CHAPTER I

INTRODUCTION
1. Background
Protein (the root of the Greek word protos meaning "most important") is
a complex organic compounds of high molecular weight which are polymers of
amino acid monomers linked together by peptide bonds. Protein molecules
containing carbon, hydrogen, oxygen, nitrogen and sometimes sulfur and
phosphorus. (Wikipedia, 2007).
Chemically, the protein is a polymer, with the amino acid as monomer
and linked by peptide bonds. Peptide bond is a bond between the carboxyl group
of one amino acid with the amino group of the amino acid next to him
(Lehninger, 1995). Amino acids are compounds that have one or more carboxyl
groups (-COOH) and one or more amino group (-NH2), one of which is located
on the right of the C atoms (Sudarmadji et al, 1989).
Protein plays an important role in the structure and function of all living
cells and viruses. In addition, the protein has a characteristic, one of which is the
solubility. Therefore, to determine the nature of the protein solubility in a
solvent, we did test the solubility of albumin.
2. Problem Formulation
How does the solubility of albumin to various solvents?
3. Purpose
Students are able to prove the solubility of albumin to various solvents.

CHAPTER II
LITERATURE

A. Solubility of Proteins
Protein is a major component in all living cells, both plants and animals. In
most tissues of the body, protein is the largest component after water.
Approximately 50% by weight consisting of the elements carbon (50-55%),
hydrogen ( 7%), oxygen ( 13%) and phosphorus (P) in small amounts (1-2%).
There are several other proteins containing metals such as copper and iron
(Sirajuddin, 2012).
Protein molecule having an amino group (-NH2) and a carboxylic group (COOH) at the ends of the chain. This causes the protein has a lot of charge
(polyelectrolyte) and is amphoteric, which can react with acids or bases. With an
acid solution or low pH, the amino group on the protein will react with H + ions,
so that the protein is positively charged. Conversely, in an alkaline solution
carboxylate ions react with OH, thus negatively charged proteins. As for the
charge on the molecule causes the protein to move under the influence of an
electric field (Sirajuddin, 2012).
Proteins are amphoteric, which can react with the acid and alkaline
solutions. Different protein solubility in water, acids, and bases. There are soluble
and there are poorly soluble. However, all proteins are not soluble in fat solvents
such as ether and chloroform. If the protein is heated or plus absolute ethanol,
then the protein will clot (coagulated). This is due to the ethanol draw water
mantle surrounding the protein molecules. The solubility of the protein in a liquid,
in fact strongly influenced by several factors, among others, pH, temperature,
ionic strength and dielectric constant of the solvent.
Based on the solubility in saline solution akueso. Proteins can also be
classified based on the shape of the whole. So, globular proteins (eg, many
enzymes) have polypeptide chains are twisted and folded solid ratio is not more
than 3-4. Protein pibrosa axial ratio greater than 10 (Roberrt)
2

B. Factors Protein Solubility

1. PH
Such as amino acids, proteins are soluble in water to form ions that
have positive and negative charges. In acidic protein molecules to form
positive ions, whereas under alkaline conditions to form negative ions. At point
isolistrik proteins have positive and negative charges are equal, so it does not
move toward the positive and negative electrodes are placed between the two
electrodes. Protein has a point isolistrik different. Protein isolistrik point is
significant because in general the physical and chemical properties are closely
related to this isolistrik pH. At pH above the point isolistrik negatively charged
proteins, whereas under isolistrik point, positively charged proteins. Isolistrik
point on albumin is at a pH of 4.55 to 4.90. The presence of amino and
carboxyl-free at the ends of chains of protein molecules, causing the protein to
have a lot of cargo (polyelectrolyte) and is amphoteric (can react with acids or
bases). Reactivity of various types of proteins to acids and bases are not the
same, depending on the number and location of the amino and carboxyl groups
in the molecule. In acid solution (low pH), the amino group reacts with H +, so
that the protein is positively charged. Conversely, in an alkaline solution (pH
high) protein molecules will react as acids or negatively charged. At pH
isolistrik charge free amino and carboxyl neutralize each other so that the
charged molecules zero (Winarno, 2002).
2. Salt Concentration
When the salt concentration increases, the majority of water molecules to
be attracted by the salt ions, which then would reduce the number of water
molecules that can interact with the hydrophobic parts of the protein. As a
result of the increasing demand for solvent molecules, the interaction between
the protein to be stronger than the interaction between the solvent and solute,
This will cause the protein molecules coagulate to form a hydrophobic
interaction with one another.

