RU2226841C1 - Protein food additive production method - Google Patents

Abstract

FIELD: food industry and biotechnology. SUBSTANCE: invention relates to production of food additives from animalorigin raw materials, in particular from secondary products of poultry-processing industry, which additive may be used in food industry and medicine. Method of invention comprises mincing meat-and-bone materials on mincer, mixing with water, defatting on centrifuge, enzymatic hydrolysis at 40-50 C and pH 6.5-7.8, centrifugal separation of sediment from hydrolyzate, heating the latter to 90-100 C and keeping it at this temperature for 10-15 min, and drying on spray drier. According to invention, raw material is preliminarily subjected to autoclaved heat treatment for 30 min at 120-123 C and pressure 0.1 MPa, mixing with water is performed at raw material-to-water ratio 1:15, and, before enzymatic hydrolysis, medium pH is adjusted to 7.0-7.2 by adding 1 N sodium hydroxide solution. Enzymatic agent utilized is collagenase solution, which is prepared by dissolving collagenase in buffer solution, pH 7.2, at collagenase-to-buffer ratio 1:12. Addition of enzymatic agent, collagenase or protosubtilin, is accomplished on the basis of 30-50 proteolytic activity units per 1 g protein in treated raw material. Treatment time is 2.5-3 h. Such procedure allows enzyme-endocrinal and meat raw material to be substituted by secondary products of poultry-processing industry. EFFECT: reduced expenses on raw materials, increased productivity, enabled control of hydrolyzate fraction composition, improved organoleptic characteristics of product, and increased biological value of the latter due to enrichment with minerals and food fibers. 14 dwg, 2 tbl, 5 ex

Description

The invention relates to biotechnological methods for producing a protein composition from animal raw materials, and in particular to methods for producing a protein supplement from secondary products of the poultry processing industry, which can be used in the food industry and medicine.

A known method of obtaining protein nutritional supplements [Russian Patent 2075944, MKI ⁶ A 23 J 1/02. A method of obtaining a protein dietary supplement / V.G. Morozov, V.Kh. Khavinson (Russia). - No. 96104838/13; Declared March 25, 1996. Publ. 03/27/97. Bull. No. 9], which provides for the extraction of prepared animal raw materials (crushed frozen organs and mammalian tissues) using 1-2.5% sodium carbonate solution at pH 10.4-10.8 for 2-4 hours with the addition of 0 as an extractant 1-0.5% magnesium chloride. The ratio of the volumes of extracted tissue and soda solution is in the range from 1: 8 to 1:20. Then the precipitate is separated by filtration or in a centrifuge, the extract is acidified to a pH of 3.8-5.5, ballast substances are removed by centrifugation, the precipitate is washed with organic solvents and dried in a stream of hot air at a temperature of 55 ° C.

The disadvantages of the known technical solutions are:

low yield of proteins and peptides from animal raw materials;

limited resources due to the use of enzyme-endocrine raw materials as a source of food protein, the production of which is associated with the difficulty of extracting resources, poor storage and feasibility of using it to obtain enzyme and hormonal preparations;

the need for additional and constant monitoring of the hormone content in the additive and the impossibility of its use in food products without appropriate tests;

significant duration and multi-stage process.

The closest in technical essence and the achieved effect is a method of producing a protein hydrolyzate from meat and meat and bone raw materials of slaughter animals [Russian Patent 2112397, MKI ⁶ A 23 J 1/10, C 12 N 9/64. A method of obtaining a protein hydrolyzate from meat and meat and bone raw materials of slaughtered animals / N.A. Baer, A.D. Neklyudov, A.N. Ivankin, V.I. Dubina, A.V. Berdutina, N.A. Bakanov (Russia) . - 97115279/13. Declared 09/18/97. Publ. 06/10/98. Bull. No. 16], which provides for the grinding of meat or meat and bone raw materials of slaughtered animals, homogenization with water in a meat grinder, degreasing in a centrifuge, hydrolysis with enzyme preparations obtained from the pancreas of slaughtered animals and protosubtin G20X by two-fold application with an interval of 2-3 hours in an amount of 3- 5 wt.% Of the enzyme preparation from the pancreas and with the same time interval of protosubtilin G20X in an amount of 1-2 wt.% For 3 hours. Enzymatic hydrolysis is carried out at a temperature of 40-45 ° C and a pH of 6.5-7.8 . Then the precipitate is separated from the hydrolyzate in a centrifuge and dried on a spray dryer, preheating the hydrolyzate to a temperature of 90-100 ° C.

