BIOL 204 Lab For Week 12

Digestive Enzymes
 Digestive Enzymes:
are used in the lumen of the GI tract to break down complex
molecules into absorbable subunits
Enzymes are biological catalysts which increase the rate of a
chemical reaction without themselves becoming part of the product:
In this experiment we will observe the activity of three
different enzymes:

Amylase: digests starch into disaccharides

Trypsin: digests proteins into peptides
Lipase: digests fats into glycerol and fatty acids
Variables tested ……
1. Incubation temperature:
How will enzyme activity compare when incubated at O o C and
37 o C?
2. Denaturization of Enzymes
How will enzyme activity be altered if the enzyme is denatured by
boiling?
Activity 1 Trypsin Activity

Trypsin an enzyme, secreted by pancreatic acinar cells,

that hydrolyzes proteins into small fragments (peptones,
peptides and proteoses)”
In this experiment, trypsin will act on an artificial
substrate to yield a yellow colored product:
Trypsin
BAPNA + H2O N-benzoyl-L-arginine + p-nitroaniline
(yellow)
1. Reaction rate will be compared at these different incubation
temperatures:

370 C and 0o C
What do you predict will happen?

2. The rate of the reaction will be determined using boiled

trypsin. What do you predict will happen?
Activity 2 Amylase Activity
Amylase is a carbohydrate digesting enzyme, produced by
salivary glands and pancreatic acinar cells, that hydrolyzes
starch into maltose (disaccharides):
Amylase
Starch + H2O maltose (disaccharides)
Indicators:
Starch: IKI (Lugol’s solution) - turns “blue/black” in the
presence of starch

Benedict’s Solution: turns green/orange/ red/brown

in the presence of reducing sugar
IKI Test For Starch in a
Spot Plate

Benedict’s Test for

reducing sugar in heat
test tubes
As in activity one, the reaction will be observed under
different conditions:

Boiled Enzyme (Amylase)

Incubation Temperature: 0oC and 37oC
Activity 3 Lipase Activity

Lipase is a hydrolytic enzyme, made primarily by pancreatic

acinar cells, that hydrolyzes neutral fats into fatty acids and
glycerol:
Lipase
Fat + H2O Glycerol + Fatty Acids
Fat Digestion:

Indicator:
Blue litmus terns pink (red) in
the presence of acid (one the
products is fatty acids)
Bile emulsifies fat….
In this activity, you will examine the effect of:
incubation temperature (0oC vs. 37oC)
and
addition of bile salts
on the reaction
Activity 4 Gastroesophageal Junction

Different microscopic
views
Histology of The Stomach Wall
Histology of the Small Intestine (Duodenum)

Micrograph
Intestinal Villus
Liver – Microscopic Anatomy
Microscopic View of the Pancreas:
Islet Cells: endocrine; secrete insulin and glucagon
Acinar Cells: exocrine; secrete digestive enzymes