RU2376893C1 - Production method of protein product equivalent for suffering from phenyl ketonuria - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to production of health food production. Product protein equivalent proposed for treating of people suffering from phenyl ketonuria production method includes preporation of protein containing biological system - concentrate of whey proteins, hydrolysis, separation of phenylalanine, condensation and drying. For separation of phenylalanine hydrolysate is processed with a ferment providing turning of phenylalanine into tyrosine for instance phenylalanine-4-hydroxylase in quantity of 0.0001-5% of protein weight. After that inactivation of ferment is performed by short-time warming up to 90°C during 3-5 seconds. Protein hydrolysate condensation free from phenylalanine is performed till dry substances content 30-50%. Drying is performed till dry substances content 14%.

EFFECT: invention allows to obtain product with improved treating effectively due to reduce of phenylalanine content in protein equivalent.

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Description

The invention relates to the food industry, namely to the production of therapeutic food products intended for patients with phenylketonuria with a low content of phenylalanine.

A known method of obtaining a food product for patients with phenylketonuria, including the preparation of the biological system, carrying out hydrolysis, filtering and drying (see AS USSR No. 634748, IPC A61K 37/18, claimed 07.29.1977, published 30.11.1978, ) However, the known method does not ensure complete removal of phenylalanine from the biological system.

There is also a method of obtaining a food product for patients with phenylketonuria, including the preparation of the biological system, hydrolysis, membrane fractionation, adsorption, drying (see Kruglik V.I. Theoretical rationale and practical implementation of milk protein hydrolysates and specialized products using them: abstract of dis. .... Doctor of Engineering Sciences: 05.18.04: defended on 02.19.08 / Kruglik Vladimir Ivanovich. - Kemerovo, 2008. - 42 p.). This technical solution is the closest to the claimed combination of essential features (prototype).

According to the prototype, a whey protein concentrate (BSC) is used as a biological system, which is subjected to hydrolysis, membrane treatment, passage of chemically modified polystyrene through porous granules, thickening and drying. The disadvantage of this method is the complex membrane treatment and low yield of the target product.

The objective of the technical solution is to improve therapeutic efficacy by reducing the phenylalanine content in the protein equivalent and reducing the cost of the final product.

The technical result is achieved through the use of a whey protein concentrate as a protein-containing biological system, which is subjected to one of the known hydrolysis methods (acidic, alkaline, enzymatic), which provides a fairly complete cleavage of the polypeptide chain of the protein molecule into free amino acids. The depth of protein cleavage, ceteris paribus, should be at least 85-98%. The obtained hydrolyzate, mainly represented by a mixture of amino acids, is treated with an enzyme that converts phenylalanine to tyrosine - phenylalanine-4-hydroxylase, then the enzyme is inactivated by short-term heating to 90 ° C for 3-5 s. As a result of appropriate processing, protein hydrolysates are completely free of phenylalaine.

The method of obtaining the protein equivalent intended for the treatment of patients with phenylketonuria is implemented as follows: the biological system (KBS is used as the protein-containing biological system) is subjected to one of the known methods of hydrolysis in order to split the polypeptide chain of the protein molecule into free amino acids. The resulting hydrolyzate is treated with an enzyme that converts phenylalanine to tyrosine - phenylalanine-4-hydroxylase, in an amount of 0.0001-5% by weight of the protein, then the enzyme is inactivated by brief heating to 90 ° C for 3-5 s. After this, the protein hydrolyzate, completely free of phenylalanine, is concentrated to a solids content of 30-50% and dried to a moisture content of 14%.

Example 1

A protein-containing biological system is prepared, which is used as a dry whey protein concentrate in an amount of 20 kg per 100 l, dissolved in 40-60 l of water at a temperature of 35-40 ° C for 30 minutes, filtered, water is added to the required volume. Then the resulting mixture is pasteurized and cooled to a temperature of 37 ± 1 ° C). As the hydrolysis using the hydrochloric acid method. To do this, prepare hydrochloric acid by distillation three times to obtain 5.7 N. solution. After the preparation of both systems, acid hydrolysis is started. For this, 18.8 liters of 5.7 n are added to the prepared biological system. hydrochloric acid solution. Hydrolysis is carried out at a temperature of 120 ° C, a duration of 4 hours, a pressure of 1.5 atm.

