ENZYMES

Virtually all reactions in the body are mediated by enzymes, which are protein catalysts that
increase the rate of reactions without being changed in the overall process. Among the many biologic
reactions that are energetically possible, enzymes selectively channel reactants (called substrates) into
useful pathways. Enzymes thus direct all metabolic events.

They have a high degree of specificity for their substrates, they accelerate chemical reactions
tremendously, and they function in aqueous solutions under very mild conditions of temperature and
pH. Few non-biological catalysts have all these properties. Enzymes are central to every biochemical
process. Acting in organized sequences, they catalyze the hundreds of stepwise reactions that degrade
nutrient molecules, conserve and transform chemical energy, and make biological macromolecules from
simple precursors. The study of enzymes has immense practical importance. In some diseases, especially
inheritable genetic disorders, there may be a deficiency or even a total absence of one or more
enzymes. Other disease conditions may be caused by excessive activity of an enzyme. Measurements of
the activities of enzymes in blood plasma, erythrocytes, or tissue samples are important in diagnosing
certain illnesses. Many drugs act through interactions with enzymes. Enzymes are also important
practical tools in chemical engineering, food technology, and agriculture.

In a normal process of reactions, some simple inorganic mixtures such as the elements of
oxygen and hydrogen will not react simultaneously towards one another, until a sequence of events that
an initial energy will be provided like a spark or heat a reaction will take place and thus form water. The
same situation of reaction will take place if platinum will be added to produce an explosion to form
water; at the end of the reaction the platinum will remain unchanged. In this type of reaction platinum
metal serves as the catalyst.

This type of substance which speeds up reactions that in the end of this reaction will remain
unchanged is known as CATALYST. This catalyst merely affects the speed and rate of reactions; it does
not alter the direction of certain reactions. In return, it will increase the amount of energy involve in
bonding, but lessens the energy needed to start with a reaction making it more favorable.

For living cells we also have this type of catalyst, but instead of inorganic catalyst such as platinum
stated above we have an organic catalyst and these are called ENZYMES. These enzymes are highly
specific in their reactions; most of these enzymes are globular and are proteins in nature. Like all
catalyst, enzymes lower the activation energy they needed to start a reaction, but they cannot make a
reaction that could not take place on its own. Due to the amino acids sequence in each enzyme,
enzymes are different from one another; they have a unique three-dimensional structure, which makes
them specific. Most enzymes can interact only with substrate that fits only to the active site.
 EXPERIMENT No. 7

ENZYME ACTIVITY

OBJECTIVE:

Determine the enzymatic activity at different pH and at a different temperature

MATERIALS:

Clean drinking water, tap

Clean drinking water, luke warm

250 beaker

PROCEDURE:

1. Rinse your mouth several times with clean drinking tap water.
2. Wash your mouth with 20 ml of luke warm water for 1 minute. (Gargle the water.)
3. Collect your washings in a beaker. This will serve as a dilute solution of amylase for the
succeeding experiments.

INFLUENCE OF THE pH ON THE ENZYMES

MATERIALS:

Salivary amylase
Tripod
0.2 molar NaCl
Wire gauze
1% starch solution
Liquid or medicine dropper
Buffer solution pH 7
2 test tubes
0.01 Molar Iodine solution
Test tube rack
Test tube brush
Water bath or 500 ml beaker
Spot plate or watch glass with white background

PROCEDURE:

