2 General Biochemistry Author Federal University of Agriculture, Abeokuta
2 General Biochemistry Author Federal University of Agriculture, Abeokuta
2 General Biochemistry Author Federal University of Agriculture, Abeokuta
DR AKINLOYE’S ASPECT
Introduction
The use of enzymes in the diagnosis of disease is one of the important benefits
derived from the intensive research in biochemistry since the 1940's. Enzymes have
provided the basis for the field of clinical chemistry.
It is, however, only within the recent past few decades that interest in diagnostic
enzymology has multiplied. Many methods currently on record in the literature are
not in wide use, and there are still large areas of medical research in which the
diagnostic potential of enzyme reactions has not been explored at all.
John H. Northrop and Wendell M. Stanley of the Rockefeller Institute for Medical Research
shared the 1947 Nobel Prize with Sumner. They discovered a complex procedure for
isolating pepsin. This precipitation technique devised by Northrop and Stanley has been
used to crystallize several enzymes.
Definition.
Enzymes are often referred to as biological catalyst that speeds up the rate of chemical
reactions by converting substrate(s) to product(s).
N.B Not all enzymes are protein because we have ribozyme that is nucleic acid in nature.
Almost all processes in biological cells needs enzyme action(s) in order to occur at
significant of appreciable rate.
Many enzymes are named by adding the suffix ‘-ase’ to the name of their substrate e.g
urease catalysis the hydrolysis of urea. However, this is not always true for all enzymes
e.g pepsin, trypsin acts on protein. The classification based on the International Union of
Biochemistry is broadly into six (6) classes thus:
CLASS 4- LYASE: This group of enzyme removes group from a particular substrate or
breaks bonds by mechanism other than hydrolysis, e.g aldolase
CLASS 5- ISOMERASE-They catalyse the conversion of one isomer to the other. They
produce optical geometric or positional isomer of substrate, e.g triose phosphate
isomerase.
CLASS 6- LIGASE- This group catalyses the linkage of two substrates together, usually
with the simultaneous hydrolysis of ATP, e.g acetyl CoA carboxylase.
(viii) Enzymes have enormous catalytic power i.e they can accelerate
reaction rate by at least a million
(ix) Enzymes are highly specific i.e highly specific both in the choice of
substrate and in reaction catalysed
(x) Activities of some enzymes are regulated i.e different kind of
regulatory mechanisms affect enzyme catalysed reaction.
(xi) Enzymes do not alter the reaction equilibria i.e enzymes do not alter
the equilibrium position, meaning that they accelerates the forward and
back ward reactions by precisely the same factor.
(xii) Enzymes decrease the activation energy e.g the lowers the activation
energy by reducing the transition state / activation complex
(xiii) Enzymes transform different kinds of energy i.e energy of reactant
could be converted into different form with high efficiency.
Enzymes can be denatured and precipitated with salts, solvents and other reagents.
They have molecular weights ranging from 10,000 to 2,000,000.
Many enzymes require the presence of other compounds - cofactors - before their
catalytic activity can be exerted. This entire active complex is referred to as the
holoenzyme; i.e., apoenzyme (protein portion) plus the cofactor (coenzyme,
prosthetic group or metal-ion-activator) is called the holoenzyme.
3. A metal-ion-activator - these include K+, Fe++, Fe+++, Cu++, Co++, Zn++, Mn++,
Mg++, Ca++, and Mo+++.
pH: The state of ionization of amino residues in the active site of an enzyme is pH
dependent. A typical enzyme has an optimum pH of activity.
Note that: the interaction of substrate and enzyme could be expressed in term of two
models namel:
(i) lock and key model
(ii) induced fit model.
Enzyme Kinetics
Michaelis and Menten derived equation for enzyme catalyzed reaction involing a single
substrate and single product thus:
S→P
as
v =Vmax X [S]/Km +[S]
where v = initial velocity
Vmax = maximum velocity
[S] = substrate concentration
Km = Michealis-Menten constant.
Note that: any enzyme that obeys M-M equation will give an hyperbolic curve when the
plot of v vs [S] is made.
ALLOSTERIC ENZYMES
These are regulatory enzymes that functions through reversible non-covalent binding of a
modulatory molecule. They usually determine the rate of overall sequence of reaction
because they catalyze the committed/slowest step. Such enzyme is usually the first in the
sequence of a multienzyme reaction system. They are known to have the following
properties:
(i) They have both catalytic and regulatory sites for binding of substrate
(ii) Generally larger and more complex than the simple enzyme
(iii) Shows deviation from classical M-M behaviour in that thet give sigmodial
curve for the plot of v vs [S].
(iv) They undergo conformational changes in binding of modulatory molecule
(v) They may be inhibited by their modulator (-ve modulator) or stimulated by
modulator (+ve modulator)
CO-ENZYMES.
These are additional non-protein part of an enzyme that is required for enzymatic
activities. Inorganic forms of coenzyme are called cofactors. Tightly forms of coenzyme
are called prosthetic group.
Different types of coenzymes, type of reaction and group transfer are given below