Laboratory Report

Enzymes and Digestion

I. Introduction
Chemical digestion could not take place without the help of digestive enzymes. An
enzyme is a protein that speeds up chemical reactions in the body. Digestive enzymes
speed up chemical reactions that break down large food molecules into small molecules
(CK-12 Foundation, 2013). Digestive enzymes are released in both anticipation of food
and in response to food. This means that just thinking about or looking at food is enough
to get your juices flowing! As we smell and eventually taste our food, the number of
enzymes that are being secreted increases. Enzymes are secreted from our salivary
glands, and then from the cells lining our stomach, pancreas, and large and small
intestines. Different types of enzymes are secreted depending on the types of foods that
we eat (Murray, Bender, Botham, Kennelly, Rodwell & Weil, 2012; Lowes &
Anderson, 2015).
Did you ever utilize a wrench to fix a jolt? You could fix a jolt with your fingers,
yet it would be troublesome and moderate. On the off chance that you utilize a wrench,
you can fix a jolt significantly more effectively and rapidly. Chemicals resemble
torques. They make it a lot simpler and more efficient for concoction responses to
happen. Like a wrench, compounds can likewise be utilized again and again. Be that as
it may, you need the proper size and state of the wrench to productively fix the jolt,
much the same as every catalyst is explicit for the response it makes a difference.

II. Objectives
1. To explain how the various digestive enzymes aid digestion relative to the stage or
phase of digestion in which they are activated.
2. To describe the importance of enzymatic activity in the physiological functioning
of a human individual’s digestive system.

III. Method
Prior to the experiment proper, the group prepared and requested for the necessary
tools and equipment for the experiment (Patton, 2005): sheets of clean paper; hot plate;
test tubes with test tube rack and tongs; thermometer; disposable 1mL measuring
droppers; beakers; distilled water; and test solutions (Benedict’s and Lugol’s reagent;
1% starch soln. and 1% maltose soln.; pancreatin; oil and bile salts; and litmus cream).
These materials are necessary for observing the gamut of reactions enzymes undergo
relative to temperature, pH and other factors. Subsequently, the group commenced with
the experiment.
The experiment initiated with the heating of the water bath and dropping of the test
solutions into their corresponding labelled test tubes. The group periodically measured
the temperature of the water bath. Subsequently, the group began heating the test
solutions respective of the temperature required for each test: 0°C; 22°C; 37°C; and
100°C. The group observed the color change in each of the test solutions. Doing so
provided the group with the necessary information for interpreting how the reactants
react at a given temperature.
 In the latter part of the experiment, the group recorded the information procured
form examining the solution color and precipitate formation from each test. Upon
finalization of data gathering, the laboratory activity concluded.

IV. Results
Table 1. Qualitative Tests of Starch and Lipids

V. Discussion
Digestion is the process of breaking down food into nutrient molecules; absorbing
these molecules; and then expels any and all unwanted digestive products. Crucial to
digestion are biologic polymers called enzymes; and are responsible for catalyzing the
gamut of chemical reactions necessary to keep the human body functioning, and in a
broader sense, make life possible. The vast majority of enzymes are proteins. These
enzymes must be complete and maintained at a balance proportion in the human body
in order for them to function properly. The foregoing enables enzymes to carry the
following major functions (Murray et al., 2012; OpenStax College, 2013):
a. Breaking down of nutrients to supply energy and chemical building blocks
b. Assembling the same chemical building blocks into proteins, DNA,
membranes, cells and tissues; and
c. Harnessing of necessary energy to power cell functionalities such as
motility, neural function, and muscle contraction.
Enzymes are able to carry out these functionalities when provided with suitable pH
and temperature. Increase in temperature and pH directly increases the kinetic energy
of enzymes; thus, speeding up enzyme-substrate binding. A drastic increase and
decrease in temperature and pH, respectively, causes enzyme denaturation,
consequently halting enzyme activity. The optimal temperature and pH for enzymes is
37°C-40°C and 7.0-8.0, respectively. However, these values differ for every enzyme
as they have different requirements (Shier, Butler & Lewis, 2002; Scanlon & Sanders,
2007).
The digestive process initiates in the mouth with the aid of the teeth for mastication
and of salivary enzymes and lingual lipase for chemical breakdown of carbohydrates
and lipids, respectively. Deglutition enables the food, now bolus, to be conducted
towards the esophagus with peristalsis aiding in propulsion of bolus towards the
stomach via the gastroesophageal sphincter. In the stomach, bolus is converted into
acid chyme via gastric acid that catalyzes activation of inactive pepsinogen (produced
by chief cells) into pepsin, which is responsible for breaking down proteins into
polypeptides and peptides. Gastric lipases break down a small proportion of fats into
glycerol and fatty acids. Lastly, hydrochloric acid (HCl) is produced by the cells of the
gastric mucosal lining in order to aid pepsin and kill certain bacteria (Widmaier, Raff
& Strang, 2019).
Acid chyme passes the pyloric sphincter and enters the duodenum with the aid of
peristalsis. Once in the duodenum: pancreatic lipase further breaks down intact fats into
glycerol and fatty acids; amylase breaks down starch into maltose; and trypsin and
chymotrypsin, which are powerful proteases, split proteins into polypeptides and
peptides. The digested acid chyme travels towards the distal jejunum and ileum and is
further broken down by maltase, sucrase and lactase, which convert disaccharides into
their component monosaccharide subunits. Also, peptidases split large peptides into
smaller ones and then the smaller peptides into amino acid units. Lastly, undigested
food enters the large intestine from the ileum of the small intestine via ileocecal valve.
Here, water and salt are absorbed by the intestinal lining thru microvilli. The residues,
along with waste pigments (bilirubin), dead cells and bacteria, are pressed into feces
and then stored for excretion via defecation (Marieb & Hoehn, 2013).
Iodine test is specifically conducted for starch. The latter, is a homopolymer of
glucose forming an α-glucosidic chain, hence a glucosan. This polysaccharide is
composed two main constituents: helical, non-branching amylose (13-20%); and
branched amylopectin (80-87%). Starch is composed of an interval of 24-30 glucoside
residues with α-1,4-glucosidic linkages in the chains an α-1,6-glucosidic linkage at the
branch points (Murray et al., 2012). Figure 9a shows the coiled structure of starch.
The amylose in starch is responsible for the formation of a blue-black color in the
presence of iodine. The iodine molecule diffuses into the amylose coil and causes this
reaction. The reagent used for this test is the Iodine-KI reagent, which is made by
dissolving iodine in water in the presence of potassium iodide because iodine has low
solubility in water. As a result, a linear triiodide ion complex is produced (LibreTexts,
2019, June 6).
Benedict’s test, as mentioned prior, is also a reduction test. As such, it is conducted
specifically to test for the presence of reducing sugars (i.e., all monosaccharides and
some disaccharides such as lactose and maltose). Moving on, Benedict’s reagent is
composed of a bluish copper sulfate solution. Upon addition and mixing of a sugar
solution and heating in a water bath, changes in color will vary depending on the
reducing properties of the sugar. Reducing sugars will gradually produce a brick-red
cuprous oxide precipitate, whereas non-reducing sugars will not produce said
precipitate and the boiled solution will remain perfectly clean. Cuprous oxide, or
copper (I) oxide (Cu2O), is formed due to the oxidation of the carbohydrate and
reduction of the Cu2+ ions to Cu+ ions (Figure 4b) and is insoluble in water; thus,
precipitated out of the solution (Biology Discussion, 2015, October 14). The color and
amount of the precipitate is dependent on the volume of the reducing sugar present –
the greater the amount of free carbonyl groups present, the greater the amount and
intensity of the brick-red precipitate.

