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CHAPTER I

INTRODUCTION
A.

Background
Protein is the most organic molecules in the cell. This substance is
found in all parts of the bodies of living and a group of the most diverse
kinds of compounds important in biology. (Pine, 1988)
Humans need energy to perform activities and everyday activities, the
energy can be obtained from a variety of foodstuffs. Protein is a polypeptide
biopolymer composed of a number of amino acids linked by peptide bonds.
Protein is a multifunctional biopolymers, namely as structural cells and
tissues and organs, as a biocatalyst enzyme, as a carrier or carrier when the
compound or substance through biomembran cells, and as a regulator.
(Hawab, 2004). Protein is contained in foods commonly consumed by
humans. As the eggs, tempeh, fish and others. In general, a protein derived
from vegetable and animal. Protein is essential for living organisms in
general, because it serves to improve the body's cells are damaged and the
supply of nutrients the body needs.
There are various ways in which the protein test to prove the
elements contained in proteins, one of which biuret test to demonstrate or
prove the peptide bonds that make up proteins.

B.

Formulation of the problem


1. How to show or prove the peptide bonds that form proteins?
2. Are in all materials tested contained the peptide bonds that form proteins?

C. Purpose
1. Students can demonstrate or prove the peptide bonds that make up proteins.

CHAPTER II
LITERATURE
The word protein comes from the Greek word which means first and
foremost, the origin of the word protos. Proteins are composed of various classes
of heterogeneous macromolecules. Protein is a natural polymer composed of
various amino acids by peptide bonds. (Hart, 1987).
Protein is a source of amino acids containing the elements C, H, O and N are
not owned by fat and carbohydrates. Terpor sugar molecules containing sulfur
proteins, and there is a type of protein that contain metals such as iron and copper.
(Winarnno, 1997).
Key thousands of different proteins in the molecular structure is cluster
development units are relatively simple proteins are built from the same basic
circuit, of 20 amino acids having a side chain specific, covalent bonds in a typical
sequence. Because each amino acid side chains which have a special that gives the
chemical nature of each individual, a group of 20 units of this development can be
considered as the alphabet of protein structure. (Lehninger, 1996).
1. Protein function
As Enzymes
Tool Carrier and Storage
Regulatory Movement
Supporting Mechanics
Body Defense or Immunization
Media Propagation Neural Impulses
Control of Growth (Lehninger, 1996)
2. Protein structure
There are four levels of protein structure are:
1. The primary structure shows the number, type and sequence of
amino acids in a protein molecule (sequence of amino acids in a
protein molecule)
2. The secondary structure showed many properties of a protein is
determined by the orientation of the molecule as a whole, form a
protein molecule (eg spiral) and spatial arrangement skeleton (a

hydrogen bond between the NH group, one amino acid residue with
a carbonyl group C = O acid residues others).
3. Tertiary structure shows a tendency circumstances polypeptide folds
combined strap (more like terlipatnya interaction framework to
form a circle).
4. quaternary structure indicates the degree of partnership units of
proteins. (Winarno, 1997).
In terms of its structure, the protein can be divided into two groups, namely:
1. Simple Protein is a protein that is only made up of amino acid
molecules
2. Protein is a protein composite that consists of protein and not the
protein group. This group is called the prosthetic group and consist of
carbohydrates, lipids or nucleic acids.
Simple proteins according to their molecular shape is divided into two
groups, namely:
1. Protein fiber. The protein molecule consists of several polypeptide
chains are elongated and connected to each other by some
crosslinking to the fiber or fibers that form stable. Protein fiber is
insoluble in aqueous solvents, either saline, acidic, alkaline or
alcohol. Large molecular weight could not be determined with
purified starch and difficult. The usefulness of this protein only to
form a network structure and materials, for example, is the keratin in
the hair.
2. globular proteins. Globular proteins are generally spherical or
elliptical and consists of polypeptide chains involved. Globular
protein / speroprotein spherical, this protein dissolved in saline
solution and dilute acid, is also easier to change under the influence
of temperature, acid concentration and dilute acid. This protein is
denatured. Many are in milk, eggs and meat. (Fessenden, 1982)
To test the protein there are several ways one biuret test means a way to give
a clear result of the compounds are believed to have a peptide bond. Biuret test is
used as a way to determine the protein compounds. (Syakur, 2012).

