RU2333663C2 - Method of peptide/amino acids production from raw material containing protein, associated products and application of products - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to methods of protein-free items production containing free amino acids and short peptides. Production method is proposed for peptide/amino acids with fat content less 0.5% mass from raw material containing protein. Method provides for raw material graining and hydrolysis in presence of endogenetic ferments. The produced mass is then subject to separating treatment including coagulation of protein residues. Amino acids/peptides produced by this method contain free amino acids in the amount of 30-60% mass, and fat less 0.5% mass. Amino acids/peptides produced by this method contain free amino acids in the amount of 5-100% mass and fat less 0.5% mass. Hydroxuapatite is produced by this method. Hydrolysis method for protein-containing raw material includes application of endogenetic ferments, heating and blending. Method of peptide/amino acid removal from hydrolysed mixture with molecular weight less 10000 dalton through filter. Application of amino acids/peptides for production of pharmaceutical/biotechnological/food/ feeding stuffs.

EFFECT: production of amino acids/peptides free of allergens and DNA traces.

18 cl, 3 dwg

Description

The present invention relates to aspects regarding:

a) a method of obtaining a protein-free product containing peptides, free amino acids and minerals from untreated animals and aquatic materials, as well as products and their use as animal feed and / or as products for the biotechnological, pharmaceutical and food processing industries. An oil product resulting from the implementation of this method is also described.

b) a method for producing a protein product enriched with free amino acids and short peptides, the resulting product and its use as animal feed and / or in veterinary medical products or in the food processing industry. An oil product resulting from the implementation of this method is also described.

c) a method for the regeneration of peptides, free amino acids and minerals from animal and aqueous raw materials.

It is known in the industry to produce peptides and amino acids by acid hydrolysis using biotechnological and / or chemical / technical, both natural and synthetic concentrated enzymes. The present invention is a method of using decomposing enzymes of natural origin from animal or water raw materials in the industrial process of obtaining a product of pharmaceutical, biotechnological, food or veterinary medical quality.

The term "pharmaceutical quality" as used refers to products for intravenous use and products that are classified as medicines for humans and animals or natural medicines.

Used the term "biotechnological quality" refers to products that can be used, for example, as a culture medium or catalysts for the cultivation of cells, bacteria, fungi and algae.

The term “nutritional quality” as used refers to products that are consumed by humans as a dietary supplement or an independent product.

Used the term "veterinary medical quality" refers to products that are classified as medicines for animals.

The present invention may optionally be used for the manufacture of food products in the form of additives or independent products.

Amino acids and peptides are well known in pharmacy, natural medicine and the veterinary medical industry as components of products such as intravenous nutrition and special nutrition to alleviate some traumatic conditions. To date, for these purposes, mainly used extracts from blood plasma and protein hydrolyzate obtained in the presence of pancreatic enzymes of pigs and calves. The present invention provides the pharmaceutical industry with the ability to obtain a source of amino acids and peptides of previously unknown quality.

Amino acids and ultrashort peptides are also used in biotechnological processes, for example, in those cases where it is desirable to produce a very powerful culture medium. For all industries where single-celled microorganisms or cell substrates from higher microorganisms are cultivated, there is a limitation associated with providing a culture medium of adequate quality. The limiting factors are environmental defects or its high cost. In addition, amino acids or peptides produced by biotechnological methods usually contain substances that inhibit growth, which can be excluded from the process using products obtained by the method according to the invention. The combination of the spectrum of natural amino acids and biological micronutrients / minerals obtained by the described method provides a unique product for obtaining a culture medium intended for the biotechnological industry. In addition, this technique allows you to recycle proteins from various cultures in a mixture of amino acids and peptides, which can be reused.

Peptides / amino acids are used in the food processing industry as binders, emulsifiers, perfumes, and the like. Such an application occupies a significant place, and its volumes are increasing. In the food processing industry, peptides and amino acids derived from soybeans and milk are most widely used. It is known that amino acids and peptides from soy and milk cause allergic reactions, which can only be avoided by using other peptide / amino acid compositions made from other sources or peptide / amino acid compositions from soy and milk that have been specially modified to suppress these reactions. In this regard, there is a great need for a method for producing a composition of amino acids and peptides, also originating from soy and / or milk, but not causing allergic reactions. Products from most animal sources do not have the necessary degree of utilization, since there is no extraction technology that preserves the functional properties of the product while removing undesirable components, such as salt and fat, that impair the quality of the product.

For the production of animal feed, many different compositions of proteins, peptides and amino acids of various origins are used. The composition of the peptides, amino acids and proteins is a very important factor in the production of animal feed, as the growth potential of the animal depends on the balanced absorption of food. Therefore, in this area there is a great need to develop a method for obtaining the desired composition, providing optimal conditions for the growth of animals.

As used below, the term “endogenous enzymes” refers to enzymes that exist within a protein product, in contrast to “exogenous” enzymes, which are extraneous enzymes that are added to protein feeds during conventional hydrolysis. One example of “exogenous” enzymes is Diterzyme APY, which is a bacterial protease (E.C. 3.4.21) that is obtained by controlled fermentation of Bacillus alcalophilus and which can be purchased from various suppliers. The term "endogenous" enzymes is also used to refer to enzymes extracted from other natural enzyme materials or raw materials, preferably from cold-blooded animals.

As used below, the term "hydrolyzate" is used to indicate the raw material to be processed, and it is understood that the hydrolyzate includes a heated mixture of raw materials and water with a set pH value. This term is particularly applicable to aspects a) and b) of the present invention.

There are a number of patents related to the field of the invention, for example, RU 2103360, which describes a nutrient medium for the cultivation of eukaryotic cells and a method for preparing a hydrolyzate from the internal organs of fish, which is obtained by proteolytic hydrolysis. The hydrolysis process is carried out at a high pH value established by sodium hydroxide using thermal inactivation, filtration and drying, and fish waste is mixed with distilled water in a ratio of 1: 1. Hydrolysis is carried out at a temperature of 40-42 ° C to achieve an amine nitrogen content of 5.5-6.5 wt.% And a free amino acid content of 50-60 wt.%.

SU 1755417 describes a method for producing hydrolysates from fish raw materials in a fermenter, to which an enzyme preparation is added, after which the resulting hydrolyzate is filtered and dried, while the raw material is periodically added to the fermenter.