Table 2. Benedict’s Test Theoretical Results

Solution-precipitate Color Reducing Carbohydrate
Concentration (g% or 1/100g)
Blue 0.0
Green ~0.5
Yellow ~1.0
Orange ~1.5
Red (brick-red) ~≥2.0

Pancreatin is an enzyme that works optimally with an ambient pH of 8.0 ± 0.1 at a

temperature of 40°C. Also called pancrelipase, this enzyme is normally produced by
the pancreas to aid in the digestion of fats, proteins and sugars. Pancreatin is a group
of pancreatic enzymes comprised of amylases, lipases and proteases (Marieb & Hoehn,
 2013). The amylases present in pancreatin speed up the catalysis of the amylose in
starch at 22 °C and 37°C (Table 1: 4a and 4b, 5a and 5b), which is closer to the optimal
40°C temperature for pancreatin activity and 32°C-37°C for amylase activity; thereby,
producing the characteristic deep blue-black solution at a faster rate. However, the
pancreatin present in tubes 6a and 6b produced a different result due to the significantly
high temperature of the water bath, causing the denaturation of pancreatin, especially
of amylases.
In test tubes 8a and 8b, pancreatin and lipases aided in the catalysis of the fats
present in litmus cream – which is a cream that contains fats and a litmus indicator –
into glycerol and fatty acids. The free fatty acids lower the pH of the solution (acidic)
and induce the conversion the stock blue color into the characteristic pink color.
However, in the case of test tube 8c, the solution remained blue because the fats in the
litmus cream were not catalyzed; thus, no free fatty acids were present to lower the pH
causing the pH to remain relatively high and the solution basic. The presence of water
induced the formation of two layers characteristic of emulsifiers.
The difference in results due to the manipulation of concomitant variables such as
temperature and pH (due to addition of other compounds or substances) is indicative
of the complexity of the digestive process. The foregoing ensures that the body is able
to utilize all of the beneficial nutrient molecules and dispose of the undesirable ones.
Apropos of this, the nutrient molecules the body absorbs are in their simplest
monomeric form. The microvilli of the large intestine are only able to absorb these
monomers and minerals because of the abundance of diverse enzymes the all-inclusive
enzyme reservoir the digestive system has.

VI. Conclusion
Given the discussion, digestion is a process that involves mechanical and chemical
catalysis. However, the abundance of enzymes along the digestive tract makes the
digestive process enzyme-driven and dependent. Enzymes are present in the mouth,
stomach, small intestine and large intestine. The abundance and variety of salivary,
gastric, intestinal and pancreatic enzymes are due to the difference in pH and
temperature requirements of each enzyme relative to the organ in which they catalyze
reactions. This entails that the human body is able to digest a gamut of food, may they
be acidic, neutral or alkaline, and utilize them for growth, energy production, muscle
contraction, and maintenance of homeostasis. Hence, enzymes and digestion are
concomitant variables that contribute to the well-being of the human individual.

VII. Bibliography
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