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Enzymology for Clinical Chemistry| questions Solved 100% Correc
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Enzymology for Clinical Chemistry|questions Solved 100% Correct
Measurement of Enzyme Activity - ✔Enzymes are present in every small quantities and are
difficult to isolate from similar compounds. Therefore, most quantitations of enzymes measure
CATALYTIC ACTIVITY or the rate of the change in absorbance and relate this to the
concentration.
Types of Measurements - ✔1. Measure decreases in concentration in substrate (decrease
in starch)
2. Measure increase in concentration of product (Increase glucose)
3. MEASURE AN INCREASE OR DECREASE IN ABSORBANCE OF NADH AT 340 NM
Coupled Reactions - ✔Enzyme reactions that do not require NAD-NADH can be coupled
with additional enzyme reactions that involve NAD.
Enzyme Progress Curve - ✔Lag: Few seconds to few minutes
Initial Rate: (Linear) rate of product formed is constant and increases linearly with respect
to time
Reduced Velocity: (Substrate Depletion) A decrease in substrate leads to first-order kinetics.
pH - ✔Most enzyme reaction require buffers to maintain a neutral pH, usually 6-8.
Extreme variations will denature enzymes.
, Temperature: - ✔Enzyme activity increases as temperature increases. The reaction rate
doubles for each 10* rise in temperature. The majority are enzyme reactions are run at 30C and
most will denature at 40-50C
Presence of Cofactors - ✔Coenzymes: organic compounds (NAD)
Activators: Inorganic ions (Mg, Cl, Zn, Ca)
Substrate Concentration: - ✔The substrate concentration must be in excess of zero-
order kinetics. In first-order kinetics, an increase in substrate will increase the reaction
rate until substrate is depleted.
Enzyme Concentration: - ✔If a reaction follows zero order kinetics, the more enzyme
present, the higher the reaction rate up to a point. Enzyme concentration should be the only
rate-limiting factor
Inhibitors - ✔The presence of a substance that decreases the rate of a reaction by binding
to the active site of the enzyme and blocking access of substrate to enzyme.
Cardiac Disorder Enzymes - ✔Ck
Ck-MB
Troponin
Myoglobin
Hepatic Disorder Enzymes - ✔Obstruction-ALP, GGT
Hepatocellular-AST, ALT
Skeletal Muscle Disorder Enzymes - ✔CK
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