The disadvantages of the known technical solutions are:

a rather high cost of the resulting hydrolyzate in connection with the use of a complex of enzymes from the pancreas of slaughter animals, the technology for which is distinguished by high costs;

the complex of enzymes from the pancreas of slaughter animals is not specific for the hydrolysis of meat proteins, especially connective tissue (collagen and elastin), and therefore its use is not economically feasible - high consumption (up to 5% by weight of raw materials) and two-fold application;

the resulting hydrolyzate has a low organoleptic rating and functional and technological properties;

maintaining a given pH of the medium (at a level of 7.8) throughout the entire course of the enzymatic hydrolysis leads to additional difficulties in organizing the technological process - the consumption of alkaline solutions, increasing their concentration, the need for their subsequent neutralization and, as a consequence, the formation of a significant amount table salt, which reduces the taste of the product, inhibits the growth of organisms and therefore requires purification from it;

there are no exact recommendations for introducing the necessary mass of the enzyme preparation, taking into account its proteolytic activity in terms of the mass fraction of protein in the processed raw materials, which makes it difficult to ensure optimal conditions for the hydrolysis process;

the time of the process of enzymatic hydrolysis is not clearly defined, since in the description of the invention a complex of proteolytic enzymes is introduced into the raw material twice with an interval of 2 to 3 hours with a total duration of enzymatic hydrolysis of 7 hours

An object of the invention is the replacement of enzyme-endocrine and meat raw materials with less-used secondary raw materials of the poultry industry, increasing the yield of the finished product, regulating the fractional composition of the hydrolyzate, reducing the time of enzymatic hydrolysis, improving the organoleptic characteristics of the product.

This object is achieved by the fact that the method for producing protein food additives provides for grinding meat and bone raw materials on a top, mixing with water, degreasing in a centrifuge, hydrolysis with enzyme preparations at a temperature of 40-50 ° C and pH 6.5-7.8, and separating the precipitate from hydrolyzate in a centrifuge, heating the hydrolyzate to a temperature of 90-100 ° C with a holding time of 10-15 minutes and drying in a spray dryer.

What is new is that secondary products of poultry processing are used as feedstock; before grinding, it is subjected to heat treatment in an autoclave at a temperature of 120-123 ° C, a pressure of 0.1 MPa for 30 minutes; mixing with water occurs when the ratio of raw materials and water is 1:15, respectively; before enzymatic hydrolysis the pH of the medium is adjusted to 7.0-7.2 by making 1 N. sodium hydroxide solution; as an enzyme preparation, a collagenase solution is used, which is prepared by dissolving in a buffer solution with a pH of 7.2 at a ratio of enzyme preparation and buffer of 1:12, respectively; the introduction of a solution of the enzyme preparation of collagenase or protosubtilin G10X is carried out at the rate of 30-50 units of proteolytic activity (PS) per 1 g of protein of processed raw materials; processing time 2.5-3 hours

The technical result is expressed not only in achieving the task, but also in improving the functionality of the additive, increasing the biological value due to enrichment with minerals, dietary fiber, rational use of secondary raw materials of the poultry processing industry, increasing the environmental friendliness and profitability of production.

