BARECUATRO, ANGELICA CLAIRE B.

BSN 1 – 11L

Learning Activity No. 13

ENZYMATIC CATALYSIS OF THE HYDROLYSIS OF FATS

(ACTION OF PANCREATIN ON BUTTERFAT)

Specific Learning Objectives: At the end of the experiment, the students can:

1. Explain the digestive function of pancreatin in the hydrolysis of fats.

2. describe the condition under which fat is hydrolyzed with pancreatin.
3. Describe the function of pancreatin in the liver
Procedures:

1. Search on the internet the actions of pancreatin on milk that contains butyric acid.

Data and Results

Observations With Pancreatin Without Pancreatin

Color white
Milk + Pancreatin = blue to pink
Odor Mild sweet
Milk + Pancreatin = soapy

Conclusion (Give explanations for all your observations)

Overall, the learner was able to identify the effect of pancreatin on milk. For its color, it produced a blue to pink
product. While for the odor, it is a soapy or vomit like since it broke down the milk molecules.

Learning Evaluation

Pre-laboratory Questions

1. Explain the function of sodium carbonate solution on the digestion of butterfat by pancreatin.

Sodium carbonate serves as a catalyst between the digestion of butterfat by pancreatin. It helps speeds up the
process of digestion. Further, if the maximum point of the sodium carbonate is reached, the process will eventually
slow down.

2. Name three enzymes in pancreatin and state the function of each.

Lipase – It is an enzyme that catalyzes the hydrolysis of triglyceride ester bonds, converting them to free fatty
acids and glycerol. Pancreatic secretions contain lipases, which take role in the breakdown and metabolism of
fat. The hepatic lipase in the liver, the hormone-sensitive lipases in the adipocytes, the lipoprotein lipase in the
endothelial cells, and the pancreatic lipase in the small intestine all play crucial roles in the transport of lipids.

Amylase - The primary job of amylases is to hydrolyze the glycosidic linkages in molecules of starch, turning
them to simple sugars. The three primary kinds of amylase enzymes—alpha, beta, and gamma—each affect the
carbohydrate molecule in a different way. Alpha-amylase is a protein that is present in microorganisms, plants,
animals, and humans. Both bacteria and plants have beta-amylase. Animals and plants both have gamma-amylase.
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

Trypsin - This enzyme mostly breaks down proteins into smaller peptides or even amino acids. The breakdown
of dietary protein in the small intestine is carried out by trypsin and other digestive proteases like chymotrypsin.
Environmental elements including pH, temperature, and the presence of trypsin inhibitors in the enzymatic
reaction media as well as the enzyme supplies have an impact on its function.

3. Write the structure of a typical molecule of butterfat and write the equation for its digestion with pancreatin.

C6H10O5 + H2O → 3CH4 + 3CO2

II. Post Laboratory Questions:

1. Where are lipids absorbed? How do they travel to the liver? What happens to them in the liver?

In order to absorb lipids, dietary fat had to be digested in the intestine's lumen, and then the products of that
hydrolysis had to be taken up by enterocytes. Re-synthesized lipids are either released with chylomicrons and
high-density lipoproteins or are retained as cytoplasmic lipid droplets after being produced in the endoplasmic
reticulum. Very-low-density lipoproteins are formed when chylomicrons enter the liver (VLDL). Triacylglycerols
(TAGs) are transported by VLDL from the liver to extra hepatic organs. When the TAGs are released to the target
tissues, the density of those tissues rises, which also raises LDL and HDL levels.

2. What is the action of pancreatin on butterfat?

Our pancreas produces pancreatic fluids called enzymes that aid in digesting. These enzymes disintegrate
carbohydrates, lipids, and fats.
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

Learning Activity No. 14

PROTEIN DIGESTION

Specific Learning Objectives: At the end of the experiment, the students can

1. explain the process of digestion of proteins in food.