RU 1559466 describes a method for the production of hydrolysates, which involves grinding fish products or their processing waste, mixing with water, heating the mixture, adding a proteolytic fermentation preparation, fermentation, filtering and drying, and the raw materials and water are mixed in a ratio of 2: 1-1: 1 and heated to a temperature of 40-45 ° C, and fermentation is carried out for 0.5-2.5 hours using an exogenous enzyme, protosubtilin G3x.

You can also refer to FR 2168259, which describes the enzymatic hydrolysis of fish proteins, which is carried out by grinding fresh fish to a fine paste without adding water. Exogenous enzymes are added to the system and the paste is hydrolyzed for 15 hours, depending on the desired solubility. The product is stabilized for 5-20 minutes at 90-100 ° C, after which it is filtered, pasteurized and centrifuged. As a result of this process, products of high nutritional value are obtained.

As shown above, various technologies are known for the isolation of proteins, peptides and amino acids from fish, which is suitable for food production. In addition, methods are also known for producing oil / fat from both plant and animal feed.

In accordance with aspect a), the purpose of the present invention is to develop a method for producing a protein-free hydrolyzate using natural enzymes without adding any substances of non-natural origin. This aspect distinguishes the present invention from other methods that use enzymes from a large number of different sources, for example, from bacterial cultures and the like.

This goal also includes the process of obtaining a product that is completely free of protein and DNA, as well as other allergenic compounds, without adversely affecting the utilization of raw materials. This method also includes reducing the amount of fat in the final product to such a low value that eliminates the disadvantages of using fish raw materials. It is intended to produce such a product, which can be used in many areas where the use of products produced by known methods is limited or impossible due to their fat content.

The purpose of the invention also provides for the complete utilization of raw materials and the minimization of environmental loads associated with production.

Thus, in accordance with aspect a), the invention relates to a method for isolating peptides / amino acids and oil / fat from protein-containing raw materials, characterized in that it comprises the following steps:

but. grinding of raw materials;

b. heating the crushed raw materials to a temperature in the range of 40-62 ° C, preferably 45-58 ° C;

from. optionally isolating the oil / fat from the feed before and / or after the heating step in order to obtain a first oil product;

d. adding water having about the same or the same temperature as the raw materials and the pH value established by the addition of calcium;

E. the hydrolysis of raw materials using endogenous enzymes in order to obtain a hydrolyzate;

f. optionally adding a pH adjuster to maintain the desired pH of the hydrolyzate carried out in the hydrolysis step;

g. heating the hydrolyzate to 75-100 ° C, preferably 85-95 ° C;

h. removal of large particles from the hydrolyzate, including non-hydrolyzed proteins that can be returned to the hydrolysis stage;

i. the optional selection of a mixture of fat / oil in order to obtain a second oil product;

j. protein coagulation;

k. removal of coagulated proteins;

l. optional isolation of the fat / oil mixture in order to obtain a third oil product;

m. optional concentration of the remaining amino acids and short-chain peptides;

n optional drying of the concentrate in order to obtain short-chain (short) peptides and amino acids.

Preferred embodiments of the method according to aspect a) of the present invention are given in Auxiliary Clauses 2-6.

The method according to the invention contributes to the production of a peptide and amino acid product with a low fat content, preferably less than 0.1%. In addition, the resulting product contains natural minerals of biological origin. The resulting product contains less than 1% salt.

In accordance with paragraphs 7-10, the invention also relates to the use of one of the methods according to the invention for the production of a biotechnological product, pharmaceutical product, food product and feed product.

As indicated in paragraph 11, the invention also includes the use of the method according to the invention for the production of hydroxyapatite.

As indicated and described in paragraph 12, the present invention also makes it possible to produce amino acids / peptides by the method of the invention.

As indicated in paragraph 13, using the method according to the invention also provides hydroxyapatite, characterized by the absence in it of any allergen and traces of DNA.

In addition, in accordance with paragraph 14, it is indicated that using the method according to the invention provides an oil characterized by the absence of allergens and traces of DNA.

On the one hand, the present invention solves the problem of manufacturing products with a wide range of quality characteristics, ranging from the possibility of their use in food products to their use in products that satisfy, for example, pharmaceutical requirements.

On the other hand, with the help of the present invention, this problem is solved by using endogenous enzymes from raw materials and by adapting technological conditions to such enzymes.

The considered method differs significantly from the known methods of hydrolysis in that it can be carried out or it is carried out:

- in the absence of any additives, such as chloroform, in order to prevent unwanted bacterial growth;

- without the addition of sodium hydroxide;

- if hot or cold regeneration of the oil / fat system is possible;

- with the ability to regulate the spectrum of free amino acids and peptides in the final product in the selected form of raw materials for the process as a result of the selection of special raw materials;

- with the ability to regulate the process results related to the amino acid and peptide composition by using the necessary technological parameters, such as temperature and pH;

- without adding acid during the hydrolysis process;

- with a flexible combination of various types of raw materials;

- using an adapted concentration step for separating product fractions;

- by removing proteins and large peptides that are not completely hydrolyzed by coagulation;

- as a result of filtration and separation of coagulated proteins,

- using adapted concentration technology,

- using a shorter hydrolysis stage,

and the result is a product containing minerals and micronutrients of biological origin.

In accordance with aspect b), the object of the present invention is to provide a method for producing a protein product containing free amino acids, as well as short and long chain peptides, based on the use of natural enzymes without the introduction of any substances of artificial origin. As a result of this, the present invention differs from other methods in which enzymes from a large number of different sources are used, for example, bacterial cultures and the like.

This method also includes reducing the amount of fat in the final product to such a low value that eliminates the disadvantages of using fish raw materials. It is intended to produce such a product, which can be used in many fields where the use of products produced by known methods is limited or impossible due to the fat contained in them.

The purpose of the invention also provides for the complete utilization of raw materials and the minimization of environmental loads associated with production.