When determining the total chemical composition of the raw materials, the following determination methods were used: mass fraction of moisture in accordance with [GOST 9793-74. Meat products. Moisture determination methods]; mass fraction of fat - by the Soxhlet method, protein, ash - [Antipova L.V., Glotova I.A., Rogov I.A. Research methods for meat and meat products. - M .: Kolos, 2001. - 376 p.]. Determination of proteolytic activity (PS) of enzyme preparations was carried out in accordance with [GOST 20264.2-88. Enzymatic preparations. Methods for determination of proteolytic activity]. Amino acid composition - by chromatographic method on an AAA-T 339 automatic amino acid analyzer (Czech Republic) according to the instructions for the device and recommendations [L. Antipova, I. Glotova, I. A. Rogov Research methods for meat and meat products. - M .: Kolos, 2001. - 376 p.].

During slaughter and primary processing, birds receive a significant share of secondary products (up to 30%), which include offal, heads, legs, wings, which are currently used extremely poorly.

Offal is mainly sold together with poultry carcass or as a soup set. They have insignificant consumer demand among the population, and also have a low shelf life. At the same time, they have a fairly high mass fraction of protein and have good functional properties. For a more rational use of these secondary products, it is necessary to use additional technological methods that reduce the strength properties of raw materials in order to isolate food protein.

For the destruction of hardened protein structures of connective tissue, it is advisable to use enzyme preparations of proteolytic action, which include collagenase and protosubtilin G10X.

The muscle and connective tissues of the secondary raw materials of the poultry industry are characterized by the predominance of various fractions of proteins in their composition. The main requirements for enzyme preparations for the directed modification of the composition and biotransformation of structural and mechanical properties are, first of all, high substrate specificity for the hydrolysis of these protein fractions, maximum activity in the neutral zone of pH 7.0-7.8 and a moderate temperature range of 40-50 ° C.

For the hydrolysis of secondary raw materials of the poultry industry, enzymatic preparations of animal and microbial origin were used: collagenase, protosubtilin G10X. The selection criterion was the level of general and specific proteolysis on the corresponding protein substrates in different zones of pH and temperature range.

The collagenase preparation is obtained by extraction from Far Eastern crab from hepatopan-creas and is produced at JSC Bioprogress in Schelkovo (TU 9158-002-11734126-94 Enzyme preparation of collagenase. Technical conditions).

Protosubtilin G10X (STP 100-02-88, introduced 23.03.88) is produced on an industrial scale by the Vyshnevolotsk enzyme preparations plant (settlement Zelenogorsk, Tver region).

The results of studies of the proteolytic activity of enzyme preparations depending on the pH and temperature of the environment are presented in figure 1-2. Curve 1 characterizes the change in the proteolytic activity of the enzyme preparation of collagenase, and curve 2 - protosubtilin G10X.

A method of obtaining a protein dietary supplement is as follows.

Feedstock - secondary products of poultry processing (legs or heads of birds) are washed with water, they are pre-heat treated in an autoclave at a temperature of 120-123 ° C, a pressure of 0.1 MPa for 30 minutes, then the raw materials are ground on a spinning top with the diameter of the lattice holes 2-3 mm, add water in a ratio of 1:15 and mix. The resulting homogenizate is degreased, separating the resulting layer of fat in a centrifuge. Then bring the pH of the medium to 7.0-7.2 by making 1 N. sodium hydroxide solution. Next, enzymatic hydrolysis is carried out by introducing a solution of the enzyme preparation of collagenase or protosubtilin G10X at the rate of 30-50 units of proteolytic activity (PS) per 1 g of protein of processed raw materials. The hydrolysis process is carried out with constant stirring and a temperature of 40-50 ° C for 2.5-3 hours. After that, the hydrolyzate is heated to a temperature of 95-100 ° C and incubated for 10-15 minutes to inactivate the enzymes. Next, the precipitate is separated by centrifugation or separation. The resulting hydrolyzate is dried on a spray dryer.

The yield of the final product (powder) is 17.9-21.2% by weight of the feedstock.

A method of obtaining a protein dietary supplement is illustrated by specific examples.

