Casein (casein protein). Meaning of casein hydrolyzate in the drug formulary

Recently, young children often suffer from allergic reactions to any food, including milk. Usually cow's milk allergy is caused by lactose intolerance, so mothers need to carefully monitor the health of the baby and his diet. In the presence of allergies, doctors prescribe hydrolyzed formulas if breastfeeding is difficult or impossible. Such mixtures are popular due to their hypoallergenic formula and the main component - protein hydrolyzate. Mixtures can be used for children who are allergic to any products, including cow's milk.

When are hydrolyzed formulas prescribed for children

Infant formula, regardless of its type, can be used as main food or complementary foods. Usually, mixtures are divided into categories: as an independent source of nutrition from birth to six months of the child, and as complementary foods from 6 months to 1.5 years.

Mixtures with protein hydrolyzate are prescribed by doctors; parents themselves cannot transfer the child to these types of mixtures. Protein hydrolyzate is a therapeutic component, therefore mixtures based on it are prescribed for children at risk or who already have any allergies. A special indication for the use of these mixtures is sensitivity and allergy to cow's milk and even soy. To understand that the child has an allergy, the mother must monitor the baby's reaction to cow's milk. Usually, allergies appear in the form of rashes, swelling and asthma. Therefore, at the first signs, the mother should urgently consult a doctor.

Also, mixtures with protein hydrolyzate are prescribed for skin diseases, gluten intolerance, dystrophic changes in the intestines and for prevention in children who are at an increased risk of allergic diseases. Instead of cow's milk, whey protein or casein is included in the mixture with protein hydrolyzate, which are much easier to digest for the child's body. Protein hydrolyzate mixtures are divided into types, depending on the degree of hydrolysis. So, there are highly hydrolyzed mixtures, partially and low hydrolyzed mixtures.

During hydrolysis, different protein sizes can be obtained, on which the degree of hydrolysis of the mixture depends. So, the protein can be large, because of this, the degree of hydrolysis will be low, and the child’s allergic reactions will not go away if this protein size does not suit him. The average protein size is obtained by partial hydrolysis of the mixture, and the smallest protein is obtained by highly hydrolyzed mixtures.

Most often, it is a high degree of hydrolysis that children with allergies need, but if the doctor prescribes a different type of mixture, this means that the allergic reaction is not strong, and the body will be able to absorb a different size of protein. When choosing a mixture, you need to consider the presence of carbohydrates and lactose, as usually An allergy to cow's milk appears due to lactose intolerance.

Rating of the best mixtures with protein hydrolyzate

Often the mixture includes beneficial prebiotics and bifidobacteria, which improve the baby's digestion. Such mixtures are used by children who have stomach and digestive disorders. In order for parents to understand which mixtures are considered the most effective for allergies in children, it is necessary to compile a rough list of the most popular mixtures with protein hydrolyzate. So, in the top three are mixtures of Alimentum, Alfare and Good Start. Next on the list is Damil Pepti, Nan and Nutramigen. Nutrilac and Nutrilon mixtures are also quite effective and useful; parents often prefer them. These types of mixtures differ from each other, but they are all hypoallergenic and contain protein hydrolyzate.

Usage: in the food industry when receiving baby food. The essence of the invention: casein or caseinate is dissolved in an aqueous medium to obtain a suspension or solution and proteolytic hydrolysis is carried out by means of a non-static pH method using one or more neutral Bacillus endoproteases at a concentration of at least 0.005 units. Ansona per 100 g of protein, one or more Aspergillus exoproteases at a concentration corresponding to at least 1000 peptidase units per 100 g of protein, one or more Bacillus alkaline endoproteases at a concentration of at least 0.005 units. Anson per 100 g of protein at a temperature of 45-60 o C to the degree of hydrolysis of 15-35%. The hydrolysis is terminated by enzyme inactivation and the product is dried. 2s. and 11 z.p. f-ly, 6 ill.