In accordance with paragraph 15, there is provided a method for regenerating a protein product containing peptides and free amino acids from one or more protein-containing raw materials, characterized in that it comprises the following steps:

but. grinding of raw materials;

b. heating the crushed raw materials to a temperature in the range of 40-62 ° C, preferably 45-58 ° C;

from. optionally isolating the oil / fat from the feed before and / or after the heating step in order to obtain a first oil product;

d. the addition of water having approximately the same or the same temperature as the raw materials and the pH value established by the addition of calcium;

E. the hydrolysis of raw materials using endogenous enzymes in order to obtain a hydrolyzate;

f. optionally adding a pH adjuster during the hydrolysis step, for example calcium, nitrogen or bone meal, to maintain the desired pH of the hydrolyzate,

g. heating the hydrolyzate to 75-100 ° C, preferably 85-95 ° C;

h. removal of large particles from the hydrolyzate, including non-hydrolyzed proteins;

i. optional release of fat / oil in order to obtain a second oil product;

j. removal of proteins and long peptides;

k. the concentration of the remaining amino acids and peptides;

l. the return of proteins and long peptides to the concentrate in order to obtain a protein product;

m. optionally drying the protein product to obtain a dry product containing proteins, free amino acids, and short and long peptides.

In the attached subclauses 16-22, preferred embodiments of the method of the invention according to aspect b) are described.

The protein product obtained by the method according to the invention is characterized in that it contains 5-95 wt.%, Preferably 30-60 wt.% Of free amino acids, while the remaining 95-5% are proteins and minerals, the minerals being natural products of biological origin .

In addition, the protein product contains fat in an amount of less than 0.5 wt.% And has a low salt content, usually less than 1 wt.%.

As indicated in the relevant paragraphs 23, 24 and 25, the present invention also includes the use of the method according to the invention for the production of a veterinary medical product, food product and feed product.

As described in paragraph 26, the invention also relates to oil, characterized in that it is the first oil product obtained by the method according to the invention, having the quality of a food product.

On the one hand, the present invention in accordance with aspect b), inter alia, solves the problem of producing products with a wide range of quality characteristics, ranging from the possibility of their use in food production to the use of pharmaceutical quality products.

On the other hand, in accordance with aspect b), the present invention solves this problem by using endogenous fish enzymes and adapting the process conditions to such enzymes.

The considered method differs significantly from the known methods of hydrolysis in that it can be carried out or it is carried out:

- in the absence of any additives, such as chloroform, to help prevent unwanted bacterial growth;

- without the addition of sodium hydroxide;

- if hot or cold regeneration of the oil / fat system is possible;

- with the ability to regulate the spectrum of free amino acids and peptides in the final product in the selected form of raw materials for the process as a result of the selection of special raw materials;

- with the ability to regulate the process results related to the amino acid and peptide composition by using the necessary technological parameters, such as temperature and pH;

- without adding acid during the hydrolysis process;

- with a flexible combination of various types of raw materials;

- using an adapted concentration step for separating product fractions;

- using adapted concentration technology, as well as

- as a result of using adapted concentration technology;

- use of a shorter hydrolysis stage;

resulting in a product containing minerals and micronutrients of biological origin.

The methods described above according to aspects a) and b) are the natural hydrolysis of proteins to produce dry end products with different compositions of short peptides and free amino acids. As a result of the process, products are obtained containing 5-100% free amino acids, optionally in the absence of proteins and peptides with a long chain (aspect (a)). In addition, such methods describe the release of oil / fat.

In accordance with aspect c), the object of the present invention is to provide a method for the production of a protein hydrolyzate based on the use of natural enzymes without the introduction of any substances of artificial origin. This approach is different from other methods that use enzymes from a large number of different sources, such as bacterial cultures, etc.

In addition, the aim of the present invention is that the method according to aspect c) provides a product that is completely free of protein and DNA, as well as other allergenic compounds, without adversely affecting the utilization of raw materials. This method also includes reducing the amount of fat in the final product to such a low value that eliminates the disadvantages of using fish raw materials. It is intended to produce such a product that can be used in many different fields where the use of products produced by known methods is limited or impossible due to the fat contained in them.

The purpose of the invention also provides for the complete utilization of raw materials and the minimization of environmental loads associated with production.

Thus, according to aspect c) of the invention, in paragraph 27, a method for regenerating peptides / amino acids and an oil / fat system from a protein-containing raw material is disclosed, characterized in that it comprises the following steps:

but. grinding of raw materials;

b. heating the crushed raw materials to a temperature in the range of 40-62 ° C, preferably 45-58 ° C;

from. optionally isolating the oil / fat from the feed before and / or after the heating step in order to obtain a first oil product;

d. the addition of water having approximately the same or the same temperature as the raw materials and the pH value established by the addition of calcium;

E. the hydrolysis of raw materials using endogenous enzymes or enzymes from similar raw sources, preferably from cold-blooded varieties, in order to obtain a hydrolyzate;

f. optionally adding a pH adjuster in the hydrolysis step to maintain the desired pH of the hydrolyzate,

g. removal of solid particles and non-hydrolyzed proteins that may return from the hydrolyzate to the hydrolysis stage;

h. periodic or continuous release of fat / oil in order to obtain a second oil product;

i. optional treatment of the hydrolyzate, preventing the growth of microorganisms, preferably treatment with UV radiation;

j. the selection of the desired molecular weight fraction of peptides / amino acids by membrane filtration, preferably transfer type;

k. the direction of the parts of the hydrolyzate that do not penetrate through the membrane filter in paragraph j back to the hydrolysis stage carried out in stage e;

l. the concentration and optional drying of the permeate in order to obtain a mixture of peptides and amino acids;

m. full or partial return of the distillate from the concentration stage to the penetrating side of the membrane filter.

In the attached subparagraphs 28-31, preferred embodiments of the method according to the invention are described.

The term “membrane filter” is used in the context of the description to mean membrane-like filters, such as membrane filters, osmotic filters, ultrafilters, electrostatic filters, cross-flow filters, and the like. Preferably, such filter media has a cut-off value of less than or equal to 10,000 daltons.

In addition, paragraph 32 discloses a method for hydrolysis of one or more types of raw materials containing protein, and the selection of amino acids / peptides, which is characterized in that the hydrolysis is carried out using endogenous enzymes of protein-containing raw material or materials, as well as the fact that the hydrolyzate is passed through a membrane filter, and the peptides / amino acids pass with the flow of permeate, while the active enzymes continuously destroy protein residues deposited on the membrane surface, as well as the fact that the enzymes together with etentatom recycled to the hydrolysis step.

As described in paragraph 33, another distinguishing feature of the invention is a method for isolating peptides and amino acids from a hydrolysis mixture, characterized in that the hydrolysis mixture containing active enzymes, amino acids, peptides and unconverted proteins is passed through a membrane filter, and the amino acids and any peptides are filtered and the active enzymes present provide the breakdown of proteins deposited on the membrane filter.