Carrying out acid hydrolysis of proteins under these conditions provides a fairly complete (85.8%) cleavage of the polypeptide chain of the protein molecule in the protein-containing biological system into free amino acids, as evidenced by the data shown in table 1.

Table 1 Sample The degree of release of amino acids from the polypeptide chain,% The degree of release of phenylalanine,% The control 0 0 Experience 85.8 ± 1 99 ± 1

Then the hydrolyzed mixture is sent for demineralization in order to free from hydrochloric acid. The column is loaded with an ion exchange resin - anion exchange resin grade AB-17-8, while the height of the filter loading layer is 0.4 m, the filter diameter is 0.2 m, and the filtration speed is 5-8 m / h. Demineralization is carried out at pH 7 ± 0.5, the duration of the process is 120 ± 5 min, at normal atmospheric pressure.

The obtained hydrolyzate is treated with an enzyme that converts phenylalanine to tyrosine, for example phenylalanine-4-hydroxylase (FAG) in an amount of 0.000016-0.8 kg, which corresponds to 0.0001-5% by weight of the protein (table 2), then inactivation of the enzyme by short-term heating to 90 ° C for 3-5 s.

table 2 Sample The content of phenylalanine, g / 100 g The content of tyrosine, g / 100 g KBS 14.1 ± 1 16.5 ± 1 KBS after acid hydrolysis 14.1 ± 1 16.4 ± 1 KBS after acid hydrolysis treated with phage 0 30.5 ± 1

After that, the hydrolyzate freed from phenylalanine in an amount of 75 ± 0.2 kg is subjected to thickening to a solids content of 30-50% and drying to a moisture content of 14%. After carrying out the process, 8.0 ± 0.1 kg of powder with a moisture content of 14% is obtained.

Example 2

A protein-containing biological system is prepared, using a whey protein concentrate (protein mass fraction 94.0%). For this, a dry whey protein concentrate in an amount of 20 kg per 100 l is dissolved in 40-60 l of water at a temperature of 35-40 ° C for 30 minutes, filtered, water is added to the required volume. Then the resulting mixture is pasteurized and cooled to a temperature of 37 ± 1 ° C). As the hydrolysis using the alkaline method. To do this, prepare 2 N. sodium hydroxide solution. After preparing both systems, alkaline hydrolysis is started. To do this, 1.8 l 2 N are added to the prepared biological system. sodium hydroxide solution. Hydrolysis is carried out at a temperature of 110 ° C, a duration of 5 hours. At the end of hydrolysis, the resulting solution is cooled, add 2.7 l of sodium citrate buffer solution with a pH of 3.3 and 1.8 l of 5.7 N. hydrochloric acid solution to neutralize sodium hydroxide.

Carrying out alkaline hydrolysis of proteins under these conditions provides a fairly complete (85.22%) cleavage of the polypeptide chain of the protein molecule in the protein-containing biological system into free amino acids, as evidenced by the data shown in table 3.

Table 3 Sample The degree of release of amino acids from the polypeptide chain,% The degree of release of phenylalanine,% The control 0 0 Experience 85.22 ± 1 99 ± 1

The resulting hydrolyzate is treated with an enzyme that converts phenylalanine to tyrosine, for example phenylalanine-4-hydroxylase (FAG) in an amount of 0.000016-0.8 kg, which corresponds to 0.0001-5% by weight of the protein (table 4), then inactivation of the enzyme by short-term heating to 90 ° C for 3-5 s.