The invention relates to a casein hydrolyzate and a method for its production. Casein hydrolysates are mainly used as baby food ingredients. There are various types of casein hydrolysates and methods for their preparation. In order to obtain the most effective results regarding the type of casein hydrolyzate and methods of its preparation, the following four factors are of greatest importance: the value of DH (degree of hydrolysis), which leads to the formation of shorter peptides in the product, and therefore to low allergenicity; low content of free amino acids, which determine the low osmolality characteristic, which is considered preferable if the product is introduced into food products; reduced bitterness; high output. Most processes for producing casein hydrolysers with good organoleptic properties give low yields. It is widely believed that it is very difficult to achieve an optimal balance between these four factors. The aim of the invention is to present a casein hydrolyzate and a method for obtaining such a hydrolyzate with optimal properties, i.e. low DH value, reduced free amino acids, slight bitter taste and high yield. In accordance with the invention, it has been found that a specific combination of specific enzymes and hydrolysis at variable pH results in a casein hydrolyzate with an optimal balance between degree of hydrolysis, free amino acids, bitterness and yields. The casein hydrolyzate according to the invention does not contain any amount of non-hydrolyzed casein and is characterized in that it completely dissolves or almost completely dissolves in an aqueous medium at a pH of 3.5 to 7, has good organoleptic properties and contains peptides in relative amounts corresponding to the following distribution according to molecular weight (MW), wt. MW > 5000 0-1 5000 > MW > 1500 15-35 1500 > MW > 500 40-60 500 > MW 15-35, and free amino acids in an amount of less than 10% and also because the average molecular weight (Mn) is 400 -650 (by quantity). 1. The principle of receiving. The sample is dissolved, filtered and injected into a liquid chromatographic system using the gel permeation chromatography method. This separation technique makes it possible to pass a liquid stream through a column filled with porous particles having a well-defined pore diameter. When a solution of peptides with different molecular sizes passes through the column, small peptides pass through the pores, while larger peptides are delayed in passing. Thus, the peptides in the solution are distributed according to the size of the molecules (by molecular weight), since larger peptides are more quickly washed out of the column than smaller peptides. A detector installed at the outlet of the column continuously measures the outlet flow. The chromatographic system is calibrated using peptides of known molecular weight. 2. Chromatographic equipment. 2.1. High performance liquid chromatography. The assay system consists of a high pressure pump, Waters M 510, flow rate 0.7 ml/min, injector, Waters WISP M 710 detector, Waters M 400 with wavelength up to 214 nm. 2.2. Column for analysis by gel permeation chromatography, 3xTSKG2000 SWXL, connected in series and operating at ambient temperature. 2.3. Waters 820 MAXIMA SIM system for processing chromatographic data, integrated into a single complex, with 810/820 GPC modifications. 3. Reagents. 3.1. Phosphate buffer, NaH 2 PO 4 2H 2 O 3.2. Ammonium chloride, NH 4 CI 3.3. Trifluoroacetic acid (TFA), CF 3 COOH 3.4. Acetonitrile, CH 3 CN 3.5. Mobile phase: 0.05 M phosphate buffer/0.5 M ammonium chloride solution containing 0.1% TFA and 25% acetonitrile. 4. Description. 4.1. Calibration. The chromatographic system is calibrated by introducing a large amount of peptides of known molecular weight. Plot the molecular weight of each standard against the marked volume of mobile phase required to elute from the column. The least squares method calculates the largest polynomial of the third degree. The curve represents the calibration curve. 4.2. Analysis. The sample is diluted/dissolved in the mobile phase to approximately 55 mg/mL. The solution is filtered through a 22 µm filter and 20 µl is used to inject into the chromatograph. Register the response of the detector (reaction) to the eluted volume. The recorded chromatogram curve reflects the actual distribution of molecular weights in the sample. To calculate the total weight distribution and average molecular weight, the chromatogram is divided into small time intervals (and elution volumes) segments, each segment is characterized by the volume of extraction of the target fraction and the area of the chromatogram over the elapsed time period. 5. Estimated data. Results are given in terms of weight and number average molecular weights.
where m w: weight average molecular weight,
m n: number average molecular weight,
A i: area of the chromatogram for each segment, measured as the overall response (response) of the detector for each time interval,
M wi is the corresponding molecular weight for each segment. Values are calculated using a calibration curve using the average volume of the recovered target fraction over a certain period of time. A preferred embodiment of the casein hydrolyzate of the present invention is characterized in that the casein hydrolyzate is obtained from precipitated rennet casein, that it contains peptides in relative amounts corresponding to the following molecular weight distribution, wt. MW > 5000 0-0.2
MW > 3000<5
1500 > MW > 500 40-60
500 > MV 15-35
as well as free amino acids in an amount of less than 10% and that the number average molecular weight (M n) is 400-650. In the embodiment represented by the invention, the molecular weight of the hydrolyzate is very different, since it contains a relatively small amount of long peptides. The absence of high molecular weight peptides reduces antigenicity. This effect is of exceptional importance in relation to the use of rennet-derived casein hydrolyzate as an ingredient in breast milk substitutes where low antigenicity is highly desirable. At the same time, the digestibility of the hydrolyzate is also improved, causing a reduction in the occurrence of colic phenomena. The subject matter of the invention thus presented is in an infant formula or breast milk substitute. A preferred embodiment of the casein hydrolyzate provided by the present invention is characterized in that the casein hydrolyzate is completely soluble in an aqueous medium at a pH of 3.5 to 7.0. In view of the complete solubility of the casein hydrolyzate, the latter is very suitable as a dietary component. In addition, the invention relates to a fully soluble dietary formulation, highly stable at low pH values, which contains the casein hydrolyzate variant according to the present invention as a protein source. Such a fully soluble dietary composition based on the subject matter of the invention will prevent the process of protein clotting in the stomach, which is a very serious problem with conventional tube feeding products. The method according to the invention for the production of casein hydrolyzate is characterized in that casein or caseinate with at least 85% protein, based on the dry product, is suspended/dissolved in an aqueous medium with a protein content of up to about 20%, preferably up to 10% suspension/solution from step ( 1) in a one-step reaction, proteolytically hydrolyze to a degree of hydrolysis of 15-35%, preferably 22-28%, with three groups of proteases, which are one or more neutral endoproteases from Bacillus at a concentration of at least 0.005 Anson units per 100 g of protein, one or more Bacillus endoproteases at concentrations of at least 0.005 Anson units per 100 g of protein and one or more Aspergillus exoproteases at concentrations corresponding to at least 1000 peptidase units per 100 g of protein at a temperature of 45 to 60 o C according to the method with variable pH, that the hydrolysis process is terminated by enzyme inactivation, and that the effluent from step 3/ is converted to a dry phase. US Pat. No. 3,761,353 describes a casein hydrolyzate that uses milk protein as a raw material. This casein hydrolyzate, however, is obtained in lower yields than according to the process according to the invention. In addition, the prototype does not use the same combination of proteolytic enzymes as specified in the present invention. EP 384 303 describes a process for the preparation of a protein hydrolyzate, which may be a casein hydrolyzate. Further, despite the fact that the resulting casein hydrolyzate is characterized by reduced bitterness, it was found that the degree of hydrolysis according to this method is 4.4%, while a similar characteristic