In accordance with paragraphs 34-37, the invention also includes the use of one of the methods according to the invention for the manufacture of a pharmaceutical product, biotechnological product, food product and feed product.

As indicated in paragraph 38, the invention also includes the use of the method according to the invention for the production of hydroxyapatite.

In addition, as described in paragraph 39, the invention relates to amino acids / peptides obtained by the method according to the invention and characterized in that they do not contain allergens and traces of DNA.

According to paragraph 40, the invention also relates to oil obtained by the method according to the invention and characterized in that it does not contain allergens and traces of DNA.

Finally, in accordance with paragraph 41, the invention relates to hydroxyapatite obtained by the method according to the invention and characterized in that it does not contain allergens and traces of DNA.

On the one hand, in accordance with aspect c), the present invention solves the problem of obtaining products with a wide range of quality, which can vary from their use in food production to their use in products that satisfy, for example, pharmaceutical requirements.

On the other hand, in the present invention, such a problem is solved by using endogenous enzymes in the feed and adapting the process conditions to such enzymes.

In accordance with aspect c), the present invention combines the use of endogenous enzymes with the method of regenerating special-sized molecules from systems from simple amino acids to large peptides with a size of slightly less than 10,000 daltons.

The present invention includes the retention of enzymes during the fermentation process, while separating the released amino acids and peptides.

In addition, the present invention provides for the possibility of a continuous hydrolysis process with the introduction of an additional amount of raw materials during the process.

The considered method differs significantly from the known methods of hydrolysis in that it can be carried out or it is carried out:

- in the absence of any additives, such as chloroform, to prevent unwanted bacterial growth;

- without the addition of sodium hydroxide;

- if possible hot or cold regeneration of protein-free and sterile oil / fat of marine animals;

- with the ability to regulate the spectrum of free amino acids and peptides in the final product in the selected form of raw materials for the process as a result of the selection of special raw materials;

- with the ability to regulate the process results related to the amino acid and peptide composition by using the necessary technological parameters, such as temperature and pH;

- without the addition of acid;

- with a flexible combination of various types of raw materials;

- as a result of using an adapted concentration stage to isolate product fractions; as well as

- when using a continuous process of enzymatic decomposition;

- without coagulation of proteins and / or peptides using an acid or base;

- as a result of the classification of the obtained peptides by size;

resulting in a product containing minerals and micronutrients of biological origin.

Thus, a method is provided for regenerating a mixture of peptides and amino acids, minerals and oil or fat from proteinaceous materials, preferably originating from aquatic organisms.

Known methods differ from the present invention in a completely different way to obtain fish proteins, which are also present in various fractions containing peptides and amino acids. Technological conditions also differ significantly from the prior art, and in the present invention are excluded by the use of exogenous enzymes. In addition, as indicated in the material cited above, the high-quality oil / fat of marine animals in the present invention is obtained by developing a special method that is not described in the prior art.

The disclosed method is a natural fermentation of proteins to produce dry end products or liquid products containing various amounts of peptides and free amino acids. This method provides the finished products, which optionally contain 5-100% free amino acids. The product in question does not contain allergens or traces of DNA. The product contains very small amounts of fat and biological micronutrients, usually less than 0.1%. The method according to the invention provides a product that is extremely useful as a culture medium for all types of cultures, including cells of higher organisms.

The present invention makes it possible to carry out the process without the use of sodium hydroxide, the presence of which creates problems in the production of amino acids and peptides on an industrial scale. In addition, the ratio of water can vary to a greater extent than in the known methods, while on a larger scale, the mass percentage of free amino acids can change.

Compared with known methods, both the crushed and the crushed material are used in the present invention, without the addition of a fermenting preparation, but using natural enzymes already present in the feed. Thus, endogenous enzymes of crude protein material are used, which ensures hydrolysis in a more stable and less expensive way. In addition, the conditions must directly adapt to the activity of endogenous enzymes, which also differs from known methods.

Another production problem that exists in this industry is that exogenous enzymes are expensive materials and can have different qualitative characteristics. In the present invention, this problem is solved by recycling endogenous enzymes.

In addition, active enzymes have a special cleaning effect. Since they are retained on the filter, such enzymes act on unfiltered proteins and peptides. Enzymes contribute to the degradation of such materials and therefore have a longer service life compared to traditional filtration methods, so far used in known hydrolysis processes. All this provides significant advantages in terms of cost, service life and filter efficiency, product quality and the degree of disposal of the system and process.

In addition, the method describes the regeneration of oils / fat, as well as solid materials. One of the solids that can be formed using the method of the present invention is hydroxyapatite. Hydroxyapatite is used, for example, in biochromatography and other processes for the separation of biological objects, in NMR and other methods of detection, and, thus, this substance is a commercially attractive by-product of the process.

The choice of method and process parameters determines the nature of the resulting final product. Thus, it becomes possible to adapt products to customer requirements.

Figure 1 depicts a first variant of the installation, which carry out the hydrolysis method according to aspect a).

Figure 2 depicts a second variant of the installation, which carry out the hydrolysis process according to aspect b).

Figure 3 depicts a third embodiment of a plant in which the hydrolysis process is carried out according to aspect c).

Embodiments of the methods according to aspects a) and b) of the invention are further explained in more detail with reference to FIGS. 1 and 2, respectively.

Raw materials are pumped from tank 101; 201 through a grinding system 102; 202, which provides the desired crushing of raw materials. Oil / fat from the feed can be released before the fermentation process begins. So, for example, a method of cold oil recovery can be carried out. You can also not allocate oil before fermentation. In this case, the feed is pumped directly to the heat exchanger 105; 205 with a bypass flow D1; D2.

Cold oil isolation is carried out using the following operations:

1. centrifugation of raw materials and separation of liquid and solid particles into two fractions, for example, in a decanter centrifuge 103; 203;

2. the selection of oil from the liquid phase stream A1; A2 using, for example, a separator 104; 204;

3. mixing the flow of the solid phase B1; B2 and the heavy phase from the separation stage and forcing the stream C1; C2 into the fermenter;

4. moreover, the oil phase from the separation stream D1; D2 can be pumped into the reservoir 110; 210 through a sterile filter 109; 209 and therefore does not require additional purification to obtain food-grade material.