This process is known as salting-out. In another discussion mentioned that

salting out occurs when the high salt concentration, the salt will be more likely
to bind water and cause aggregation. So that protein molecules undergo
precipitation.
Usually in pure water, sparingly soluble proteins. With the addition of
salt, protein solubility will increase. It is caused by a perfectly hydrated
inorganic ions will bind to the surface of the protein and prevent incorporation
(aggregation) of protein molecules. This is called salting in.
3.

Protein denaturation
Denaturation of proteins can be interpreted a change or
modification of the secondary structure, tertiary and quaternary protein
molecule without breaking the covalent bonds. Therefore, denaturation can
mean a process of breaking up the hydrogen bonding, hydrophobic
interactions, salt bond and open folds of protein molecules.
Denaturation of proteins include disruption and damage that
may occur in the secondary and tertiary structure of proteins. Since it is known
denaturation reaction was not strong enough to break the peptide bond, where
the primary structure of proteins remained the same after denaturation process.
Denaturation due to interruption of the secondary and tertiary structure of
proteins. At the tertiary protein structure, there are four types of interactions
that form a bond in the side chain such as; hydrogen bonds, salt bridges,
disulfide bonds and hydrophobic non-polar interactions, which may be
impaired. Denaturation commonly encountered is the process of precipitation
and coagulation proteins.
Denaturation, coagulation and redenaturasi can be distinguished as
follows. Denaturation of proteins is a state has been a change in the protein
structure that includes changes in the shape and folds of molecules, without
causing disconnection or damage to the crease between the amino acids and the
primary structure of proteins. Coagulation is a denaturation of proteins by heat
and alcohol. Redenaturasi is denaturation of proteins that take place in
4

reveresibel. Heat can be used to disrupt hydrogen bonding and hydrophobic

non-polar interactions. This happens because the high temperatures can
increase the kinetic energy and causes the building blocks of protein molecules
move or vibrate so fast that disrupt the molecular bonds. Protein denaturation
and coagulated egg experienced during cooking. Some of the food is cooked to
denature the proteins contained in order to facilitate the digestive enzymes to
digest the protein. Warming will make denatured protein material so the ability
to bind water decreases. This happens because the heat energy will lead to the
dissolution of the non-covalent interactions that exist in the natural structure of
the protein but not sever ties kovalennya form peptide bonds. This process
usually takes place in a narrow temperature range.
The nature of the protein solubility is very dependent on the type of
protein. In addition, the type and kind of a suitable solvent also plays a role.
For example, albumin is soluble in water, acids, bases and aqueous salt
solution, can be coagulated by heat and can be precipitated by saturated salt
(ammonium sulfate), eg, serum albumin, lactalbumin (milk) and ovalbumin
(the egg).

CHAPTER III
RESEARCH METHODE
A.

Materials and Tools

Materials

1. Albumin solution 2%
2. NaOH 0,2%
3. NaCO3 0,2%
4. HCL solution 0,2%
5. Aquades
Tools
:

B.

1.

Reaction tube

2.

Pipettes

3.

Reaction tube rack

4.

Tweezers

5.

Measure glass

Procedure
1.
2.

3.

Preparing 4 reaction tubes, adding 1 ml albumin solution 2% on each test

tube.
Then, adding on test tube :
Tube 1 : 1 ml aquades
Tube 2 : 1 ml NaOH solution 0,2%
Tube 3 : 1 ml HCL solution 0,2%
Tube 4 : 1 ml NaCO3 solution 0,2%
Shaking each tube with tweezers until 1-2 minutes, let a while and
observe the result.

