RATIFICATION PAGE

Complete Report of Basic Biology with the tittle “The influence of pH on enzim
Activity” that made by :
name : Nur Ummu Pratiwi Arifuddin
ID : 1414442007
class :A
group : III
after checked and approved by Assistant and Assistant Coordinator.

Makassar, January 14th 2015

Assistant Coordinator, Assistant,

Djumarirmanto S.Pd Muhammad Nur Arsyad

ID. 091404158

Known by,
Responsibility Lecture

Drs.H.Hamka L,MS
ID.1921231 198702 1 005
 CHEPTER I
INTRODUCTION

A. Background
Biology is part of science that studies the science of matter and energy
associated with living organisms and life processes. Living things is identical to
the processes in they life. Starting from their birth, until they can sustain life. In
everyday life there is a process that occurs in the body of an organism, which
such existing process that takes place in the complex. For example, in the process
of digestion, food eaten by a living creature certainly can not directly be digested,
will remain through the process first. Sure there are catalysts that facilitate
making such food easier to digest. This catalyst can be either enzyme. In the
body of a living creature there are various kinds of enzymes that regulate
processes in the body of a living being.
The enzyme is a biological catalyst produced by living cells and can help
speed up the various biochemical reactions. Without the enzyme, the reaction in
the human body is slow and uncontrollable. Catalysts although in small amounts
has the ability to speed up chemical reactions without enzyme was unchanged
after the reaction is complete. The catalyst also showed specificity, meaning that
a particular catalyst will work on just one particular type of reaction. Although all
of the enzyme was originally produced in the cells, some in eksresikan cell wall
and can function outside the cell.
In the known digestive enzymes, Reviews such as amylase enzyme amylase
enzyme when hydrolyzed would convert starch into sugar. Not only that, but
there are many more examples of enzymes, and every enzyme has a different
function. Amylase enzymes or called diatase was instrumental in the formation of
sugar in the living body. Amylase enzymes can be found in grains or nuts.
Therefore, it takes some practice to discuss this further. addition to the theory we
can also prove the practice done.
Enzymes are very influential, in terms of the reaction. In learning we get there
some factors that can affect the work. One is the degree of acidity or pH. We can
takes the examples ptialin enzymes in our mouth, if the high acidity level what
 will be happen? will be useful whether or even harmful? and how to raise and
lower its pH?
        Students can answer these questions by doing this experiment "Influence
of pH on enzyme activity". Although in this experiment we took samples of the
enzyme amylase, at least we can see what the effect of the acidity level

B. Purpose
Proving the influence of pH on enzyme activity amilase
C. Benefit
With the existence of this experiment, students can see firsthand evidence.
The purpose of the practicum is to prove the effect of pH on the activity of the
enzyme amylase. that pH influence on the activity of the enzyme, such as amylase.
 CHEPTER II
PREVIEW OF LITERATURE