Materials can be pumped through the built-in continuous heat exchanger 105; 205 or a batch heat exchanger into the fermentation tank 106; 206. The fermentation tank can also be used for heating, if this operation is not carried out in the heat exchanger before pumping materials. A stream of warm water E1; E2 with a set pH value is added to the feed, which is heated by passing through a heat exchanger 108; 208 to approximately the temperature of the fermentation. It is preferable to adjust the pH by passing water through a filter medium that releases calcium 107; 207.

During the process in the fermenter using sensors 111,111 '; 211,211 'or other methods control the temperature and pH. The desired pH during the process is set using bone meal or calcium, which are added from the store 112,212. Nitrogen 113,213 can also be used to adjust the pH during the process.

After completion of the fermentation, the hydrolyzate is preferably heated by passing through heat exchangers 114; 214, resulting in the activation of enzymes.

If the hydrolyzate contains bones or other solid particles, they are preferably removed using a sieve device 115; 215. Particulate matter, flow F1; F2, may be separated into two or more fractions by flotation 116; 216. The heavy fraction 117; 217 consists of bones (hydroxyapatite), which can be dried and / or used before the pH adjustment stage, flow H1; H2. The light fractions, the G1; G2 stream, are mainly composed of proteins not hydrolyzed.

The light fraction G1 can be returned to the crusher 102, as shown in figure 1, and is used as raw material for the new hydrolysis process or is discharged as a by-product.

According to aspect b) of the present invention shown in FIG. 2, the light fractions G2 can be passed separately and mixed with the concentrate from the enrichment plant 222. On the other hand, they can be dried as a separate product.

The fat remaining after the first separation is isolated using, for example, a three-phase separator 118; 218. The oil / fat stream I1, I2 is filtered on a filter 119; 219, and it can be fed into a reservoir 120; 220 for optional subsequent further processing, etc.

As shown in FIG. 1, the hydrolyzate stream J1 is sent to a mixer 121 to which acid 122 (preferably phosphoric acid) is added to pH <5. After this, calcium 123 is added, resulting in agglomeration of long peptides and proteins. The proteins and the calcium / calcium phosphate system are separated using a centrifuge 124 and sent to the reservoir 125. Such a by-product can be dried at high temperature, resulting in the burning of residual protein.

The protein-free hydrolyzate stream K1 is preferably concentrated in a vacuum evaporator 126. The condensate stream L1 can be used as an additional water stream E1. The concentrate stream M1 may be dried in a spray dryer 127, preferably in a Filtermate® type apparatus. The concentrate can also be extracted as a stream N1 of a liquid product.

As shown in FIG. 2, the hydrolyzate stream J2 can be processed using a centrifuge 221, which separates the stream N2 from proteins and long peptides. This material is mixed with a concentrate, an M2 stream consisting of free amino acids and short peptides from a concentrator 222. A condensate stream D2 can be used as an additional water stream E2. A stream of M2 concentrate mixed with proteins and long peptides can be dried in a spray dryer 223, preferably a Filtermate® device.

Unless otherwise specified, all percentages given in the text are given in mass terms.

The following is a more detailed summary of the invention.

1) Raw materials

The raw materials for the described method may consist of protein material, preferably fish, fish products, shellfish, crustaceans and by-products of the fishing / fishing industry, for example, waste and other marine organisms living in both fresh and salt water. Different types of raw materials can be used separately or as a combination of products containing "enzyme material" and "protein material". "Enzyme material" is a raw material containing endogenous enzymes in sufficient quantity and satisfactory quality. "Protein material" is a raw material that does not contain a sufficient amount of endogenous enzymes of satisfactory quality, which therefore must be supplemented with an enzyme product in order to ensure enzymatic processing. In some cases, the enzyme material may be identical to the protein material. Known methods describe a combination of waste and protein material in a 1: 1 ratio. The method disclosed in the present invention makes it possible to change this ratio in order to achieve the desired result in relation to the final product.

Raw materials meet the established requirements for the starting substances for the production of food products. Previously, raw materials were classified in accordance with legislation and terminology as waste. Thanks to good logistics and technical programs, in this case, raw materials can be recognized as a food product. This allows you to carry out production on an industrial scale and use the product in the food and / or pharmaceutical industry.

2) Pre-treatment of raw materials:

Raw materials are pumped from the tank through a crushing system that provides the desired grinding of materials. Crushing provides a large working surface for enzymes and accelerated isolation of raw enzymes. Oil / fat from the feed may be released before the fermentation process begins. Cold oil regeneration may be used.

Cold oil recovery includes the following stages:

1. centrifugation of raw materials and separation of liquid and solid particles into two different fractions;

2. the selection of oil from the liquid phase;

3. mixing the solid phase and the heavy phase from the separation stage and pumping the mixture into the fermenter;

4. additional processing of the oil phase from the separation stage with the formation of a finished product, acceptable to the consumer, not requiring additional purification to obtain the quality of the food product.

Materials can be pumped in-line through a continuous heat exchanger or through a batch heat exchanger to a fermentation tank. The fermentation tank can also be used for heating, if such an operation is not carried out in the heat exchanger before pumping the materials. If it is desirable to mix different types of raw materials in a certain ratio, these products can be pumped and mixed at the same time. The number of different types of raw materials can be controlled using a flow meter and / or level control in the fermenter.

In addition, it is possible that the raw material does not grind or different types of raw materials should not be mixed at the same time, in which case a known amount of raw material A is pumped into the mixer, after which a known amount of raw material B.

It is desirable that the feed is heated to a temperature conducive to the manifestation of maximum activity by the enzymes during hydrolysis. Usually use a temperature of 40-62 ° C. In most cases, the optimal temperature range is 45-58 ° C. The use of different temperatures provides the formation of various enzymes and regulates the composition of amino acids.

The present invention provides the possibility of regeneration / separation of fat during the process after hydrolysis to obtain the final product with a low fat content.

Fat can be removed before and / or after protein coagulation. Typical dry peptide / amino acid end products contain less than 0.1% fat.

Well-known methods are used to separate fat, such as decantation, separation and / or chemical methods.

