BIOCHEMISTRY - LABORATORY
EXPERIMENT 3A  Salivary amylase hydrolyzes the
ENZYMES
A. PEPSIN
 Pepsin is a hydrolase (EC
3.4.23.1), more specifically a
peptidase found in the stomach
with optimum pH of 1.6-2.5
 Inactive form pepsinogen is
converted to pepsin; this
occurs only at low pH
which is due to the
secretion of HCl by parietal
cells in the stomach
 Optimum temp: 37°C
(humans)
 Pepsin has broad specificity with a
preference for peptides containing
linkages with aromatic or
carboxylic L-amino acids. It
preferentially cleaves C-terminal to
Phe and Leu and to a lesser extent
Glu linkages. The enzyme does not
cleave at Val, Ala, or Gly.
 Pepsin cleaves long
polypeptide chains into a
mixture of smaller peptides.
 In the experiment, addition of HCl
will cause the breakdown of the
proteins in the egg white
B. SALIVARY AMYLASE
 Salivary α-amylase or ptyalin
(EC 3.2.1.1) is a hydrolase
MIDTERMS
internal (α 1→4) glycosidic
linkages of starch, producing short
polysaccharide fragments or
oligosaccharides.
 Optimum: pH 7, human body temp
 In the experiment, a negative I2KI
reaction (left test tube) means
amylase has digested starch in the
solution
C. CATALASE
 Catalase is an oxidoreductase (EC
1.11.1.6)
 Optimum pH: 4.0-8.5; 6-8
for plant catalases
 Optimum temp: human
body temp (in humans);
varies in plants but can
work around 0-40°C
 Converts reactive oxygen species
(such as H2O2) to harmless
products, such as in the reaction
below:
2 H 2 O2 →2 H 2 O+O2
 The production of oxygen is seen
by the formation of bubbles
C1 C2 C3
3% HAc 3% NaHCO H O
3 2 2
BIOCHEMISTRY - LABORATORY MIDTERMS

C.EFFECT OF ENZYME
CONCETRATION

EXPERIMENT 3B
A. EFFECT OF TEMPERATURE
 Enzymes in the human body work
within a range of temperatures.
 Enzymes such as catalase, work
best at optimum temperatures.
 In humans, catalases work
at an optimum of 37°C
 Enzyme velocity of D. EFFECT OF SUBSTRATE
catalase, however, has been CONCENTRATION
found to increase slightly
with temperature
 In the experiment, the highest
volume of bubbles was produced
by the potatoes that were unboiled.