BIOCHEMISTRY - LABORATORY
EXPERIMENT 3A B. SALIVARY AMYLASE
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
MIDTERMS
• Salivary α-amylase or ptyalin
(EC 3.2.1.1) is a hydrolase
• Salivary amylase hydrolyzes
the 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 𝐻2 𝑂2 → 2 𝐻2 𝑂 + 𝑂2
• The production of oxygen is
seen by the formation of
bubbles
BIOCHEMISTRY - LABORATORY MIDTERMS

C1 C2 C3

3% HAc 3% NaHCO HO
3 2 2

EXPERIMENT 3B
A. EFFECT OF TEMPERATURE C.EFFECT OF ENZYME CONCETRATION
• 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
catalase, however, has
D. EFFECT OF SUBSTRATE
been found to increase
CONCENTRATION
slightly with temperature
• In the experiment, the highest
volume of bubbles was
produced by the potatoes that
were unboiled.