To determine the presence of protein or peptide bonds including protein


hydrolysis as metaprotein, proteosa, peptone, amino acid polypeptide except the
biuret test. In alkaline conditions, CuSO4 react with compounds containing two or
more peptide bonds to form a purple complex. Biuret is a compound with two
peptide bonds formed on heating two urea molecules.
(NH2) 2C = O + (NH2) 2C = O ----> H2N-CO-NH-CO-NH2 + NH3. The
positive reaction occurs in the presence of a color change to purple or pink due to
the compounding of reserves of peptide N and O of the water. Colors that occur
depending on the length of the peptide bond. When the peptide bond length in
purple, otherwise if short color to pink.
Biuret reagent containing ions of copper (II) under alkaline conditions which
will react with the polypeptide or peptide bonds that make up proteins form a
complex compound purple (violet). Positive biuret reaction against two or more
peptide bonds, but negative for the free amino acid or dipeptide. The reaction was
positive for compounds containing two groups -CH2NH2, -CSNH2, -C (NH) NH2
and -CONH2.

CHAPTER III
METHODS
A. Tools and materials
Tools :

1.
2.
3.
4.
5.
6.

Test tube
Pipette drops
The test tube rack
Clamp the test tube
Measuring cup
vortex

5 pieces
3 pieces
1 piece
1 piece
1 piece
1 piece

Materials :
1.
2.
3.
4.

10% NaOH
CuSO4 0.01m
albumin solution
7 kinds of protein test :
Tempe
Tofu
Chicken liver
Chicken
Meat fish
Peanuts
Green beans
B. Step work
1. Prepare 7 kinds of protein solution and 1 solution of albumin.
2. Insert each protein solution into a test tube, each 3 ml.
3. Add 1 ml of 10% NaOH solution into each reaction tube containing any
type of protein solution, homogen with vortex.
4. Insert 3 drops of 0.01 M CuSO4 solution, then stir. If not arise color, add
1-2 drops of CuSO4.
5. Observe the changes that occur.
BAB III
RESULT AND DISCUSSION
A. Result
No.
1

Procedure
Albumin + NaOH
10% + CuSO4
0,01 M

Result
Before
Albumin color:
Colorless
NaOH: Colorless
CuSO4 : Blue

After
Albumin + NaOH :
Colorless
After mixed with vortex :
colorless
Albumin + NaOH

Extract tempe +

Extract tempe

+CuSO4:
Purple + blue sediment
Tempe Extract + NaOH:

NaOH 10% +
CuSO4 0,01 M

color : cream

Yellow

NaOH: Colorless
CuSO4 : Blue

After mixed with vortex :


yellow
Tempe Extract + NaOH
+CuSO4: dark Purple, No
sediment.

Extract tofu +

Extract tahu

Tofu extract + NaOH =

NaOH 10% +

color:
White

yellow wisk

NaOH: Colorless
CuSO4 : Blue

After mixed with vortex :

CuSO4 0,01 M

white
Tofu extract + NaOH +

Extract fish meat + Extract fish meat

CuSO4=purple (+)
Fish meat extract +

NaOH 10% +

NaOH= cream

CuSO4 0,01 M

color : white
NaOH: Colorless
CuSO4 : Blue

muddy,white sediment
After mixed with vortex :
Muddy yellow
Fish meat extract + NaOH
+ CuSO4= muddy

Extract chicken

Exract chicken

meat + NaOH

meat color:
Soft pink

10% + CuSO4
0,01 M

NaOH: Colorless
CuSO4 : Blue

purple,purple sediment
chicken meat extract +
NaOH= cream
After mixed with vortex :
Cream
chicken meat extract +
NaOH + CuSO4= muddy

Extract chicken

Extract chicken

purple(++)
Chicken heart extract +

heart + NaOH

heart

NaOH 10% + CuSO4

10% + CuSO4

color:maroon

0,01 M : deep brown

0,01 M

NaOH: Colorless
CuSO4 : Blue

After mixed with vortex :


Deep brown

Extract peanut +

Extract peanut

Peanut extract + NaOH =

NaOH 10% +

color : white

yellow white muddy

CuSO4 0,01 M

muddy solution
NaOH: Colorless
CuSO4 : Blue

After mixed with vortex :


Yellow white
Peanut extract + NaOH +

Extract mungbean

Extract

CuSO4 = muddy purple


Mungbean extract +

+ NaOH 10% +

mungbean color:

NaOH:

CuSO4 0,01 M

white

light yellow

NaOH: Colorless
CuSO4 : Blue

After mixed with vortex :


Light yellow
Added CuSO4 :
Blackish purple, blue
precipitate.

B. Anallysis
From the data table above, the biuret test the addition of NaOH
solution and CuSO4 in setap materials tested. Color albumin and NaOH
solution is colorless, to color CuSOA 0.01 M solution is blue, as well as for
color 7 containing material that is extracted soybean protein: cream, tofu
extract: white, fish meat extract color: white, Exract chicken meat color:
soft pink, Extract chicken heart color: maroon, peanut Extract color: white
muddy solution, mungbean Extract color: white.
When Albumin + NaOH: Colorless, After mixed with vortex:
colorless and after Albumin CuSO4 + NaOH + Purple + Blue sediment
yield.

