Topic 1: Biological Molecules

Carbohydrates and Lipids

Carbohydrates

Carbohydrates are long chains of sugar molecules (the technical term is ‘polysaccharides’) formed
from lots of individual sugar molecules (monosaccharides) joining together. When two
monosaccharides react to form a disaccharide, a condensation reaction occurs. During the
condensation reaction, a molecule of water is removed (from a hydroxyl group on one sugar and a
hydrogen on another) and a glycosidic bond forms. When polysaccharides are broken down during
digestion, a hydrolysis reaction occurs in which a water molecule is added to break the glycosidic
bond.

Carbohydrates are made up of carbon, hydrogen and oxygen only. Monosaccharides such as
glucose and fructose consist of a ring structure with the general formula CH2O. Glucose is a hexose
sugar, which means it is made up of six carbon atoms. It can exist as two different forms, alpha and
beta which differ in the position of the hydrogen and hydroxyl groups on the right-hand carbon. An
easy way to tell the difference between them is:
Alpha - the hydrogen atom is above the carbon.
Beta - the hydrogen atom is below the carbon.
Ribose is another example of a monosaccharide - it’s a component of RNA molecules. It is a
pentose sugar with five carbon atoms in the ring.

Polysaccharides

The polysaccharides found in plants are starch and cellulose. Starch is broken down by the plant
when it needs energy and cellulose is the major component of plant cell walls. Starch exists in two
different forms: amylose and amylopectin. In animals, the energy storage carbohydrate is glycogen.

• Amylose: unbranched spiralling chains of alpha-glucose molecules. Its coiled structure means
that it is very compact so lots of amylose can be packed into a cell.
• Amylopectin: branched chains of alpha-glucose molecules. Its branches increase its surface
area which means that enzyme can quickly break it apart when glucose is needed for
respiration.
• Cellulose: long unbranched chains of beta-glucose molecules. Multiple chains are linked
together by hydrogen bonding to form strong structures called micofibrils. The strong
microfibrils in the cell wall help to give plant cells their shape and structural support.
• Glycogen: branched chains of alpha glucose, similar to amylopectin but with more side-
branches. This gives it a large surface area for enzyme action to release glucose when energy is
needed. It is more compact that amylopectin, which means more can be stored in a cell.

Lipids

Triglycerides, phospholipids and cholesterol are

all types of lipid. Triglycerides and phospholipids have
similar structures as they are both made up of
glycerol connected to fatty acids molecules through
ester bonds. However, triglycerides contains three
fatty acid ‘tails’ whereas phospholipids only have
two. Phospholipids also possess a phosphate group
which is absent in triglycerides. The fatty acid tails of
triglycerides or phospholipids can be saturated (only
single carbon-carbon bonds) or unsaturated (contain
at least one double carbon bond).
 The synthesis of a phospholipid or triglyceride from glycerol and fatty acids involves the formation of
an ester bond. Just like we saw for the formation of a glycosidic bond, the synthesis of ester bonds is
a condensation reaction in which a water molecule is released. Breaking apart a triglyceride or
phospholipid involves the addition of water in a hydrolysis reaction.

Different types of lipids have different functions in living organisms:

• Triglycerides: used as an energy store. A lot of energy is released when the ester bonds are
hydrolysed - around twice as much compared to the breakdown of carbohydrates. Inside cells,
triglycerides group together into lipid droplets where the hydrophobic tails face inwards and the
hydrophilic heads face outwards. These insoluble droplets make good energy storage molecules
since they do not affect the osmotic potential of the cell.
• Phospholipids are the main component of cell membranes. They form a phospholipid bilayer with
the hydrophobic tails facing each other and the hydrophilic heads facing outwards, forming a barrier
to prevent any polar molecules from entering or leaving the cell.
• Cholesterol is also found in cell membranes and helps to strengthen the membrane. Cholesterol
pushes the hydrophobic tails of phospholipids closer together, making the membrane more rigid.

Testing reducing sugars using Benedict’s solution

A reducing sugar is any sugar that can act as a reducing agent because of its aldehyde or ketone
group. All monosaccharides are reducing sugars, along with some disaccharides (such as maltose
and lactose). If the sugar you are testing is a reducing sugar, it will change colour from blue to
green/yellow/orange/brick red, depending on the concentration of sugar you are testing. Low
concentrations of reducing sugar will cause a green or yellow precipitate to form whereas a reducing
sugar of higher concentration will appear orange or brick red.

