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.