5. lap.pengaruh ph terhadap aktivitas enzim
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APPROVAL SHEET
Complete report of general biology with title “The Influence of pH On Enzyme
Activity”, created by :
Name : Ummi Qalsum
Reg. Number : 091204174
Class : B (ICP)
Departement : Physics
Group : V (Five)
After checked by Assistant and Assistant Coordinator, so this is report accepted.
Makassar, December 2009
Assistant Coordinator Assistant
(Djumarirmanto, S.Pd) (Zaidah . R)
Nim.061404026
Known By
Lecturer of Responsibility
(Ir. Muh. Wiharto, M.Si)
NIP.132 006 81
CHAPTER IINTRODUCTION
A. Background
Enzyme is one or some polypeptide bunchs ( protein) what is functioning
as catalyst ( compound quickening process reaction of without pot is clean reacts) in
a chemical reaction. Enzyme works by the way of patching at surface of reacting
matters molecule and thereby quickens reaction process. Acceleration happened
because enzyme to reduce activation energy which by itself will water down the
happening of reaction. Most of enzyme works characteriscally, with the meaning
every enzyme type can only work for one kinds of compound or chemical reaction.
This thing is caused by chemical perbedaanstruktur every enzyme having the
character of permanent. For example, enzyme amylase can only be applied at
alteration process of extract becomes glucose.
There is two enzyme factions, that is functioning digestion enzyme as
catalyst, and metabolism enzyme which accountable for compiling, improve;repairs
and forms again cells in body. There is many factors influencing active job activity
an enzyme. namely temperature, concentration and hydrogen ion exponent. Substrate
concentration ( Enzyme activity increases with substrate concentration, more
collisions between substrate molecules and the enzyme); Temperature (Enzyme
activity increases with temperature, warmer temperatures cause more effective
collisions between enzyme and substrate, however, hot temperatures destroy
enzyme); pH ( Most enzymes are optimized for a particular pH)
Main digestion enzyme consisted of protease enzyme ( unloads protein),
lipase enzyme ( unloads fat) and amylase enzyme ( unloads charcoal hydrate). In this
practicum will be studied is special worked is active from amylase enzyme. From
theorys before all is known that amylase enzyme works is active from pH 4,5 - 4,7, to
know pH influence to active mode of action of amylase enzyme can be done attempt
about pH influence to enzymatic activity at unit V practicum of General Biology.
B. Experiment’s Purpose
The students can know how the influence of pH on enzyme activity .
C. Experiment’s Benefit
Students will be able to explain how the influence of pH on Amylase
enzyme activity .
CHAPTER IIPREVIEW OF LITERATURE
Enzymes
Protein molecules that function as catalysts. The reactants of an enzymatically
accelerated reaction are called substrates. Each enzyme accelerates a specific
reaction. Each reaction in a metabolic pathway requires a unique and specific
enzyme. End product will not appear unless ALL enzymes present and functional
(Sylvia S. Mader,_____,______ )
Enzymes Energy of Activation
Reactants often “reluctant” to participate in reaction. Energy must be added to
at least one reactant to initiate the reaction. Energy of activation. Enzyme Operation:
Enzymes operate by lowering the energy of activation . Accomplished by bringing
the substrates into contact with one another (Sylvia S. Mader,____,____ )
Enzyme is protein is in the form of orbicular needed to all chemical reactions
taking place in body. Partly small enzymes is produced in salivary gland in mouth
part. But most digestion enzyme produced by pancreas gland. There is two enzyme
factions, that is functioning digestion enzyme as catalyst, and metabolism enzyme
which accountable for compiling, improve;repairs and forms again cells in body.
Main digestion enzyme consisted of protease enzyme ( unloads protein), lipase
enzyme ( unloads fat) and amylase enzyme ( unloads charcoal hydrate) (Anonim,
2009).
Digestion of carbohydrate have been started since food to come into mouth;
food is munched to be broken to become small parts, so that number of surfaces of
broader food contacted with digestion enzymes (Anonim, 2009).
Allosteric enzyme change their structure in response to binding of effectors.
Modolation can be direct, where the effector binds directly to binding sites in the
enzyme, or indirect, where the effector binds to other proteins or proteins or protein
subunits that interact with the enzyme and thus influence catalys activity (Poejadi,
1984).
In mixed food mouth with spit containing Enzyme Amilase ( ptyalin).
Amylase Enzyme works breaks long chain carbohydrate like amilum and dextrin,
will be decomposed to become molecule which more maltose simple. While spit
good for smoothing out food that easier to be swallowed. Only partly small amilum
earning in cema in mouth, because of food just just resided in in buccal cavity.
