CHEM 301— LEC | BIOCHEMISTRY PRELIMS

POWERPOINT PRESENTATION | AUGUST 28, 2023 A.Y. 2023- 2024

WEEK 4: ENZYMES

ENZYMES Group Catalyze one Hexokinase

type if adds a
ENZYMOLOGY reaction for phosphate
similar group to
• Study of enzymes
substrates hexoses
o Activity of enzymes
Linkage Catalyze one Chymotrypsin
o Chemical reactions it catalyzes type of catalyzes the
o Clinical use reaction for a hydrolysis of
ENZYMES specific type peptide bonds
of bond
• From the Greek work “en” meaning “in”
and “zyme” meaning “yeast”
DEFINITION OF TERMS
• Biologic catalysts
o Hasten chemical Reactions 1. Substrate: Acted upon by the enzyme
• Not consumed during the reactions and specific
• Not undergoing a chemical change after 2. Isoenzyme: Different form but with the
the reactions same action
3. Cofactor: Non-protein molecule of a
conjugated enzyme
a. Activator/Metal ions: Inorganic
cofactor
b. Coenzyme: Organic cofactor
4. Apoenzyme: Polypeptide portion of a
conjugated enzyme and inactive enzyme
5. Holoenzyme: Apoenzyme + Cofactor

PROENZYME
ENZYME SPECIFICITY
• Also known as zymogen
• Enzymes may recognize and catalyze:
o A single substrate • Inactive form or precursor of enzyme
o A group of similar substrates • It is then converted, usually by
o A particular type of bond proteolysis, to the active form when it has
reached the site of its activity
Type Reaction Example
Absolute Catalyze on Urease
type of catalyzes only
reaction for a the hydrolysis
single of urea
substrate

JOHN ANDRIE S. ALBENTO, BSMLS 2A 1

CHEM 301— LEC | BIOCHEMISTRY PRELIMS
POWERPOINT PRESENTATION | AUGUST 28, 2023 A.Y. 2023- 2024

WEEK 4: ENZYMES

ENZYME NOMENCLATURE TRANSFERASES

1. Substrate • Transfer of functional groups other than
2. Reaction hydrogen from one substrate to another
3. Enzyme Commission Nomenclature o Transaminases: transfer of amino
group
ENZYME NOMENCLATURE
o Kinases: transfer of phosphate
1. SUBSTRATE + -ASE group from ATP to ADP +
a. Lipid= lipase phosphorylated product
b. Ester= esterase • E.C. 2.6.1.1: L-Aspartate: 2-
c. Protein= protease Oxaloglutarate Aminotransferase
2. REACTION IT CATALYZES o Aspartate Aminotransferase
a. Oxidation= oxidase
b. Reduction= reductase
c. Hydrolysis= hydrolase
d. Remove hydrogen atoms=
dehydrogenase
e. Remove carboxyl groups=
decarboxylase
3. ENZYME COMMISSION
NOMENCLATURE
a. E.C. 1.1.1.27
b. 1st digit- class
c. 2nd digit- subclass
d. 3rd and 4th digit- serial number
CLASSIFICATION OF ENZYMES
HYDROLASES
1. Oxidoreductases
2. Transferases • is an enzyme that catalyzes a hydrolysis
3. Hydrolases reaction in which the addition of a water
4. Lyases molecule to a bond causes the bond to
5. Isomerases break
6. Ligases • Central to the process of digestion
o Carbohydrase: breaking of
OXIDOREDUCTASE glycosidic bonds in
oligosaccharides and
• Oxidation and reduction
polysaccharides
o Oxidation – removal of H ion
o Proteases: breaking of peptide
o Reduction – accept H ion
linkages in proteins
• E.C. 1.1.1.27: L-lactate NAD+
o Lipases: breaking of ester
Oxidoreductase linkages in triglycerides
o Lactate Dehydrogenase
• E.C. 3.2.1.1: 1,4-D-Glucan
Glucanohydrolase
o Alpha-Amylase

JOHN ANDRIE S. ALBENTO, BSMLS 2A 2

CHEM 301— LEC | BIOCHEMISTRY PRELIMS
POWERPOINT PRESENTATION | AUGUST 28, 2023 A.Y. 2023- 2024

WEEK 4: ENZYMES

LYASES ENZYME KINETICS

• is an enzyme that catalyzes the addition of • Michaelis-Menten Theory

a group to a double bond or the removal of o E + S → E-S → P + E
a group to form a double bond in a
manner that does not involve hydrolysis or
oxidation
• any member of a class of enzymes that
catalyze the addition or removal of the
elements of water (hydrogen, oxygen),
ammonia (nitrogen, hydrogen), or carbon
dioxide (carbon, oxygen) at double bonds
o Dehydratase: removal of the
components of water from a
double bond
o Hydratase: addition of water to a
double bond
• Examples: Carbonic Anhydrase and
Citrate Lyase

