BIOCHEMISTRY

LECTURE
ENZYMES
DEFINITION
Enzymesarebiologicalcatalystsproducedbythecaptivityofliving
organismsassuchasplants,animals,andmicroorganisms-andwhich
modifythespeedofreactionwithoutbeinguseduporappearingasone
oftheendproducts.
CHEMICAL NATURE OF ENZYMES
So far, all enzymes isolated and studied were proven to be proteins.
In general, they are soluble in water, glycerol and dilute alcohol. They
are precipitated by protein precipitants like concentrated alcohol,
ammonium sulfate and trichloroacetic acid. In solution, they are
colloidal in nature and are non-dialyzable.
BIOLOGICAL IMPORTANCE OF ENZYMES

Most chemical reactions characteristic of living matter are

accelerated by enzymes. It is estimated the a given cell could
contain a thousand different enzymes. Practically for every organic
compound which occurs in nature there exists also an enzyme
capable of reacting with it. These enzymes are responsible for the
different reactions in living matter- synthesis, oxidation,
hydrolysis, tautomeric changes- necessary during digestion,
metabolism, respiration, energy release, and energy transfer in all
metabolic reactions. It is not an exaggeration to say that life is
impossible without enzymes.
NOMENCLATURE (NAMING) OF ENZYMES

The present convention of naming enzymes is based on:

A.Upon the substance they act upon. The suffix “ase” is
added to the name of the substance acted upon. Some
examples are:
1. Amylase acts on starch
2.Maltase acts on maltase
3.Cellulose acts on cellulose
4.Urease acts on urea
5.Lipase acts on fats
B. Upon the reaction enhanced. The suffix “ase” is
also added to the type of chemical reaction
activated. Examples are:

1.Oxidasefor oxidation
2.Decarboxylase for removal of CO2
(decarboxylation)
3.Transaminase for transamination
4.Hydrolase for hydrolysis
5.Hydrase for hydration
C. The first enzyme discovered and studied
were given names that do not indicate the
substance acted on or reaction enhanced; at
that time there was no system of
nomenclature yet. Among these enzymes are
pepsin, ( in gastric juice), trypsin (in
pancreatic juice), ptyalin (in saliva), rennin
(the milk-curdling enzyme). etc.
CLASSIFICATION OF ENZYMES

A. Hydrolases - enzymes that bring about hydrolysis. Ex. Estrases, peptidases,

amidases, glycosidase, etc.
B.Desmolases or Lyases - enzymes that add or remove specific groups from
substances. Ex. Hydrases, dehydrases, decarboxylases, deaminases
C.Transferases - enzymes that transfer specific groups from one compond to
another.
Ex. Transamidases, transaminases, transpeptidases
D. Oxido-reductases - enzymes that bringabout oxidation and reduction Ex.
Oxidases, reductases, dehydrognases, hydroxylases
E.Ligases - enzymes that catalyze bond formation between two substrate
molecules. Ex. Synthetases, carboxylases,
F. Isomerases - enzymes that catalyze intramolecular arrangement. Ex.
Epimerases, mutates
TERMINOLOGIES IN ENZYME CHEMISTRY

1.Substrate- the term used to denote the substance acted

upon by the enzyme. Starch is the substrate of amylases
2.Zymogen (proenzyme) - this is the inactive form of
enzyme. Some enzymes are secreted in this form. They
remain inactive until another substance acts on them. Ex.
Pepsin (active form) is secreted as pepsinogen in the
stomach. Trypsin is secreted as trypsinogen in the small
intestines.
3.Activator- this term is applied to substances that convert a
proenzyme or zymogen (inactive form) into the active enzyme.
Activators may be inorganic substances HCl in the activator in
converting pepsinogen to pepsin) or organic in nature
(enterokinase converts trypsinogen to trypsin. Organic activators
are also called KINASES.
4.Coenzyme-organic substances which may exist free or loosely
attached to the enzyme (protein part) that help or assist in the
activation of a chemical reaction. They are necessary or required
for the proper functioning of some enzymes. Without their
presence , these enzymes will not function. Chlorides are needed
for amylase activity.
5.Apo-enzyme- the protein part to which the co-enzyme
attaches itself or is attached in the enzyme-coenzyme
systems.
6. Holo-enzyme - the combination of the Apo-enzyme
and coenzyme.
7.Anti-enzyme- substances that inhibit enzyme activity.
Ex. Is the intestinal parasite ascaris contains anti-trypsin
which prevent the enzyme trypsin from digesting the
worm. AntiTypsin has been found in soy beans, raw egg,
and pancreas.
MECHANISMS OF ENZYME ACTION