1. Set up 3 test tubes each containing 5 ml of 1 % starch solution, 1 ml of 0.2 M NaCl.

2. To test tube No.1 add the 2 ml pH 7 buffer solution.
3. To test tube No. 2 add the 2 ml pH 5 buffer solution.
4. To test tube No. 3 add the 2 ml pH 9 buffer solution.
5. Place all the 3 test tubes in a water bath heated at 370C.
6. Add 3 ml of amylase to each of the 2 test tube rapidly. Mix well and note the time. Get the pH of
3 test tube using the pH paper.
7. Take note and record your results.
8. Immediately at 1-minute interval, take 1 drop from each of the tube and place on a spot place or
a watch glass with a white background on which drops of iodine solution have been placed.
9. Repeat the withdrawal of the samples from each test tube and note for each test tube the time
in minutes at which the blue color is no longer obtained.
10. Count the number of drops you place in the spot plate or watch glass with a white background,
until no blue color is obtained.
11. Plot this test in a graph (time against pH).
12. Note and record all your result and observations.
13. Repeat procedure from No. 1 to No. 12 but this time instead of 37˚C use 80˚C.
14. Note and record all your results and observations.
 Name: CHARISMA J. ANTONIO Course/Year/Section: ________________
Instructor: _________________________Group No.:________ Date: _________ Score: _______
_______

EXPERIMENT NO. 7

ENZYME ACTIVITY

POST LABORATORY DATA/RESULTS:

Temperature Time pH Level

Test Tube Number 1 5˚C 14min
Test Tube Number 2 37˚C 4min
Test Tube Number 3 50˚C 10min

POST LABORATORY GUIDE QUESTIONS:

1. Why do you have to place the test tubes in water heated to about 37˚C?

To be able to determine the enzyme because at this optimum temperature, the enzyme is most
active and hence, takes less time to digest the starch.

2. What is an enzyme? Give an example.

Enzymes are substance that, in living things, acts as a catalyst to control the rate at which
chemical reactions happen without changing itself.
Examples are
Lipases: This group of enzymes help digest fats in the gut.
Amylase: In the saliva, amylase helps change starches into sugars.
Maltase: This also occurs in the saliva, and breaks the sugar maltose into glucose.
Trypsin: These enzymes break proteins down into amino acids in the small intestine.
Lactase: Lactase breaks lactose, the sugar in milk, into glucose and galactose.
Acetylcholinesterase: These enzymes break down the neurotransmitter acetylcholine in nerves
and muscles.
Helicase: Helicase enzymes unravel DNA.
DNA polymerase: These enzymes synthesize DNA from deoxyribonucleotides.
 3. What are the factors that affect enzyme activity?

Enzyme activity can be affected by a variety of factors, such as temperature, pH, and
concentration. Enzymes work best within specific temperature and pH ranges, and sub-optimal
conditions can cause an enzyme to lose its ability to bind to a substrate.

4. How is time related to the enzymatic activity of the amylase?

At a lower temperature, the enzyme salivary amylase is deactivated and at the higher
temperature, the enzyme is denaturated. Therefore, more time will be taken by an enzyme to
digest the starch at lower and higher temperatures.

5. Illustrate the structural formula of amylase. In the human body, amylase is predominantly
produced by the salivary glands and the pancreas. Although salivary and pancreatic
amylases are similar, they are encoded by different genes (AMY1 and AMY2,
respectively) and show different levels of activity against starches of various origins.

CLINICAL GUIDE QUESTIONS:

1. What is the importance of enzymatic activity in our body in terms of digestion and metabolism?
Our bodies' biochemical reactions are facilitated by enzymes. They support digestion and
breathing among other functions. Problems with one's health can result from consuming an
enzyme in excess or insufficiently. Enzyme supplements may be necessary for some people with
chronic conditions to help their bodies function normally.

2. Can you diagnose a patient’s disease by means of any enzymatic activity? Give an example of a
particular disease?
Yes,you can diagnose a patients disease.An enzyme marker is a blood test to measure enzymes,
proteins in your blood that can indicate tissue damage or disease. Lysosomal enzyme tests are
available to diagnose essentially all LSDs identified to date. Gaucher disease is one of the most
common lysosomal storage disorders (LSDs). LSDs are inherited disorders resulting from a lack of
specific enzymes that break down certain lipids (fats) or carbohydrates (sugars) in the body cells.

3. If a patient has fever, what is its relationship to an enzyme activity? Why?

As temperature increases, enzyme reactions increase. But if the temperature gets too high, the
enzyme stops working. That's why a high fever can disrupt bodily functions.

4. Show your graph by plotting (time against pH).