Experiment No.

6
POWER OF ENZYME!
INTRODUCTION:

For any reaction to occur at the appreciable rate, proper conditions are required. The reactants
called substrate in the enzymatic reaction must collide in proper orientation, & with the enough
energy for the bond breakage or formation necessary to produce products. Most reactions in the
cells would not occur at rates sustainable to life without the catalytic effect of enzymes. Catalysts
are molecules that speed up chemical reactions, and almost all catalysts in biological systems are
proteins. These organic catalysts are called Enzymes.

OBJECTIVE:
To observe the Enzyme Catalysis.

PROCEDURE:
Explore by doing the activity (follow the steps in pre-recorded video found in the internet) and
experience the power of enzyme.

MATERIALS:

QUESTIONS:
1. Based on your observation, what caused the difference in the activity of the catalase
enzyme in each cup? What factors do you think consider?
  The amount of hydrogen peroxide added to the solution determines how active the
catalase enzyme is in each cup. The reaction rate will rise as more substrate
molecules collide as more hydrogen peroxide is added to the solution. This is due
to the fact that when catalase is introduced to hydrogen peroxide, a fast evolution
of oxygen occurs that lasts for around two minutes, depending on the peroxide
concentration. After that, oxygen is released at a constant rate that gradually drops
over the course of an hour. The drop in rate is almost certainly due to enzyme
breakdown, and numerous kinetic models have been devised to account for it. The
quick development and constant pace, on the other hand, are fundamental
characteristics of peroxide decomposition.

2. Explain why increasing enzyme concentration promote enzyme activity?

 As long as there is a substrate to bind to, increasing the enzyme concentration will
speed up the process. Because there will be nothing for more enzymes to bind to
once all of the substrate has been bound, the process will slow down. The rate of
reaction speeds up as the substrate concentration rises. This is due to the fact that
there will be more substrate molecules colliding with enzyme molecules, resulting
in the formation of more product. The concentration of enzymes and the activity
of enzymes have a direct proportional connection.

3. Compare & contrast the 4 cups. Take a picture.

 When comparing cup 1 to cups 2, 3, and 4, the enzymatic response appears to be

the same. If you recall the experiment, we used the same amount of dishwashing
detergent in each cup, but we didn't use even a little bit of hydrogen peroxide in
cup 1, which resulted in nothing compared to the other three cups. 1/2 tablespoon
 3 percent hydrogen peroxide, 1 tablespoon hydrogen peroxide, and 2 teaspoons
hydrogen peroxide in cups 2, 3, and 4. In this experiment, we discovered that as
the concentration of hydrogen peroxide rises, so does the pace of reaction.
Because it created oxygen gas quickly, more concentrated hydrogen peroxide
produced more oxygen bubbles and the reaction rate was quicker. As may be seen
in the second image. Cup 1 has the same outcome as cup 3 since the same amount
of hydrogen peroxide is used. This is because, as long as there is substrate to bind
to, increased enzyme concentration will speed up the process. The process will no
longer accelerate up once all of the substrate has been bound, as there will be
nothing for new enzymes to bind to. The rate of reaction increases as the substrate
concentration rises. This is because more substrate molecules will collide with
enzyme molecules, resulting in the formation of more product.

4. Enumerate some factors affecting the enzyme activity, discuss each.

 Concentration of Enzyme:
As the concentration of the enzyme is increased, the velocity of the reaction
proportionately increases (Fig. 66.1). In fact, this property of enzyme is made use
in determining the activities of serum enzymes for diagnosis of diseases.
 Concentration of Substrate:
Increasing Substrate Concentration increases the rate of reaction. This is because
more substrate molecules will be colliding with enzyme molecules, so more
product will be formed.However, after a certain concentration, any increase will
have no effect on the rate of reaction, since Substrate Concentration will no longer
be the limiting factor. The enzymes will effectively become saturated, and will be
working at their maximum possible rate.

 Effect of Temperature:
Velocity of an enzyme reaction increases with increase in temperature up to a
maximum and then declines. The optimum temperature for most of the enzymes is
between 40°C-45°C. However, a few enzymes (e.g. venom phosphokinases,
muscle adenylate kinase) are active even at 100°C. In general, when the enzymes
are exposed to a temperature above 50°C, denaturation leading to derangement in
the native (tertiary) structure of the protein and active site are seen. Majority of
the enzymes become inactive at higher temperature (above 70°C).

 Effect of pH:
Increase in the hydrogen ion concentration (pH) considerably influences the
enzyme activity. Each enzyme has an optimum pH at which the velocity is
maximum. Most of the enzymes of higher organisms show optimum activity
 around neutral pH (6-8). There are, however, many exceptions like pepsin (1-2),
acid phosphatase (4-5) and alkaline phosphatase (10-11) for optimum pH.
 Effect of Product Concentration:
The accumulation of reaction products generally decreases the enzyme velocity.
For certain’ enzymes, the products combine with the active site of enzyme and
form a loose complex and, thus, inhibit the enzyme activity. In the living system,
this type of inhibition is generally prevented by a quick removal of products
formed.
 Effect of Activators:
Some of the enzymes require certain inorganic metallic cations like Mg 2+, Mn2+,
Zn2+, Ca2+, Co2+, Cu2+, Na+, K+ etc. for their optimum activity. Rarely, anions are
also needed for enzyme activity e.g. chloride ion (CI–) for amylase.

5. Are enzymes considered as reagents? Why?

 A reagent is a "substance or compound that is supplied to a system in order to
bring about a chemical reaction or to check whether a reaction has happened or
not" in chemical science. This type of reaction is used to confirm the existence of
another material. The design of diagnostic reagents incorporates several elements
of an enzyme-catalyzed reaction. An enzyme's specificity can be used to assess a
substrate or to eliminate interferents from another process. The usage of the
proper enzyme can be used to assess compounds that function as cofactors,
inhibitors, or activators. In clinical chemistry, enzymes are used as reagents. For a
clinical biochemist, enzymes are crucial analytical tools. Enzymes are used as
reagents in three ways: to help generate a signal, to recognize an analyte, and to
serve as a label in a number of immunoassay procedures.

6. What do you think would happen if an enzyme in your body was not working
properly?
 An enzyme is a kind of protein found within a cell. Enzymes catalyze chemical
reactions in the body. By accelerating the rate of a chemical process, they help to
maintain life. The body's enzymes assist in the completion of important processes.
Building muscle, getting rid of pollutants, and breaking down food particles are
just a few. When an enzyme stops working. This has ramifications for the
biological functions that the enzyme aided. For example, enzymes are required for
proper digestive system function. The bulk of digestive enzymes are produced by
the pancreas, stomach, and small intestine. Salivary glands produce digestive
enzymes even when chewing to begin breaking down food molecules. Without
these enzymes, we wouldn't be able to digest our food correctly.