Experiment 12

 Biological catalyst
 Proteins
 Undergo all the reactions of proteins
 Denatured/coagulated by heat,
alcohol, strong acids and alkaloidal
reagents
 The names of most enzymes end with the
suffix “-ase” like peptidase, lipase, and
hydrolase
 Highly increases the rate of the
reaction (106 to 1012 times faster),,
without themselves being changed in
the overall process
 Very specific with the reaction it
catalyzed
 Formation of side product is rare
 Have very complex structure, hence
are capable of being regulated
 Lowers the activation energy ( energy
required for a chemical reaction to
occur)
 Provides unique binding surface called
ACTIVE SITE.
Reactant molecule called SUBSTRATE
binds to the enzyme’s active site
 Catalytic activity of some enzyme
depend only on the interaction
between the active site and the
substrate
 Other enzymes require non-protein
component for their activity, called
enzyme cofactors which are
a. Metal activators ( Fe+2, Cu+2, Na+ )
b. Coenzymes (organic molecule
needed for an enzyme-catalyzed
reaction to occur)
ex. NAD+ is the cofactor (coenzyme)
that oxidizes lactate to pyruvate
with the aid of the enzyme
lactate dehydrogenase:
•Coenzyme FAD (flavin adenine
dinucleotide) is an oxidizing agent as
well.
 Terms:
❑ Apoenzyme – an enzyme (protein
part) that lacks an essential
cofactor
❑ Coenzyme + apoenzyme →
holoenzyme
❑ Holoenzymes – intact enzymes
with their bound cofactors
❑ Zymogens -(proenzymes) are an
inactive form of an enzyme that can be
converted to the active form when
needed.
A. Activity of Amylase
Saliva + Starch + I2 solution → blue
color formation
(indicates presence of starch)
Disappearance of blue color after
some time indicates that hydrolysis
of starch took place:
Saliva-Starch-I2 Solution mixture +
Fehling’s A & B → brick red ppte
( indicates presence of red. sugar)
B. Activity of Catalase

Catalase – enzyme extracted from

potato; occurs in plants, cells and
blood
Factors affecting the rate of enzyme-
catalyzed reaction
1.substrate and enzyme concentration
2. temperature
Optimum temp.- temp. at which the
rate of enzyme’s activity is at
maximum.
3. pH
Optimum pH – pH at which the rate of
enzyme’s activity is at maximum