The enzyme is a biological catalyst produced by living cells and can help
speed up the various biochemical reactions. Without the enzyme, the reaction is slow
and uncontrollable in the person body. Although in small Amounts catalysts has the
ability to Accelerate the chemical reaction without enzyme was unchanged after the
reaction is complete. The catalyst also showed specificity, meaning that a particular
catalyst will work on just one particular type of reaction. Although all of the enzyme
was originally produced in the cells, some in excretion cell wall and can function
outside the cell (Ristiati, 2000).
Enzymes can be found both in animals and plants. One of the enzymes found
in plants is amylase. The enzyme can hydrolyze starch into sugar. Amylase produced
by the leaves or seeds germinating. Amylase activity is affected by inorganic salts,
pH, temperature, and light. pH optimuum amylase according to Hopkins, Cole and
Green (Miller, 1938) is 4.5 to 4.7 (Tim Pengajar, 2014).
Starch found in many plant sources that are rich in carbohydrates, for
example: rice, cassava, corn and so on. Shape and large starch depending on the
species. Starch consists of two there are, amylase and amylopectin. The difference
between the two lies in its chain, one straight, the other branched, as well as the
shorter amylopectin chain. Starch hydrolysis using glucose result in acid, whereas
when using the enzyme produces maltose. With iodine Amylase to give a blue color,
whereas amylopectin gives a purple color. Missing color when heated. Once cool
colors recur. With the colors disappear because alkali alkali binding iodidum. Starch
does not reduce and do not form osazon. At the time of hydrolyzed by enzymes or
acid will form the intermediate results, is amylodekstrine, erytrodekstrine,
akhrodekstrine, and finally into maltose which eventually can be affected enzyme
glucose after maltase (Saptasari, 2000).
Some enzymes require a cofactor that is not unusual binding protein and a little
loose with the enzyme. The cofactor called coenzyme. While coenzyme binds tightly
to the enzyme called prosthetic groups. Many enzymes require metal cofactors such
as Mn++, Fe++, Mg+, etc. In the process of isolation sometimes enzyme cofactors that
bind to the enzyme irrespective of loose causing enzyme activity decline or even
disappear Altogether. Part of the enzyme protein called apoenzyme, while the overall
enzyme called holoenzim (Girindra,1986).
According by Campbell (2010), enzyme activity-how efficient enzyme function
is influenced by environmental factors, such as :
1. Effect of Temperature and pH
Temperature and pH is an important environmental factor in the enzyme.
Every enzyme has an optimal temperature, is when the temperature of the reaction
rate is high. Without denaturation of the enzyme, this Allows collision sushu most
molecules and molecular conversion of reactants into products that most large
quickly in part human enzyme has an optimum temperature of about 35-400C. As
each enzyme has an optimum temperature, the enzyme also has a pH when he was
most active. The optimal pH value for most of the enzyme is in the range of pH 6-
8, but there are some exceptions. for example, the wasp, a type of digestive
enzymes in the human stomach, works best at pH 2. Such acidic environment
denature most enzymes.
2. Cofactorsr
Many enzymes require helper nonperotein to carry out the catalytic activity.
This additional supplemental-called cofactors, can bind to the enzyme as a
permanent embelan, or may bind loose and forth along the substrate. Some
enzyme cofactors are inorganic, such as metal atoms, zinc iron and copper in the
form of ions. Cofactor function in a variety of ways, but in all cases the use of
cofactors, these molecules perform a crucial function in the catalyst.
3. Enzyme Inhibitors
Certain chemical substances selectively inhibit the action of a specific
enzyme, and we have learned many things about the function of the enzyme to
assess the effects of these molecules. If inhibitors (inhibitors) is attached to the
enzyme by covalent bonds, inhibition that occurs usually can not and forth.
However, many enzyme inhibitors bind to the enzyme through weak interactions,
which means that nature can and forth.
a. Competitive inhibitor, lowers productivity by means of blocking the enzyme
substrate enters the active site.
 A noncompetitive inhibitor, did not compete directly with the substrate to bind
to the active site of the enzyme. Instead, this type inhibitors interfere with the
enzymatic reaction by binding to other parts of the enzyme. The enzyme activity was
heavily influenced by the state of temperature and pH. Each of enzyme can work
effectively at a certain temperature and pH and its activity is reduced in a state below
or above the point of the enzyme pepsin digestion of proteins are most effective at pH
1-2 while the other proteolytic enzymes, trypsin, on the pH becomes not active, but
very effective at pH 8. Since we now have understood and about :
1) The essential role of tertiary struktus, is in the form of enzyme function.
2) The role of weak power such as hydrogen bonds and ions in the formation of
the tertiary structure.
Hydrogen bonds are easily damaged by menaikkkan temperature. This in turn will
damage the parts of the tertiary structure of the enzyme that is essential for substrate
binding. Changes in pH, change the state of ionization of charged amino acids (is acid
aspartiat. Lisnina) which may have an important role in substrate binding and
catalytic processes. Without the group - COOH of Glu - 35 unionized and groups -
COO of ASP-52 ionised, the catalytic process of lysozyme would be halted (Kimball,
1983).
A lot of enzymes will not work without an additional nonprotein substance called
cofactors. Cofactors can be either a metal ion such as Zn ++ (a cofactor carbonic
anhydrase), Cu++, Mn++, Mg++, K+, Fe++, or Na++. Or cofactors may be a small organic
molecule called a coenzyme. Vitamin B group such as thiamine (B1), riboflavin (B2)
and functions as a coenzyme nicotinamide. Kooenzim can tightly bound (covalent
bonds) in the protein portion of the enzyme as a prosthetic group, the other can be
loosely or even only temporarily at the time of these enzymes perform its catalytic
function (Kimball, 1983).
There are a number of mechanisms that play a role in order to work enzymes that
efficiently and coordinated. For enzymes such as proteinase, which can remodel
substabsi cell itself, we find that it works hampered if they are inside the cell. For
example proteinase pepsin is formed in a cell in an inactive form is pepsinogen
(Kimball, 1983).
Substrate complex concept enzim- first proposed by Emil Fisher, an organic
chemist in 1884. Part enzyme fused to a substrate is referred to as the active side
(active side). On the active side of this enzyme substrate in the reorganized into
products. If the active side of rigid and specific to a particular substrate, the reverse
reaction can not occurred because the molecular structure of the compound has a
different product with a product originally had different compounds with compounds
of origin so as not "recognized" by the enzyme. In contrast to the active side of the
rigid concept of Fisher, Daniel E. Koshland forward the concept that the enzyme
active site can dissuaikan the substrate or product structure after the molecules
approaching the enzyme active site. Thus, the incorporation of the enzyme with the
substrate becomes more fit. This concept is now known as the hypothesis of
“stimulated - to – fit” (Lakitan, 1993).
Ph medium can affect the activity of the enzyme. Generally there is an
optimum pH of an enzyme that can serve a maximum and decreases enzyme activity
at pH higher or lower. Represented by a bell-shaped curve, but for other enzymes may
be relatively flat curve is sometimes the image relationship with the pH of the enzyme
activity is represented by a bell-shaped curve, but for other enzymes may curve is
relatively flat, often in the range of pH optimum between pH 6 to pH 8 (Lakitan,
1993).
 CHEPTER III
OBSERVATION METHOD