If the test for reducing sugars is negative and you want to confirm the presence of a non-reducing
sugar, such as sucrose, you need to break down the sample into monosaccharides using
hydrochloric acid which breaks the glycosidic bonds. The sample is neutralised with sodium
hydrogencarbonate and the Benedict’s test can be carried out. Since sucrose will have been broken
down into glucose and fructose (both reducing sugars), you should now see a positive result if your
original sample contained sucrose.

If you want to accurately measure the amount of reducing sugar in a sample, you can use
calorimetry. Calorimetry works by measuring how much light is able to pass through the solution.
The less light that passes through, the higher the absorbance of the sample and the higher the
concentration. Before measuring the unknown sample, you first need to measure the absorbance
readings of a series of solutions of a known concentration. You would plot a calibration curve of
absorbance against concentration then measure the absorbance of the sample of unknown
concentration. You can read off your graph to find the concentration which corresponds to this
particular absorbance reading.
Testing for starch using iodine

Starch is the storage carbohydrate in plant cells. If you want to test a part of a plant (such as a potato)
for starch, you need to add iodine dissolved in potassium iodide solution. A positive result occurs
when the solution changes colour from orange/brown to blue/black.

Testing for lipids using the Emulsion test

To test for the presence of lipid in a sample, you need to add ethanol to a sample and mix thoroughly
by shaking. Add an equal volume of water and if lipid is present a milky white suspension should
form. If the solution remains colourless, no lipid was present in the sample.
Topic 1: Biological Molecules
Nucleic Acids and DNA Replication
Nucleic acids

Nucleic acids are long chains (polymers) of lots of nucleotide monomers joined together by
phosphodiester bonds. Each nucleotide is made up of three components:

1. Pentose sugar (either deoxyribose or ribose),

2. Nitrogenous base (guanine, cytosine, adenine, thymine or uracil)
3. Phosphate

DNA consists of two nucleic acid strands bonded together by complementary base pairing, the
strands twisted around each other to form a double helix. The two strands have the same sequence
running in opposite directions so we say they are anti-parallel. The bonding between bases is
predictable, since guanine always pairs with cytosine with three hydrogen bonds holding them
together. Adenine always bonds with thymine (in DNA) or uracil (in RNA) with two hydrogen bonds
between them. Adenine and guanine are purines, which have a large double ring structure.
Cytosine and thymine are pyrimidines, which are smaller as they are composed of a single ring.
Notice that a purine always base pairs with a pyrimidine - this happens because it’s the only way DNA
can keep a regular shape and not become too bulky or narrow.
There are some important differences between a molecule of DNA and RNA:

1. They contain different sugars - the pentose sugar in DNA is deoxyribose and in RNA is ribose.
The difference is that deoxyribose has one less oxygen atom.
2. RNA contains uracil instead of thymine
3. DNA is double-stranded whereas RNA is single-stranded
4. DNA has hydrogen bonds between the two complementary strands

During DNA and RNA synthesis, nucleotides are connected through the formation of phosphodiester
bonds (a type of covalent bond) between the phosphate group on one nucleotide and the pentose
sugar of the next nucleotide. It is a condensation reaction which means that water is formed during
the reaction and it is catalysed by an enzyme called DNA polymerase or RNA polymerase
depending on whether a DNA or RNA strand is being synthesised. Breaking phosphodiester bonds
requires the addition of a water molecule, so it is a hydrolysis reaction.

Semi-conservative DNA replication

During cell division, cells need to make a complete copy of their genetic information. When DNA is
replicated, the new DNA molecule is made up of one strand of the original DNA whereas the other
strand is made of freshly made DNA. Since half of the DNA is preserved from the previous round of
DNA replication, we describe the process as semi-conservative. It takes place in the following
stages:

• DNA helicase unwinds the double helix, breaking the hydrogen bonds between
complementary base pairs to separate the strands. One of the strands will act as a template for
synthesis of the other strand.
• Complementary nucleotides will attach to the template strand by hydrogen bonding.
• DNA polymerase catalyses the formation of phosphodiester bonds between nucleotides,
forming a complementary strand alongside the template parent strand.
• Two daughter DNA molecules are formed, each containing half of the original DNA molecule.