when working according to the present invention is 15-35%. In addition, in accordance with according to the method proposed in the patent, it is necessary to maintain a constant pH during hydrolysis (see page 6, line 35), while the hydrolysis in accordance with the invention is carried out at a variable pH. EP 223 560 describes a process for the preparation of a protein hydrolyzate, which may be a casein hydrolyzate, by sequential hydrolysis. Hydrolysis according to the method of the invention can be carried out as a one-step reaction and, moreover, the prior art does not provide for the specific combination of proteolytic enzymes that is used in the method of the invention. The patent describes a protein hydrolyzate obtained by using a different combination of proteolytic enzymes than the corresponding combination in the process of the invention. In addition, milk protein hydrolyzate is used as an emulsifier, while casein hydrolyzate according to the invention is used as a food additive (closest analogue). It is understood that the term "neutral endoprotease from Bacillus" means a neutral endoprotease derived from Bacillus. In addition, endoproteases are identical to the group of enzymes that are obtained by cloning in other recipients. The same interpretation is used for similar terms, such as "exoprotease from Aspergillus". A typical example of a neutral endoprotease from Bacillus can be considered "Neutraza" manufactured by Novo Nordic A / S, typical examples of alkaline endoproteases from Bacillus are alkalase, esperase and savinase of the same production, and typical examples of exoprotease from Aspergillus are "Novozym-515", also manufactured by Novo Nordic A/S. No upper limits are given for the concentration of the three enzymes, but it is clear that the upper limits differ in the amount of enzyme that detracts from the organoleptic properties of the product or that makes an uneconomical process undesirable. In accordance with the method of the invention, the treatment with charcoal is carried out to improve the organoleptic properties. Such processing can be carried out either as a separate stage, or at one of the specified stages of production. When using the ultrafiltration processing method, the carbon can be added optionally before ultrafiltration, and the used carbon is automatically separated from the reaction mixture at the ultrafiltration stage, since the desired product is the filtrate. If the ultrafiltration method is not used, the activated carbon treatment is carried out as a separate step. Enzyme inactivation (step 3) can be carried out by lowering the pH, preferably around 4.5, whereby the final product is immediately acceptable for use as a thirst inducing agent, such as orange juice, and/or by raising the temperature. When carrying out inactivation by lowering the pH value, it was found that charcoal treatment is not necessary to improve the organoleptic properties of the product. A preferred embodiment of the process according to the invention is characterized in that the three groups of proteases in the second step are:
1) one or more neutral endoproteases from Bacillus subtilis,
2) one or more alkaline endoproteases from Bacillus lichenformus,
3) one or more exoproteases from Aspergillus oryzae. It has been established that the casein hydrolyzate obtained in accordance with the invention has higher organoleptic properties. The preferred embodiment of the claimed method also differs in that the mixture from stage (2) before the hydrolysis is completed in stage (3) or after the hydrolysis is completed in stage (4) is separated by processing on ultrafiltration or microfiltration equipment to obtain a filtrate containing casein hydrolyzate. Thus, a completely soluble casein hydrolyzate is obtained. Since ultrafiltration membranes with separation values below 5000 are very rare, and because only one percent of casein hydrolyzate shows a molecular weight above 5000, the separation values of the ultrafiltration membrane used in this embodiment of the invention are in principle irrelevant. Higher separation values are preferred, although associated with higher flow. A preferred embodiment of the claimed process is also characterized in that the hydrolysis in step (2) is carried out for less than 6 hours. With this embodiment, it is not necessary to filter the raw material to ensure its microbiological stability. Preferably, the inactivation of the enzymes is carried out by treatment with a food grade acid, preferably chlorine or citric acid. This variant is simple in itself and does not involve the use of activated carbon for cleaning purposes and furthermore makes it possible to obtain a product exhibiting pH values in the range of 3.5 to 7.0 when dissolved in water. Preferably, the enzyme inactivation is also carried out by a heat treatment method, and the liquid stream leaving stage (3) is treated with activated carbon, which is then removed, while the liquid stream purified from activated carbon in stage (4) is converted into a solid phase. Such a casein hydrolyzate is particularly suitable as an ingredient in baby food. A preferred embodiment provides that step (4) is carried out using a combination of hyperfiltration and/or evaporation followed by spray drying. Hyperfiltration is most beneficial at a concentration of 20-30 o B. In addition, unwanted salts can be removed. Spray drying produces an end product that is easy to process. According to another preferred embodiment of the method, acid-precipitated casein is used as the starting material, and this casein is dissolved using a base. This embodiment makes it possible to use the cheapest raw materials. This preferred embodiment is further characterized in that the acid-precipitated casein is dissolved with Ca(OH) 2 . Thus, a casein hydrolyzate having excellent organoleptic properties can be obtained. According to another preferred process variant, rennet-precipitated casein is used as starting material and this casein is dissolved with sodium phosphate or sodium carbonate. In this embodiment, the molecular weight of the hydrolyzate is very different because the product contains relatively low amounts of long peptides. This effect is particularly important in relation to the use of rennet casein hydrolyzate as an ingredient in breast milk substitutes, where low antigenicity is particularly desirable. The final product was found to have the following molecular weight distribution, wt. MW > 5000 0-0.2
MW > 3500<5
1500 > MW > 500 20-60
500 > MV 15-35,
contains free amino acids in an amount of less than 10% and that the amount of number average molecular weight is 400-600 (Mn). A preferred embodiment of the process according to the invention is characterized in that the rennet-precipitated casein is dissolved with sodium phosphate or sodium carbonate. In this case, rapid hydrolysis occurs and high yields of the product are obtained. The present invention is illustrated by the following examples. Example 1 Calcium caseinate produced by MD-Foods Denmark (Miprodan 40) containing about 87% protein was used as a raw material for hydrolysis. The caseinate is suspended in deionized water at 50° C. at concentrations of 8% protein. Record pH value, osmotic pressure characteristic and Brix: pH 6.96, osmotic pressure characteristic 24 mosmol/kg and 0 Brix equal to 7.20. Add enzyme:
- Neutrase 0.5 l containing 2.0% protein,
- alkalase 2.4 l containing 0.5% protein,
- Novozyme 515 containing 0.8% protein. Hydrolysis is carried out for 6 hours at a temperature of 50 o C, controlling the pH value, osmotic pressure and Brix. In the final hydrolysis step, the pH is 5.91, the osmotic pressure is 222 mosmol/kg, the Brix is 12.00 o V and the degree of hydrolysis is respectively 26.2% The hydrolysis process is completed by heat treatment for 3 minutes at 85 o C. Separation of the hydrolyzed mixture carried out by processing on PCI ultrafiltration equipment with FP 100 membranes installed on it (molecular weight separation is 100,000). The yield after ultrafiltration is > 93% relative to the amount of protein in the crude product. The product is concentrated using a PCI nanofiltration processing system fitted with AFC30 membranes. The overall yield after completion of this processing step is 90.7%. The concentrate is obtained by spray drying. The resulting product is a completely soluble powder containing 91% protein in dry matter, the molecular weights of which are distributed in accordance with Fig.1. Number average Mn=516, free amino acids 7% The product is completely soluble at pH 3.5 to 7.0. The pH value in a 5% solution of the product is 6.45. Example 2 Calcium caseinate produced by MD-Foods Denmark (Miprodan 40) containing about 87% protein was used as a feedstock for hydrolysis. Caseinate suspended in deionized water at 50 o C to a concentration of 8% protein. The pH value is recorded, the osmotic pressure and Brix:pH is 6.96, the osmotic pressure characteristic is 24 mosmol/kg, o Brix is 7.20. Add enzyme:
