RU2245078C1 - Method of producing dry elastin hydrolyzate - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: elastin hydrolyzate is intended for use as additional animal protein source or food consistence-influencing additive. For the first purpose, elastin is subjected to enzymatic treatment to transfer it into its soluble form. When further dried, hydrolyzate is converted to dry powder for use as food additive.

EFFECT: enabled production of soluble form of elastin.

1 tbl, 3 ex

Description

The invention relates to the meat industry, and the resulting product can be used in the food industry as an additional source of amino acids and additives that affect the consistency of the product.

Recently, much attention has been paid to the problems of rational use of secondary raw materials and the attraction of new types of natural protein raw materials and the creation on their basis of products for mass consumption.

The reserve protein raw material of animal origin is secondary raw materials - connective tissue. The main proteins of connective tissue are collagen and elastin. The similarity between them is that they are complex fibrillar proteins, the amino acid composition of which is represented mainly by non-polar amino acids, they also have a large number of transverse covalent cross-links. These are strong proteins that are poorly broken down under the action of solutions of acids and alkalis. However, these are different proteins each with their own properties. Their widespread use in the food industry is limited by their complex structure and strength.

The structure and properties of collagen have been studied well, and collagen-containing raw materials are used in the food industry.

Known methods for producing collagen substances from animal tissues (ANTIPOVA L.V., GLOTOVA I. Production and properties of collagen substances from animal tissues. - "Biotechnology", 1999, No. 5, p. 47-53). As a source of collagen, we used the edge sections of animal skins, mezra, veins, tendons, and fascia. For fermentation of feedstock, industrial enzyme preparations were used: megaterin and protosubtilin G20x. As a result, the resulting collagen masses were used to form model collagen films.

The meat industry also uses dry protein semi-finished product obtained by drying the broth after boiling the bones of young cattle, the mass fraction of protein in which is at least 86%, it is mainly collagen.

Various meat products have been developed using secondary sources of food protein of animal origin, so a meat product for baby food (patent RU 2165723, 04/27/2001). The meat product consists of meat, blood, collagen-containing offal, milk whey. In the preparation of minced meat, a protein emulsion made from pig skin is added to sausages, which is ground and aged in 3-5% sodium chloride solution (patent RU 2166854, 05.20.2001).

A disadvantage of the known products is the use of only collagen-containing raw materials.

The second most common protein is elastin; its chemical and amino acid composition is currently known, but the protein structure has not been fully established. There are only assumptions, hypotheses. Elastin-containing raw materials are used in industry for the production of only feed flour. Also known is the possibility of using elastic tissue to obtain iodized food additives (patent RU 2142723, 20.12.1999). However, we are talking about elastic tissue (vyjna ligament).

The closest set of essential features to the proposed method for producing a dry elastin hydrolyzate is a method for isolating elastin during its sequential extraction with acetone, ether and alkali. The method of isolation of elastin according to the known VFS technique is as follows.

Exhausted ligaments of cattle, cleaned of fat and other tissues, finely chopped, chopped in a meat grinder and filled with acetone, the operation is repeated several times until the turbidity of acetone disappears. Then carry out the treatment with ether. After removal of the ether by drying in air in a fume hood, the material is treated with a solution of caustic soda by boiling. Subsequently, the precipitate is washed with water, basified to pH 7.0. The precipitate is dehydrated, dried in air, crushed and sieved (VFS 42-2285-97 Elastolitin).

A disadvantage of the known method for producing elastin is its insolubility, which limits the possibility of use.

The problem solved by the invention is the use of biochemical methods to create a soluble form based on connective tissue protein - elastin.

The technical result of the invention is to obtain a soluble form of elastin in the form of a dry hydrolyzate.

The technical result is achieved by the fact that in the method for producing a dry elastin hydrolyzate, it is characterized by the fact that they isolate protein - elastin from the digging ligament obtained by trimming the cervical cut of cattle as elastic tissue, which is stripped from the remains of muscle and fatty tissue by sequential extraction with acetone, ether and alkali, followed by drying and grinding, according to the invention, the isolated elastin is subjected to enzymatic treatment with pepsin under conditions: pH 1.60-1.65, temperature 37 ± 2 ° C for 20- 22 hours, then the resulting soluble elastin hydrolyzate is subjected to atmospheric freeze-drying at a temperature of -10 ° C ...- 12 ° C for 44-48 hours and ground to a powder state.

Comparison of the invention with other technical solutions known from the level of technology made it possible to establish the following:

- the basis of the powder is protein of secondary raw materials of animal origin - elastin after enzymatic treatment and freeze-drying;

- elastin obtained by the proposed method is soluble;

- elastin due to its unique structure is able to recover after drying;

- dry protein hydrolyzate can be used in the food industry as an additional source of amino acids.

A method of obtaining a dry elastin hydrolyzate is as follows.

Half-carcasses of cattle are subjected to cutting, deboning and venation. As a result of the venation of the cervical cut, elastic tissue is released - the ligament of the neck.

