RATIFICATION PAGE

Complete report of Basic Biology observations with the title The Effect of
pH on Enzyme Activity. Which made by:
Name
: Muhammad Tri Prasetia Nua
ID
: 1512441003
Class
: ICP of Physics Education
Group
: 6th
After checked by assistant and assistant coordinator, so this report accepted
Makassar, January 2016
Assistant Coordinator

Assistant

Rusdianto Nurman
ID: 1214041002

Kamaliah Guntur
ID : 1314440012

Known,
Lecture of Responsibility

Drs. H. Abdul Muis, M.Si

ID: 196409131990111001

CHAPTER I
INTRODUCTION
A. Background
Enzyme is protein has function as catalisator for chemist reaction in
biology system. Enzyme is substance have work if combined only one so
its a little time. When combined with reactor substance, enzyme will
down the energy barrier from reaction, so reaction happen quickly.
Without enzyme, there are not life that we know. As a biocatalysator that
controlling all of the speed of physiology process, enzyme was holding the
main actor in the healthy and disease.
Although in the healthy condition all of the physiology process will be
passed off with the step that was formed in the temporary. While the
homeostasis will be defended, but the homeostasis condition can get the
difficult trouble in the pathology condition.
To studying about the enzyme, there are know some of terminology there
are holoenzyme, apoenzyme, cofactor, prostatic group, coenzyme, dan
substrat. Enzyme we can found in the animal and plant. One of examples
of enzyme that we can found in the plant is amylase. The other name of
amylase is dilatase. That enzyme can hydrolysis the amylum become
sugar. Amylase we can found it in the leaf or seed.
In this practicum about influence pH of enzyme activity the university
student will know and try to evidence the influence the PH of enzyme
activity.
B. Purpose
The purpose of this practicum, to showing influenced pH about amylase
enzyme activity.
C. Benefit
Based on this practicum the benefit of this practicum the university student
will know about influenced pH about amylase enzyme activity.

CHAPTER II
REVIEW OF LITERATURE
When

enzyme

was

first

discovered

they

were

given

various

unsystematic names by their discoverers, such as pepsin, trypsin, ptialin, etc. in

the more recent times enzymes have been designated be the suffix ase,
preceded by aterm which indicated either the general nature of the substrate, the
actual name of substrate, the type of reaction catalyzed, or a combination of
several of these facts (Cantarow, 1962).
Enzyme is a protein which is the function as a catalysator for the chemical
reactions in the biology system. Catalysator to make fast the chemical reaction.
Although the catalysator follow in the reaction, it will be back into the first
condition if the reaction was finish. The part of the reaction wasnt catalyst by the
enzyme (Indah, 2004).
There are know some of terminology there are holoenzyme, apoenzyme,
cofactor, prostatic group, coenzyme, dan substrate. Apoenzyme is an enzyme
which is the all of was formed from protein, whereas the holoenzyme is an
enzyme that has some of protein group and non-protein group. The group that is
not the protein was knowing with cofactor. In the cofactor there are bide by in the
protein and it so difficult to loosely that was say coenzyme. In the prostetic and
coenzyme, the two of them that is a part that make it possible the enzyme has
working in the substrate. Substrate is the substance is a substance which is can be
change and reacted by the enzyme (Anonym, 2010).
Different with the non-protein catalysator (H+, OH-, or the metalloid ion),
every enzyme was catalyzing small size of reaction, oftentimes only one. So
enzyme is catalystor which is the specify reaction because all of the biochemist
reaction needed to catalyzed by enzyme. Actually for all of the organic compound,
there are one enzyme on some living organism that can be reaction with and
catalyzing some of chemist change. The reactions like hidrolisis and oxidation
was happened with fast on the living cells in the PH around the netral and in the
body temperature. It can be happened because there is enzyme. Enzyme was

