test tube
3) Heat in a water bath at 60-70°C for 5 minutes
4) Take test tube out and record the colour
● If glucose is present the solution will turn brick red
● If glucose is not present that the solution will remain blue
Test for starch
1) Pipette the sample solution into wells or on a tile
2) Add drops of iodine solution and leave for 1 minute
3) Record any colour change
● If starch is present the solution will turn blue-black
● If starch is not present the solution will remain brown

Test for fat (lipids)

1) Add 2cm3 of ethanol to the test solution
2) Add 2cm3 of distilled water
3) Leave for 3 minutes and then record the colour
● If fat is present a milky white emulsion will form
● If fat is not present that the solution will remain colourless
Test for protein
1) Add the sample solution into a test tube
2) Add drops of Biuret solution into the test tube
3) Leave for 1 minute and then record the colour
● If protein is present the solution will turn purple
● If protein is not present that the solution will remain blue
Enzymes
Enzymes are biological catalysts (a substance that increases the rate of reaction without
being used up). They are protein molecules and the shape of the enzyme is vital to its
function. This is because each enzyme has its own uniquely shaped active site where the
substrate binds.
Enzyme activity is affected by:
- Temperature
- pH
- Enzyme and substrate concentration
- Competitive and non-competitive Inhibitors

Temperature
changing temperature changes the rate of an enzyme catalysed reaction, a higher
temperature increases the rate at first, the enzymes and substrates have more kinetic
energy so they move more so it is more likely for them to collide. If the temperature gets
too high, the bonds holding enzymes will break, this changes the shape of the active site
so the substrate will not fit anymore. This enzyme is said to be denatured. All enzymes
have an optimum temperature.

pH
if the pH is too high or too low, it interferes with the bonds holding the enzymes together.
This changes the shape of active site so the substrate will not fit anymore, the enzyme is
said to be denatured. All enzymes have an optimum temperature that they work best at.
Substrate concentration
Enzymes will work best if there is plenty of substrate. As the concentration of the
substrate increases, so does the rate of enzyme activity. However, the rate of enzyme
activity does not increase forever. This is because a point will be reached when the
enzymes become saturated and no more substrates can fit at any one time even though
there is plenty of substrate available.

As the substrate concentration increases so does the rate of enzyme activity. An

optimum rate is reached at the enzyme’s optimum substrate concentration. A continued
increase in substrate concentration results in the same activity as there are not enough
enzyme molecules available to break down the excess substrate molecules.

Competitive Inhibitors
They are molecules with a similar shape to the substrate. They fit into the active site,
stopping the substrate from entering. However, this is temporary – the inhibitor can leave,
and the substrate can take its place. Hence it is called a competitive inhibitor as the
enzyme and substrate are competing for the active site.
The inhibitor has a similar shape to the substrate and therefore is complementary in
shape to the enzyme so it can bind to the active site. The enzyme acts as the lock and
the substrate is the key, as per the lock and key theory.
Non-competitive Inhibitors
They do not have a shape like that of the substrate. They do not attach to the active site
but to the other parts of the enzyme. When they attach, they change the shape of the
whole enzyme including the active site. The active site can hence no longer receive the
substrate, so the reaction slows down. However, unlike competitive inhibitors they are not
able to catalyse the reaction even if the concentration of the substrate is increased as
the substrate cannot fit into the active site. The relative concentration of inhibitor and
substrate do not affect the rate of reaction.
It binds at a place on the enzyme other than the active site so it changes the shape of
the active site so that the substrate cannot bind to the enzyme. It is an irreversible
process.

Immobilised Enzymes
An immobilised enzyme is advantageous over a dissolved enzyme as:
1. The enzymes are more stable at high temperatures and are less likely to denature
2. The enzymes are more resistant to changes in pH
3. The enzymes are less likely to be broken down by organic solvents
4. The products are uncontaminated by enzyme and can be collected more easily
5. The enzyme can be kept and re-used
6. An industrial process can use columns of immobilised enzyme, allowing large scale
production.
Enzymes can be immobilised in several ways, such as by attaching the molecule the
surface of a material such as porous glass or cellulose, or by trapping the enzyme
molecules in a permeable membrane such as nylon or in a polymer such as alginate.
Three uses of immobilised enzymes are:
1. Producing lactose-free milk
People who are lactose-intolerant cannot digest lactose as they cannot produce the
enzyme lactase. However, the lactose in milk can be broken down by the use of
immobilised enzymes. Lactase is immobilised in porous beads such as calcium alginate
and are held in a column. Milk passes down the column as the lactose is broken by
down the lactase. Lactase is an expensive enzyme and therefore it is beneficial as
lactase can be re-used.
Immobilised lactase is found in porous beads such as calcium alginate and is held in a
column. Milk is poured down the column containing the immobilised lactase and
lactose binds with lactase to produce glucose and galactose. The milk collected at the
end of the column does not contain lactose therefore lactose-intolerant people can
drink it. It is beneficial as lactase can be re-used (lactase is expensive)
2. Breakdown of sucrose
The enzyme invertase is produced by yeast cells and can break down sucrose to
fructose and glucose. Invertase is used to break down sucrose and produce ‘invert
syrup’ which is used for sweetening products in food.
Enzyme invertase is produced by yeast cells. Invertase binds to sucrose and breaks
down sucrose to glucose and fructose. The invertase produces ‘invert syrup’ which
is used for sweetening products in food.

3. Testing for glucose in blood or urine

Diabetes results in higher than normal levels of glucose in blood and triggers the
appearance of glucose in urine. People can be tested for diabetes by using test strips
which makes use of immobilised enzymes.
Test strips consists of two immobilised enzymes.
The first enzyme is glucose oxidase. This catalyses the oxidation of glucose to
gluconic acid and hydrogen peroxide.
Glucose + O2 gluconic acid + H2O2 (Reaction 1)
The hydrogen peroxide produced in reaction 1 oxidises a
colourless organic substance to its coloured form, X, by removing hydrogen.
XH2 + 2H2O2 2H2O + X
Reaction 2 is catalysed by another enzyme called peroxidase. The
amount of coloured substance X produced is a direct measure of the concentration of
glucose present in the sample of blood or urine. The colour change on the strip gives
an approximate measure of the concentration of glucose.
Immobilised enzymes are bound to a carboard strip and urine is passed over the strip
including the enzyme. If glucose is present it binds to the immobilised enzyme to
produce hydrogen peroxide which causes a colour change.