CHAPTER IV
RESULT AND DISCUSION

4.1

Result
Protein test
Protein
NUM.
Materials
Tested
1.
1 ml solution

Activities

Before

Vort

Albumin :

of albumin

eks

2% + 1 ml

during 1-

colorless
Aquades :

aquades

Experiment Result

2minutes

Let

After

Soluble
Colorless,th
ere is lumb in

colorless

surface (++)

a
moment
an
2.

1 ml solution

observe

Vort

of albumin

eks

2% + 1 ml

during 1-

solution of

2minutes

Let

NaOH 0,2 %

Albumin :
colorless
NaOH :

Soluble
Colorless,th
ere is not lumb

colorless

in surface

a
moment
an
3.

1 ml solution

observe

Vort

of albumin

eks

2% + 1 ml

during 1-

solution of

2minutes

Let

HCL 0,2 %

Albumin :
colorless
HCL : colorless

Soluble
Colorless,th
ere are many
lumb in surface
(+++)

moment
an
observe
7

1 ml solution

of albumin

eks

2% + 1 ml

during 1-

solution of

2minutes

Let

NaCO3 0,2

Albumin :

Vort

colorless
NaCO3 :

Soluble
Colorless,th
ere is little lumb

colorless

in surface (+)

moment
an
observe
Protein Extract Tested

NUM.
1

Protein Material Tested

Peanuts

Activity

Peanuts extract
+ aquades 1 ml

Vorte

Experiment Tested
Before

After

Peanuts

Soluble
White

ks during

extract :

1-

white

muddy

2minutes

Let a

muddy

solution
There is

moment

Aquades :

an observe

solution

no lumb

Vorte

colorless
Peanuts

+ HCL 0,2 % 1

ks during

extract :

ml

1-

white

muddy

muddy

solution
There is

Peanuts extract

2minutes

Let a
moment
an observe

solution
HCL :

Vorte

colorless
Peanuts

+ NaOH 0,2%

ks during

extract :

1 ml

1-

white

Peanuts extract

Soluble
White

no lumb

Soluble
Light
Yellow
muddy

2minutes
Let a

muddy
solution

moment

solution
There is
no lumb

an observe NaOH :
colorless
Peanuts extract

Vorte

Peanuts

Soluble
White

+ NaCO3 0,2

ks during

extract :

% 1 ml

1-

white

muddy

muddy

solution
There is

2minutes

Let a

solution

moment

no lumb

an observe NaCO3 :
colorless

Mung bean
2.

Mung
Bean

Vorte

Mung

White,

extract +

ks during

bean

there is

aquades 1 ml

1-

extract :

white

2minutes
Let a

white

sedime

Aquades :

nt

moment

Mung bean
extract + HCL
0,2 % 1 ml

colorless
an observe

Vorte Mung

ks during

bean

1-

extract :

2minutes
Let a

white

moment

colorless

ks during

bean

1-

extract :

2minutes

Let a
9

sediment

HCL :

an observe

Vorte Mung

Mung bean
extract +
NaOH 0,2% 1
ml

White, no

white
NaOH :

White, no
sediment

moment
Mung bean
extract +
NaCO3 0,2 %
1 ml

an observe

Vorte Mung
bean

color,

1-

extract :

white

white

sediment

moment
Tofu
extract

Tofu extract +

NaCO3 :
colorless

an observe

Vorte Tofu

aquades 1 ml

White

ks during
2minutes

Let a

3.

colorless

White

ks during

extract :

color (no

1-

white

lumb)

2minutes

Let a

Aquades :
colorless

moment
Tofu extract +

an observe
Vorte Tofu

HCL 0,2% 1

ks during

extract :

ml

1-

whit

2minutes

Let a
moment
Tofu extract +

HCl :
colorless

an observe

Vorte Tofu

NaOH 0,2% 1

ks during

extract :

ml

1-

white

2minutes

Let a

White
color
Have
white
lump

White
color
No lumb

NaOH :
colorless

moment
Tofu extract +

an observe

Vorte Tofu

NaCO3 0,2% 1

ks during

extract :

ml

1-

white

10

White
color
Have

2minutes
Let a

NaCO3 :

white

colorless

lumb

moment
4.

