EXPERIMENT #6:

Enzymes

POST-LABORATORY QUESTION AND RESULTS:

Group: 1 March 13, 2020

Group Members:

ABESAMIS, BLANCA,
CABRERA, GUERRA,
MAGSALAY, MISSION, RUEL

Section:​ 8 Instructor: ​Prof. Jenny Rose Aggabao

A. DATA AND RESULTS

Oxidase Sample Color Change Time when color change

took place

Potato Light yellow to brown. 10 minutes

Banana Light yellow to brown. 9 minutes

Apple Light yellow to brown. 7 minutes

B. POTATO OXIDASE/CATECHOLASE
a. Observations:
- The color changed into brown slowly from the edge then to the
middle.

b. What changes do you think take place as the color changes upon the
addition of Phenol, Pyrogallol or Catechol and Guaiac solution to the
potato extract?
- Potatoes contain catechol oxidase which acts on colorless catechol
and converts it to brown-colored melanin. The browning that occurs
when you cut and expose these items to air is a result of this
reaction.
-
C. POTATO PEROXIDASE/CATALASE
a. Observations:
- There is a bubbling reaction when the potato extract was placed
under boiling water for 10 minutes.
b. What changes do you think take place as the color changes upon the
addition of Phenol, Pyrogallol or Catechol and Guaiac solution to the
potato extract?
- Catechol oxidase was seen to be present in potato extract because
there is a change in color. There was an appearance of color
reaction in which a brown pigment color showed.

D. QUESTIONS:
a. What are enzymes? Cofactors? Apoenzyme? Prosthetic group?
Holoenzyme?
- Enzymes acts as a catalyst to bring about a specific biochemical
reaction.Cofactors can be metals or small organic molecules and
they assist in enzyme activity. Following, holoenzymes are the
active forms of apoenzymes. And prosthetic group is a tightly
bound, non-polypeptide unit required for the biological function of
some proteins; may be organic or inorganic like vitamin or metal ion
but they must not be composed of amino acids.
b. What are the factors that influence enzymatic activity?
- Several factors can affect enzymatic activity which are temperature,
pH level, enzyme concentration, substrate concentration and the
presence of any inhibitors or activators.
c. What is denaturation? How does heat/pH affect denaturation?
- Denaturation is a process where proteins or nucleic acids lose the
quaternary, tertiary and secondary structure by application of some
external factors such as heat or pH level.
- The hotter it is the more rapidly molecules are vibrating and
moving; collisions of water molecules with enzymes could undo its
folded structure that is held in position by hydrogen bonds and
when this happens, the molecule is considered to have been
denatured.
d. What are the substrates of Catecholase? What is the color of the
product produced? Write the reaction. Correlate this with the
browning of fruits​.
- The substrates of catecholase are catechol and oxygen. The
substrates react with one another within the active site of the
 enzyme. The products formed by this reaction are benzoquinone
and water; since benzoquinone has a brown color, you can see that
the reaction has taken place.
e. What is the reaction of peroxidase?
- Peroxidase reacts by mechanisms similar to catalase, but the
reaction catalyzed is the oxidation of a wide variety of organic and
inorganic substrates by hydrogen peroxide.
f. What is blanching and what is its purpose?
- Blanching is used to destroy enzymatic activity in vegetables and some
fruits prior to other processing like freezing or dehydration or canning or
thermal processing. It is a pretreatment by mild heat for a specific time
followed by rapid cooling or passing immediately to the next processing
stage.
The purpose of blanching is to achieve several objectives.
● To soften the tissue to facilitate packaging.
● To avoid damage to the product.
● To eliminate air from the product.
● To preserve the natural colour.
● To destroy or retard certain undesirable enzymes.
● To help preserve natural flavour.
- The major purpose of blanching is frequently to inactivate enzymes, which
would otherwise lead to quality reduction in the processed product.
g. What factors affect the rate of enzymatic browning in
fruits/vegetables?
- Enzymatic browning is a chemical process which occurs in fruits and
vegetables by the enzyme polyphenoloxidase, which results in brown
pigments.
- The most important factors that determine the rate of enzymatic browning
of vegetables and fruits are the concentration of both active PPO and
phenolic compounds present, the pH, the temperature and the oxygen
availability of the tissue.

REFERENCES:
http://ecoursesonline.iasri.res.in/mod/page/view.php?id=3489
https://ucanr.edu/datastoreFiles/608-361.pdf