It is important that DNA polymerase accurately copies the template strand to avoid placing the
wrong DNA nucleotide in the incorrect position. To avoid this, DNA polymerase ‘proofreads’ the
complementary strand as it moves along the DNA. If it detects a mismatch, it can ‘snip out’ the wrong
nucleotide and replace it with the right one. DNA polymerase has an accuracy rate of about 99%,
which means that mistakes do occur every once in a while. A mistake results in a change to the DNA
base sequence, which is known as a mutation. DNA mutations can have detrimental effects to the
organism, since an altered base sequence can change the sequence of amino acids in a protein,
causing it to fold differently and possibly lose its function.
Meselson and Stahl’s Experiment

The evidence that DNA replication is semi-conservative comes from a pretty clever experiment carried
out by Matthew Meselson and Franklin Stahl. Before this, scientists were unsure whether DNA
replication was conservative or semi-conservative. If DNA replicated conservatively, the original DNA
strands would remain intact and the newly synthesised DNA would consist of two freshly-made
strands.

To figure this out they used a heavy isotope of nitrogen (N-15) which has an extra neutron
compared to the normal, lighter form of nitrogen (N-14). They grew bacteria in the presence of the
heavy nitrogen and any new DNA that the bacteria made would incorporate this isotope and so
would weigh heavier. If DNA replicates conservatively, Meselson and Stahl knew that they’d see
some DNA made of just heavy nitrogen with the rest made of only light nitrogen, but if it replicated
semi-conservatively it would be a mixture of the two isotopes. Here’s how they carried out the
experiment in more detail:

• Two populations of bacteria were grown - one in a solution containing heavy nitrogen and the
other in a solution containing light nitrogen. As the bacteria grow and reproduce, they
incorporate the nitrogen into their DNA.
• DNA was extracted from the bacteria and centrifuged to separate the DNA according to its
weight. The DNA from the bacteria grown in N-15 separated at a higher density (a band lower
down the test tube) compared to the DNA from the bacteria grown in N-14.
• The scientists then took the bacteria that had been growing in N-15 (which now just contains
heavy DNA) and grew them in the light isotope N-14. Again, they extracted their DNA and
separated it using centrifugation.
• After the first round of DNA replication, they saw just one band of DNA which was an
intermediate weight between 14-N and 15-N. This indicates that the DNA was made up of both
types of nitrogen isotopes (i.e. one old strand and one newly synthesised strand).
• After the second round of DNA replication, there are now 2 bands of DNA. One band has an
intermediate weight but further newly synthesised DNA is now only being made using the
lighter isotope. This proved that DNA is replicated semi-conservatively.
• If DNA replication is conservative, Meselson and Stahl would have seen two bands of DNA (one
heavy and one light) after both the first and second rounds of replication.
Topic 1: Biological Molecules

Proteins and Enzymes

Proteins

Proteins are polymers of amino acids joined together

by peptide bonds. There are 20 different amino acids
that our body needs to stay healthy, all of which have
the same general structure. They consist of a central
carbon atom attached to four different groups: an
amine group, a hydrogen atom, a carboxyl group
and an 'R’ group which is different in each amino acid.
The identity of the R group will influence how the amino
acid interacts with other amino acids, therefore influencing protein folding. For example, lysine has
a charged R group and is able to form ionic bonds with negatively charged amino acids.

Peptide bond formation involves the removal of water in a condensation reaction. The water is
formed by removing a hydrogen atom from the amine group and a hydroxyl (-OH) group from the
carboxylic acid. What’s left behind is a peptide bond (-CONH) which can be broken with the addition
of water in a hydrolysis reaction.

Once a long polypeptide chain is synthesised from the formation of numerous peptide bonds between
adjacent amino acids, the polypeptide undergoes multiple stages of folding. These stages of protein
origami are defined as the primary, secondary, tertiary and quaternary structures which describe
a protein from its most unravelled to its final compact structure.
• Primary structure: peptide bonds have formed between amino acids to form a long, straight chain
(polypeptide).
• Secondary structure: hydrogen bonds form between
nearby amino acids (from the amine group on one
amino acid to the carboxyl group of another) to form
either an alpha helix or a beta pleated sheet. Proteins
which form neither of these two structures will form a
random coil.
• Tertiary structure: more bonds form between the
different R groups to give the protein a 3D structure.
R-group interactions involve hydrogen bonds,
disulfide bonds, ionic bonds and polar interactions.
If proteins are made of a single polypeptide chain, this is
their final overall structure.
• Quaternary structure: this is the structure formed from
the interaction of multiple polypeptide chains held
together by bonds. Haemoglobin is an example of a
protein with quaternary structure. It consists of four
polypeptide chains (two alpha chains and two beta chains)
bonded together. Each chain surrounds an iron-containing
haem group. The haem group is referred to as a prosthetic
group - non-protein components which are required for
protein function.
Globular vs fibrous proteins

