Etiket Arşivleri: rennin
Isolation of casein from milk
hydrolyze to yield only amino acids e.g.: albumins , globulins.
is a protein that functions in interaction with other chemical groups attached by covalent bonds or by weak interactions.
Some examples of conjugated proteins are lipoproteins, glycoproteins, phosphoproteins, hemoproteins
Phosphoproteins are proteins that are physically bonded to a substance containing phosphoric acid. Ionized phosphate groups by calcium ion increases hydrophobic interaction, which leads to precipitation.
Whole milk contains vitamins (vitamins A, D, and K), minerals (calcium, potassium, sodium, phosphorus,), proteins (which include all the essential amino acids), carbohydrates (chiefly lactose), and lipids.
The only important elements in which milk is seriously deficient are iron and Vitamin C.
Infants are usually born with a storage supply of iron large enough to meet their needs for several weeks. Vitamin C is easily secured through an orange juice supplement.
There are three kinds of proteins in milk:
3.lactoglobulins. All are globular
Structure of casein
Is a phosphoprotein, which has phosphate groups attached to some of the amino acid side chains. Mostly these amino acid are serine and threonine.
casein is a mixture of at least three similar proteins, which differ primarily in molecular weight and amount of phosphorus they contain (number of phosphate groups).
Casein is made up of the main 3 types of proteins are α-casein, β-casein, and κ-casein.
all casein proteins have different hydrophobic and hydrophilic regions along the protein chain.
α-Caseins are the major casein proteins. Its containing 8-10 phosphate groups,
β- casein contains about 5 phosphate residues,
β- casein it is more hydrophobic than α-caseins and κ-casein
Because α-caseins and β-caseins are highly phosphorylated, they are very sensitive to the concentration of calcium salts, that is, they will precipitate with excess Ca2+ ions
Unlike other caseins, κ-caseins are glycoproteins, and they have only 2 phosphate group.
Hence, they are stable in the presence of calcium ions, and they play an important role in protecting other caseins from precipitation and make casein more soluble forming casein micelles.
Neither the α nor the β casein is soluble in milk, singly or in combination.
If κ casein is added to either one, or to a combination of the two, however, the result is a casein complex that is soluble owing to the formation of the micelle
Carbohydrate that found k casein :
The main carbohydrate present in milk is the sugar lactose.
Lactose is a disaccharide containing the monosaccharaides galactose and glucose.
when bacteria get into milk, they digest the lactose and form the acid lactic acid. This causes lowering the pH and cause a precipitation of the protein casein.
The word “micelle” is a chemical term. It is used to describe the structure that certain very large molecules
Very large molecules are considered to be too large to be truly soluble in water.
Instead, these large molecules will form structures that allow them to remain suspended in water
The dispersion of these large structures in water is known as a colloidal suspension.
The structures that allow large molecules to remain colloidally suspended in water are termed micelles.
In the case of casein, the parts of the casein molecules that have an affinity for water form the outside of the casein micelle.
Conversely, the parts of the casein molecule that are hate the water form the inner core of the micelle spheres
Casein micelles consist of water, protein, and salts.
Casein is present as a caseinate, which means that it binds primarily calcium and magnesium.
The pI of most proteins ranges between the pH 4 to 6.
So pI for casein is 4.6 and the pH of milk is 6.6 at this pH the casein is have negative charge and soluble by bacteria action or by adding acid that lowering the PH cause ppt of casein
Casein can be precipitated by rennin
Renin enzyme :
Found in calves and gout
Is hydrolysis enzyme (peptidase) so cleave the peptide bond between amino acid
1.Enzyme attack the k- casein and breaking the peptide bond and release small part of peptide bond
2.This destroy cause left k-casein the α and B casein which protect the casein from ppt and ppt it as Para kappa casein
3.Unlike kappa the Para kappa casein cannot prevent the casein from ppt in the present of calcium ion
If calcium removed from milk the renin cannot form ppt to milk
nWhen the pH level decreases the milk forms Curds or whey.
nCurds are a dairy product obtained by ppt of milk with renin or an acidic substance and then draining off the liquid portion.
Whey or milk serum is the liquid remaining after milk has been coagulation and remove curds
1.Place 20 ml (20 g) of milk into a 125 ml flask and heat at 40 oC in a water bath.
2.Add 5 drops of glacial acetic acid and stir for about 1 min.
3.Filter the resulting mixture through filter paper held in a funnel and gently squeeze out most of liquid.
4.Remove the solid (casein and fat) from the cheesecloth, place it into a 100 ml beaker and add 10 ml of 95% ethanol.
5.Stir well to break up the product. Pour off the liquid and add 10 ml of 1:1 ether-ethanol mixture to the solid.
6.Stir well and filter through filer paper
7.Let the solid drain well, then scrape it into a weighed filter paper and let it dry in the air.
8.Calculate the casein percentage in milk as follows:
% Casein = (grams of casein \ grams of milk) x 100
Normal Range 3-5 %
Cheese is defined as the fresh or matured product obtained by draining the whey after coagulating casein, the major protein in milk. The casein is coagulated by acid from selected m/o and / or by milk-clotting enzymes added to the milk. The resulting curd is cubed, cut, heated, drained and salted. Fresh and uncured cheese, such as cottage and cream, can be eaten immediately.
Ripening or uncuring of the knitted or streched curd includes exposure to a temperature humıdıty controlled environment for a specified length of time. Changes during curing are bought about by specially selected enzymes, bacteria, mold, yeast or combinations of these added curing agents in or on the cheese. During ripening nutrients such as protein fat and carbohydrate (lactose) are changed to simpler compounds thatproduce the chacteristic flavour and affect the final texture of the cheese.
We made the cheese using two different methods. One of both is using lactic acid, other is using rennin. In rennin method, we applied pasteurization method except boiling. Also after added rennin we incubated the milk. However we didn’t apply incubation process to the milk added lactic acid. In lactic acid method we observed coagulum, while adding lactic acid to the
There was not big difference in yield between these two methods. But the yield obtained obtained from milk was lower than the others for our sample. This is because of the quality of milk and changes from milk to milk. In acidity, there are some contradictions. In lactic acid method, lactic acid level in whey was bigger than curd’s. But in rennin method, L.A level of whey was smaller than curd’s. On the other hand, the pH level in lactic acid method was smaller than rennin’s. Also pH of whey was smaller than curd in these both methods. All cheese dishes should be cooked at a low temperature for a short time. Excessive heat and overcooking cause fat seperation, stringing, and the tougheningof the cheese.