Therefore is better if food munched to be longer, to give opportunity of more
resolvings of amilum in buccal cavity. With mechanic process, food is swallowed by
through throat and hereinafter will enter stomach (Anonim, 2009).
The seed is a remarkable structure that enables seed plants to survive
unfavorable condition. Each seed contains an embriyo that can grow into a mature
plant. In addition, the seeds of flowering plants have structure called cotyledons. The
cotyledons store food use by the embryo in the form of starch. Strach is long chains
of glucose molecules. The embryo needs to break these chains, forming sugars it can
use for energy. It does this enzyme action, by first testing dry bean seed for the
presence of glucose, the testing a been seed that has germinated (Dwijoseputra,
1994).
Enzyme Insuffiency
If body experiences lacking of enzyme, stomach easy to rebel when
consuming certain victuals. According to dr. Ari Fahrial, “ Lack of one enzyme types
generally accompanied by lack of other enzyme. Trouble lacking of chronic enzyme
can cause patient to experience malagizi ( less gizi), what causes body weight to
decrease and body endurance also declines.” (Anonim, 2009).
Obtainable amylase from various sources like crop, animal and
microorganism. now a number of amylase enzymes has been produced
commercially. Usage of mikrobia assumed to be more prosepektif because easy to
grow, quickly yields and condition of manageable area (Anonim, 2009).
Produce of amylase enzyme can apply various source of carbons. Example of
source of cheap carbons is chaff, molasses, corn flour, corn, tapioca waste etcetera. If
it is applied waste as substrate, hence the waste can be enriched its(the nutrition to be
optimal produced enzyme. Source of carbon that is serve the purpose of supplement
between laian: extract, sucrose, lactose, maltose, dekstyrosa, fructose, and glucose.
Sources of nitrogen as supplement of inter alia: peptone, tripton, flesh extract, yeast
extract, ammonium sulphate, soya bean meal, urea and sodium nitrate (Anonim,
2009).
pH
pH is a measure of the acidity or basicity of a solution. It is defined as the
cologarithm of the activity of dissolved hydrogen ions (H+). Hydrogen ion activity
coefficients cannot be measured experimentally, so they are based on theoretical
calculations. The pH scale is not an absolute scale; it is relative to a set of standard
solutions whose pH is established by international agreement (Anonim, 2009).
pH Indicator
A pH indicator is a halochromic chemical compound that is added in small
amounts to a solution so that the pH (acidity or basicity) of the solution can be
determined visually. Hence a pH indicator is a chemical detector for hydronium ions
(H3O+) (or Hydrogen ions (H+) in the Arrhenius model). Normally, the indicator
causes the color of the solution to change depending on the pH. At 25 degrees
Celsius, considered the standard temperature, the pH value of a neutral solution is
7.0. Solutions with a pH value below 7.0 are considered acidic, whereas solutions
with pH value above 7.0 are basic. As most naturally occuring organic compounds
are weak protolytes, carboxylic acids and amines, pH indicators find many
applications in biology and analytical chemistry. Moreover, pH indicators form one
of the three main types of indicator compounds used in chemical analysis. For the
quantitative analysis of metal cations, the use of complexometric indicators is
preferred, whereas the third compound class, the redox indicators, are used in
titrations involving a redox reaction as the basis of the analysis (Anonim, 2009).
For optimal accuracy, the color difference between the two species should be
as clear as possible, and the narrower the pH range of the color change the better. In
some indicators, such as phenolphthalein, one of the species is colorless, whereas in
other indicators, such as methyl red, both species confer a color. While pH indicators
work efficiently at their designated pH range, they are usually destroyed at the
extreme ends of the pH scale due to undesired side-reactions (Anonim, 2009).
pH
measurement with indicator paper
Universal indicator and Hydrion papers are blends of different
indicators that exhibits several smooth color changes over a very wide range
of pH values (Anonim, 2009).
pH in living systems
Compartment pH
Gastric acid 0.7
Lysosomes 4.5
Granules of chromaffin cells 5.5
Urine 6.0
Neutral H2O at 37 °C 6.81
Cytosol 7.2
Cerebrospinal fluid (CSF) 7.3
Blood 7.34 – 7.45
Mitochondrial matrix 7.5
Pancreas secretions 8.1
CHAPTER IIIOBSERVATION METHOD
A. Place and Date
This experiment’s is done at:
Day and Date : Wednesday, December 9th 2009
Time : at 13.30-15.00
Place : Laboratory of Biology
Faculty Mathematis and Science
Makassar State University
(at the 2nd east floor part)
B. Tools and Materials
1. Tools
a. Centri fuge and tube centrifuge
b. Mortar and pistilum
c. Test tube
d. Pipette
e. Small funnel
f. Tube rack
g. Lights spritus
h. Filter paper
i. pH Paper (pH indicator)
2. Materials
a. Green Bean Sprouts
b. Starch Solution
c. Fehling solution A and B
d. JKJ solution
e. Dilute HCl
f. NaOH solution
g. Aquades
C. Work Procedure
1. Taking a handful of mung bean sprouts. Entering into the mortar and
grinding, adding 30 ml with aquades crushed.
