Biological Molecules

Biology

Contents: ~Specification 2(c) + CGP p.6,7,13,14

-​ Biological molecules - Food Tests
-​ Investigating enzymes activity - Enzymes

Enzymes
Enzymes are biological catalysts, proteins that speed up chemical reactions without being changed or
used up in the reaction. While reactions could happen faster if temperature was raised, there’s a limit
to how long this relationship can remain positive. At one point, cell’s will start getting damaged and
unwanted reactions will be sped up too.
Enzymes reduce the need for high temperatures and they are only
used to speed up useful reactions, also called metabolic reactions.
As they are proteins, which are molecules made up of chains of
different amino acids, the chains can be folded up into different
unique shapes, thus making lots of different enzymes in the
process.

In most chemical reactions, reactants are being either split apart or

joined together. The substances changed in the reaction
(reactants) are also called substrates.
Every enzyme has an active site, which is the part where
substrates join onto to form ES complexes. Everyone is unique to a single reaction as substrates
must have a distinct shape to fit into the active site. This is called the “Lock and Key” model. Enzymes
whose active site fits into a substrate are complementary to that substance.
Enzymes and substrates don’t find each other as they don’t have a conscience, they just collide
randomly.

Thousands of enzymes in the human body exist to perform around 5,000 different functions. A few
examples include:
Lipases: This group of enzymes help digest fats in the gut.
Amylase: In the saliva, digests starch (a polysaccharide of glucose) into maltose (a disaccharide of
glucose)
Maltase: This also occurs in the saliva, and breaks the sugar maltose into glucose (monosaccharide).
Trypsin: These enzymes break proteins down into amino acids in the small intestine.
Catalase: breaks down hydrogen peroxide into water and oxygen.

You could speed reactions further by increasing temperature. Increased temperatures affect the rate
of an enzyme-catalysed reaction. At first, increasing the temperature would transfer more thermal
energy to the enzymes and the substrates, which would be transformed into kinetic energy, allowing
them to move more and be more likely to collide quicker.
However, if temperature rises past the optimum temperature ( which is the temp the enzymes work
best at), the energy would break the bonds holding the enzymes together, changing its shape and
therefore its active site, so the substrate would not fit in it anymore. The enzyme becomes denatured.
pH also affects enzyme activity. If it’s not around the optimum pH, it interferes with the bonds,
altering the active site’s shape and denaturing the enzyme. The optimum pH is often 7, so neutral, but
not always.

Investigating temperature’s effect on enzyme activity

You could measure how fast a product appears in the breakdown of hydrogen peroxide, you can
measure how temperature affects catalytic activity of catalase.

1) Use a pipette to add a set amount of hydrogen peroxide to a boiling tube in a water bath at a set
temperature.
2) Add a source of catalase, e.g. potato to the hydrogen peroxide and quickly attach the bung to it.
3) Record how much oxygen is produced in the first minute and repeat various times, then calculate
the mean.
4) Repeat this process for several temperatures to be compared
amongst them and be sure to control any variable like pH, size or
potato used… to avoid systematic errors and make it a fair test.

…or how fast a substrate disappears → The quicker the reaction is

completed, the faster the enzyme is working.

Method
1)Add 5cm3 starch solution to a test tube and heat to a set
temperature using beaker of water with a Bunsen burner
2)Add a drop of Iodine to each of the wells of a spotting tile
3)Use a syringe to add 2cm3 amylase to the starch solution and
mix well
4)Every minute, transfer a droplet of solution to a new
well of iodine solution (which should turn blue-black)
5)Repeat this transfer process until the iodine solution
stops turning blue-black (this means the amylase has
broken down all the starch)
6)Record the time taken for the reaction to be completed
7)Repeat the investigation for a range of temperatures
(from 20°C to 60°C)

Results and Analysis

This investigation shows:
At the optimum temperature, the iodine stopped turning
blue-black the fastest. This is because the enzyme is
working at its fastest rate and has digested all the starch
At colder temperatures (below optimum), the iodine took
a longer time to stop turning blue-black. This is because
the amylase enzyme is working slowly due to low kinetic
energy and few collisions between the amylase and the
starch.
At hotter temperatures (above optimum) the iodine turned
blue-black throughout the whole investigation. This is
because the amylase enzyme has become denatured
and so can no longer bind with the starch or break it
down