Fish
meat
extract

an observe

Vorte Fish meat

Fish meat
extract +

ks during

extract :

aquades 1 ml

1-

muddy

2minutes
Let a

cream

moment

Soluble
Muddy
yellowish

Aquades :
colorless

an observe

Vorte Fish meat

extract + HCL

ks during

extract :

Soluble
Muddy

0,2% 1 ml

1-

muddy

white

Fish meat

2minutes

Let a
moment

Fish meat

cream
HCL :
colorless

an observe
Vorte Fish meat

extract +

ks during

extract :

NaOH 0,2% 1

1-

muddy

ml

2minutes

Let a

cream
NaOH :

Yellowish
muddy
Found
residue on
the liquid

moment

colorless
an observe

Vorte Fish meat

Fish meat
extract +

ks during

extract :

NACO3 0,2%

1-

muddy

1 ml

2minutes

Let a

Yellowish
muddy
Found

cream

residue on

NaCO3 :

the liquid

moment

5.

Soybean cake
11

colorless
an observe

Vorte Soybean

Soluble

extract +
Soybean
aquades 1 ml
cake

ks during

cake

1-

extract :

2minutes

Let a

white

moment

Aquades :

an observe
Soybean cake

Vorte

cake

0,2% 1 ml

1-

extract :

2minutes
Let a

white

an observe
Soybean cake

Vorte

muddy (+)

muddy
HCl :
colorless
Soybean

Soluble
Brown

ks during

cake

NaOH 0,2% 1

1-

extract :

muddy (+

white

+++)

2minutes

Let a
moment
an observe

Soybean cake

Vorte

muddy
NaOH :
colorless
Soybean

extract +

ks during

cake

NaCO3 0,2% 1

1-

extract :

2minutes
Let a

white

moment

NaCO3:

ml

an observe
Chicken
heart
extract

Soluble
White

extract +
ml

6.

colorless
Soybean

ks during

moment

muddy (++)

muddy

extract + HCL

White

Chicken heart

Vorte

Soluble
White
muddy

muddy
colorless
Chicken

Soluble
Coffee

extract +

ks during

heart

aquades 1 ml

1-

extract :

brown

2minutes

maroon

color

12

Let a
moment

Chicken heart
extract + HCl
0,2% 1 ml

Aquades :
colorless

an observe
Vorte Chicken
ks during

heart

1-

extract :

2minutes

Let a

Solub
le
Coffe
e brown

maroon

color

HCl :

moment

Chicken heart

colorless
an observe

Vorte Chicken

Soluble
Coffee

extract +

ks during

heart

NaOH 0,2% 1

1-

extract :

brown

maroon

color

ml

2minutes

Let a

NaOH :

moment

Chicken heart

colorless
an observe

Vorte Chicken

extract +

ks during

heart

NaCO30,2% 1

1-

extract :

ml

2minutes

Let a

Solub
le
Coffe
e brown

maroon

color

NaCO3 :

moment

7.

Chicken
Meat
extract

Chicken meat

colorless
an observe

Vorte Chicken

extract +

ks during

meat

aquades 1 ml

1-

extract :

2minutes

Let a

Solub
le
Yello
w (+++)

Aquades :
colorless

moment
Chicken meat
extract + HCl
13

an observe

Vorte Chicken

Solub

0,2% 1 ml

ks during

meat

1-

extract :

2minutes

Let a

le
Yello
w+

HCl :
colorless

moment
Chicken meat
extract +
NaOH 0,2% 1
ml

an observe

Vorte Chicken
ks during

meat

1-

extract :

2minutes

Let a

Solub
le
Mudd
y

NaOH :
colorless

moment
Chicken meat

an observe

Vorte Chicken

extract +

ks during

meat

NaCO3 0,2% 1

1-

extract :

ml

2minutes

Let a

NaCO3 :

Solub
le
Yello
w ++

colorless

moment
an observe
B. Analysis
The results of the lab that we do that albumin dissolved in acidic
conditions, bases, salts and neutral. While the results for ekstract materials
containing proteins are as follows:

Peanuts extract become white muddy, and soluble in aquades, HCl,

NaOH and NaCO3 solution. And there is no lumb in solution

Mung bean extract become white color, in aquades and NaCO3 there is

white sediment. In HCl and NaOH there is no sediment.