3) Hydrolysis process:

The heated feed mixture is pumped into hydrolysis tanks or fermentation tanks. Warm water with a set pH value is added to the mixture, having a temperature close to the temperature at which fermentation is carried out. The amount of water may vary depending on the nature of the feed and the desired result. In known methods, the amount of water was 50% of the total amount of material, i.e. used 50% of raw materials and 50% of water. The method according to the invention allows the use of less water, which is associated with the optimization of the used enzyme, protein material, as well as temperature and pH. In this method, add 10-40% of water. The optimal amount of added water is 20-30%. A decrease in the amount of added water means that the hydrolyzate obtained after hydrolysis contains increased amounts of short peptides and free amino acids, which reduces the cost of the process and energy costs.

The hydrolyzate in the fermentation tank (s) is under constant stirring and pumping. This is done to improve the performance of the hydrolysis process. Violations during the mixing and pumping of the hydrolyzate can lead to insufficient control of pH, temperature and the process itself. Different parts of hydrolysis can be hydrolyzed in various ways, which can lead to the loss of enzymes.

To determine the depth of hydrolysis, an analysis is carried out for amino-bound nitrogen. Such an analysis can be carried out directly in the fermenter using automatic equipment or in a technological laboratory using well-known methods such as titration with formaldehyde and the like. The duration of the process can be 1-4 hours. The hydrolysis process is stopped after the cessation of the increase in the concentration of free amino acids in the hydrolyzate. This is done in order to avoid the formation of undesirable ammonia, which leads to a decrease in the degree of utilization of the feedstock.

A necessary condition for the considered method is the use of alkaline enzymes. For this, it is necessary that during hydrolysis a pH> 7.00 be maintained. The range of operating pH is 7.00-8.50. Optimum hydrolysis is carried out at pH 7.60-8.20. At pH> 8.1, but <8.4, tryptophan disappears from the amino acid spectrum during the whole process. On the contrary, if during the whole process 7.6> pH> 7.4, the amount of tryptophan increases to the maximum possible value that can be isolated, which is determined by the type of raw material. If during the whole process the temperature is maintained <46 ° C, but> 44 ° C, and the pH is less than 7.8, but more than 7.7, then collagen does not noticeably dissolve, and it is obtained in the form of solid particles.

Various bases can be used to adjust the pH of the hydrolyzate, for example bone meal from pre-isolated fish bones, calcium and nitrogen / nitrogen gas, but not sodium hydroxide according to aspect a).

In the method according to the invention do not add HCI. The reason for this is the possibility of the formation of undesirable salts. In addition, the cost of production may increase. Known methods also describe the use of chloroform as a means of preventing the growth of bacteria. In the method according to the invention, such an operation is not carried out due to the short hydrolysis time. On an industrial scale, the use of chloroform is not desirable, but possible.

To stop the hydrolysis using an elevated temperature, preferably above 70 ° C. The temperature increase is preferably carried out using built-in heat exchangers.

The coagulation of the remaining proteins is carried out at a low pH. To reduce the pH to the desired value, preferably 3.2-5.5, phosphoric acid can be added to the system. After the addition of phosphoric acid, optionally in combination with heating / cooling, it becomes possible to remove proteins and peptides from the hydrolyzate. Other methods can be used to denature proteins and peptides. So, for example, you can use the method of electrodenaturation.

Known filtration methods can be used to separate free amino acids and short peptides from denatured proteins and long peptides. The separation of these materials is also facilitated by various masses and various chemical properties of the fractions. The implementation of this process is facilitated by the use of calcium phosphate, calcium hydroxide and calcium chloride. These substances cause joint “bonding” of proteins and peptides. As a result, their separation is facilitated by increasing density. The amount of added chemicals may vary depending on the content of proteins and peptides, as well as other buffering agents, in the solution.

4) Concentration

The finished hydrolyzate is preferably subjected to concentration. This operation is carried out to remove water before the drying stage, as a result of which the productivity of the process maximizes. Prior to crystallization, a preliminary concentration can be carried out before drying to a dry matter content (DM) of 70%.

For this purpose, distillation processes such as vacuum evaporation are particularly suitable, however, any other types of concentration devices can be used. A vacuum evaporator provides liquid concentration at low temperature, which prevents damage to peptides or amino acids.

Evaporation is carried out at a temperature of 50-85 ° C. The optimum temperature range is 65-70 ° C. In addition, the hydrolyzate can be fed directly to the drying stage (see paragraph 5), bypassing the concentration stage, or the concentration can be carried out by methods other than boiling or vacuum evaporation. For the implementation of this stage can also be used various types of installations: filtration, membrane and osmotic types.

5) Drying / granulation

After concentration, if desired, the product may be dried, but the product may also exist in liquid form or any intermediate state between the dry and liquid material. Drying increases the stability of the product during storage and simplifies the processes of logistics and processing. The method of drying the product is an important factor for the final result. The resulting peptide / amino acid product may be highly hygroscopic material, which may become a problem for the method under consideration. To facilitate processing of the product, it is desirable to deal with its granular form.

In accordance with one embodiment of the present method, drying and granulation are carried out in two stages, although one-step processes are possible. The first stage includes drying to a powder state in a spray dryer or similar device, together with a cooling stage, after which granulation is carried out by adding a liquid hydrolyzate.

The granulation is carried out in such a way that the granulate is “constructed” as a result of the intense movement of the powder / product system, which is carried out using mechanically rotating blades that ensure the fluidized nature of the product. After that, the resulting mass is sprayed with a mixture of concentrate / hydrolyzate. This process provides the gradual formation of granular material. All of these operations are carried out in a continuous manner. At the end of the granulation process, dry cold air is passed through the granulate or on its surface. The granulate is sieved and the desired fractions are taken. The remaining fine particles are recycled to the stage of additional granulation. Excessively large granules are ground and again passed through a sieve.

As noted above, the described drying and granulation processes correspond to one embodiment of the invention that can be used to dry and granulate the products of the method of the invention, and it should be apparent to one skilled in the art that other suitable methods may be used to obtain a similar result.

Various additives can also be added to the product, preferably at the granulation stage.

The method of the invention according to aspect c) is described in more detail below. The reference numbers in FIG. 3 correspond to the following installation elements:

301 = fermenter

302 = separation device for removing particulate matter, preferably a sieve

303 = separation device, preferably a centrifuge, such as a decanter

304 = flotation tank for the separation of proteins and hydroxyapatite

305 = oil separation device, preferably a centrifuge

306 = filter device for sterile filtration

307 = reservoir for oil / grease system

308 = microorganism reduction unit

309 = membrane filtration module

310 = hub

311 = drying unit

312 = grinding equipment

313 = centrifuge preferably decantation type

314 = oil filter

315 = oil recovery tank before the hydrolysis step

316 = heat exchanger for heating raw materials

317 = calcium dosing device

318 = heat exchanger for heating water

319 = bone meal feeder

320 = nitrogen supply

321 = capacity for regenerated bone fraction

In addition, figure 3, together with the description of the installation, shows an embodiment of the method of the present invention according to aspect c), using the following flow designations:

A3 = a stream comprising proteins, enzymes, oil / fat, peptides and free amino acids.

