GROUP 4:

ESMAEL, BUNGABAI KISSES SAMPULNA, JEFF JALIL

BITUANAN, SITTIE MAIZAM DELA CRUZ, JANADELLE
JOVELLANO, FRANCHESKA FELLON, REY MARTHIN
MASUKAT, AYESSA REIJN ABO, MARGARET NICOLE

EXPERIMENT 4: ENZYMES AND DIGESTION

OBJECTIVES:
 To identify the factors that affect the rate of enzyme activity;
 Describe the role of enzymes in the digestive process; and
 Use chemical tests to identify the products of digestion of carbohydrates, fats, and proteins of various sites in the digestive tract.

TEST SOLUTIONS AND EXPECTED ACTUAL RESULT REMARKS

REAGENTS USED RESULT
Effect of Saliva Dark blue sol’n Ranking: (with 1
Temperature - Served as the - Presence of for being colorless
enzyme since it starch (no signifying greatest
is made up of digestion digestion and 3
amylase occurred) which means least
Starch Violet or red sol’n or no digestion)
- Served as the - Partial 1 – C1
substrate of digestion 2 – B1
amylase Colorless sol’n 3 – A1
Iodine sol’n - Complete
- Used for digestion Note:
detection of A1- ice bath
starch B1- 37 degrees
centigrade
C1- boiling water
Effects of PH Tube 1 (pH 4) Tube 1 (pH 4) Tube 1 (pH 4) Components in
test - Iodine solution - Dark Blue - The sample indicated partial digestion saliva help keep
solution resulting to a violet coloured solution. the pH in your
mouth between
Tube 2 (pH 7) Tube 2 (pH 7) Tube 2 (pH 7) 6.5 and 7 so that
- Iodine solution - Violet or red - The sample indicated partial digestion the enzyme
solution resulting to a red coloured solution. salivary amylase
can start to
Tube 3 (pH 10) Tube 3 (pH 10) Tube 3 (pH 10) break down
- Iodine solution - Colorless - The sample indicates presence of starch carbohydrates.
solution resulting to blue coloured solution. That is why at
pH 4 the sample
shows a dark
blue color
because of the
presence of
starch which the
saliva cannot
breakdown at
that pH level.
Effect of Saliva - The tubes with the All tubes
Enzyme Contains the enzyme, smaller showed a
Concentration amylase. concentration of smaller
enzymes concentration of
1% Starch solution - remained a blue enzymes for all
Serves as the substrate black color longer. tubes remained
of the enzyme amylase. a blue black
The higher coloration, but it
Iodine solution - concentration of can be observed
To detect and test the enzyme amylase, the in tube 1 that it
presence of starch in a lighter has a slightly
solution. the color of the starch higher
will appear. concentration of
enzyme
The faster the blue compared to the
color of starch is lost, other tubes, for
the faster the having a lighter
enzyme amylase is shade of blue
working. black on top.

Effects of Benedict’s Solution By using Benedict’s

Substrate reagent, a brick-red
Concentration -A solution that uses the precipitate indicates a
change of color (blue to complete digestion of
red) to detect the the starch. A green to
presence of digestion. yellow to orange
indicates a partial
digestion of the
starch. At higher
substrate
concentration the rate
of reaction becomes
constant, showing no
further increase as
substrate
 concentration
increases.

Effect of Saliva On vial 6: Both tubes 4 and

Metal-ion - Contains the - Partial 6 have not
Poisons on fresh amylase digestion will resulted to what
Enzyme Distilled water occur due to is expected. No
Activity - Dilute the the fact that digestion
enzyme that the silver occurred. This
1% silver nitrate nitrate act as signifies that
- Heavy metal an inhibitor possible human
1% starch solution On vial 4: error was made.
- The substrate - Digestion will
Iodine sol’n take place
- Used to the because the
starch to obtain solution used
the color blue is distilled
solution water. Hence,
it is pure and
not an
inhibitor
Note:
Complete digestion
- Colorless
Partial digestion
- Violet or red
No digestion
- Dark shade of
blue
Pancreatic Benedict’s Reagent Benedicts Test Benedict’s Test The Barfoed
Amylase - a test reagent - Yellow-green Test yield
that is used for precipitate - Tube 1 (Saliva)- Yellow-green precipitate negative
simple (positive) (positive) because of
carbohydrates, it (positive) possible
identifies - Tube 2 (pancreatin and Sodium Carbonate)- presence of
reducing sugars Yellow-green solution (positive) disaccharide
 (monosaccharide - positive of (maltose) and
and some presence of Barfoed’s Test the need for
disaccharides) reducing further digestion
which have free sugar - Tube 1 (Saliva)- Blues solution (negative) of maltose
ketone or (maltose)
aldehyde Barfoed Test - Tube 2 (pancreatin and Sodium Carbonate)-
functional - Yellow Blue solution (negative)
group. precipitate
Barfoed’s Reagent (ppositive
- a test reagent - Positive
used to because of
distinguish presence of
monosaccharide reducing
from reducing sugar
disaccharides, (glucose)
obtaining a brick
red color due to
formation of
cuprous oxide.
Sodium carbonate
- is used to
activate the
pancreatic
enzymes and to
raise the pH of
the solution.

Pancreatic Egg white solution Test tube 1: After TUBE1: The solution turned dark violet
Proteases immersing it at 37 TUBE 2: The solution turned light brown with
1% CuSO4 degrees Celcius water precipitate
- used to denature bath, an intense violet TUBE 3: The solution remained violet
proteins in egg solution will appear
white/albumen with denatured egg
white submerging at
3M NaOH the bottom since it did
 - crucial for the not underwent protein
alkalinity of the denaturation
solution
Test tube 2: After
3N HCl immersing it at 37
- provides degrees Celcius water
appropriate pH bath, the complex of
for the enzyme the solution will turn
to act lighter. Egg white
will dissolve by far
Pepsin the most.
- effects only
partial Test tube 3: After
degradation of immersing it at 37
proteins into degrees Celcius water
smaller units bath, violet complex
called peptides solution will remain
which are either and the rate of
absorbed from dissolution will be
the intestine into much slower because
the bloodstream of the absence of
or are broken pepsin.
down further by
pancreatic
enzymes

Pancreatin
- used to replace
digestive
enzymes when
the body does
not have enough
of its own
Pancreatic Test tube 1 - There should be a The Pancreatic lipase
lipase and bile - 20 drops of decreased in pH and bile test have
Vegetable oil when hydrolyze resulted into something
- 3.0mL of
Pancreatin
that is not expected
- 3.0mL of Distilled from the task mainly
water because of human
errors such as not being
Test tube 2 able to pay much
- 20 drops of attention to the tubes
Vegetable oil
and the removal of the
- 3.0mL of Sodium
choleate tubes from the
- 3.0mL of Distilled waterbath is either late
water or too early.

Test tube 3
- 20 drops of Test tube 1
Vegetable oil - Initial pH: 11
- 3.0mL of - Final pH: 11
Pancreatin
- 3.0mL of Sodium Test tube 2
choleate - Initial pH: 4  9
- Final pH: 10

Test tube 3:
- Initial pH: 11
- Final pH: 11