- Neutrase 0.5 l containing 2.0% protein,
- alkalase 2.4 l, in the amount of 0.5% protein,
- Novozyme 515 with 0.8% protein. The hydrolysis is carried out for 6 hours at 50° C. while controlling the pH value, the osmotic pressure and the Brix value. The final hydrolysis step is pH 5.91, osmotic pressure 222 mosmol/kg, Brix 12.00° B and degree of hydrolysis 26.2% The hydrolysis process is completed by heat treatment for 3 minutes at 85° C. 12EW) in the amount of 4% B o . Separation of the hydrolyzed mixture is carried out by processing on PCI ultrafiltration equipment with FP 100 membranes installed on it (molecular weight separation is 100,000). The yield after ultrafiltration treatment is > 93% relative to the protein in the crude product. The resulting product is concentrated using for this purpose PCI nanofiltration equipment with AFC30 membranes installed on it. The overall yield after completion of this processing step is 90.7%. The concentrate is obtained by spray drying. The resulting product is a fully soluble powder containing 91% protein on a dry matter basis, having the molecular weight distribution shown in FIG. Average value (Mn 564). The content of free amino acids is 7%. The taste of the product is described as milder than the taste of the product according to example 1. The product dissolves completely at a pH of 3.5 to 7.0. The pH value in a 5% solution of the product is 6.38. EXAMPLE 3 A product manufactured by MD-Foods Denmark (Miprodan 26) containing about 87% protein was used as a raw material for hydrolysis. Casein suspended in deionized water at 75 o C to a concentration of 8% protein. To dissolve the casein, 2% disodium diphosphate and 1% monosodium diphosphate are added per amount of protein. Complete dissolution occurs after 60 minutes. The mixture is cooled to 50° C. The pH value, osmotic pressure and Brix value are recorded: pH 6.95, osmotic pressure is 40 mosmol/kg and o Brix is 11.4. Add enzyme:
- Neutrase 0.5 l, with 2% protein
- Novozyme 515 with 0.5% protein
The hydrolysis is carried out for 6 hours at a temperature of 50 o C, controlling pH, osmotic pressure and Brix value. At the end of the hydrolysis process, the pH is 6.15, the osmotic pressure is 211 mosmol/kg, and the Brix value is 12.00 o V. The complete hydrolysis process is completed by heat treatment for 3 minutes at 85 o C. Separation of the hydrolyzed mixture is carried out by processing on PCI ultrafiltration equipment with FP 100 membranes installed on it (molecular weight division is 100,000). Yield after ultrafiltration treatment is > 80% for protein in the crude product. The product is concentrated using for this purpose a PCI nanofiltration processing system with installed AFC30 membranes. The overall yield after this treatment step is 77.5%. Activated carbon (Picatif 120EW) is added at 4% B o to the concentrate, after which it is filtered on a filter plate and spray dried. The resulting product is a fully soluble powder containing 91.3% protein on a dry matter basis with the molecular weight distribution shown in FIG. The number average molecular weight Mn is 496. The content of free amino acids is 5%. The taste of the obtained product is more pleasant than that of the product from example 1. The product dissolves completely at a pH of 3.5 to 7.0. The pH value of a 5% solution of the product is 6.50. Example 4 Na-caseinate produced by MD-Foods Denmark (Miprodan 30) containing about 87% protein was used as the material for hydrolysis. Caseinate suspended in deionized water at 50 o C to a concentration of 8% protein. Add enzyme:
- Neutrase 0.5 l with the amount of protein 2.0%
- alkalase 2.4 l with 0.5% protein
Hydrolysis is carried out for 6 hours at 50° C. Parallel hydrolysis is carried out under the same conditions. Only instead of the above species, calcium caseinate (Miprodan 40) is used. Analysis of the mixture after hydrolysis, heat treatment and filtration shows that the hydrolyzate obtained from calcium caseinate is significantly lower in taste than the hydrolyzate obtained from sodium caseinate. The product dissolves completely at pH 3.5 to 7.0. Example 5 In order to study the effect of solubility on hydrolysis efficiency, the degree of hydrolysis of rennet casein as a starting material was examined. As a starting material for hydrolysis, a product manufactured by MD-Foods Denmark (Miprodan 26) containing about 87% protein is used. Casein suspended in deionized water at 75 o C to a concentration of 8% protein. To dissolve the casein, 2% disodium diphosphate and 1% monosodium diphosphate are added. Complete dissolution occurs after 60 minutes. The mixture is cooled to 50 o C. Hydrolysis is carried out for 4 h, controlling the increase in osmotic pressure. Under the same conditions, but without phosphate, parallel hydrolysis is carried out. The degree of hydrolysis is much weaker, as shown in the data shown in Fig. 4. By dissolving rennet casein with phosphate prior to hydrolysis, the yield increases accordingly. Example 6 Calcium caseinate produced by MD-Foods Denmark (Miprodan 40) containing about 87% protein was used as a material for hydrolysis. Caseinate suspended in deionized water at 50 o C to a concentration of 8% protein. The pH value, osmotic pressure and Brix value are recorded: pH 6.86, osmotic pressure equal to 25 mosmol/kg and Brix value equal to 8.40. Add enzyme:
- Neutrase 0.5 l with 2% protein
- alkalase 2.4 l with 0.5% protein
- Novozyme 515 with 0.8% protein
The hydrolysis is carried out for 6 hours at 50° C. while controlling the pH value, the osmotic pressure and the Brix value. At the end of the hydrolysis process, the pH is 5.92, the osmotic pressure is 212 mosmol/kg, Brix is 11.40 o V and the degree of hydrolysis is 26.1% 3 min at 75° C. Separation of the hydrolysed mixture is carried out by processing on a PCI ultrafiltration system with FP100 membranes mounted on it. (molecular weight division is 100,000). The yield after ultrafiltration is >84.7% based on the amount of protein in the crude product. The indicated output is obtained without diafiltration. The product is concentrated by processing in PCI nanofiltration equipment fitted with AFC30 membranes. The overall yield after this processing step is 79.7%. The concentrate is spray dried. The resulting product is completely soluble and is a powdery substance containing 90% protein in the dry product with the distribution of molecular weights shown in Fig.5. Number average molecular weight Mn=500, free amino acid content 7% The pH value in a 5% solution of the product is 4.67. Example 7 Calcium caseinate produced by MD-Foods Denmark (Miprodan 40) containing about 87% protein was used as a raw material for hydrolysis. Caseinate suspended in deionized water at 50 o C to a concentration of 8% protein. The pH value is recorded, the osmotic pressure and Brix:pH is 6.86, the osmotic pressure is 25 mosmol/kg and the Brix value is 8.40. Add enzymes:
- Neutrase 0.5 l with the amount of protein 2.0%
- alkalase 2.4 l with 0.5% protein
- Novozyme 515 with 0.8% protein
The hydrolysis is carried out for 6 hours at 50° C., controlling pH, osmotic pressure and Brix. At the end of the hydrolysis process pH 5.92, osmotic pressure equal to 212 mosmol/kg, Brix value equal to 11.40 o V and degree of hydrolysis 26.1% followed by thermal treatment for 3 minutes at 75° C. Separation of the hydrolysed mixture is carried out by treatment on PCI ultrafiltration equipment fitted with FP100 membranes (molecular weight separation 100,000). Yield after ultrafiltration treatment > 84.7% based on the amount of protein in the dry product. The specified output is obtained without additional diafiltration. The product is concentrated by PCI nanofiltration treatment with the additional installation of AFC30 membranes. The overall yield after this processing step is 79.7%. Activated carbon (Picatif 120EW) is added to the concentrate at 4% o B, after which it is filtered on a filter plate and spray dried. The resulting product is a fully soluble powder containing 89.6% protein on a dry matter basis with the molecular weight distribution shown in FIG. The average number of molecular weights Mn 541. The content of free amino acids is equal to 7% The pH value of a 5% solution is 4.64. The palatability analysis (triangle test) showed no significant difference between this product and the product from the previous example, where no activated charcoal treatment was applied.

