BIOCHEMISTRY LECTURE Classification of Enzymes

ENZYMES ❖ When early workers isolated certain

enzymes, whimsical names were given.
❖ Berzelius in 1835 showed hydrolysis of
Some of these, such as Pepsin, Trypsin,
starch by malt extract and put forward the
Chymotrypsin, etc. are still used.
theory of enzyme catalysis.
❖ Later, it was agreed to call the enzymes
❖ Willy Kunhe, in 1878 coined the word
by adding the suffix "-ase" to the
enzyme, which in Greek means "in yeast “
substrate.
❖ Eduard Buchner (Nobel prize 1907) showed
❖ Thus, enzyme Lactase acts on the
that cell-free extract of yeast could catalyze
substrate lactose, and the products
the fermentation of sucrose to ethanol. He
glucose and galactose are formed.
named this active principle as Zymase
❖ Enzymes that hydrolyse starch (amylose)
❖ Sir Arthur Harden in 1897 (Nobel prize
are termed as amylases; those that
1929) showed that Zymase is a complex
dehydrogenate the substrates are called
mixture of enzymes, each catalyzing a
dehydrogenases. These are known as the
separate step in the degradation of sucrose
trivial names of the enzymes.
❖ In 1926, James Sumner (Nobel prize 1946)
❖ IUBMB System of Classification
was the first to crystallize the enzyme
urease.
Six Major Classes of Enzymes
❖ In 1930, John Northrop (Nobel prize, 1946)
1. Oxidoreductase
crystallized a number of proteolytic enzymes
from gastrointestinal tract and proved that ➢ an enzyme that catalyzes an oxidation-
they are all proteins. reduction reaction.
➢ this enzyme will catalyze oxidation of one
substrate with simultaneous reduction of
another substrate or co-enzyme
➢ Transfer of hydrogen or addition of oxygen
Subtypes:
a. Oxidases - use oxygen as an electron acceptor
but not incorporate it into the substrate.
Characteristics of Enzymes
❖ Almost all enzymes are proteins. Enzymes b. Dehydrogenases – use molecules other than
follow the physical and chemical reactions oxygen as an electron acceptor
of proteins.
c. Oxygenases- directly incorporate oxygen into the
❖ They are heat labile.
substrate
❖ They are water-soluble.
❖ They can be precipitated by protein- d. Peroxidases - use hydrogen peroxide as an
precipitating reagents (ammonium sulfate or electron acceptor
trichloroacetic acid).
❖ They contain 16% weight as nitrogen. e. Reductases - reduction of a substrate
2. Transferase
➢ an enzyme that catalyzes the transfer of a
functional group from one molecule to
another.
Subtypes:
a. Methyltransferases - transfer one carbon unit
between substrates
b. Aminotransferases/ transaminases - transfer
NH2 from amino acid to keto acid
c. Kinases - transfer PO3 from ATP to a substrate
d. Phosphorylases - transfer PO3 from inorganic
phosphate to a substrate
3. Hydrolases
➢ an enzyme that catalyzes a hydrolysis
reaction in which the addition of a water
molecule to a bond causes the bond to break.
➢ this class of enzyme can hydrolyze ester,
ether, peptide or glycosidic bonds by adding
water and then breaking the bond.
Subtypes:
a. Phosphatases - remove PO3 from the substrate
b. Phosphodiesterases - cleave phosphodiester
bonds such as those in nucleic acids
c. Proteases - cleave amide bonds such as those in
proteins
d. Lipases - hydrolysis of ester linkages in lipids
e. Carbohydrases - hydrolysis of glycosidic bonds
in carbohydrates
4. Lyase
➢ an enzyme that catalyzes the addition of a
group to a double bond or the removal of a
group to form a double bond that does not
involve hydrolysis or oxidation.
Subtypes:
a. Decarboxylases - produce CO2 via elimination
reaction
b. Aldolases - produce aldehydes via elimination
reactions
c. Synthases - link two molecules without
involvement of ATP
d. Dehydratases – removal of H20 from a substrate
e. Deaminases - removal of NH3 from a substrate
f. Hydratases - addition of H20 to a substrate
5. Isomerase
➢ an enzyme that catalyzes the isomerization
of a substrate in a reaction.
➢ this enzyme can produce optical, geometric
or positional isomers of substrates.
Subtypes:
a. Racemases - interconvert L and D stereoisomers
b. Mutases - transfer groups between atoms within
molecules
6. Ligase
➢ an enzyme that catalyzes the bonding
together of two molecules into one with the
participation of ATP.
Subtypes:
a. Carboxylases - use CO2 as a substrate
b. Synthetases - link two molecules via an ATP
dependent reaction
 Enzyme Structure
➢ can be divided into two general structural
classes:
1. simple enzyme - an enzyme with only one
protein.
2. conjugated enzyme - an enzyme with a non-
protein part in addition to a protein part.
B. Lock and Key Hypothesis
➢ This is the most accepted of the theories of
enzyme action.
➢ This theory states that the substrate fits
exactly into the active site of the enzyme to
form an enzyme-substrate complex.
➢ This model also describes why enzymes are
so specific in their action because they are
specific to the substrate molecules.