A. Time and Place

Day / Date : Wednesday, January 7th 2015

Time : 4.30 pm until 5.30 pm
Place : Biology Laboratory of third floor west,
Mathematic & Science Faculty State University of
Makassar
B. Tools and Materials
1. Tools
a. Mortar and pistilum
b. 10 pieces of Reaction Tube
c. small funnel
d. 1 piece of Reaction Tube Rack
e. 1 piece of Spiritus Lighter
f. tube clamp
2. Materials
a. Bean sprouts (Arochis hypogea)
b. Dilute of HCl
c. NaOH solution
d. pH paper
e. Amylum
f. A and B Fehling
g. Matches

C. Work Procedures
1. Take a handful of mung bean sprouts. Put it in a mortar and then crushed. Add
30 ml of distilled water while crushed.
2. Strain the liquid obtained from the number 1, enter into a centrifuge tube. Play
centrifuge for 15 minutes at medium speed.
3. Pour the supernatant fluids (lymph) obtained in a test tube.
4. Prepare 5 pieces of large test tube and insert into each of these tubes 1 ml of
starch solution, then give the table I - IV.
5. Enter the sprouts extract obtained from number 3 to the first tube, check pH
and record. Furthermore, for the liquid into 3 small test tube, give the table a,
b, c. After 10 minutes, add JKJ or Fehling's solution A and B into a tube. After
15 minutes, add the same substance into the tube b; and after 15 minutes,
include the same substances into the tube c. Record the color.
6. In the second tube add 1-2 drops of dilute HCI, check pH and record. Then
add 1 ml extract of number 3. Further treatment such as number 5.
7. In the third tube add 1 drop of NaOH solution, check pH and record. Then add
1 ml of extract of number 3. Further treatment such as number 5.
8. In the fourth tube add 10 drops of a solution of JKJ and note the color. At V
tube add 10 drops of Fehling's solution A and B, heat for 2 minutes, observe
and record the color change.
9. Compare the color that occurs on the tube I - V, create tables and conclude.
 CHEPTER IV
OBSERVATION AND RESULT

A. Result
Long Storage
Change Before After heated pH

Tube I
Added A 12
and B
fehling

Tube II
Added 9
NaOh
solution

Tube III
Added
HCl 3
solution

Tube IV
Added A
and B 12
fehling

Note :
 1. Tube IA, IIA and IIIA tubes was heated in water for 5 minutes.
2. Tube IB, IIB and IIIB tubes was heated in water for 10 minutes.
3. Tube IC, IIC, and IIIC tubes was heated in water for 15 minutes.