Claim

1. Casein hydrolyzate of good organoleptic quality, containing free amino acids in an amount of less than 10%, characterized in that the casein hydrolyzate is completely or almost completely soluble in an aqueous medium with a pH level of 3.5 to 7.0 and contains peptides in relative amounts corresponding to the following distribution molecular weights (MM), wt. MM > 5000 Less than 1
MM 1500 5000 15 35
MM 500 1500 40 60
MM< 500 15 35
and the average MM is 400 650. 2. Casein hydrolyzate according to claim 1, characterized in that it is obtained from casein precipitated by rennet, and peptides with MM > 5000 contain less than 0.2 wt. and MM > 3000 less than 5 wt. 3. A method of obtaining a casein hydrolyzate, involving the suspension or dissolution of casein or caseinate with a protein content of at least 85 wt. based on dry matter in an aqueous medium to obtain a suspension or solution with a protein content of up to 20%; Ansona per 100 g of protein and one or more Aspergillus exchoproteases at a concentration corresponding to at least 1000 peptidase units per 100 g of protein, stopping hydrolysis by enzyme inactivation and drying, characterized in that proteolytic hydrolysis is carried out to a degree of hydrolysis of 15-35% using additional one or more Bacillus alkaline endoproteases at a concentration of at least 0.005 units. Anson per 100 g of protein at 45 60 o C. 4. The method according to p. 3, characterized in that at the stage of suspension / dissolution, a suspension or solution is obtained with a protein content of up to 10% and proteolytic hydrolysis is carried out to a degree of hydrolysis of 22 - 28%
5. The method according to claim 3 or 4, characterized in that the hydrolysis is carried out with one or more neutral Bacillus sublitis endoproteases, and during the hydrolysis one or more Bacillus licheniformis alkaline endoproteases and one or more Aspergillus oryzae exoproteases are also present. 6. Method according to one of claims 3 to 5, characterized in that before or after enzyme inactivation, the product consisting of casein hydrolyzate is isolated from the suspension or solution as a filtrate by means of ultramicrofiltration. 7. The method according to one of claims 3 to 6, characterized in that the hydrolysis step is carried out for less than 6 hours. 8. The method according to one of claims 3 to 7, characterized in that the enzymes are inactivated by treatment with food acid, preferably hydrochloric or lemon. 9. The method according to one of claims 3 to 7, characterized in that the inactivation of enzymes is carried out by heat treatment and the hydrolyzate is treated with activated carbon before drying, which is then removed. 10. Method according to one of claims 3 to 9, characterized in that before drying the hydrolyzate is concentrated by hyperfiltration and/or evaporation, and drying is carried out by spraying. 11. The method according to one of claims 3 to 10, characterized in that acid precipitation casein is used from caseins, its dissolution is carried out with a base. 12. Method according to claim 11, characterized in that the acid-precipitated casein is dissolved with calcium hydroxide (Ca(OH) 2). 13. The method according to one of claims 3 to 10, characterized in that from caseins, casein precipitated by rennet is used, and it is dissolved with phosphate or sodium carbonate.