synthetic into the cell, but after extraction in outside of cell still has activity.
Enzyme working very specific. The enzyme can be catalyzing some of reaction
but sometimes only one reaction. It is one of the important characteristic of
enzyme. There is some enzyme that can be catalyzing the kind of the same
reaction for example to moving the phosphate, oxidation-reduction, etc. so there is
some specificity (Indah, 2004).
Enzyme, being protein, cannot withstand the action of strong acid or base.
However, even over the PH range in which inactivation doesnt occur, enzyme
exhibit optima in their activity. In the case of an enzyme which attack non-ionic
substrates. The optimum pH is constant for the several substrates. The optimum in
the curve, then, must correspond to some particular configuration of electrical
charges on the reactive surface of the enzyme protein, in which condition the
action on the substrate is most efficient. As in the case of the temperature effect,
the very existence of an optimum in the curve suggest the action of two opposing
forces. In the case of pH effect. It has been suggested that at least two ionizable
group are involved at the active site, having different pH, an idea which is in
agreement with recent theory concerning the chemical groups at the active site.
Ionized substrate will themselves very in electrical properties over the pH range,
so that enzymes attacking such substance often show pH optima which differ from
one substrate to another. In these examples the peak in the curve must be a
resultant, or compromise, between the most efficient, charged states of the enzyme
and substrate (Cantarow, 1962).
The transferring enzymes catalyze the transfer or shift, from one molecule
to another, of a chemical group which is not present in the free state during a
transfer. The nature of the group transferred forms the basis for subclassification
of these enzyme. Although, strietly speaking, the transfer of hydrogen atoms or
electrons can be included in this group, such reaction have complexities and
peculiarities all their own, and for convenience are best classified separately
(Cantarow, 1962).
If the enzyme activities were measured in the different pH, so the bigger
part of enzyme inside the body will show the optimum activity between pH 5,0-

9,0, except some of enzyme for example pepsin (optimum pH = 2). Its cause by:
1. In the low or high pH, enzyme will be denaturasi, 2. In the low or high pH,
enzyme or substrate can be change the electricity cause the chage of enzyme
activities. For examples some enzyme reaction can be work if that enzyme has the
negativity electricity and the substrate has the positif electricity (Indah, 2004).
Biological oxidation is chiefly dehydrogenation or the de-electronetions. The
enzyme catalyzing such reaction almost all require coenzyme, or contain
prosthetic groups or heavy metals. One system of classification is based on the
nature of this non-protein substance which is essential to the reaction (Cantarow,
1962).

CHAPTER III
OBSERVATION METHOD
A. Time and Place
Day/date

: Monday/December 28th 2015

Time

: 01.10 am 03.10 am

Place

: Laboratory of Biology, Mathematics and Science Faculty,

State University of Makassar.

B. Tools and Materials

1. Tools
a. Centrifuge tube
b. Mortar
c. Reaction tube
d. Pipette
e. Small funnel
f. Tube rack
g. Spiritus
2. Material
a. Sprout of rice plant, corn and greenpeal
b. Amylum solution
c. Fehling solution A and B
d. KJ solution
e. HCl solution (10%)
f. NaOH solution (1%)
g. PH paper
h. Filter paper
i. Aquades
C. Work Procedure
1. Took the sprout of rice plant, corn and green peal. Entered into the mortar
the polished
2. Filtered the liquid that we get from number 1, entered into centrifuge tube.
Rolled it in the centrifuge during 15 minute normally speed.
3. Poured the clean liquid that we get into the reaction tube.