Table 4 Sample The content of phenylalanine, g / 100 g The content of tyrosine, g / 100 g KBS 14.1 ± 1 16.5 ± 1 KBS after alkaline hydrolysis 14.0 ± 1 16.4 ± 1 KBS after alkaline hydrolysis treated with phage 0 30.4 ± 1

After that, the hydrolyzate freed from phenylalanine in an amount of 75 ± 0.2 kg is subjected to thickening to a solids content of 30-50% and drying to a moisture content of 14%. After carrying out the process, 8.0 ± 0.1 kg of powder with a moisture content of 14% is obtained.

Example 3

A protein-containing biological system is prepared, using a whey protein concentrate (protein mass fraction 94.0%). For this, a dry whey protein concentrate in an amount of 20 kg per 100 l is dissolved in 40-60 l of water at a temperature of 35-40 ° C for 30 minutes, filtered, water is added to the required volume. Then the resulting mixture is pasteurized and cooled to a temperature of 37 ± 1 ° C). An enzymatic method is used as hydrolysis. To do this, prepare an enzyme preparation, for example pancreatin, in an amount of 0.08 kg, which corresponds to 0.5% by weight of protein. After the preparation of both systems, enzymatic hydrolysis is started. For this, pancreatin is introduced into the prepared protein-containing biological system at a temperature of 37 ± 1 ° C, incubated for 24 ± 0.2 hours, then the enzymes are inactivated by short-term heating to 90 ° C for 3-5 s.

Enzymatic hydrolysis of proteins under these conditions provides a fairly complete (91.3%) cleavage of the polypeptide chain of the protein molecule in the biological system into free amino acids, as evidenced by the data shown in table 5.

Table 5 Sample The degree of release of amino acids from the polypeptide chain,% The degree of release of phenylalanine,% The control 0 0 Experience 91.3 ± 1 99 ± 1

The obtained hydrolyzate is treated with an enzyme that converts phenylalanine to tyrosine, for example, phenylalanine-4-hydroxylase (FAG) in an amount of 0.000016-0.8 kg, which corresponds to 0.0001-5% by weight of the protein (table 6), then inactivation of the enzyme by short-term heating to 90 ° C for 3-5 s.

Table 6 Sample The content of phenylalanine, g / 100 g The content of tyrosine, g / 100 g KBS 14.1 ± 1 16.5 ± 1 KBS after enzymatic hydrolysis 14.0 ± 1 16.4 ± 1 KBS after enzymatic hydrolysis treated with phage 0 30.4 ± 1

After that, the hydrolyzate freed from phenylalanine in an amount of 75 ± 0.2 kg is subjected to thickening to a solids content of 30-50% and drying to a moisture content of 14%. After carrying out the process, 8.0 ± 0.1 kg of powder with a moisture content of 14% is obtained.

Thus, using one of the known methods of hydrolysis, a sufficiently complete cleavage of the polypeptide chain of the protein molecule in the biological system into free amino acids is provided, and due to a specific enzyme, such as phenylalanine-4-hydroxylase, phenylalanine is converted into tyrosine - phenylalanine-4- hydroxylase, which results in the complete removal of phenylalanine from the biological system, improving therapeutic efficacy by reducing the content of phenylalanine in protein equivalent Alente and the possibility of expanding the already uniform diet of the patient, as well as increasing the nutritional and biological value and reducing the cost of the final product.

Claims (1)

A method of obtaining a protein equivalent of a product intended for the treatment of patients with phenylketonuria, including preparing a protein-containing biological system, hydrolysis, removal of phenylalanine, thickening, drying, characterized in that a whey protein concentrate is used as a protein-containing system, which is treated with an enzyme after hydrolysis to remove phenylalanine, converting phenylalanine to tyrosine, for example phenylalanine-4-hydroxylase, in an amount of 0.0001-5% by weight of protein, then the enzyme is inactivated by short-term heating to 90 ° C for 3-5 s, after which the protein hydrolyzate, completely free of phenylalanine, is concentrated to a solids content of 30-50% and dried to a moisture content of 14%.