➢ apoenzyme is the part of the conjugated

C. Induced Fit Model
protein
➢ cofactor is the non-protein part of a ➢ This is similar to the lock and key
conjugated enzyme hypothesis.
➢ holoenzyme is the biochemically active ➢ the shape of the enzyme molecule changes
conjugated enzyme produced from an as it gets closer to the substrate molecule in
apoenzyme and a co-factor such a way that the substrate molecule fits
➢ cofactor could be: simple metal ions and exactly into the active site of the enzyme.
small organic molecules
➢ simple metal ions cofactors: Zn,Mg, Fe, Cu
➢ small organic molecules are the co enzymes
that are synthesized in the human body
How Do Enzymes Work?
A. Reaction Mechanism
➢ In any chemical reaction, a substrate (S) is
converted into a product (P)
➢ In an enzyme-catalysed reaction, the Enzymes Specificity
substrate first binds to the active site of the
➢ the extent to which the enzyme’s activity is
enzyme to form an enzyme-substrate
restricted to a specific substrate, a specific
(ES)complex, then the substrate is converted
group of substrates, a specific type of
into product which is attached to the
chemical bond, or a specific type of
enzyme, and finally the product is released,
chemical reaction.
thus allowing the enzyme to start all over
➢ determined by the active site
again.
 Types of Enzyme Specificity
1. Absolute specificity
➢ the enzyme will catalyze only one reaction
2. Group specificity
➢ The enzyme will act only on molecules that
have a specific functional group
3. Linkage specificity
➢ the enzyme will act on a particular type of
chemical bond
4. Stereochemical specificity
➢ the enzyme will act on a particular
stereoisomer
Factors that affect Enzyme Activity
➢ enzyme activity is the measure of the rate at
which an enzyme converts substrate to
products in a biochemical reactions.
Temperature
➢ Enzymes have an optimum temperature at
which they work fastest.
➢ Up to the optimum temperature the rate
increases geometrically with temperature
pH
➢ Enzymes are very sensitive to changes in the
pH and work in a very small window of
permissible pH levels.
➢ Below or above the optimum pH level, there
is a risk of the enzymes disintegrating and
thereby the reaction slows down.
Substrate Concentration
➢ Increasing Substrate Concentration increases
the rate of reaction. This is because more
substrate molecules will be colliding with
enzyme molecules, so more products will be
formed.
➢ However, after a certain concentration, any
increase will have no effect on the rate of
reaction, since Substrate Concentration will
no longer be the limiting factor. The
enzymes will effectively become saturated,
and will be working at their maximum
possible rate.
Enzyme Concentration
➢ Increasing Enzyme Concentration will
increase the rate of reaction, as more
enzymes will be colliding with substrate
molecules.
➢ However, this too will only have an
effect up to a certain concentration,
where the Enzyme Concentration is no
longer the limiting factor.
 HOW ARE ENZYMES REGULATED? E. Isoenzyme
A. Feedback Control ➢ enzyme that performs the same function but
have different combinations of subunits and
➢ an enzyme regulation process in which the
thus different quaternary structures
formation of a product inhibits an earlier
➢ lactate dehydrogenase catalyzes the
reaction in the sequence.
oxidation of lactate to pyruvate and vice
➢ the reaction product of one enzyme may
versa is an example.
control the activity of another.
Enzyme Inhibition
B. Proenzymes/ Zymogens
➢ an enzyme inhibitor is a substance that slows
➢ the inactive form of enzymes
or stops the normal catalytic function of an
➢ trypsin is manufactured in the pancreas as
enzyme by binding to it.
the inactive molecule trypsinogen (a
zymogen). When a fragment containing six Reversible Competetive Inhibition
amino acid residues is removed from the N-
➢ a competitive enzyme inhibitor is a molecule
Terminal end, the molecules become a fully
that resembles an enzyme substrate in shape
active trypsin molecule.
and charge distribution that it can compete
C. Allosteric Enzymes with the substrate for occupancy of the
enzyme’s active site.
➢ all allosteric enzymes have quaternary
structures; they are composed of two or Reversible Noncompetitive Inhibition
more protein subunits
➢ a non competitive inhibitor is an enzyme
➢ all allosteric enzymes have two kinds of
that decreases enzyme activity by binding to
building sites; those for substrate and those
a site on an enzyme other than the active site
for regulators
➢ substrate can still occupy the active site , but
➢ active and regulatory binding sites are
the presence of the inhibitor causes a change
distinct from each other in both location and
in the structure of the enzyme that catalytic
shape.
groups at the active site from properly
➢ binding of molecule at the regulatory site
effecting their catalytic action
causes change in the overall three
➢ heavy metal ions of Pb, Ag and Hg are
dimensional structure of the enzyme ,
examples of noncompetitive inhibitors.
including structural changes at the active site
❖ regulators are substances that bind at the Irreversible Inhibition
regulatory sites of an allosteric enzymes
➢ irreversible enzyme inhibitor is a molecule
D. Protein Modification that inactivates enzymes by forming a
strong covalent bond to an amino acid side
➢ a process in which enzyme activity is
chain group at the enzyme’s active site.
altered covalently modifying the structure
of the enzyme through attachment of a
chemical group to or removal of a chemical
group from a particular amino acid within
the enzyme’s structure
➢ an activation or inhibition of enzymes by
phosphorylation is the best-known example.
 Medical Uses of Enzymes