1. SimpleEnzyme Action: (enzyme-substrate combination. This

theory was proposed by Michaelis and Menten. The reaction
is presented as follows:
The above diagram may be explained as follows:
a.There is a temporary union between the enzyme and
the substrate forming an activated complex. The
union is brought about by the presence of a reactive
 site on the surface of the enzyme into which the
substrate can fit.
b.The bond in the substrate is then strained to the point
of rupture .
c.The reaction products are then formed and the
enzyme is regenerated thus available for another
catalytic action.
2. Complex Enzyme Action (enzyme-coenzyme systems) - The different steps involved may be
explained as follows:
a. The substrate and the Apo enzyme (with an appropriate site)
combine to form an activated complex, but in the presence of
the co-enzyme.
b. The bond in the substrate is strained, one of the cleavage
products (B) is transferred to the coenzyme (which has also
an appropriate site).
c. Product (A) dissociates from the Apo enzyme
d. Product (B) dissociates from the coenzyme; it may be
liberated as such or may be passed to other enzymes for
 additional changes. The Apo enzyme and the coenzyme are
thus liberated for another action.
CHARACTERISTICS OF ENZYME ACTION

A.HighDegreeofSpecificity

This is one of the most noteworthy properties of

enzymes. This means that a given enzyme will act only on
a particular substance or close-related substances. This is
explained by the fact that enzymes act and attack a
 compound at specific reactive site and definite type of
linkage. Proteoses can act only on proteins but not on fats
and carbohydrates; carbohydrases on carbohydrates; as
lipases act only on lipids.
Classification of Specificity:

1.Absolute specificity- when the enzyme acts on a one and only one
substrate. Ex., arginase on arginine,
2.RelativeSpecificity- when an enzyme can act on morethanone
substrateofrelated structure but at different intensities. Ex.,
 pancreatic lipase can hydrolyze fats with great rapidity but of simple
esters at a slower rate.
3.StereochemicalSpecificity- when an enzyme can act on only a
substance of specific configuration. Ex., maltase can act on alpha
glucosides but not on beta glucosides; emulsin can act only on
betaglucosides but not on alpha glucosides.
A very interesting proof o specificity is the enzymatic hydrolysis of the
trisaccaride raffinose as shown in the diagram below
 Emulsin

Saccharase

It shows that emulsin specifically attack the glucose-

galactose linkage while saccharase acted on the glucose-
fructose linkage.
B.ReversibilityofEnzymeAction
 As the products of the reaction increase in amount, the
reaction approaches the point of equilibrium and therefore
the rate decreases. There is tendency of the products of
the reaction to recombine although the rate will be slower,
This aspect of enzyme action is significant in that it
provides moderation of the rate of activity in cells thus
permitting the cells to adjust to a constantly changing
environment.

AB + enzyme AB-enzyme A + B + enzyme

C.EnzymaticActivityismaximalatanoptimumtemperature:
 There is an optimum temperature for each enzyme . Enzyme
of animal origin have an optimum temperature orf37 - 40 0C,
some plant enzymes have higher optimum temperature e.g.
Papain (enzyme from papaya) requires 650C for maximal
activity. Enzyme action is decreased at low temperatures. This
explains how foods are preserved by refrigeration (spoilage of
food is under enzyme control). A rise in temperature increases
the rate of activity until a point is reached where the enzyme is
irreversibly destroyed. Usually, above 600C, they begin to be
inactivated. Being proteins, they are denatured.
D.HaveoptimumpHformaximalactivity
 Each enzyme requires a definite pH zone for
activity. A change in the pH on either side of the
zone will gradually decrease its activity and an
extreme change will cause inactivation and
destruction of the enzyme. Pepsin requires an
acid pH (pH 2), trypsin requires a very alkaline
pH (pH
8).
FACTORS THAT AFFECT THE RATE OF ENZYME
ACTION.