All proteins can be organised into one of two broad categories:

globular proteins and fibrous proteins. Globular proteins are
spherical and arranged with their hydrophobic amino acids
tucked inside and the hydrophilic amino acids exposed on the
outside. This means globular proteins are soluble and can be
transported easily from one part of the cell to another. They
perform functional roles and include things like enzymes
(such as amylase), hormones (such as insulin) or proteins like
haemoglobin. Globular proteins unravel and denature when
the temperature or pH deviates from optimum levels. Globular proteins with prosthetic groups
attached (such as haemoglobin) are referred to as conjugated proteins.

Fibrous proteins are long and thin and their primary structure consists of a repetitive sequence of
amino acids. They perform structural roles so they are strong and insoluble. Examples of fibrous
proteins include collagen, keratin and elastin. Collagen consists of three polypeptide chains
wrapped tightly around each other to form a stable quaternary structure held together by numerous
hydrogen bonds. Collagen is found in connective tissue, such as skin, muscle and bone. Fibrous
proteins tend to be less sensitive than globular proteins to changes in temperature and pH.

Testing for proteins using Biuret reagent

The Biuret test involves adding sodium hydroxide to a sample followed by copper sulfate solution
(together these compounds are referred to as Biuret reagent). A colour change from blue to purple
indicates a positive result.

Enzyme function and structure

Enzymes are biological catalysts - they speed up the rate of chemical reactions happening inside
our body. They work by reducing the activation energy of a reaction. Activation energy is defined as
the minimum amount of energy needed for a reaction to happen. If less energy is needed, then
reactions can take place as lower temperatures than would be needed without an enzyme. Without
the enzymes in our bodies, the reactions that happen inside of us would not be possible at normal
body temperature. Remember that enzymes are unchanged at the end of a reaction which means
they can be reused.

Enzymes can be classed as either intracellular if they catalyse reactions inside cells e.g. RNA
polymerase, or extracellular if they catalyse reactions outside of cells e.g. amylase. All enzymes are
globular proteins and have regions called active sites. The active site of an enzyme has a specific
shape and allows the substrate to bind. Other enzymes may have regulatory regions where an
inhibitor can bind, which we refer to as the allosteric site.

Mechanisms of enzyme action

Scientists have two ideas to explain the way in which enzymes work: the ‘lock-and-key’ model and
the ‘induced-fit’ model. They are models because they are our best-accepted theories based on the
evidence we have available.

Lock and Key model

The lock and key model is the simpler of the two theories of enzyme action. This model suggests
that the substrate fits into the enzyme’s active site in the same way in which a key fits into a lock. The
shape of the substrate and the active site are perfectly complementary to each other. Catalysis
happens in the following stages:

1. The substrate binds to the enzyme’s active site, forming an enzyme-substrate complex (ES
complex).
2. The enzyme converts the substrate into product, forming an enzyme-product complex (EP
complex).
3. The product is released from the enzyme’s active site.
The Induced Fit model

The induced fit model suggests that the shapes of the enzyme’s active site and its substrate are not
exactly complementary, but when the substrate enters the active site, a conformational change
(change of shape) occurs which induces catalysis. The induced fit model can be broken down into
the following stages:

1. The substrate enters the enzyme’s active site, forming an ES complex.

2. The enzyme undergoes a conformational change which causes the conversion of substrate into
product, forming an EP complex.
3. The product is released from the enzymes active site.