After passing through the microorganism reduction unit 308, the stream does not contain solid particles and when using separation devices 302 and 303 has a significantly reduced content of non-hydrolyzed proteins and fat;

A31 = flow if block 302 is used; after passing block 302, the stream no longer contains particulate matter;

A32 = flow if block 303 is not used;

B31 = stream including solid particles removed by a sieve;

B32 = flow comprising solid particles separated by centrifuge;

C3 = stream comprising an oil / fat system;

D3 = a stream of distillate or similar material designed to drain the permeate from the membrane filter;

E3 = stream of concentrated peptide-amino acid solution sent to the drying unit;

E31 = a stream of concentrated peptide-amino acid solution sent to the packaging step as a liquid product;

F3 = feed stream to fermenter 301;

F31 = feed stream, when no oil separation is carried out before the hydrolysis steps;

G3 = non-hydrolyzed protein stream returned to stream A3;

H3 = stream of added water;

I3 = flow of oil regenerated before the hydrolysis step.

Unless otherwise indicated, all percentages are given in mass terms.

The following is a more detailed description of the method:

1) Raw materials

The feed for the described process may consist of protein material, preferably fish, fish products, mollusks, crustaceans and by-products of the fishing / fishing industry, such as waste and other aquatic organisms living in both fresh and salt water. Different types of raw materials can be used separately or as a combination of products containing "enzyme material" and "protein material". "Enzyme material" is a raw material containing endogenous enzymes in sufficient quantity and satisfactory quality. "Protein material" is a raw material that does not contain satisfactory quality endogenous enzymes in sufficient quantities and which therefore needs to be supplemented with an enzyme product in order to ensure enzymatic processing. In some cases, the enzyme material may be identical to the protein material. The prior art describes a combination of waste and protein material in a 1: 1 ratio. The method disclosed in the present invention makes it possible to change this ratio in order to achieve the desired result in relation to the final product.

Raw materials meet the established requirements for the starting substances for the production of food products. Previously, raw materials were classified in accordance with legislation and terminology as waste. Thanks to good logistics and technical programs, in this case, raw materials can be recognized as a food product. This allows you to carry out production on an industrial scale and use the product in the food and / or pharmaceutical industry.

2) Pre-treatment of raw materials:

Raw materials are pumped from the tank through a crushing system 312, which provides the desired grinding of materials. Crushing provides a larger work surface for enzymes and faster isolation of raw enzymes.

The first oil / fat system may be released from the feed prior to the start of the fermentation process. In this case, for example, cold oil recovery may be used.

Cold oil recovery includes the following stages:

1. centrifugation of 313 raw materials and separation of liquid and solid particles into two different fractions;

2. the selection of oil from the liquid phase, see 314 and 315;

3. mixing the solid phase and the heavy phase from the separation stage and pumping the mixture into the fermenter 301;

4. additional processing of the oil phase from the separation stage with the formation of a finished product, acceptable to the consumer, not requiring additional purification to obtain the quality of the food product.

Raw materials added to the fermenter 301 may be stored in one or more buffer vessels after grinding. For protein and enzyme materials, different capacities can be used.

Materials can be pumped through the heat exchanger 316 to the fermentation tank 301. The fermentation tank 301 can also be used for heating if such an operation is not performed in the heat exchanger before pumping the materials.

The conditions in the fermenter 301 are monitored continuously, automatically, or by sampling. The introduction of various additives is carried out in such a way that the most constant fermentation conditions are maintained within the ranges of values that are optimal for the product being produced.

3) Fermentation process:

Heated raw materials or unheated raw materials in the form of protein and enzyme materials were pumped into one or more fermentation tanks. Warm water with a fixed pH value was added to the mixture, having a temperature at which fermentation takes place. The amount of water can vary according to the type of raw material and the desired result, depending on the optimization of the available enzyme and protein material. The pH adjustment is carried out, for example, by adding gaseous nitrogen or bone meal.

Further in the text, a heated mixture of raw materials F3 and water H3 with a set pH value is referred to as "hydrolyzate". The hydrolyzate is kept in a fermentation tank 301, optionally under conditions of constant and intensive mixing. The purpose of this operation is to intensify the fermentation process. The hydrolyzate is continuously pumped through the removal system of the desired amino acids and peptides.

To maintain constant fermentation conditions in the fermentation tank 301, the amount of amino-bound nitrogen, the amount of total protein and pH are regularly monitored.

If necessary, protein and enzyme material is added during the process.

A necessary condition for the considered method is the use of alkaline enzymes. For this, it is necessary that during fermentation a pH> 7.00 be maintained. The range of operating pH is 7.00-8.50. Optimum fermentation is carried out at a pH of 7.60-8.20. At pH> 8.1, but <8.4, tryptophan disappears from the amino acid spectrum during the whole process. On the contrary, if during the whole process 7.6> pH> 7.4, the amount of tryptophan increases to the maximum possible value that can be isolated, which is determined by the type of raw material. If during the whole process the temperature is maintained <46 ° C, but> 44 ° C, and the pH is less than 7.8, but more than 7.7, then collagen does not noticeably dissolve, and it is obtained in the form of solid particles.

To adjust the pH value, various bases can be added to the hydrolyzate, such as bone meal from a previously obtained batch, calcium and nitrogen, see 319 and 320.

In accordance with aspect c) of the present invention, it is possible to continuously monitor the fermentation process using various parameters in order to maintain the technological conditions at an optimal level.

At the end of the process, enzymatic activity should be limited by thermal inactivation or another way to prevent ammonification.

4) Control the growth of microorganisms:

In the prior art it is indicated that chloroform is added to prevent the growth of microorganisms, however, this material is preferably not used in the present method. In the method according to the invention for the destruction of bacteria and fungi, UV irradiation or another suitable method is used that does not lead to coagulation of enzymes. This operation is performed to prevent significant growth of microorganisms due to the consumption of released short peptides and free amino acids in the formation of new proteins. It should also be borne in mind that on an industrial scale, chloroform is preferably not used.