2. Fluid filter has been crushed and put into a centrifuge tube. Then play
a centrifuge for 15 minutes at medium speed.
3. At first tube takes three test tubes and fills it with starch counted 1 ml
at every tube, then gives name with IA ( for observation of 5 minute),
IB ( for observation of 10 minutes), IC ( for observation of 15
minutes), enters sprout extract which has been filtered, cheque the pH,
then adds fehling A and B, and heated third the tube so boiling and
observing as according to time each tube.
4. At second tube takes and fills it with starch counted 1 ml at every
tube, then gives name with IIA ( for observation of 5 minute), IIB
( for observation of 10 minutes), IIC ( for observation of 15 minutes),
enters sprout extract which has been filtered, cheque the pH, adds
fehling A and B and solution of HCl, cheque the pH, and heated third
the tube so boiling and observing as according to time each tube.
5. A third tube takes three test tubes and fills it with starch counted 1 ml
at every tube, then gives name with IIIA ( for observation of 5
minute), IIIB ( for observation of 10 minutes), IIIC ( for observation
of 15 minutes), enters sprout extract which has been filtered, cheque
the pH, adds fehling A and B and solution of NaOH, and heated third
the tube so boiling and observing as according to time each tube.
6. At fourth tube, enters solution of starch counted 1 ml and adds
solution of JKJ.
7. At fifth tube, entered solution of starch counted 1 ml and adds fehling
A and B solution.
8. Observes change happened in every tube.
CHAPTER IVOBSERVATION RESULT
A. Observation Result
From the experiment, we can get the result such as :
a. Observation Table
Nomor pH Warna
Tabung Awal Akhir Awal Akhir
I A 6 9 GreenRather Yellow
Green
I B 6 9 GreenRather Yellow
Green
I C 6 9 GreenYellower
Green
II A 6 2 Transparent White of Milk
II B 6 2 TransparentWhite of
Greenness
II C 6 2 TransparentMore Green
White
III A 6 10 Dark GreenOld Dark
Green
III B 6 10 Dark GreenOld Dark
Green
III C 6 10 Dark GreenOld Dark
Green
IV 6 - Dark Blue -
V 6 -Transparent
Purple-
A B C (5 minutes) (10 minutes) (15 minutes)
Green Bean Sprouts
Starch Solution
Fehling A and B
A B C (5 minutes) (10 minutes) (15 minutes)
Green Bean Sprouts
Starch Solution
HCl Solution
Fehling A and B
b. The Pictures of Tube’s Situation
1st Tube
2nd Tube
D.
E.
F.
Starch Solution
Fehling A and B
Starch Solution
JKJ Solution
3rd Tube
4th Tube 5th Tube
G.
A B C
(5 minutes) (10 minutes) (15 minutes)
Green Bean Sprouts
Starch Solution
NaOH Solution
Fehling A and B
B. Discussion
1) 1 st Tube
At first tube, initial pH which in measuring is good at crest IA, IB, IC was 6,
while pH finally is 9.
1. Tube colour IA, after addition of condensation of starch, green peanut
extract, and fehling A and B, then is heated is green and after 5 minute
observation of its the colour turns into chartreuse
2. Tube colour IB, after addition of condensation of starch, green peanut
extract, and fehling A and B, then is heated is becoming green and after 10
minutes observation of its the colour turns into chartreuse
3. Tube colour IC, after addition of condensation of starch, green peanut
extract, and fehling A and B, then is heated was becoming green is young
and after 15 minutes observation of its the colour turns into rather green
brass
From result of attempt to there was no enzyme which can work optimum pH
pad because its the pH wa s very height, ought to at first tube, its the enzyme can
work carefully but because pH got too height that was 9 so that enzyme at tube
firstly cannot work carefully
2) 2 nd Tube
At second tube, initial pH was 6 and pH finally after addition of HCl was 2.
