2.

4 EXPLORING ENZYMES
Enzymes
• Enzymes are biological catalysts that
speed up the rate of biochemical
reactions.
• They are protein in nature.
• They are neither used up nor changed at
the end of the reaction.
• Enzymes catalyse various
reactions in the body such as
digestion, respiration etc.
 Properties/Characteristics of enzymes
1. They are globular proteins and are synthesized within cells.
2. They are biological catalysts - they speed up the chemical reaction
3. They catalyse chemical reactions by lowering the energy required to start the reaction
(activation energy).
4. They do not alter the properties of the end products of the reaction they catalyse.
5. They are very efficient – only a small amount of enzyme is required to produce large
amounts of substrate.
6. Enzymes remain unchanged at the end of the reaction.
• Enzymes are specific. Enzymes possess active sites where reaction takes place. These
active sites have specific shapes which are complementary to a specific substrate.
• Enzyme activity is affected by pH, temperature, substrate concentration, and enzyme
concentration.
• Enzymes work best at an optimum temperature and pH.
• Enzymes are denatured by high temperatures and extremes of pH.
• Some enzymes work in conjunction with cofactors.
• The catalysed reaction is reversible.
 Mode of Action of Enzymes
1. Lock and key Model (hypothesis) of enzyme action - Enzymes are globular proteins
having a tertiary structure, that is, the polypeptide is folded on itself and the structure is
maintained by hydrogen bonds, ionic bonds, disulphide bonds and hydrophobic
interactions. The enzyme has on its surface a specialized molecule configuration known
as the active site. The shape of the active site is specific and allows substrates with a
complementary shape to fit in and form an enzyme-substrate complex. The enzyme
represents the lock and the substrate represents the key. The substrate binds to the
active site of the enzyme and is held in place by 2 hydrogen bonds. Reaction takes
place and the substrate is broken down into products which then leave the active site.
The enzyme remains unchanged and can be used over again.
2. Induced fit hypothesis - The enzyme’s active site is not initially an exact fit to the
substrate molecule. However, the enzyme molecules are more flexible and can change
shape slightly as the substrate enters the enzyme. This means that the enzyme
molecule will undergo conformational changes as the substrate combines with
enzyme’s active site, forming the enzyme-substrate complex. The reaction proceeds,
products are formed. The products leave the active site and the enzyme regains its
initial shape.
 Activation Energy
• It is the energy required in any chemical reaction to break the bonds in reactant
molecules so that new bonds are formed to make the product.
• An enzyme lowers the activation energy required for the reaction.
• However, overall energy released during reaction is maintained.
• Enzymes speed up the rate of a reaction by lowering the activation energy of a
reaction, they do this by holding the substrate in such a way that their molecules can
react more easily.
 Effects of Enzymes
The effect that enzymes have on the rate of reactions can be measured in two ways:
1. By measuring the amount of product accumulated over a period of time.
Rate of reaction = volume of product formed/ time
E.g.
Enzyme Catalase breaking hydrogen peroxide
to water and oxygen
2. By measuring the rate at which the reactants disappear from the reaction mixture, the
effect of the enzyme on the rate of reaction can be determined. Eg: measuring the rate
at which starch disappears when the enzyme amylase is added.
Initially, there are a large number of substrates
(starch) and every enzyme (amylase) has
a substrate in its active site. The rate at
which the reaction occurs depends only on
how many enzymes there are and the
speed at which the enzyme can convert
the substrate into product, release it,
and then bind with another substrate.
However, overtime, there are fewer
substrates to bind with enzymes; the
reaction gets slower, until it eventually
stops. The rate of an enzyme-controlled
reaction is always fastest at the beginning.
 Intracellular Enzymes
• The enzymes that act within the cells in which they are produced are called
intracellular enzymes or endoenzymes.
• As these enzymes catalyze most of the metabolic reactions of the cell, they are also
referred to as metabolic enzymes.
• Most of the enzymes in plants and animals are intracellular enzymes or
endoenzymes.
• Intracellular enzymes usually break down large polymers into smaller chains of
monomers.
• All intracellular enzymes undergo intracellular digestion during cell death.
 Extracellular Enzymes
• The enzymes which are liberated by living cells and catalyze useful reactions outside
the cell but within its environment are known as extracellular enzymes or
exoenzymes.
• Exoenzymes act chiefly as digestive enzymes, catalyzing the breakdown of complex
macromolecules to simpler polymers or monomers, which can then be readily
absorbed by the cell.
• These mostly act at the end of polymers to break down their monomers one at a time.
• Exoenzymes are enzymes found in bacteria, fungi, and some insectivores
like Drosera and Nepenthes.
• Extracellular enzymes, unlike intracellular enzymes, undergo external digestion
during cell death.
 Economic Importance of Enzymes
1. ELISA:
• It is enzyme based detection of diseases like AIDS.
2. Endonucleases:
• They are enzymes used in breaking DNA at specific sites. DNA fragments are
employed in genetic engineering.
3. Alcoholic Beverages:
• Enzyme complex zymase obtained from yeast is used in brewing or fermentation of
alcoholic drinks.
4. Detergents:
• They contain protease for brighter washing of clothes and amylase for dish washing.
5. Baby Foods:
• Trypsin is added to partially pre-digest baby foods.
6. Streptokinase:
• The enzyme is used in clearing blood clots inside blood vessels.
7. Digestive Aids:
• Diastase and other enzymes are used regularly by patients with deficient digestive
juices.
8. Cheese:
• Rennet or rennin tablets are used for preparation of cheese. Lactase and lipase are
employed to provide proper consistency and flavour to cheese.
9. Pectinase:
• It is used for clearing fruit juices, retting of fibres and preparation of green coffee.
10. Protease:
• The enzyme is employed for chill-proofing of beverages, degumming of silk,
cleaning of hides, softening of bread and meat.
Apoenzymes and Holoenzymes
• Some enzymes have a non-protein chemical compound or metal called a cofactor to
function.
• When an enzyme combines with its cofactor, its referred to as holoenzyme.
• The enzyme (protein) part alone is called apoenzyme.
Coenzymes
• They are organic compounds required by an enzyme for catalytic activity.
• They are often vitamins or derivatives of vitamins. E.g CoA, NAD, FAD, NADP etc.
• The difference between cofactors and coenzymes is that a coenzyme is required for
an enzyme activity while a cofactor is not.