2. write the chemical equation involved in the digestion of protein.
3. investigate the digestive functions of pepsin and trypsin.
4. explain the effect of different kinds of pepsin on the digestion of proteins

Data and Results

Time (min) OBSERVATION

Pepsin Acidified Basic Distilled water Acidified pepsin
pepsin Pancreatin + HgCl2
Start No digestion No digestion Digestion takes No digestion Digestion takes
process process place process place
10 No digestion Digestion takes Digestion takes No digestion Digestion takes
process place place process place
20 No digestion Digestion takes Digestion takes No digestion Digestion takes
process place place process place
30 No digestion Digestion takes Digestion takes No digestion No digestion
process place place process process
40 No digestion Digestion takes Digestion takes No digestion No digestion
process place place process process
50 No digestion Digestion takes Digestion takes No digestion No digestion
process place place process process
60 (1 Hr) No digestion Digestion takes Digestion takes No digestion No digestion
process place place process process

Conclusion (Give explanations for all your observations)

Overall, the learner was able to observe and differentiate the effects of pepsin, acidified pepsin, basic pancreatin,
distilled water, and acidified pepsin with mercury (II) chloride. First, no digestion took place with the pepsin only.
According to Heda et al., 2022, for protein digestion, pepsin requires an acidic environment. As a result, it works
best at pH levels ranging from 1.5 to 2. Pepsinogen can cleave itself and form active pepsin when the pH is low.
Because of this, the explanation leads to the second results which is for the acidified pepsin. From the table, it
can be see that protein digestion took place since the pepsin was acidified by the addition of hydrocholoric acid
(HCl). Next, as for the basic pancreatin, the results show that digestion took place. Pancreatin are composed of
bicarbonates and enzymes such as trypsin, chymotrypsin, elastase, and carboxypeptidase. Further, pancreatin also
works best on alkali or basic medium. Hence, protein digestion happened since the medium used was a basic
pancreatin. Fourth, no digestion took place with the distilled water since it is considered as a control variable.
Lastly, protein digestion happened with acidified pepsin with HgCl2. This medium had the fastest reaction among
all the media used since HgCl2 was utilized as a catalyst. Meaning, it is increased the rate of reaction or digestion
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

of protein. In addition, it can also be observed that no digestion took place after 20 minutes indicating that the
maximum rate was reached within the reaction.

Learning Evaluation

Pre-Laboratory Questions
1. Arrange the following structures form the most complex to the least: amino acids, polypeptides, proteins.

Proteins → Polypeptides → Amino Acids

2. Compare and contrast pepsin and trypsin. What are their digestive functions?

DIFFERENCES SIMILARITIES
• Pepsin is produced by the stomach (gastric • The only thing that pepsin and trypsin have in
gland), while Trypsin is produced by the common is that they both break down proteins
pancreas by disrupting the links that exist between
• Pepsin is a component of gastric juice, while amino acids.
Trypsin is a component of pancreatic juice
• Pepsin is activated by HCl, while Trypsin is
activated by enterokinase
• Pepsin is an aspartic protease which uses a
catalytic aspartate in its active site, while
trypsin is a serine protease employing the
serine residue in active site
• Pepsin is an enzyme which acts only in acidic
medium, trypsin is an enzyme which acts in
alkaline medium
• Pepsin hydrolyzes peptide bonds between
large hydrophobic amino acid residues,
whereas trypsin hydrolyzes peptide bonds at
the C-terminal side of lysine or arginine
• Pepsin acts on proteins and converts them into
peptones, while trypsin converts peptones into
polypeptides.

3. Give the effect of pH and the presence of mercury salt on the hydrolysis of protein?

pH: Acidic enzymes most likely increases the rate of protein digestion/hydrolysis. As showcased in the
experiment, protein most likely digested on the acidic pepsin whereas the pepsin only did not exhibit any process
of digestion.

Mercury: Salts of heavy metals like mercury increases the disruption rate of the proteins. As shown in the
experiment, HgCl2 was used as a catalyst. Hence, the hydrolysis reaction of proteins was fast since an acidic
pepsin and a catalyst was used. However, the reaction stopped or slowed down during the 30 min mark indicating
that the reaction already reached its maximum point.
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

Post Laboratory Questions:

1. Can the body store carbohydrates? Fat? Protein? If so, in what form?

Yes, the body can store carbohydrates, fat, and protein. Carbohydrates are the starches and sugars found in foods.
During digestion, sugars and starches are broken down or converted into Glucose. Glucose is the chief fuel of our
body and unused glucose are stored in the liver and muscles as glycogen. Next, although people said fats are
harmful to our body, it is an essential component of our body. Fat represents the most concentrated form of energy
available. It delivers 9 calories per gram- more than twice that of carbohydrates and proteins. Adipocytes, or fat
cells, are found throughout the body and store fat. Body fat functions differently than food fat. Body fat is required
to surround and protect your important organs. It also acts as an insulator against cold. Fats can also be stored as
cholesterol. Cholesterol is instrumental in the production of sex hormones, vitamin D, and the protective sheath
around nerve fibers as well as cell walls. Lastly proteins are nutrients that help build and maintain body tissues.
Proteins found in enzymes, hormones, and antibodies also aid in the regulation of numerous bodily functions.
Enzymes are chemicals that aid in the regulation of hundreds of biomechanical events in the cells of your body.
Hormones control responses. Antibodies aid in the identification and destruction of bacteria and viruses that cause
illness in the body.