And to test materials containing protein 7. First, Tempe Extract +


NaOH: Yellow, After mixed with vortex: yellow and after Tempe Extract +
NaOH + CuSO4 produce dark color Purple, No sediment. In the extract
tofu, when Tofu extract + NaOH = yellow wisk, After mixed with a vortex:
white And after Tofu extract + NaOH + CuSO4 produce color purple (+).
To extract fish meat, when Fish meat extract + NaOH = cream muddy,
white sediment, After mixed with a vortex: Muddy yellow, and after Fish
meat extract + NaOH + CuSO4 produce muddy color purple, purple
sediment. In the test chicken meat, when chicken meat extract + NaOH =
cream, After mixed with vortex: Cream, and after chicken meat extract +
NaOH + CuSO4 produce muddy purple color (++).
In chicken liver extract test when Chicken liver extract + 10%
NaOH + 0.01 M CuSO4: deep brown, mixed with a vortex After: Deep
brown. To test the peanut extract when Peanut extract + NaOH = yellow
white muddy, After mixed with a vortex: Yellow white, Peanut extract +
NaOH + CuSO4 produce muddy purple color. In the last test protein bhan
ie extract mungbean, when mungbean extract + NaOH: light yellow, After
mixed with a vortex: Light yellow, CuSO4 Added produce color Blackish
purple, blue precipitate.
In all test produces a peptide bond, and if at the end of the results
when adding 0.01 M CuSO4 solution produces purple, peptide bond length,
but of some of the material tested produce a dense purple color, meaning
the longer the peptide bond.
C. Discussion Of The Result
To determine the presence of protein or peptide bonds including
protein hydrolysis as metaprotein, proteosa, peptone, amino acid
polypeptide their biuret test. In alkaline conditions, CuSO4 react with
compounds containing two or more peptide bonds form a purple complex.
Biuret is a compound with two peptide bonds formed on heating two urea
molecules.
(NH2) 2C = O + (NH2) 2C = O ----> H2N-CO-NH-CO-NH2 + NH3.
Positive reactions occur with a change in the color purple or pink for
backup compounding peptide N and O of the water. Colors that occur

depending on the length of the peptide bond. When the length of the
peptide bond in purple, otherwise if short color to pink.
And the results of the materials tested, all were found to contain a
peptide bond but on every test produces a peptide bond different, as in
soybean extract produces dark color Purple meaning peptide bond is very
long, the tofu and fish meat extract extract produces the color purple mean
bond length but not along in soybean extract. That is because the protein
content in each test is different, because the protein in soybean
literaturekandungan indeed very much.
But the chicken liver extract produces deep brown color that should if
formed their peptide bonds when the length of the peptide bond in purple,
otherwise if short color to pink. Probably within experimental apparatus
and the solution was contaminated with other materials to produce a deep
brown color. Biuret test on chicken liver showed negative results. It shows
the lack of peptide bonds.
D. Discussion
1. Can the biuret test is used to determine the hydrolysis of such
proteins has been completed?
Biuret test may be used to determine the protein hydrolysis has been
completed or not. Hydrolysis of proteins characterized by a color
change to blue keunguan.Warna purple indicates that the protein has
an alkaline and biuret test itself reacted positively to the formation
of complex compounds of Cu on -CO and -NH group of the peptide
chain under alkaline conditions. Biuret test is done to determine the
presence of protein or peptide bond. including the results of
hydrolysis of proteins such as metaprotein, proteosa, peptone,
except the amino acid polypeptide.

CHAPTER V

CLOSING
A. Conclusions
Protein solution has a kind of bond that is different, there are composed
of a peptide bond length and there are composed of a short peptide bonds. In
alkaline conditions, CuSO4 react with compounds containing two or more
peptide bonds form a purple complex. Biuret is a compound with two peptide
bonds formed on heating two urea molecules.
(NH2) 2C = O + (NH2) 2C = O ----> H2N-CO-NH-CO-NH2 + NH3.
Positive reactions occur with a change in the color purple or pink for backup
compounding peptide N and O of the water. Colors that occur depending on
the length of the peptide bond. When the length of the peptide bond in purple,
otherwise if short color to pink.
In soybean extract produces dark purple color, the tofu and fish meat
extract produces purple color, the chicken meat extract produce muddy purple
color ++, to extract the beans produce a muddy color purple, to extract
mungbean yield Blackish color purple, blue precipitate. Which indicates that
the presence of the peptide bond panjangt. But the chicken liver extract
produces deep brown color that shows on chicken liver showed negative
results. It shows the lack of peptide bonds.
B. Suggestions
As for suggestions to improve the quality of research:
a. It should be more careful in mixing the ingredients so that the results
agree with the theory that has been taught.
b. Tools and materials need to be added Because The test process must
use a considerable reaction tube, so do not spend a lot of time.
c. Maintain hygiene before and after use laboratory.
d. The existence of material preparation before entering the study