ISOMERASES
FACTORS THAT INFLUENCE ENZYMATIC
• is an enzyme that catalyzes the REACTIONS
isomerization (rearrangement of atoms)
1. SUBSTRATE CONCENTRATION
of a substrate in a reaction, converting it
• First-order reaction
into a molecule isomeric with itself
• Second-order reaction
• Only one product and one reactant
(substrate) • Zero-order reaction
• Example: Phosphoglycerate mutase SUBSTRATE CONCENTRATION

LIGASES
• is an enzyme that catalyzes the bonding
together of two molecules into one with
the participation of ATP
• First-order reaction
o Are those which proceed at a rate
exactly proportional to the
concentration of one reactant
o A ------------------------------P
o Rate of reaction is exactly
proportional to the rate of
disappearance of A or the
appearance of P
• Second-order reaction
o Are those in which the rate is
proportional to the product of the
concentration of two reactants
o A+B---------------------P

JOHN ANDRIE S. ALBENTO, BSMLS 2A 3

CHEM 301— LEC | BIOCHEMISTRY
POWERPOINT PRESENTATION | AUGUST 28, 2023
WEEK 4: ENZYMES
o Rate of reaction is exactly
proportional to the rate of
disappearance of A or the
appearance of P
• Zero-order reaction
o The reactions are zero-order with
respect to the reactants
o Rate of reaction depends on the
concentration of catalysts or on
some factor other than the
concentration of the molecular
species undergoing reaction.
FACTORS THAT INFLUENCE ENZYMATIC
REACTIONS
2. ENZYME CONCENTRATION
• The higher the enzyme level, the
faster the reaction will proceed
3. pH
• pH = 7.0 – 8.0
• Changes in pH may denature the
enzyme
• Protein in nature
4. TEMPERATURE
• 37 degrees Celsius (normal)
• Denaturation at 40-50 degrees
Celsius
• assay temperatures: 25, 30, or 37
ºC
5. COFACTORS
• Nonprotein entities that must bind
to particular enzymes before a
reaction occurs
• Metallic or nonmetallic / organic or
inorganic
6. ACTIVATORS
• Proper substrate binding
• Linking substrate to the enzyme or
coenzyme
PRELIMS
A.Y. 2023- 2024
• Undergoing oxidation or reduction
7. INHIBITORS
• Interfere with enzyme reaction
o Competitive- (closely
resembling the substrate)
o Noncompetitive- (binds to
a site on an enzyme that is
not the active site)
o Uncompetitive
o Irreversible- (molecule that
forms a covalent bond to a
part of the active site)
MEASUREMENT OF ENZYME ACTIVITY
• Increase in product concentration
• Decrease in substrate
concentration
• Decrease or increase in coenzyme
concentration
• An increase in the concentration of
the altered enzyme
ENZYMATIC ASSAYS
COUPLED-ENZYMATIC ASSAY
• Substances other than substrate or
coenzyme are necessary and must be
present in excess
• NAD+ or NADH
FIXED-TIME (ENDPOINT) ASSAY
• Reactants are combined
• Reaction proceeds for a designated time
• Reaction is stopped
• Measurement is made of the amount of
reaction has occurred
JOHN ANDRIE S. ALBENTO, BSMLS 2A 4
CHEM 301— LEC | BIOCHEMISTRY PRELIMS
POWERPOINT PRESENTATION | AUGUST 28, 2023 A.Y. 2023- 2024

WEEK 4: ENZYMES

CONTINUOUS-MONITORING (KINETIC) ASSAY

• Multiple measurements
• Absorbance change
o Increasing in absorbance
o Decreasing in absorbance
• Time intervals
CALCULATIONS OF ENZYME ACTIVITY

• INTERNATIONAL UNIT (IU)

o Amount of enzyme that will
catalyze the reaction of 1
micromole of substrate per minute
under specified conditions
• INTERNATIONAL UNIT PER LITER
(IU/L)
o Enzyme concentration
• KATAL UNIT (MOLE/S)
o Amount of enzyme that will
catalyze the reaction of 1 mole of
substrate per second under
specified conditions

JOHN ANDRIE S. ALBENTO, BSMLS 2A 5