Example 1. A protein supplement from poultry legs is prepared as follows. The legs of the bird are washed with water, they are pre-cooked in an autoclave at a temperature of 120-123 ° C, a pressure of 0.1 MPa for 30 minutes, then the raw materials are ground on a spinning top with a grid diameter of 2-3 mm, water is added in the ratio 1: 15 and mix. The resulting homogenizate is degreased, separating the resulting layer of fat in a centrifuge. Then bring the pH to 7.0-7.2 by making 1N. sodium hydroxide solution. Next, enzymatic hydrolysis is carried out by introducing a solution of the enzyme preparation of collagenase at the rate of 30-50 units of proteolytic activity (PS) per 1 g of protein of processed raw materials. The hydrolysis process is carried out with constant stirring and a temperature of 40 ° C for 2.5-3 hours. After that, the hydrolyzate is heated to a temperature of 95-100 ° C and incubated for 10-15 minutes to inactivate the enzymes. Next, the precipitate is separated by centrifugation or separation. The resulting hydrolyzate is dried on a spray dryer.

The yield of the final product is 17.9-21.2% by weight of the feedstock, which is from 83 to 88% protein conversion. The resulting product is a yellow or cream powder.

Example 2. Similar to example 1, characterized in that the raw materials used are the heads of birds.

The yield of the final product is 13.8-14.8% by weight of the feedstock, which is from 86 to 94% protein conversion. The resulting product is a yellow or cream powder.

Example 3. Similar to example 1, characterized in that the protosubtilin G10X is used as an enzyme preparation. The temperature of the enzymatic hydrolysis process is 50 ° C.

The yield of the final product is 16.8-20.4% by weight of the feedstock, which is from 81 to 86% protein conversion. The resulting product is a powder of dark yellow or cream color.

Example 4. Similar to example 2, characterized in that the protosubtilin G10X is used as an enzyme preparation. The temperature of the enzymatic hydrolysis process is 50 ° C.

The yield of the final product is 12.8-13.4% by weight of the feedstock, which is 84 to 90% of the protein conversion. The resulting product is a powder of dark yellow or cream color.

Example 5. Similar to example 1, characterized in that the enzymatic hydrolysis was carried out previously untreated raw materials in an autoclave. The duration of the process was 3.5-4 hours. The yield of the final product was 15.4-18.6% of the mass of the feedstock, which is from 81 to 85% of the protein conversion. The duration of the process is 1 hour higher compared with example 1, the yield of the final product is 1.7-2.5% less than the pre-processed raw materials. The resulting product is a powder of dark yellow or cream color.

Studies of the chemical, amino acid composition of the feedstock and protein food additives obtained in examples 1-4 are presented in table. 1-2. The dynamics of enzymatic hydrolysis of the legs and heads of a bird by a change in the protein concentration in the hydrolyzate, the accumulation of hydrolysis products by enzyme preparations with collagenase or protosubtilin G10X is presented: in Figs. 3-8 by enzymatic preparation collagenase, in Figs 9-14 by protosubtilin G10X. Curve 1 characterizes the processes of enzymatic hydrolysis of secondary raw materials of the poultry processing industry - the legs of the bird, curve 2 - the head of the bird.

The advantages of using the proposed enzyme preparations of collagenase and protosubtilin G10X are: their production on an industrial scale by domestic enterprises; The optimum pH of their action is in the neutral zone, which is very important when receiving food additives; the optimum temperature is 40-50 ° C (Fig.1-2).

As can be seen from figure 3, the hydrolysis of the raw material took place with an increase in the concentration of soluble protein for both the heads and legs of the bird. The maximum protein content was noted at 4 hours of hydrolysis and amounted to 7.6 and 8.2 mg / cm ³ . From 4 to 5 hours, hydrolysis of soluble protein to low molecular weight products was observed, and from 5 hours to the end of the process, the concentration of soluble protein remained at the same level - 6.9 and 7.5 mg / cm ³ .

