Kelarutan Albumin INDRIE

CHAPTER IINTRODUCTION1. 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 IILITERATUREA. 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)

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 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 IIIRESEARCH 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

:

1.Reaction tube2.Pipettes3.Reaction tube rack4.Tweezers5.Measure glassB.

Procedure1. Preparing 4 reaction tubes, adding 1 ml albumin solution 2% on each test tube.2. 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%

3. Shaking each tube with tweezers until 1-2 minutes, let a while and observe the result.

CHAPTER IVRESULT AND DISCUSION

4.1Result

Protein test

NUM.Protein Materials TestedActivitiesExperiment Result

BeforeAfter

1.1 ml solution of albumin 2% + 1 ml aquades Vorteks during 1-2minutes

Let a moment an observe Albumin : colorless

Aquades : colorless Soluble

Colorless,there is lumb in surface (++)

2.1 ml solution of albumin 2% + 1 ml solution of NaOH 0,2 % Vorteks during 1-2minutes


NaOH : colorless Soluble

Colorless,there is not lumb in surface

3.1 ml solution of albumin 2% + 1 ml solution of HCL 0,2 % Vorteks during 1-2minutes


HCL : colorless Soluble

Colorless,there are many lumb in surface (+++)

41 ml solution of albumin 2% + 1 ml solution of NaCO3 0,2 % Vorteks during 1-2minutes


NaCO3 : colorless Soluble

Colorless,there is little lumb in surface (+)

Protein Extract Tested

NUM.Protein Material TestedActivityExperiment Tested

BeforeAfter

1PeanutsPeanuts extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Peanuts extract : white muddy solution

Aquades : colorless Soluble White muddy solution

There is no lumb

Peanuts extract + HCL 0,2 % 1 ml Vorteks during 1-2minutes


HCL : colorless Soluble

White muddy solution

There is no lumb

Peanuts extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes



Light Yellow muddy solution

There is no lumb

Peanuts extract + NaCO3 0,2 % 1 ml Vorteks during 1-2minutes

Let a moment an observe Peanuts extract : white muddy solution NaCO3 : colorless Soluble

White muddy solution

There is no lumb

2.Mung BeanMung bean extract + aquades 1 ml Vorteks during 1-2minutes Let a moment an observe Mung bean extract : white

Aquades : colorless White, there is white sediment

Mung bean extract + HCL 0,2 % 1 ml Vorteks during 1-2minutes

Let a moment an observe Mung bean extract : white

HCL : colorless White, no sediment

Mung bean extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes


NaOH : colorless White, no sediment

Mung bean extract + NaCO3 0,2 % 1 ml Vorteks during 1-2minutes


NaCO3 : colorless White color, white sediment

3.Tofu extractTofu extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Tofu extract : white

Aquades : colorless White color (no lumb)

Tofu extract + HCL 0,2% 1 ml Vorteks during 1-2minutes

Let a moment an observe Tofu extract : whit

HCl :

colorless White color Have white lump

Tofu extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes

Let a moment an observe Tofu extract : white NaOH : colorless White color No lumb

Tofu extract + NaCO3 0,2% 1 ml Vorteks during 1-2minutes

Let a moment an observe Tofu extract : white

NaCO3 : colorless White color Have white lumb

4.Fish meat extractFish meat extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Fish meat extract : muddy cream


Muddy yellowish

Fish meat extract + HCL 0,2% 1 ml Vorteks during 1-2minutes


HCL : colorless Soluble Muddy white

Fish meat extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes


NaOH : colorless Yellowish muddy Found residue on the liquid

Fish meat extract + NACO3 0,2% 1 ml Vorteks during 1-2minutes


NaCO3 : colorless Yellowish muddy Found residue on the liquid

5.Soybean cake Soybean cake extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Soybean cake extract : white muddy

Aquades : colorless Soluble White muddy (++)

Soybean cake extract + HCL 0,2% 1 ml Vorteks during 1-2minutes


HCl : colorless Soluble White muddy (+)

Soybean cake extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes


NaOH : colorless Soluble Brown muddy (++++)

Soybean cake extract + NaCO3 0,2% 1 ml Vorteks during 1-2minutes


NaCO3: colorless Soluble White muddy

6.Chicken heart extractChicken heart extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Chicken heart extract : maroon


Coffee brown color

Chicken heart extract + HCl 0,2% 1 ml Vorteks during 1-2minutes


HCl : colorless Soluble Coffee brown color

Chicken heart extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes


NaOH :

colorless Soluble Coffee brown color

Chicken heart extract + NaCO30,2% 1 ml Vorteks during 1-2minutes


NaCO3 : colorless Soluble Coffee brown color

7.Chicken Meat extractChicken meat extract + aquades 1 ml Vorteks during 1-2minutes

Let a moment an observe Chicken meat extract :


Yellow (+++)

Chicken meat extract + HCl 0,2% 1 ml Vorteks during 1-2minutes


HCl : colorless Soluble

Yellow +

Chicken meat extract + NaOH 0,2% 1 ml Vorteks during 1-2minutes



Muddy

Chicken meat extract + NaCO3 0,2% 1 ml Vorteks during 1-2minutes


NaCO3 : colorless Soluble

Yellow ++

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. 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 soluble and yellow ++.C.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 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 zeroCHAPTER 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.BIBLIOGRAPHYhttp://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 13.50http://haiyulfadhli.blogspot.com/2011/06/kelarutan-protein.html diakses pada tanggal 27 november 2014 pukul 13.55http://budikolonjono.blogspot.com/2010/12/protein-uji-kualitatif-indetifikasi-uji.html diakses pada tanggal 27 november 2014 pukul 14.202

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Kelarutan Albumin INDRIE