2. What is the normal daily requirement of protein? What can affect this amount?

Protein's Recommended Dietary Allowance (RDA) is a modest 0.8 grams per kilogram of body weight. The RDA
is the quantity of a nutrient that is required to fulfill your basic nutritional needs. In other ways, it's the bare
minimum you need to avoid being ill — not the exact quantity you should consume every day (Harvard Health,
2022)
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

Learning Activity No. 15

QUALITATIVE TESTS FOR NORMAL CONSTITUENTS OF URINE

Specific Learning Objectives: At the end of the experiment, the students can

1. identify the constituents in urine that are normally present.

2. perform the different qualitative tests for normal urine constituents.
3. explain the importance of each constituent in urine.

Data and Results

Test for Observations

Specific gravity

Urea Present

Uric Acid Present

Indican Not Present

Creatinine Present

Conclusion (Give explanations for all your observations)

Overall, urea, uric acid, and creatinine are the components of urine. For the specific gravity, the normal value for
a well hydrated urine is less than 1.010. Next, Urea, also known as carbamide, is a waste product produced by
many living organisms and is the most abundant organic component in human urine. This is due to its location at
the end of the sequence of events that break down the amino acids that comprise proteins. Uric acid is a waste
product that may be detected in blood. It is formed when the body breaks down purine compounds. The majority
of uric acid dissolves in the blood, travels through the kidneys, and excretes in the urine. Foods and beverages
rich in purines raise uric acid levels. Third, Creatinine is a chemical substance produced by the energy-generating
systems in your muscles. Creatinine is removed from the blood by healthy kidneys. It is excreted from the body
as a waste product in the urine. Lastly, the urine indican test is a chemical analysis of urine that uses iron,
hydrochloric acid, and chloroform to determine the level of toxic by-products produced by the protein rotting
process in the intestinal tract. A positive test always indicates pathogenic activity of anaerobic bacteria in the
bowels. A healthy person's indican test is usually negative (o). A positive indican test may be light (+), medium
(++), strong (+++), or extremely strong (++++). Every second or third degree positive reaction should be
considered carefully and handled (Marđetko, 2017).

Learning Evaluation
 BARECUATRO, ANGELICA CLAIRE B.
BSN 1 – 11L

Pre-Laboratory Questions

Modified true or false - Write true on the provided blank if the statement is correct; if not correct, change the
underlined word to make the statement true.

true
________ 1. The primary organ of excretion is the kidney.
true
________ 2. The primary organ excretes water, water-soluble, and water-insoluble compounds in a
mixture called urine.
true
________ 3. The actual composition of urine varies according to a wide variety of factors including
diet, exercise, and infections.
other
________ 4. The normal constituents of urine include the following inorganic ions: chloride,
phosphate, sulfate, sodium, potassium, calcium, and ammonium.
creatinine
________ 5. The three principal organic constituents of urine are urea, uric acid, and indican.

Post Laboratory Questions:

1. What are the principal excretory organs of the body? What additional functions do they have?

The principal excretory organs of the body are the kidney, ureter, urinary bladder, and urethra. Under the control
of hormones, the kidneys filter blood in nephrons and secrete or reabsorb electrolytes. It also controls blood
pressure by manufacturing renin, increases bone marrow red blood cell formation by releasing erythropoietin,
and kidney cells convert vitamin D to its active form. The ureters then transport urine from the kidneys to the
bladder. Peristalsis causes smooth muscle layers in their walls to contract and push urine. Once in the bladder,
tiny valvelike folds of bladder mucosa that cover the ureter apertures prevent urine from returning to the ureters.
The urinary bladder is a smooth, collapsible, muscular sac that briefly holds pee. Finally, the urethra is a thin-
walled tube that transports pee from the bladder to the exterior of the body via peristalsis. The male urethra
transports both urine and sperm (in semen) out the body, but never simultaneously. As a result, the urethra in
males is part of both the urine and reproductive systems.

2. What is diuretic? Name two.

Diuretics, also known as water pills, are drugs that help kidneys get rid of excess water and salt in our body
through urine. They are also used to treat disorders such as high blood pressure, edema (extra fluid in the tissues),
and others. Two examples of this are spironolactone and bumetanide.