B. Discussion

In this experiment, there are ten test tubes were used. Tenth of the tube, was
given indicators are different from each other. The following:
1. Tube I
In the first tube, there are three tubes were used and treated namely
Tube IA, IB Tube, Tube IC with observation time of 5 minutes, 10 minutes,
and 15 minutes. In each tube was added 1 ml of starch and 2 drops of Fehling
A and B. At this time, the solution was dark blue. Then heated and allowed to
stand at a different time each tube. After an interval of 5 minutes, a yellow-
brown solution. After 10 minutes, the solution became clear orange. And hose
15 minutes the solution became dark orange color. And there is no sediment in
the third tube. The pH obtained is 12.
2. Tube II
In the second tube, there are three tubes are in use, the treated namely
Tube IIA, IIB Tube, Tube IIIB. The time used is 5 minutes, 10 minutes and 15
minutes. In this tube was added 1 ml of starch and 2 drops of NaOH. At that
time, the yellow color of the solution became turbid. Then heated and allowed
to stand at a different time each tube. After an interval of 5 minutes, the color
will be a clear yellow tube. After 10 minutes the color became clear yellow.
And at the time of 15 minutes the yellow color remains clear that, where each
solution in the tube does not form a precipitate. measured pH is 9.
3. Tube III
On the tube III also uses three tubes, namely tube IIIA, IIIB, IIIC with
observation time of 5 minutes, 10 minutes and 15 minutes. In this tube was
added 1 ml of starch, then added HCl. At this time the color becomes clear.
Then heated and allowed to stand at a different time each tube. After an
interval of 5, 10, and 15 minutes of the third color in the tube becomes
 translucent white. IIIA and IIIB to the tube there are deposits, while the tube
IIIC no sediment. Obtained by the final pH obtained 3.
4. Tube IV
At this stage, only one tube were observed. Where treatment is, added
starch then Fehling A and B, at that time the color changes from clear to blue.
Then after heated and silenced the color changes to green cloudy.

When compared with existing theory by Saptasari, Murni (2000). Hydrolysis

of starch using acid produces glucose, whereas when using the enzyme produces
maltose. With iodine amylase gives blue color, whereas amylopectin gives a
purple color. when heated in the color disappeared, after cold colors recur. Starch
does not not reduce, when hydrolyzed by the enzyme will produce amylodextrine,
and finally did become maltose which can finally be sugar.
Here in the experiment known use of starch to test whether the solution is
alkaline or acidic. Where, can be seen from the number of its pH. Each enzyme
requires an optimum temperature and pH different because enzymes are proteins,
which can change shape when the temperature and acidity different. From the
results of experiments conducted, where the addition of HCl pH number is 3
proved to be acidic, and the addition of NaOH, pH 9 which indicates alkaline. The
materials we use are water and lighter spirits which serves to heat the solution
resides in the tenth small tube, Fehling Fehling's A and B serves to identify the
sugar contained in the mixture and extract the starch solution. And the function by
HCl solution and NaOH solution is to give the nature of acids and bases in the
solution. The usefulness of phenolphtalein indicator is to measure the acid or
alkaline solution.
 CHEPTER V
CLOSING

A. Conclussion

From the experiments it can be concluded that, the enzyme can worked
optimally on neutral pH. Under acidic conditions or alkaline action of the enzyme
is inhibited or may not worked optimally, and the structure will be damaged. H+ ion
concentration or pH of the solution greatly affect enzyme activity. The function of
HCl is used to test the enzymes that worked on trait the acid. And the function of
NaOH is used to test worked on trait the enzyme alkaline or acid And the function
of Fehling's solution A and B or JKJ is to help change the colors that occur in
lerutan or as an indicator (which test the glucose in the solution).

B. Suggestion

1. To Apprentice : to be more careful in adding the solutions, also counting the

time.
2. To Assistant : in order to better control the way the work done by the their
apprentice.
3. To Laboran : in order to provide better preparation, so that the learners
can clearly see the parts.
 BIBLIOGRAPHY

Campbell. 2002. Kimia Kehidupan. Erlangga. Jakarta.

Campbell. 2010. Biologi. Erlangga: Jakarta.
Girindra, Aisjah. 1986. Biokimia I. Gramedia: Jakarta.
Kimball, John W. 1983. Biologi. Erlangga. Jakarta.
Lakitan, Benyamin. 1993. Dasar-Dasar Fisiologi Tumbuhan. Grafindo. Jakarta.

Ristiati, Ni Putu. 2002. Pengantar Mikrobiologi Umum. Jakarta: Proyek

Pengembangan Guru Sekolah Menengah

Saptasari, Murni. 2000. Petunjuk Praktikum Botani Tumbuhan RendahBiokimia.

Malang: Universitas Negeri Malang.

Tim Pengajar. 2014. Penuntun Praktikum Biologi Dasar. Laboratorium Biologi:

FMIPA UNM. Makassar.
 Answer of Question :
1. Usefulness of Fehling's solution A and B or JKJ is to help change the colors
that occur in lerutan or as an indicator (which test the glucose in the solution).
2. In the enzyme extraction of seeds need dicentrifuge because to make the
supernatant fluid (lymph) which will be added with some kind of solution to
prove the effect of pH of the solution.
3. The function of HCl is used to test the enzymes that worked on trait the acid.
And the function of NaOH is used to test worked on trait the enzyme alkaline
or acid.