Tofu extract become white color. In aquades and NaOH there is no lumb,
in HCl and NaCO3 have white lumb.

14

Fish meat extract become soluble and muddy yellowish in aquades

solution, while in HCL become soluble and muddy white, in NaOH
become yellowish muddy and found residue and in NaCO3 become

yellowish muddy, found residue on the liquid.

Soybean cake become soluble in all solution (acid, base, salt and netral).
In aquades become white muddy (++), in HCl become white muddy (+),

in NaOH become brown muddy, the last in NaCO3 become white muddy.
Chicken heart extract soluble in all solution (acid, base, salt and netral).
In aquades become coffee brown color, in HCl and NaOH become coffee

brown color, and in NaCO3 become brown color.

Chicken meat extract soluble in all solution (acid, base, salt and netral).
In aquades become soluble and yellow +++, in HCl become soluble and
yellow + and NaOH become oluble and muddy, and in NaCO3 become

C.

soluble and yellow ++

.
Discussion Based on Data
In the results table above shows that the solubility properties of the
protein depends on the type of protein. In addition, the type and kind of a
suitable solvent also plays a role. In the observations are that most of the test
material is not dissolved. For example in mungbean in the addition of HCl,
NaOH, and distilled NaCO3 color changes to white and cause sediment,
besides mungbean extract is the extract fishmeat the addition of NaOH and
NaCO3 also produce sediment.
In a test that produces sediment that may already dissolved but in early
extract made if left for a moment will produce smooth deposits. And the
differences are probably at the time of observation or experiment we are less
careful in adding a solution, and also in the process of vortex less than perfect.

D. Discussion Based on Question

Why is the nature of the protein solution depends on the type of protein as well as
the type and kind of solvent?
Because basically proteins are amphoteric, ie soluble in acid or alkaline
but not the protein soluble in fat solvents. Such as ether and chloroform ..
However, the solubility of the protein varies depending on the amount and
15

location of the amino and carboxyl groups in the molecule. Protein solubility
also depends on the solution PHN. In acid solution (low pH), the amino group
reacts with H +, so that the protein is positively charged. Conversely, in an
alkaline solution (pH high) protein molecules will react as acids or negatively
charged. At pH isolistrik charge free amino and carboxyl neutralize each other
so that the charged molecules zero

16

CHAPTER V
CLOSING
A.

Conclusion
Albumin is soluble in acidic conditions, alkaline, neutral or salt. This is
evidenced by the absence of sediment or separation of albumin solution when
mixed with acid solvents, bases, salts and neutral. However, the solubility of a
protein varies depending on the type of protein, pH of the solution, and the type
of solvent.

B.

Suggestion
In this experiment we still containing an human errors. Therefore, it is
expected for the next experiment we can be more accurate in observation,
especially observation of color change. In addition, before experiment, tools
provided should in a clean and dry condition so that no contamination.

17

BIBLIOGRAPHY
http://nazirwanabdi.blogspot.com/2012/12/laporan-praktikum-biokim-protein.html
diakses pada tanggal 27 november 2014 pukul 13.00
http://www.slideshare.net/fitriasaid/laporan-tetap-biokim-1-reaksi-uji-terhadap-protein
diakses pada tanggal 27 november 2014 pukul 13.50
http://haiyulfadhli.blogspot.com/2011/06/kelarutan-protein.html diakses pada tanggal
27 november 2014 pukul 13.55
http://budikolonjono.blogspot.com/2010/12/protein-uji-kualitatif-indetifikasi-uji.html
diakses pada tanggal 27 november 2014 pukul 14.20

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