Limitations - There are several different ways in which the

temperature could be controlled. The method described
above is not very precise, an improvement would be to
use water baths kept at each temperature. The starch
and amylase solutions that need to be used should be
placed in a water bath and allowed to reach the
temperature (using a thermometer to check) before being
used
A colorimeter can be used to measure the progress of the
reaction more accurately

pH can also be investigated. You’d do this by adding a

buffer solution (solution that can resist pH change upon
the addition of an acidic or basic component) with
different pH levels to a series of different tubes containing
the ES mixtures.
Biological Molecules
There are different biological molecules we need in life. They are usually long, complex molecules
formed from smaller basic units. They are: Carbohydrates, Lipids and Proteins

Carbohydrates are the body’s primary source of energy. They

Molecules Elements
are obtained from oxidising a sugar called glucose. They can
be found in mostly sugary foods and bread, beans, rice, etc..
Carbohydrates CHO
Starch (plant cells) and glycogen (animal cells) are large,
Lipids C H (O) *
complex and insoluble (good for storing) carbohydrates which
are formed by smaller units like glucose and maltose joined
Proteins CHON
together in a long chain.
(S-ulphur)
Starch is a polymer/polysaccharide
(many units) broken down by the
enzyme amylase to get maltose which is a disaccharide (2 units). This is
broken down by maltase into glucose, a monomer(monosaccharide).
Glucose stores energy and is used in respiration (C⁶H¹²O⁶)
Cellulose is a polymer of glucose. It’s found in the cell wall of plant cells.
Humans cannot digest it and therefore can’t get energy from it but it is still
key for us.

Lipids are found in foods like beef and lamb. Most in the human body
appear as triglycerides (polymer) though their basic unit (monomer) is one
glycerol molecule chemically bonded to three fatty acid chains varying in
size and structure.Lipids are divided into fats (solids at room temperature)
and oils (liquids at room temperature)
*Lipids contain a much smaller portion of oxygen than carbohydrates

Proteins are formed from long chains of amino acids.

There are 20 different amino acids. When amino acids
are joined together a protein is formed. Amino acids
can be arranged in any order, resulting in hundreds of
thousands of different proteins
Examples of proteins include enzymes, haemoglobin,
ligaments and keratin.
A peptide bond is a short chain of amino acids.

Food Tests
In order to perform a food test, you need a food sample. To do it follow these steps:
1)​ Get a piece of food and break it up into smaller bits using a pestle and a mortar.
2)​ Transfer the ground up food into a beaker and add some distilled water.
3)​ Give the mixture a good stir with a glass rod to dissolve some of the food.
4)​ Filter the solution out using a funnel lined with filter paper to get rid of the solid bits of food.

Benedict’s test 4 Glucose

Glucose is found in biscuits, cereals and bread. You can test for
glucose through this test.
1)​ Prepare a food sample and transfer 5cm3 to a test tube.
2)​ Prepare a water bath set to 75ºC
3)​ Add Benedict’s solution to the test tube using a pipette
(approx. 10 drops)
4)​ Place the test tube in the water bath (use a test tube holder).
Leave it for 5 min. The tube needs to point away from you.
5)​ If the sample contains glucose, the solution will no longer be
blue coloured, instead it may turn green or yellow in low
concentrations (of glucose) and red in high concentrations.
Iodine test 4 Starch
This test is used to test starch from a sample. As starch is a carbohydrate, it is mainly
found in many grains and vegetables like wheat, pasta, potatoes and rice.

1) Make a food sample and transfer 5cm³ of your sample to a test tube
2) Then add a few drops of iodine solution onto the sample and gently shake the tube to
mix the contents.
3) If the sample contains starch, the colour of the solution will change from
browny-orange to black or blue-black.

Biuret test 4 Proteins

Protein is found in foods like meat and cheese. To test for it:
1)​ Prepare a sample of your food and transfer 2cm³ of your sample to a test tube
2)​ Add 2cm³ of biuret solution to the sample and mix the contents of the tube by gently
shaking it.
3)​ If the food sample contains protein, the solution will change from blue to pink or
purple. If there is no protein present, the solution will stay blue

Sudan III test 4 Lipids

Found in foods like olive oil, margarine and milk, lipids are tested for like this:
1)​ Prepare a sample of food that you are testing (Not filtered) - Transfer about 5cm³ into a test tube.
2)​ Use a pipette to add 3 drops of Sudan III stain solution to the test tube, then gently shake it.
3)​ The solution stains lipids. If the sample contains lipids, the mixture will separate into two layers
(top layer - red). If there are no lipids, the mixture will not separate or turn red.

Some Advice…
CORMS questions…