1. Tube colour IIA, after addition of condensation of starch, green peanut
extract, fehling A and B, and HCl then is heated was transparent and after
5 minutes observation of its the colour turns into white of milk
2. Tube colour IIB, after addition of condensation of starch, green peanut
extract, fehling A and B, and HCl, then is heated was white and after 10
minutes observation of its(the colour turns into white of greenness
3. Tube colour IIC, after addition of condensation of starch, green peanut
extract, fehling A and B, and HCl, then is heated was transparent and after
15 minutes observation of its(the colour turns into rather white greenness
This t u be, the colour was white greenness and haves the character of acid
that was pH 2, this thing indicates the condensation there was glucose content
although in number a few mean enzymes changes amilum to become glucose
though slow as result of pH that was not optimumkarena amylase enzyme cannot
work for sour situation .
3) 3 rd Tube
At third tube, initial pH was 6 and pH finally after addition of NaOH was 10
1) Tube colour IIIA, after addition of condensation of starch, green peanut
extract, fehling A and B, and NaOH, then is heated was becoming green
stripper and after 5 minute observation of its the colour turns into rather
old green.
2) Tube colour IIIB, after addition of condensation of starch, green peanut
extract, fehling A and B, and NaOH, then is heated old adalahHijau and
after 10 minutes observation of its(the colour turns into dark green.
3) Tube colour IIIC, after addition of condensation of starch, green peanut
extract, fehling A and B, and NaOH, then is heated was becoming green
stripper and after 15 minutes observation of its(the colour turns into rather
dark green.
This tube , the colour rather basic and dark green that was pH 10, This thing
indicates that there was glucose content in condensation but in number slimmer ,
enzyme at this third tube doesn't work optimumly. Existence of the glucose
content meaned enzyme has worked but in slow job(activity because amylase
enzyme cannot work for situation of alkaline
4) 4 th Tube
At fourth tube, pH initially was 6 and after condensation of starch is added
with JKJ the condensation colour turns into dark blue. This indicates that at
condensation of the starch contains amilum because turning colour to become
dark blue.
5) 5 th Tube
At fifth tube, pH initially was 6 and after condensation of starch packed into
the tube and in adding fehling A and B and after heated its the colour was trans-
parent purple. Function of fehling A and B to know existence of glucose content
but at the tube doesn't turn colour to become sorrel. This indicates that
condensation of starch doesn't contain glucose.
CHAPTER VCONCLUSSION AND SUGGESTION
A. Conclussion
From result of attempt, inferential that:
Enzyme works well at optimum pH between 4,5-4,7
Enzymatic activity pursued at sour situation or alkaline
Ruddles brick to indicate existence of glucose in condensation with tested
to applies condensation fehling A and B.
Prussion blue until black indicates condensation to contain amilum and
tested by using condensation JKJ.
B. Suggestion
For practican, in doing attempt that more accurately and applies time
efficiently in order not to pursue observation on trial and in observing colour to see
truly colour happened in each tube.
For the assistant, help the practican in observing the change of the tubes.
And for the laboratory to prepare equipment and materials to be used properly, such
as the tools and materials ready to use.
BIBLIOGHRAPY
Anonim. 2009. Enzyme. http://en.wikipedia.org/wiki/Enzyme
Anonim. 2009. pH. http://en.wikipedia.org/wiki/PH
Anonim. 2009. pH. http://www.answers.com/topic/ph
Anonim. 2009. Laporan Praktikum Fotosintesis. http://smartbekantan.blogspot.com
Anonim. 2009. Pengaruh pH terhadap aktivitas enzim. http://amaulianty.blog.friendster.com/
Anonim. 2009. Peran Enzim Amilase Pada Tubuh Manusia ."http://www.w3.org
Dwijoseputra. 1994. Pengantar Fisiologi Tumbuhan. Universitas Indonesia: Jakarta
Mader, Sylvia S. 1982. Metabolism, Photosyntetic & Enzyme Biologi Ninth Edition.________:________
Poejadi. 1984. Biologi. Sinar Buku: Jakarta
APPENDIX
QUESTION
1. What is the function of condensation fehling A and B and JKJ solution?
2. Why at enzyme extract from seed needs in centrifuge ?
ANSWER
1. Reactant Fehling consisted of two parts, that is Fehling A and Fehling B. fehling
A is condensation CuSO4, while Fehling B is condensation mixture NaOH and
potassium sodium tartrate. Reactant Fehling is made by mixing both the
condensations, causing is obtained a prussion blue condensation. In reactant
Fehling, ion Cu2+ there is as complex ion. Reactant Fehling can be considered
to be condensation CuO. Reaction Of Aldehyde with reactant Fehling yields
sediment of sorrel from Cu2O, on trial this function of condensation fehling A
and B is to determine existence of glucose content in supernatant liquid
2. Sprout extract which has been grind needs in centrifuge that obtainable of pure
supernatant liquid and transparent which will be applied as component of
attempt.