Enzymology Part 1

ENZYMOLOGY PART 1 o “even in low substrate concentration, the
substrate can readily bind with free enzyme”

ENZYMES
o Enzymatic Reaction can be:
ESSENTIAL TO PHYSIOLOGIC FUNCTIONING:  First-Order Kinetic : the rate of reaction is
dependent to the substrate concentration
o Hydration of CO2  Zero-order kinetic : the rate of reaction is
o Nerve conduction dependent to the enzyme concentration
o Muscle contraction 2. PH -
o Nutrient degradation o Enzymes are protein that carry a net molecular
o Energy use charge
o Present inside the cell o Optimal pH for enzyme reaction is pH of 7-8
o Speeds up biochemical reactions
3. TEMPERATURE
o Reactants: o Temperature coefficient - in every 10 °C increase
o Enzyme in the temperature, it will result to two-folds
 Active Site - the area where the increase in the activity of enzyme
substrate binds to the enzyme o 40-50*C  denaturation of enzyme
o 60-65*C  inactivation of enzyme
 Allosteric Site - primarily made o 378C (25 or 30*C)  optimum temperature for
up of water
enzyme reaction
o Substrate
o Forms the ES complex 4. CO-FACTORS - non-protein entities that must bind
to an enzyme for a reaction to occur
 Fast chemical reaction
o Activators  inorganic cofactors
 metallic Ca, Fe, Mg, Mn, Zn, K
CATALYTIC MECHANISM OF ENZYMES  non-metallic Br, Cl
 alter the spatial configuration of enzyme
o A chemical reaction may occur spontaneously if the
o Coenzyme  organic cofactors
free energy or available kinetic energy is higher for
o serve as a second substrate to enzyme reaction
the reactant than the products (lower energy).
o e.g. vitamins, NAD, nucleotide phosphatase
o Activation Energy  reactants have enough energy
to break their bond and collide to form new bond
5. INHIBITORS - interferes with the enzyme reaction
ENZYMES ARE HIGHLY SPECIFIC: o Types of Inhibition
o Absolute Specificity  the enzyme combines with ● Competitive Inhibition – targets the
only one substrate and catalyze a single reaction active site
(strictest) ● Non-Competitive Inhibition - targets the
o Group Specificity  the enzyme combines with all allosteric site
substrates with a particular chemical group ● Uncompetitive Inhibition - targets the
ES complex
o Bond Specificity  the enzyme combines with all
6. STORAGE
substrates with a particular chemical bond
o 20*C  long term preservation
o Stereoisometric Specificity  the enzyme combines
o 2-8*C  general temperature for preservation
with all substrate with a specific optical isomer
(most used)
FACTORS THAT INFLUENCE ENZYMATIC REACTION o RT  for preservation of cold labile enzymes
(Ex. LD4 & LD5)
1. SUBSTRATE CONCENTRATION/ENZYME o Cold temperature - can cause reversible
CONCENTRATION inactivation of enzymes.
o Follows the hypothesis of Michaelis & Menten  o Repeated thawing - causes inactivation of
enzyme
NOMENCLATURE
7. HEMOLYSIS
Enzymes that are affected: Developed by the E.C. (Enzyme Commission)
K - Potassium 1. Oxido-reductase - catalyze an oxidation-reduction

L - Lactate dehydrogenase reaction between two substrates
A - Acid phosphatase, aspartate transferase,
aldolase 2. Transferase - catalyze the transfer of a group other
M - Magnesium than hydrogen from one substrate to another
P - Phosphorus
3. Hydrolase - catalyze hydrolysis of various bonds
8. LACTESCENCE/MILKY SPECIMEN – decrease in
4. Lyase - catalyze removal of groups from substrates
concentration (caused by chylomicrons)
without hydrolysis; the product contains double
ENZYMATIC REACTIONS bonds
● measures enzyme based on their activity not by

5. Isomerase - catalyze the interconversion of
their absolute value
geometric, optical, or positional isomers
2 Methods in measuring the enzymatic reactions:
o Fixed time (end point)  Single measurement of 6. Ligases - catalyze the joining of two substrate
enzyme activity molecules, coupled with breaking of the
o Continuous-Monitoring/Kinetic Assay  Multiple pyrophosphate bond in adenosine triphosphate
measurement of enzyme activity (ATP) or a similar compound
ENZYMES IS A PROTEIN (AMINO ACID)
o Primary Structure  specific sequence of amino acid

o Secondary Structure  polypeptide chain twist
o Tertiary structure  folding of the polypeptide
chains
o Quaternary structure  combination of tertiary
structure
ISOENZYMES
● enzymes with same catalytic function but

different physical properties
● CK has 3 isoenzyme
● CK1 –CKBB (Brain)
● CK2
● CK3 – CKMM (Muscle)
Differentiated by:
o Electrophoretic mobility
o Solubility
o Resistance to inactivation

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Enzymology Part 1

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ENZYMOLOGY PART 1 o “even in low substrate concentration, the

substrate can readily bind with free enzyme”

K - Potassium 1. Oxido-reductase - catalyze an oxidation-reduction

● measures enzyme based on their activity not by

ENZYMES IS A PROTEIN (AMINO ACID)

o Primary Structure  specific sequence of amino acid

● enzymes with same catalytic function but

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