Comparing the two models of enzyme action

The advantage of the lock-and-key model is that it explains why most enzymes display such high
specificity to their substrates. Each enzyme will catalyse only a certain type of reaction and will only
bind to a single specific substrate out of the millions of different molecules that are floating around our
bodies. However, not all enzymes catalyse a single chemical reaction. For example, lipase exhibits
broader specificity and can bind to a variety of lipids, which only the induced fit model is able to
explain. In addition, the induced fit model is better able to explain how catalysis actually occurs. A
conformational change, which would place stress on the bonds within the substrate, can explain
how bonds would break in order for the products to form. This makes the induced fit model the
more widely accepted model of the two.
Factors which affect rate of reaction

Enzyme concentration

As enzyme concentration increases, the rate of

reaction increases since more active sites will be
available to bind to substrate molecules. This means that
there will be more frequent collisions between the
enzyme and substrate, so there will be more formation of
enzyme-substrate complexes. However, a point will be
reached when increasing enzyme concentration does not result in further increases in reaction rate.
At this point, something else has become a limiting factor, such as the availability of substrate.

Substrate concentration

As substrate concentration increases, the rate of

reaction increases since there are more substrate
molecules to fill the enzyme’s active sites. There will
be more frequent collisions so more formation of
ES complexes. At some point a ‘saturation’
point is reached where all of the enzyme’s active
sites are occupied with substrate molecules, so
the addition of more substrate molecules will have
no effect on the rate of reaction. At this point, the
reaction is proceeding as fast as possible, which is
referred to as Vmax. The only way the reaction can go any faster is by increasing enzyme
concentration.

Temperature

At low temperatures, the rate of reaction will be

slow because the enzyme and substrate have low
amounts of kinetic energy. This means that there
won’t be many collisions so there will be reduced
formation of ES complexes. As the temperature is
increased, the number of collisions increases,
increasing the formation of ES complexes and
increasing the rate of reaction. If the temperature
becomes really high, hydrogen bonds will begin to
break within the protein, causing it to unravel and become denatured. If enzymes are denatured, they
lose the shape of their active sites which means they cannot bind to their substrate, decreasing the
rate of reaction.

pH

Each enzyme has its own optimum pH at which it

works best. Pepsin, the enzyme which digests protein
in the stomach, works best in acidic environments
whereas the enzymes responsible for the digestion of
carbohydrates work better at a more neutral pH.
Deviations from the optimum pH change the charge on
the enzyme, which affects ionic bonding within its
structure. Deviations in pH also break hydrogen
bonds. This causes it to change shape and become
denatured, decreasing the rate of reaction as pH deviates from the enzyme’s optimum conditions.

Competitive vs non-competitive inhibitors

Competitive inhibitors are ones which bind to

the active site of the enzyme, blocking the
substrate from binding - they compete with the
substrate for access to the enzyme’s active site.
The effect of a competitive inhibitor can be
reduced by increasing substrate concentration.
If there is way more substrate compared to
inhibitor then the substrate is much more likely to
collide with the enzyme’s active site.

Non-competitive inhibitors are ones which bind

to a site on the enzyme away from its active site.
This region is known as its allosteric site. The effect of a non-competitive inhibitor cannot be
reduced by increasing substrate concentration.
Investigating the rate of an enzyme-controlled reaction

There are two ways of measuring the rate of an enzyme controlled reaction: By measuring the speed
at which product is formed or reactant is removed.

Measuring product formation using catalase

A good example of a reaction in which product formation is measured is the reaction catalysed by
catalase, which breaks down hydrogen peroxide into oxygen and water.

Method:

1. Add a set volume/concentration of hydrogen peroxide to a test tube along with a set volume
of buffer solution to control pH.
2. Place a bung on top of the test tube and connect to an inverted measuring cylinder using
delivery tubing.
3. Using a pipette, add catalase the test tube. Quickly re-insert the bung and delivery tube.
4. Measure the volume of oxygen produced in the measuring cylinder every ten seconds for the
first minute of the reaction (using a stopwatch to record the time).
5. Repeat two more times. Calculate the mean volume of oxygen produced at each ten second
time interval.

Control variables: volume of catalase, volume and concentration of hydrogen peroxide solution, pH,
temperature.

Plot a graph with time on the x-axis and volume of oxygen on the y-axis. You can determine the initial
reaction rate by drawing a tangent to the curve at t=0. Draw a straight line against the curve right at
the beginning of the reaction and calculate the gradient by dividing the change in y by the change in x.
You can repeat the experiment with different concentrations of catalase to determine the effect of
changing enzyme concentration on the initial rate of reaction.

Measuring reactant removal using amylase

Amylase is the enzyme which hydrolyses starch into maltose. You can measure decreasing starch
concentration as the reaction progresses by adding iodine, which turns blue/black in the presence of
starch.