Meaning of casein hydrolyzate in the Medicines Formulary

Casein hydrolyzate

KA3EIN HYDROLYZATE (Hydrolysatum Caseini).

A product obtained by acid hydrolysis of milk protein - casein. Contains a solution of amino acids and simple peptides. The content of total nitrogen is 0.7 - 0.95%; amino nitrogen is 40 - 60% of total nitrogen, the content of tryptophan is not less than 15 mg per 100 ml.

Transparent liquid of yellow-brown color with a specific smell; pH 5.7 - 6.7.

Used as a source of protein for parenteral nutrition.

Indications for use, methods of administration and contraindications are the same as for hydrolysine solution.

Release form: in hermetically sealed bottles of 400 ml.

Storage: at temperatures from - 10 to + 23 "C.

Reference book of medicines. 2012

Which hydrolyzate is "better"?
Algorithm for choosing a mixture for an allergic baby

Postovalova Anastasia,nty @ ngs . en

Before introducing any mixture to a child, it is necessary to consult a doctor. This is a common truth for every mother. And indeed it is.

However, the baby was found to have a food allergy, the issue of choosing a mixture is especially acute. After all, an adapted breast milk substitute is the most important food artificial until he is 1 year old. The mixture should represent at least half of the child's diet (including breast milk, if the mother has it).

Allergy is a serious problem for the health of the baby and a huge burden on the shoulders of the mother. Looking at the blushing cheeks of your child, you want to do anything, pay any price to stop the allergy. But the peculiarity of this disease is that it is never known for certain what kind of price to pay, what to do, what mother's actions are right and which are not ... a feeling of complete chaos ...

So the doctor prescribed a hydrolyzate. "If only it would help!" Mom thinks as she buys an expensive can of new formula.

Really, will it help?

This article is devoted to this problem. On the one hand, she can help her mother by the fact that she can independently understand the features of mixtures - hydrolyzates and understand the logic of the pediatrician who prescribed this or that mixture to the child. On the other hand, if the doctor's prescriptions do not fit into this mixture selection algorithm, compiled on the basis of the recommendations of the specialists of the baby nutrition department of the State Research Institute of Nutrition of the Russian Academy of Medical Sciences, as well as the recommendations of the mixture manufacturers themselves, then the mother will be able to competently ask the pediatrician and / or resort to the help of other specialists.

Let's start with the most important.

In what cases are mixtures based on the hydrolysis of milk protein generally prescribed?

1. In the event that the child has symptoms of a food allergy against the background of the use of 2-3 mixtures from different manufacturers. At the same time, the mixture is the only product that the child receives, an allergy to drugs that the baby drank or drinks (for example, AquaDetrim), an allergy to various complementary foods, if it has already been introduced, must be excluded.

It is very useful (even before deciding to replace the mixture) to consult a dermatologist to confirm the allergic nature of the rash. After all, you need to finally make sure that this, for example, is not contact dermatitis ("local" skin allergy to washing powder, diapers, etc.) or other skin diseases. In addition, a dermatologist will help you choose the right treatment for your baby's skin. As for the general strategy for treating a child, it should be agreed with an allergist.

There are cases when an infant has pronounced allergic reactions to one or more modern maximally adapted mixtures (the ingredient and chemical composition of which is quite close to each other), but he tolerates a less adapted domestic mixture well. This means that the child is not allergic to cow's milk protein, but to other components of the adapted mixtures. The cases of "miraculous" healing of a child when transferred to whole cow's or goat's milk are also explained. But still, the decision on the possibility of using partially adapted mixtures should be made by the doctor - taking into account the severity of the allergy in a small patient. If there are a lot of rashes, they disturb the child, then such experiments are unacceptable. The age of the child also matters. And it is absolutely impossible to feed babies with whole cow or goat milk.

It happens that even before the transfer to the hydrolyzate, the baby has already managed to try a mixture based on soy protein. According to Russian data, sensitization to soy mixtures is observed in 15-20% of cases.

In some cases, the problem of allergy to cow's milk protein helps to solve the introduction of a mixture based on goat's milk "Nanny". However, it should be remembered that this mixture is partially adapted. It is based on whole goat's milk, not fortified with whey. Therefore, the use of "Nanny" up to 6 months is highly undesirable.

2. Mixtures based on partial hydrolysis of cow's milk protein are prescribed as the first mixture prescribed for a baby if the mother or father of the child and even his older brothers and sisters suffer from allergies.

3. Also, mixtures with partially or completely cleaved protein are prescribed for various forms of intestinal malabsorption(that is, intestinal absorption), intestinal colic, malnutrition - due to their high digestibility.

Well, we figured out in which cases the doctor can (or should) prescribe a hydrolyzate. Now let's move on to what kind of hydrolyzate, which brand is right for your child?

Further you will find several steps, depending on the severity of the allergic disease. Start reading from the very beginning and stop at the stage that corresponds to the condition of your child. Just in case, read about the next steps as well to make sure you make the right choice.

Stage 1

The lightest.

NAN GA, Humana GA

Here we will talk about a very popular group of mixtures called "partial hydrolysates" or "hypoallergenic" mixtures.

