TUDEJA, ASHLEY ANNE J.

BIO 024: BIOCHEMISTRY (LABORATORY)

UI-FA1-BSN1-5 SAS # 8 November 25, 2022

Guide Questions:

1. Compare the stability of oxidase and peroxidase to heat.

- is more stable than oxidase, very heat stable. the heat stability is apparent from the fact
that if the enzyme is 83 degrees celsius, half the original activity remains after 32 minutes.
peroxidases are known to self-regenerate due to reversible degeneration.

2. Differentiate the action of oxidase, peroxidase, and catalase.

- a. Oxidase

Polyphenol oxidase is an oxygen transferring enzyme. It activates molecular oxygen and

enhances oxidation. It catalyzes oxidation involving molecular oxygen. Oxygen as E- acceptor
and reduces to H20 or H202.

It is a metalloprotein that contains either Cu/Fe. It is inhibited by Cn and H2S (combing with
Cu/Fe).

b. Peroxidase

It is well suited for the preparation of enzyme conjugated antibodies, due in part to its
ability to yield chromogenic products, and in part to its relatively good stability characteristics. It
is used in the transfer of oxygen from H2O2 to substrate. It leads to oxidation of a substrate by
H2O2.

c. Catalase

Antioxidant enzyme naturally produced in the body. Converts hydrogen peroxide into
hydrogen and oxygen, preventing the formation of carbon dioxide bubbles in the blood. It also
uses H2O2 to break down potentially harmful toxins in the body, including alcohol, phenol, and
formaldehyde.

It is found in tissues of aerobic cells; contain ferric ions; activated by CN and H2S.

3. Why should peeled potatoes not be exposed to air longer than necessary?

When peeled potatoes are exposed to air, the color of the exposed surface changes. The
brown color is the result of a chemical catalyzed by several enzymes, one of them is
polyphenol oxidase or phenolase/tyrosinase. The polyphenol oxidase oxidizes catechol to o-
diphenol to the presence of water. The phenol oxidases are copper containing proteins
which can oxidize phenol derivatives to quinenes. There are 3 groups of enzymes involved in
the browning of potatoes: Monophenol Oxidase (tyrosinase), Polyphenol Oxidase ( catechol
oxidase), and Cytochrome Oxidase.
TUDEJA, ASHLEY ANNE J. BIO 024: BIOCHEMISTRY (LABORATORY)
UI-FA1-BSN1-5 SAS # 8 November 25, 2022

4. What are the optimum conditions for enzyme activity in the human body?

Enzymes can only work in certain conditions. Most enzymes in the human body
work best at around 98.6-degrees Fahrenheit (F) (37°C), which is the body’s typical
temperature. At lower temperatures, they may still work but much more slowly.
If the temperature is too high or if the environment is too acidic or alkaline, the enzyme
changes shape; this alters the shape of the active site so that substrates cannot bind to
it.

5. Give other factors which influence enzyme activity and state the effect of each factor.

Factor 1: Concentration of Enzyme

As the concentration of the enzyme is increased, the velocity of the reaction

proportionately increases. This property is used for determining the activities of serum
enzymes during the diagnosis of diseases.

Factor 2: Concentration of Substrate

In the presence of a given amount of enzyme, the rate of enzymatic reaction increases
as the substrate concentration increases until a limiting rate is reached, after which
further increase in the substrate concentration produces no significant change in the
reaction rate. At this point, so much substrate is present that essentially all of the
enzyme active sites have substrate bound to them.

In other words, the enzyme molecules are saturated with substrate. The excess
substrate molecules cannot react until the substrate already bound to the enzymes has
reacted and been released (or been released without reacting).

Factor 3: Effect of Temperature

The protein nature of the enzymes makes them extremely sensitive to thermal changes.
Enzyme activity occurs within a narrow range of temperatures compared to ordinary
chemical reactions. As you have seen, each enzyme has a certain temperature at which
it is more active. This point is called the optimal temperature, which ranges between 37
to 40C°.

Factor 4: Effect of pH

A change in the pH value, either acidic or basic, affects the ionization of amino acid
residues, leading to changes in the three-dimensional structure of the enzyme. The
alteration in the enzyme conformation affects its interaction with its substrate, thus
reducing its activity. In acid-base and covalent catalysis mechanisms, pH change can
hinder or suppress catalytic activity. In extreme cases, it can denature the enzyme,
destroy its three-dimensional structure, and render it permanently non-functional.
TUDEJA, ASHLEY ANNE J. BIO 024: BIOCHEMISTRY (LABORATORY)
UI-FA1-BSN1-5 SAS # 8 November 25, 2022

Factor 5: Effect of Inhibitors

Inhibitors may bind to the enzyme or its substrate to inhibit the ongoing
enzymatic activity and prevent successive catalytic events. The effect on enzyme activity
is irreversible when the inhibitors form strong bonds to the enzyme’s functional group,
leaving the enzyme permanently inactive.