4. Drew up 5 pieces of reaction tube and filled the tube with amylum 1 ml,
and then give label I-IV.
5. I tube entered the green peal extract that had got from the No. 3, check the
PH and wrote down. And then divide the liquid into the 3 pieces small
reaction tube, gave the label a, b, c. a tube after 10 minutes added JKJ
solution or fehling A and B. b tube after 15 minutes done it same with the
a tube. Wrote down the color.
6. On the II added 1 2 HCL liquid, check that PH and wrote down. And
then added 1 ml extract from the no. 3. The done it like no 5.
7. On the III added 1 drop NaOH solution and wrote down the color, check
that PH and wrote down. And then added 1 ml extract from the no. 3. The
done it like no 5.
8. On the IV added 10 drop of JKJ solution and wrote down the color, On the
V added 10 drop of fehling A and B. heated during 2 minutes, observed the
change of color and wrote down.
9. Compared the color that happen on the I-V make the table and make the
conclusion.

CHAPTER IV
OBSERVATION RESULT
A. Result
Tube

pH

Ia
Ib
Ic
IIa
IIb
IIc
IIIa
IIIb
IIIc

II
III

Color Change
Initial
Green
Green
Brown
White
White
White
Blacklish Brown
Blacklish Brown
Gray Brown

Final
Crimson
Green
Crimson
Crimson
White
Light Blue
Black
Reddish Brown
Crimson

B. Discussion
In the tube 1 in the tube given amylum, added fehling A and B, added extract
green peal and we heated the tube 1a. 1b, and 1c with different time and color
was changed in the tube 1a changed from green became cimson, tube 1b from
green to green, and tube 1c changed from brown to brick red (crimson).
In the tube 2 in the tube given amylum, added fehling A and B, added extract
green peal and 1-2 drop HCL and we heated the tube 2a. 2b, and 2c with
different time and the color was change in the tube 2a changed from white
became crimson, tube 2b changed from white into white, and the 2c changed
from white into light blue.
In the tube 3 in the tube given amylum, added fehling A and B, added extract
green peal and 1 drop NaOH solution pH and we heated the tube 3a, 3b and
3c with different time and the color was change in the tube 3a changed from
blacklish brown became black, tube 3b changed from blacklish brown into
reddish brown, and tube 3c changed from gray brown into crimson.
CHAPTER V
CLOSING

A. Conclusion
Based on the observations that have been done, conclude that The pH of the
substrate it can be evidence the enzyme activity which is enzyme function is
to make fast the reaction.
B. Suggestions
Based on the result of practicum and the conclusion, the suggestion as
follows:
1. Suggestion for Laboratory
I hope for next practicum about the influence the PH of enzyme activity.
The quality of the tool that we use in the influence the PH of enzyme
practicum activity must be improve.
2. Suggestion for Assistant
The Assistant who was guide for this practice is good. So, I hope for the
next practicum the assistant can guide excellently and the university
student will know about how to use the microscope correctly.
3. Suggestion for Friend
I hope for my friend in the next practicum, we can work together again
more than in this practicum and working seriously to get a good result to
get a good result.

Bibliography
Campbell, Neil A. 2009. Biology. California: Pearson.
Cantarow, Abraham. 1962. Biochemist. USA: W.B. Saunders Company.
Indah, Mutiara. 2004. Enzim. Sumatera Utara: Fakultas Kedokteran Universitas
Sumatera Utara.
Tim Penyusun. 2015. Penuntun Praktikum Biologi Dasar. Makassar: Universitas
Negeri Makassar.

ATTACHMENT
Question:
1. What to Fehling's solution A and B and JKJ?
2. Why does the enzyme extract from the seeds need to be in a centrifuge?
3. What is the function of HCL and NaOH in experiments conducted?
Answer:
1. Function Fehling Fehling's solution A and B is a solution or an indicator to
prove the content of glucose in solution experiments. JKJ function is as a
solution or an indicator to prove the existence of a protein in solution
experiments.
2. The enzyme extract from the seeds need to obtain a supernatant liquid
dicentrifuge purer than ever because of the centrifuge in the screening
function to precipitate the fibers or dirt from the supernatant fluid is a liquid.
3. HCL solution serves to make the acidic nature of the extract, and a solution of

sodium hydroxide to make alkaline properties in the extract to prove the pH

level in accordance with the action of the enzyme.