A.PhysicalFactors:
1.Concentrationofthesubstrate- Other conditions
being kept constant, an increase in the
concentration of the substrate will increase the rate
of activity.
2.ConcentrationoftheEnzyme- Using a highly
purified enzyme, the velocity of the reaction is
directly proportional to the concentration of the
enzyme.
3.EffectofpH-aspreviouslymentionedabove
4.EffectofTemperature-aspreviouslymentioned
5.ConcentrationofCo-factors- The absence or diminution in the
amount of co-factors (coenzymes, activators, inorganicions) for
enzymes requiring their presence for activity will therefore decrease
the rate. In some cases however, an excess of the co-factors will
inactivate the enzyme.
6.ProductsoftheReaction-as the products of the reaction increase in
amount the rate of enzyme action decreases. This is due: a.
Consumption of substrate
b. Reaction reaches equilibrium
c. Changes in pH by products formed
d. Inhibition of enzymes by products formed.

7.Time- the rate of enzyme action may

decrease as the reaction proceeds. This factor
is dependent also on the same factors
influencing products of reaction.
8.Radiation-in general, enzymes tend to be
inactivated by red and green lights. Ultra-
violet light is very destructive it has a
denaturing effect on proteins. The beta and
gamma rays of radium emanations have also a
destructive effect.
A.ChemicalFactors presenceofenzymeactionmavbe
classifiedinto:
1.CompetitiveInhibition- This is brought about by the
presence of substances closely resembling the substrate in
structure or having a similar group or radical in their
molecule, so that they unite with the enzyme, thus
competing with the natural or real substrate. The substrate
is therefore prevented from entering the enzyme complex,
rendering it inactive.. Ex., malonic acid is antagonist to
 succinic acid for succinic dehydrogenase in carbohydrate
metabolism.
2.Non-competitiveInhibition-this is brought about by the presence of
substances that bring about chemical changes in the enzyme. Examples
are:
a. Oxidation-reduction effect - the sulhydryl radical in many enzymes
when oxidized to disulfide (-S-S-) by the removal of hydrogen ,
renders the enzyme inactive.
b. Formation of substances resulting from the action of the inhibitors on
cofactors of the enzyme. This means that the inhibitor reacted with the
coenzyme prosthetic group of the enzyme or ion activator.
Decarboxylases are inhibited by cyanides, hydrazine and semi-carbamide.
 Heavy Metals - salts of heavy metals like those of Ag, Pb, Hg are
c.
inhibitors. They probably precipitate the enzymes thus rendering them
inactive.
ENZYMES OF THE KIDNEYS

If an individual’s blood pressure drops, as in the case of

hemorrhaging or in hypokelemia, the kidneys secrete the enzyme
renin (sometimes considered a hormone) into the bloodstream. The
following reaction occur a:
Renin converting enzyme
Angiotensinogen angiotensin l angiotensin 1l
Angiotensinlis a decapeptide while angiotensinllis an octapeptide,
the most powerful vasoconstrictor known. It increases the force of the
heartbeat and constricts the arteioles, thus causing an increase in blood
pressure.
Angiotensinogenis an alpha globulin produced in the liver.

Chemotherapy - is the use of chemicals to destroy infectious

microorganisms and cancerous cells without damaging the
host’s cells. These chemicals function by inhibiting certain
cellular enzyme reactions. Among the therapeutic agents are:
Antibiotics - are compounds produced by one microorganism
that are toxic to another microorganism. They function by
inhibiting enzymes that are essential to bacterial growth.
Common antibiotics are penicillin G and tetracycline
antimetabolites- are chemicals that have structures
closely related to those of the substrates enzymes act on,
thus inhibiting enzyme activity.

Isozymes-orisoenzymes, are enzymes with the same

function but slightly different structural features.
AllostericEnzymes- are enzymes whose activity can be
changed by molecules other than those of the substrate.