Exhausted ligaments of cattle, cleaned of fat and other tissues, are finely chopped, chopped in a meat grinder and poured with 5 volumes of acetone, kept for 4 hours, after which acetone is separated and the operation is repeated three times, changing acetone after 6-12 hours. Each time when acetone is added, they are continuously stirred for the first 20 minutes, and then periodically. The end of dehydration is determined by the disappearance of the turbidity of acetone. Then a double treatment is carried out with 3 volumes of medical ether, changing after 6-12 hours. After removal of the ether by drying in air, the material is treated with 6 volumes of 0.1 M sodium hydroxide solution at boiling for 1 hour and cooled to room temperature. The precipitate is separated off, washed twice with water, and then washed with stirring with water, setting pH 7.0 in the washing water by adding a 1 M hydrochloric acid solution and washed twice with water, alkalized to pH 7.0. The precipitate washed from salts is separated, squeezed out, suspended in 5 volumes of water and homogenized for 10 minutes. Then it is heated to a boil, cooled and filtered at a temperature of 60-70 ° C. The precipitate is dehydrated with 3 volumes of acetone with its triple change, dried in air, ground in a mill and sieved.

After isolation of elastin from the digestive ligament by sequential extraction in acetone, ether and alkali, it is treated with a 0.2% solution of the enzyme, pepsin. An industrial preparation of the company “MERKK" (Germany) was used, isolated from the mucous membrane of the pig’s stomach. Fermentation is carried out in a thermostat at a temperature of 37 ± 2 ° C, the pH of the medium is 12.60-1.65 for 20-22 hours. The temperature of thermostatting corresponds to the optimum temperature of the enzyme, since at a temperature below the indicated pepsin is inactive, and if the temperature is 40 ° C or higher, then pepsin will denature. A pH of 1.60-1.65 is optimal for pepsin isolated from pig gastric mucosa. The duration of the fermentation process (20-22 hours) is determined by obtaining a clear solution, that is, the protein passes from a solid state to a liquid one. The transition of elastin from insoluble to soluble form is associated with the specificity of the action of pepsin, which is expressed in the predominant hydrolysis of peptide bonds formed by amine groups of phenylalanine and tyrosine. Subsequently, the pH of the medium is neutralized with an alkali solution to 7.0-7.2.

After fermentation, the hydrolyzate solution is frozen at a temperature of -18 ° C ...- 20 ° C for 20-22 hours. A sharp freezing of an object at low temperatures of -18 ° С ...- 20 ° С leads to the formation of small ice crystals evenly spaced throughout the substance. This subsequently leads to more uniform drying of elastin.

The next step in obtaining a dry elastin hydrolyzate is freeze-drying at a temperature of -10 ° C ...- 12 ° C, for 44-48 hours. The drying temperature is selected with the position of ensuring the preservation of the structure of the product, while eliminating unnecessary energy costs. The readiness of the powder is determined by moisture not more than 10%. The resulting powder is dried in a thermostat at a temperature of 40 ± 2 ° C for 25-30 minutes. The readiness of the powder is determined by moisture - 5%.

The parameters of the obtained powder are shown in the table

Table
Powder parameters No. p / p Name Indicators 1. Moisture contents, % 5 2. The protein content,% 92-94 3. Ash content,% 1.8-2.2 4. Slope angle, degrees thirty 6. Bulk weight, g / ml 0.65 7. Particle diameter mm 0.7 8. Color light yellow 9. Taste neutral 10. Toxicity non toxic

Examples confirming the possibility of producing a dry elastin hydrolyzate.

Example 1. Elastin isolated from an elastic tissue (excision ligament) during sequential extraction: acetone, ether, alkali, dried at room temperature and crushed, was fermented with a 0.2% pepsin solution for 20 hours, at a pH of 1.6 and a temperature of 37 ° C. The resulting protein hydrolyzate solution was subjected to atmospheric freeze-drying at a temperature of -10 ° C. The duration of sublimation was 44 hours.

Example 2. Elastin isolated from an elastic tissue (excision ligament) during sequential extraction: acetone, ether, alkali, dried at room temperature and crushed, was fermented with a 0.2% pepsin solution for 21 hours, at a pH of 1.63 and a temperature of 39 ° C. The resulting protein hydrolyzate solution was subjected to freeze-drying at -12 ° C. The duration of sublimation was 46 hours.

Example 3. Elastin isolated from elastic tissue (cords) during sequential extraction: acetone, ether, alkali, dried at room temperature and crushed, was fermented with 0.2% pepsin solution for 22 hours, at a pH of 1.65 and a temperature of 35 ° C. The resulting protein hydrolyzate solution was subjected to atmospheric freeze-drying at a temperature of -10 ° C. The duration of sublimation was 48 hours.

The obtained elastin can be used for food purposes in industry, as an additional source of animal protein.

Claims (1)

A method of obtaining a dry elastin hydrolyzate, characterized in that the protein is isolated - elastin from the digging ligament obtained by trimming the cervical cut of cattle as elastic tissue, which is cleaned from the remains of muscle and fatty tissue by sequential extraction with acetone, ether and alkali, followed by drying and grinding, and the isolated elastin is subjected to enzymatic treatment with pepsin under conditions of pH 1.60-1.65, temperature (37 ± 2) ° C for 20-22 hours, then the resulting soluble elastin hydrolyzate according to subjected to atmospheric freeze-drying at a temperature of -10 ...- 12 ° C for 44-48 hours and crushed to a powder state.