Banks with such promising names are the first to catch the eye of mothers who meet them on store shelves. They are often prescribed by pediatricians. Unfortunately, it is not always reasonable. The fact is that this group of mixtures actually has a rather narrow scope. Based on the characteristics of their composition, they could not be called " hypoallergenic"but rather" slightly less allergenic than ordinary milk formulas. "That's right," a little less allergenic". After all, in order for the mixture to really lose its antigenic properties, it should not contain peptides with a molecular weight of more than 5000-6000 daltons at all. And in "partial hydrolysates" some of the peptides do have a molecular weight of less than 5000 daltons, but another part of the peptides has a high molecular weight, and therefore can retain their antigenic properties!It is this circumstance that significantly limits the range of application of mixtures of this group.They should not be administered to children with proven intolerance cow's milk protein, except for very light forms.But even in these cases, success is not guaranteed.After all, partial hydrolysates officially contain a certain amount of allergens.If the immune system has already developed antibodies to cow's milk proteins, then an allergic reaction to high molecular weight peptides will be natural , regardless of whether there are many or few such peptides!

"Why do we need such mixtures?" - you ask. In fact, " hypoallergenic mixtures" are needed in order to administer them at the maternity hospital.

"Partial hydrolysate" should be the very first mixture in the life of a child at risk. It is effective until antibodies to cow's milk protein in the child's body have not yet been developed. If, in the maternity hospital, a potential allergy sufferer was given a regular milk formula, then the opportunity to feed a child with a fairly tasty partial hydrolyzate may be lost forever.

Now let's list in what cases hypoallergenic formulas are eligible to be administered to your child.

1. In the maternity hospital, as the first mixture in life, if the mother, father or older brothers / sisters of the child suffer from allergies.

If you have already encountered this problem when feeding your first child, but are planning to give birth to a second child, then remember that it would be advisable to ask shortly before the birth what the children are fed in your maternity hospital, is it possible to introduce " hypoallergenic" mixtures. You may be given the opportunity to purchase it yourself and bring it to the hospital (for this, it is better to buy the mixture at a state pharmacy and keep the receipt).

2. With mild forms of allergies, accompanied by "cosmetic" rashes on small areas of the child's skin. But even in this case, the possibility of using " hypoallergenic mixtures" should be agreed with a specialist.

3. If the mixture is given to a child for the first time, he is prone to allergic reactions, and was previously breastfed. But in this case, it should be understood that it is generally undesirable to introduce a mixture to such children.

4. Sometimes the results of a blood test for allergens show the presence of a reaction to casein, but there is no reaction to albumins. " Hypoallergenic"The mixtures of casein do not contain, so in this case they can be prescribed, especially if the child is older than 6-8 months, he has already been introduced to complementary foods, and the mother, in general," agrees "with the results of the analysis. That is, he showed a reaction to the actually introduced in krokhin diet products.

But if the test results are doubtful for some reason, and rashes and other allergy symptoms bother the child, then the doctor still has the right to recommend a mixture based on completely hydrolyzed proteins.

Above are the recommended brands of HA mixtures, the composition of which is optimal. There are also less preferred options, such as Nutrilon GA. The disadvantage of this formula is its high osmolarity.

Stage 2

"Whey hydrolysates"

Frisopep , Nutrilon Pepti MSC, Alphare

These mixtures are truly hypoallergenic. They do not contain peptides with a molecular weight of more than 5000-6000 daltons.

The use of this group of mixtures in case of allergy to cow's milk protein is most preferable.

Conscious mixtures based on split whey proteins are very physiological, highly digestible. Their only drawback is the bitter taste, however, fortunately, most children get used to it.

In terms of composition, the mixtures of this group differ from each other. So Alfare and Nutrilon Pepti MSC do not contain lactose at all (the carbohydrate component is fully represented dextrinmaltose), and as one of the source of fats they include medium chain triglycerides. Medium chain fatty acids are able to dissolve in the aqueous phase and be absorbed directly into the portal vein system, bypassing the lymphatic, which significantly increases their digestibility, which is often impaired in infants with food allergies.

Children with unimpaired pancreatic function and normal a-priori fat absorption are shown Frisopep, which contains a small amount of lactose and does not contain medium chain triglycerides. True, in practice it happens that Frisopep causes allergic reactions, and then the child successfully switches to Nutrilon Pepti MSC. However, this circumstance should not "compromise" Frisopep to some extent, this is a fairly high-quality product, in addition, it is considered "the most delicious" among hydrolysates.

So, in what cases are 100% whey hydrolysates eligible to be prescribed to your child.

1. Proven intolerance to cow's milk protein.

2. Other forms of intestinal malsorption, malnutrition.

Step 3

"Casein hydrolysates"

Frisopep AS, Nutromigen

What if whey hydrolyzate doesn't work?

In this case, it is possible to use "casein" hydrolysates. "Wait a minute - experienced mothers will say - casein is such a substance, which" the less, the better "". Well, yes, the protein composition of a modern adapted mixture cannot contain more than 40% casein. Then why do we need casein hydrolysates at all? The fact is that casein hydrolysates contain a large number of peptides with a minimum molecular weight, that is, 3500 daltons. Therefore, it is believed that allergic and anaphylactic reactions during their use practically do not occur. At the same time, casein hydrolysates are indeed less physiological than whey hydrolysates and, unfortunately, have an even worse taste.

When are casein hydrolysates prescribed?

1. A severe form of allergy in a child.

2. Whey hydrolysates did not fit. Some doctors generally prefer to bypass the "serum" stage, and immediately prescribe casein hydrolysates, arguing that they are less antigenic. This is not correct, whey hydrolysates are effective in many cases and are more physiological than casein hydrolysates.

Instead of a conclusion

Well, we have considered all types of mixtures based on hydrolyzed proteins.