Prescription Drugs that Inhibit Enzyme Activity

❖ ACE Inhibitors
➢ an Angiotensin Converting Enzymes
➢ an octapeptide hormone used to treat high
blood pressure
➢ angiotensin is present in the body in an
inactive form as the zymogen
angiotensinogen which is a decapeptide
➢ ACE converts the inactive
angiotensinogen decapeptide to an active
form by cleaving two amino acids from
zymogen structure
➢ ACE inhibitor medications block the
action of ACE in converting
angiotensinogen to angiotensin
➢ lisinopril, a compound that is prescribed
for treatment of moderately elevated blood
pressure condition

❖ Sulfa Drugs
➢ the first antibiotics in the medical field
➢ sulfanilamide inhibits bacterial growth
that is structurally similar to PABA
(p-aminobenzoic acid)
➢ sulphanilamide acts as a competitive
inhibitor to enzymes in the biosynthetic
pathway for converting PABA to folic acid

❖ Penicillins
➢ inhibit transpeptidase, an enzyme that
catalyzes the formation of peptide cross-
link between polysaccharide strands in
bacterial cell walls.
➢ highly specific in binding to the active site
of transpeptidase
Ramos, Annyzha Mhaize A.
➢ acts as a very selective competitive
inhibitor BSN 1-B