MIDTERMS TUBE 1 TUBE 2 TUBE 3

4ml 1%
ENZYMES 5ml 5 ml 1%
pepsin
solution
distilled pepsin
+ 1ml
water) solution
Enzymes are substances that operate as a 0.4% HCl
catalyst for biological processes by solution
determining a lower activation energy path. PEPSIN (-) no (+) slight (++) fast
Each enzyme contains an active site where digestion digestion digestion
chemical reactions occur. One striking feature
of enzymes is their great specificity; they only
work on a single substrate or group of
substrates. SALIVARY AMYLASE
 a.k.a ptyalin
 a hydrolase
PEPSIN  hydrolyses the internal glycosidic linkages
 A hydrolase, specifically, a peptidase of starch, producing short polysaccharide
found in the stomach fragments or oligosaccharides
 Inactive form pepsinogen is converted to  Optimum pH: 7
pepsin  Human body temp
 Occurs only at low pH which is due to the  A negative I2Kl reaction (left test tube)
section of HCl by parietal cells in the means amylase has digested starch in the
stomach solution
 Optimum pH: 1.6-2.5
 Optimum temp: 37° (humans)
 Has a broad specificity with a preference
for peptides containing linkages with
aromatic or carboxylic L-amino acids
 Cleaves C-terminal to the 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
 Addition of HCl will cause the breakdown
of the proteins in the egg white
TEST TUBE 1 TEST TUBE 2
with saliva
AMYLASE Blue Yellowish
(-) no (+) slight
digestion digestion

TEST TEST TEST CATALASE

 An oxidoreductase
 Optimum pH: 4.0 – 8.5 / 6 – 8 for plant
catalases
 Optimum temp: human body temp, 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:
Amylases' main function is to hydrolyze the
glycosidic bonds in starch molecules,
converting complex carbohydrates to simple
sugars. Amylase belongs to the glycosylase
family of hydrolases, which hydrolyze glycosyl
molecules. Carbohydrates in the diet enter the
body as disaccharides, starches (amylose and
amylopectin), and glycogen

2 H 2 O2 →2 H 2 O+O2
CATALASE
Catalase is an oxidoreductase enzyme that
catalyzes the breakdown of hydrogen
peroxide into water and oxygen. Catalase is
used in the textile industry to remove
hydrogen peroxide from fabrics during the
bleach cleaning process to ensure that the
TEST TEST TUBE TEST material is peroxide-free. This enzyme is in
TUBE 1 2 TUBE 3 charge of neutralizing hydrogen peroxide by
breakdown, hence maintaining an optimal
1ml 3% 1ml 3% 3ml 3% amount of the molecule in the cell, which is
Acetic sodium hydrogen also required for cellular signalling activities.
acid bicarbonate peroxide

CATALAS (-) no (+) slight (++) fast

E digestion digestion digestion
FACTORS AFFECTING
ENZYME ACTIVITY

FUNCTIONS
a. TEMPERATURE
PEPSIN
Pepsin is an endopeptidase that breaks down
dietary proteins reaching the stomach into
amino acids. It functions by digesting peptide
bonds, the predominant chemical bonds
found in proteins. In response to various
stimuli, small basophilic cells in the deeper
layers of gastric glands, known as Chief cells,
produce pepsinogen. Notably, acetylcholine,
gastrin, and low pH directly stimulate chief
cells to secrete pepsinogen. Acetylcholine is a
neurotransmitter released from vagal
parasympathetic nerve terminals in the
"cephalic phase" of food digestion.
 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 been found to
increase slightly with temperature
 In the experiment, the highest volume
of bubbles was produced by the
potatoes that were unboiled.

AMYLASE
 NUCLEIC ACIDS
b. pH
 Each enzyme has an optimum pH DNA
range. Changing the pH outside of this
- stores the genetic information of an
range will slow enzyme activity.
organism and transmits that information
Extreme pH values can cause enzymes
to denature. from one generation to another

c. ENZYME CONCENTRATION RNA

 Increasing enzyme concentration will - translates the genetic information
speed up the reaction, as long as there contained in DNA into proteins needed for
is substrate available to bind to. Once all cellular function.
all of the substrates are bound, the
reaction will no longer speed up, since NUCLEIC ACIDS
there will be nothing for additional - an unbranched polymer in which the
enzymes to bind to. monomer units are nucleotides.
- All nucleic acid molecules are unbranched
d. SUBSTRATE CONCENTRATION polymers.
 Increasing substrate concentration
also increases the rate of reaction to a
certain point. Once all of the enzymes STRUCTURE
have bound, any substrate increase
will have no effect on the rate of
A nucleotide is composed of:
reaction, as the available enzymes will
a. Nitrogen-containing bases (amine)
be saturated and working at their
maximum rate. b. Sugar
 Ribose for RNA
 Deoxyribose for DNA
FACTORS VARIATIONS c. Phosphate
TEMPERATURE A1 Initial: 10ml
Final: 14ml EXPERIMENT
A2 Initial: 10ml
Final: 10.5  BLENDER – crush cell wall
A3 Initial: 10ml  HOMOGENIZING SOLUTION
Final: 10.1ml  Dishwashing liquid – removes the
pH B1 Yellow Solution (-) lipids and push DNA to the salt
B2 Colorless solution (-) solution
B3 Blue solution (+)  Sodium Chloride – increases
separation of DNA from
ENZYME C1 10ml – 12.5ml hydrophobic layer; aggregates
DNA, removes the proteins that
CONCENTRATION C2 8ml – 10ml
are bound to DNA
C3 6ml – 7ml
 COLD ETHANOL – promotes
SUBSTRATE D1 9ml – 10.5ml
aggregation of DNA and wash the DNA
CONCENTRATION D2 6ml – 7ml
from the salt
D3 4ml – 4.5ml
DISCHE DIPHENYLAMINE TEST
- For DNA
- Test for the deoxyribose in DNA
- Reagents – diphenylamine, Sulfuric acid
- Positive result – light blue to blue
depending on the amount of DNA
- Negative – clear

THEORETICAL RESULTS