Accordingly, the content of low molecular weight products of protein hydrolysis (mainly amino acids) slightly increased in the first four hours - 800 and 1200 μg / cm ³ (figure 5). Further, during hydrolysis of soluble protein from 4 to 5 hours, a smooth increase in amino acids to 1380 and 1500 μg / cm ³ was observed.

The effect of the enzyme preparation of collagenase on the studied raw material was accompanied by hydrolysis of peptide bonds formed with the participation of tyrosine (Fig. 7). A characteristic feature of the action of this drug was the increase in free tyrosine in the hydrolyzate in the first hours of hydrolysis while increasing the content of soluble protein. Tyrosine accumulation occurred during the cleavage of polypeptide chains of proteins, and was not the result of hydrolysis of peptides to amino acids.

From figure 4, 6, 8 it follows that during heat treatment all the processes of accumulation of hydrolysis products are intensified over time. So, from figure 4 it is seen that the maximum accumulation of soluble protein occurs already at 3 hours of hydrolysis (which is 1 hour less than untreated raw materials) and the curves are more gentle. The increase in the concentration of amino acids took place, unlike untreated raw materials, from the first minutes of hydrolysis and reached 1530-3600 μg / cm ³ .

The nature of the tyrosine amino acid accumulation curves remained unchanged, as in the case of raw materials without heat treatment. Apparently, there is a manifestation of the substrate specificity of the enzyme preparation of collagenase - the ability to hydrolyze polypeptide chains of proteins by peptide bonds formed with the participation of the tyrosine amino acid. And, obviously, tyrosine peptide bonds in low molecular weight products (peptides) are not available for this enzyme preparation.

Similar results were obtained with heat treatment (FIGS. 10, 12, 14) and without it (FIGS. 9, 11, 13) of poultry heads and legs, followed by enzymatic hydrolysis with protosubtilin G10X.

Thus, the results obtained indicate that preliminary heat treatment of the animal feed accelerates the process of its enzymatic hydrolysis with a significant yield of amino acids, especially for feeds such as poultry legs (from 1500 to 3600 μg / cm ³ when treated with collagenase and from 1420 to 2300 μg / cm ³ when treated with protosubtilin G10X).

The advantages of the proposed technical solution:

1) the use of secondary raw materials poultry industry;

2) the yield of finished products is 17.9-21.2% of the mass of feedstock, which is 1.5-2.5% higher compared to the prototype;

3) the ability to control the fractional composition (protein, peptides, amino acids) of the finished product depending on the time of hydrolysis;

4) reduction of the time of enzymatic hydrolysis by 57-64%;

5) reduction in the dosage of enzyme preparations by 30-40%;

6) the biological and nutritional value is higher due to the enrichment of minerals and dietary fiber;

7) increase the organoleptic characteristics of the finished product;

8) increase in the percentage of essential amino acids (or individual amino acids).

Claims (1)

A method of obtaining a protein dietary supplement, comprising crushing meat and bone raw materials on a top, mixing with water, degreasing in a centrifuge, hydrolysis with enzyme preparations at a temperature of 40-50 ° C and pH 6.5-7.8, separating the precipitate from the hydrolyzate in a centrifuge, heating the hydrolyzate to a temperature of 90-100 ° C with an exposure of 10-15 minutes and drying on a spray dryer, characterized in that the secondary products of poultry processing are used as feedstock; before grinding, it is subjected to heat treatment in an autoclave at a temperature of 120-123 ° C, a pressure of 0.1 MPa, for 30 minutes; mixed with water at a ratio of raw materials and water of 1:15, respectively; before enzymatic hydrolysis the pH of the medium is adjusted to 7.0-7.2 by making 1 N. sodium hydroxide solution; as an enzyme preparation, a collagenase solution is used, which is prepared by dissolving in a buffer solution with a pH of 7.2 at a ratio of enzyme preparation and buffer of 1:12, respectively; the introduction of a solution of the enzyme preparation of collagenase or protosubtilin G10X is carried out at the rate of 30-50 units of proteolytic activity per 1 g of protein of processed raw materials; processing time 2.5-3 hours

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