Method:

1. Place a cuvette containing iodine solution in a colorimeter with a red filter. Zero the machine.
2. In a separate cuvette, pipette a given volume (e.g. 0.5 ml) of starch solution and a set volume
of iodine solution. Mix together – the solution should turn blue/black. Place in the colorimeter
and record the absorbance.
3. Add amylase solution of a set concentration/volume into the same cuvette and starch the
stopwatch.
4. Record the absorbance of the solution every ten seconds.
5. Repeat the whole process two more times and calculate a mean absorbance for each ten
second interval.

Control variables: concentration of iodine solution, volume of starch, pH, temperature.

Plot a graph with time on the x-axis and absorbance on the y-axis. You can repeat the experiment
with difference concentrations of starch to determine the effect of changing substrate
concentration on initial reaction rate.
Topic 1: Biological Molecules
Water, ATP and Inorganic Ions
Water

Water is a polar molecule - this means that the

electrons are shared unequally within the bonds that
hold a water molecule together. Oxygen is greedy
when it comes to electrons and is able to pull the
electrons closer towards it (we say it is more
electronegative). This gives oxygen a slight negative
charge and hydrogen a slight positive charge. A
hydrogen bond forms between the slightly negative
oxygen atom of one water molecule and the slightly positive hydrogen atom on another molecule. The
ability of water molecules to hydrogen bond gives it some special properties:

1. It is an excellent solvent: water is a good solvent because it can form hydrogen bonds with
other polar molecules or charged ionic compounds. Water dissolves more substances than any
other liquid so is referred to as the ‘universal solvent’. This is handy for humans, since our
blood consists mostly of water which is able to dissolve hydrophilic molecules such as glucose,
ions and some amino acids.
2. It has a high latent heat of evaporation - this means that a lot of energy is used to convert
water from a liquid to a gas. This is why we feel cooler when we sweat, since the evaporation of
water from our skin’s surface takes a lot of heat energy with it.
3. It has a high specific heat capacity - all those hydrogen bonds within water are great at
absorbing energy, which means you have to add a lot of it to heat water up. This is really useful
for marine life since it means that bodies of water are fairly resistant to changes in
temperature, making it a stable habitat in which to live.

Adenine triphosphate (ATP)

ATP is a phosphorylated nucleotide. Its structure consists of a ribose sugar and adenine attached to
three phosphate groups. Hydrolysis of ATP to ADP removes one of the phosphate groups and
releases energy in a single reaction. It also releases energy in small, manageable quantities which
means less is wasted.
 • When ATP is hydrolysed, it is converted into adenine diphosphate (ADP) and inorganic
phosphate (Pi). This reaction is catalysed by the enzyme ATP hydrolase.
• As well as releasing energy, the phosphate can be attached to other molecules to make
them more reactive. Enzymes and other proteins can be phosphorylated to convert them from
an inactive to an active form.
• ATP is re-synthesised in a condensation reaction, joining ATP and Pi to reform ATP. This
reaction is catalysed by the enzyme ATP synthase.

Inorganic ions

There is an array of cations (positive ions) and anions (negative ions) which are necessary for many
biological processes:

• Calcium ions, Ca2+: required for the transmission of action potentials, the formation
of bone, the release of insulin from the pancreas and as an enzyme cofactor.
• Sodium ions, Na+: required for the generation of action potentials, muscle
contraction and for maintaining blood pressure.
• Potassium ions, K+: required for the generation of action potentials, muscle contraction,
maintaining blood pressure and for activating photosynthesising enzymes in plants.
• Hydrogen ions, H+: important role in respiration and photosynthesis reactions and
influences pH of a cell by increasing acidity.
• Ammonium ions, NH4+: an important part of the nitrogen cycle. It is absorbed by plants and
incorporated into amino acids for protein synthesis.
• Nitrate ions, NO3-: an important part of the nitrogen cycle. It is absorbed by plants and
incorporated into amino acids for protein synthesis.
• Hydrogencarbonate ions, HCO3-: maintains constant pH in the blood by acting as a buffer.
• Chloride ions, Cl-: maintains constant pH in the blood through the process of 'chloride
shift' and acts as a cofactor for the enzyme amylase.
• Phosphate, PO43-: used to synthesise biological molecules such as nucleic
acids and phospholipids and is added to enzymes and other proteins to regulate
their activity.
• Hydroxide, OH-: changes the pH of a cell by making the area more alkaline.