But having decided on the choice of mixture, we will also understand, how to administer hydrolyzate to a child? After all, if a decision was made to use it, then the previous mixture is not suitable. child and causes allergies. Therefore, many mothers generally try at once replace the old formula with hydrolyzate. It is not right. It's better to do it gradually though. Let not in 7 days, but in 5 or even 3 days, but not immediately. The fact is that by introducing a new mixture according to the scheme, we, first of all, adapt, adapt the child to it. It is necessary that the child's body "is not afraid" of the new composition, does not develop antibodies to it, but calmly accepts the new formula. Therefore, it is still better to introduce gradually

How long to wait for improvement? Usually a week or two or three. Up to a month.

When can complementary foods be introduced (after the introduction of the hydrolyzate)? After you have already understood that the hydrolyzate has approached, the skin has cleared, other allergy symptoms have disappeared, it is better to wait a couple more weeks. You need to let your body recover. And provided that the child has already reached the age of 6 months.

If we consider milk as a curd mass and whey, then casein protein is a curd mass. It is a dietary protein, capable of gel formation, to some extent slowly digestible, due to a decrease in intestinal motility and the presence of gel-forming fiber; when mixed with water, a pudding is formed.

basic information

Casein protein is one of two types of milk protein (the other is whey protein). It is considered a digestion-slowing component of whey protein.

Other names: Casein

Not to be confused with: Whey protein, whey protein

Note Casein protein is not a stimulant, but contains enough calories for energy production. Variety:

Protein supplements

Doesn't mix well with:

Casein: instructions for use

As with other types of protein, casein protein is dosed according to how much dietary protein is needed and how much dietary protein is ingested from other foods. The amount of dietary protein is individual for each person, but there are general indications:

If you are athletic and lead a very active lifestyle, and also strive to reduce body weight and maintain quality muscle mass, the daily recommended dosage of casein protein is 1.5-2.2 g / kg body weight (0.68-1 g / 500 g of body weight).

If you are athletic and have a very active lifestyle or are looking to lose weight and maintain quality muscle mass, the daily recommended dosage of casein protein is 1-1.5 g/kg body weight (0.45-0.68 g/500 g body weight).

The dosage of casein protein should be within the range of the above recommendations and should be based on the dietary protein intake. If the dietary protein intake has already reached the required level, the intake of casein protein is optional. People suffering from obesity (excess fat - 20/30% in men and women, respectively, or when the body mass index is greater than 30, with no muscle mass), should not resort to the above recommendations for use. In this case, you need to do the calculations yourself.

Sources and composition

Sources

Casein is a protein found in many types of milk; casein consumed by humans is mainly obtained from cow's milk. Casein is the insoluble component of milk, while whey is the soluble component; the content of casein in standard whey protein is approximately 80%, the content in breast milk depends on the period of lactation.

Components

Casein protein, like other types of protein, is a source of dietary amino acids. Since casein is an animal product, it contains all the essential amino acids within reasonable limits for humans and the minimum recommended amount of protein. Casein protein also contains various bioactive peptides. These peptides are partially digested, and before being converted to amino acids, they exhibit various effects in the intestine (before absorption). These effects are diverse and are described below.

Casein production

standard production method

The standard way to produce casein is to separate two milk proteins (casein and whey). Whole whey protein is bound to a "coagulant", usually chymosin, which coagulates the casein fragments. This process has a special place in the preparation of cheese (which requires the presence of casein to impart hardness), and is also important in the separation of whey and casein (cottage cheese). Because casein fragments are denser, whey fragments (in liquid form) separate from casein in a process called syneresis. The degree of syneresis depends on what the end product is intended to be (usually cheese), and syneresis can be done through salt, acidity, enzymatic processing, and physical manipulation, along with other techniques that affect the taste and shape of the cheese.

Micellar casein

Micellar casein is produced by microfiltration, which allows the micellar segments to be concentrated in the casein.

Peptides found in casein

Casein contains various bioactive peptides, which are a sequence of amino acids that are found in dietary proteins and partially change their properties in the stomach. Such sequence is provided by some biological effects that occur in the intestine prior to digestion, or digestion may occur by peptide transport.

Peptide alphaS-1

In women, when taken at 150 mg daily, alphaS-1 peptide promotes sedative effects. Two doses of 200 mg can reduce the effects of stress. Sedation improves the quality of sleep in people with high levels of anxiety. Like the c12 peptide, the alphaS-1 peptide can also cause ACE inhibitor-like effects and also lower blood pressure. The alphaS-1 peptide can cause allergies, so in people who are allergic to dairy products, this peptide (also in dietary supplements) can cause allergies.

Peptide c12

Glycomacropeptides

Glycomacropeptides are part of casein in whole milk products, but due to the fact that they are soluble, they are separated from casein during the initial stages of chymosin treatment. They are, of course, related to casein because they are kappa-casein molecules, but they are not part of the casein dietary supplement.

Casoxins and casomorphins

Casoxins and casomorphins are peptides that act on the opioid system, which affects the rate of digestion. Casomorphins are opioid agonists (activators) and casoxins are opioid antagonists. Casein has a high content of casoxins and casomorphins, which explains the reduced post-consumption capacity (because activation of the opioid system reduces intestinal motility).

Casein Hydrolyzate (Hydrolyzed Casein)

Neurological effects

At rest and under neuropsychic stress, casein protein showed better relaxing effects compared to maltitol, which may be due to a specific tryptic hydrolyzate in animal casein. AlphaS-1 peptide is able to reduce stress in women at a dose of 150 mg daily.

:Tags

List of used literature:

Belloque J, Ramos M. Determination of the casein content in bovine milk by 31P-NMR. J Dairy Res. (2002)

Kumar A, et al. Chymosin and other milk coagulants: sources and biotechnological interventions. Crit Rev Biotechnol. (2010)