5) Removal of solid particles:

The present invention contemplates that solid particles larger than a certain size can be continuously or periodically removed from the hydrolyzate by a sieve / filter system. The sieve system 302 may not be used or located in a bypass line if the sump system 303 in the next process stage removes all solid phase, or when the product A3 processed in this situation does not contain solid particles suitable for sieving.

Then, the removed B32 particles can be separated according to their density using the flotation process 304, whereby the G3 protein residues can be returned to the fermentation tank 301. The proteins float and can be removed mechanically or manually from above. Heavier material is collected at the bottom of the fermentation tank.

6) Decantation of the hydrolyzate:

Sump 303 can be used in the system in question up to an oil separator 305 and a membrane filter 309. This operation is intended to simplify the separation of fat from the hydrolyzate and is carried out, for example, in a three-phase separator. As a result, the load on the next membrane filter is reduced.

The separator is not able to operate in the optimal mode if the solids content is too high, which implies the presence of a large volume of sludge phase. The sump is a device designed to separate solid particles whose density is higher than the density of the liquid of which they are a part. In the present invention, such particles are mainly proteins that are subject to separation, and therefore, it is desirable to direct them back to the fermentation tank for additional fermentation. The separated solid material may float in accordance with the same principles used in the screening system. If necessary, a flotation device may be used. Sump 303 may not be used or disconnected from the system if the already mentioned sieve device provides the desired separation, or in the case where the products to be processed do not form protein residues that can be removed using the sump.

7) Separation

The hydrolyzate is separated using a three-phase separator 305 or another centrifugation method suitable for separating the lighter fat fraction from the hydrolyzate. The separation of fat is carried out continuously or batchwise depending on the amount of fat in the processed raw materials. A particularly important distinguishing feature of the present invention is that a very pure, high-quality fat fraction can be isolated, since separation can be carried out continuously throughout the process so that the extracted fat is not oxidized more than necessary when the lipoproteins are decomposed by hydrolysis. Hydrolysis under alkaline conditions also helps to maintain high quality fat, especially when nitrogen is used to adjust pH.

8) Membrane filtration:

The present invention provides for pumping the hydrolyzate through a device, preferably equipped with a membrane filter 309, acting so that only molecules of a certain size, preferably less than 10,000 daltons, penetrate the membrane. The filtration is carried out in such a way that the hydrolyzate is pumped through many tubular membranes or passes through many flat membranes.

Membrane transport is based on the principles of osmosis. As a result of the concentration of unfiltered free amino acids and peptides, distilled water is formed as a by-product, and part of it is returned to the filter at approximately the same pressure as the hydrolyzate on the other side of the membrane. Maintaining a lower concentration of amino acids and peptides on the permeable side of the membranes, penetration through them under the influence of osmosis can be carried out. The flow of the hydrolyzate along the membrane surface mechanically cleans it from the sediment of protein residues and peptides, larger than those that are able to penetrate through the membrane.

9) Concentration:

After obtaining the hydrolyzate filtrate, a concentration step follows. This operation is designed to remove water before carrying out the drying process, as a result of which the capacity of the drying stage is fully used or the level of concentration of incoming amino acids and peptides that is desirable for the liquid product is achieved.

A distillation process such as vacuum evaporation is best suited for this purpose, but any other form of concentration device can be used to remove the desired peptides and amino acids from the liquid in which they dissolve during membrane filtration. A vacuum evaporator 310 concentrates the liquid at a low temperature, as a result of which peptides / amino acids are not damaged. A prerequisite for optimal membrane filtration in the early stages is the ability of the concentration device 310 to return as pure distillate as possible to the system. In this case, optimal osmosis through the membrane is realized. Evaporation can be carried out in the temperature range of 50-85 ° C. The optimum temperature range is 65-70 ° C. There is a risk that the condensate temperature is too high to pass back to the filter or return to the fermentation tank. In this case, a heat exchanger should be used to lower the temperature to the desired value.

10) Drying:

If necessary, after concentration, the product can be dried, however, the resulting material may be in the form of a liquid or is in an intermediate state. Drying, see 311, increases the stability of the product during storage and simplifies the logistics and processing processes. The method of drying the product is an important factor for the final result. The resulting peptide / amino acid product may be a highly hygroscopic material, which may be a problem for the method under consideration. The high temperature of the drying process can cause the product to become even more hygroscopic.

Drying / granulation is carried out in two stages. First, the product is dried to a powder state in a spray dryer or similar device with a cooling step, after which the product is granulated. The granulation is carried out in such a way that the granulate is “constructed” as a result of maintaining the powder / product system in a state of intense movement using mechanical fluidization means. Then, the concentrate / hydrolyzate is sprayed on the resulting mass, resulting in the gradual formation of granulate. All of these operations are implemented in a continuous process. By the end of the granulation process, dry, cold air is passed through / through the granulate. As a result, the material becomes harder and more soluble. After that, the granulate is sieved and the desired fractions are taken. Too small particles (fines) are returned for additional granulation, while particles larger than the nominal size are subjected to grinding and sieved again. The newly formed small particles are returned to the granulation stage.

Conventional spray drying may be used, but in this case a fine-grained powder with a large particle surface is obtained. This means that the material obtained is highly hygroscopic, which makes it difficult to handle large packages, storage, etc.

During the granulation process, various additives can be introduced into the product. Products can be produced that do not granulate, as in the case of those products that have not been dried, but simply concentrated to the desired degree.

Detailed description of the membrane filter

Filter 309 may be constructed of flat or tubular filter elements. The filtration system is designed in such a way that the filtrate, consisting of a protein hydrolyzate from the fermenter, from which solid particles and fat are removed, but which contains the enzyme complex, is able to circulate freely outside the holding side of the membranes (cross-flow filtration). Thus, a stream is formed along the surface of the membranes, which mechanically minimizes the risk of the formation of a blocking deposit on the filter from retentate deposited on the membranes. The filtrate circulating on the retentate side of the filtering membranes contains enzymes that break down proteins and large peptides that are retained on the membranes but are too large to penetrate the membranes from the penetrating side. Enzymes do not penetrate the membrane if it is selected in such a way that the maximum size of the permeate molecules is 9800 daltons. Thus, undecomposed proteins are blocked, and a protein-free sterile product is formed.

The choice of a membrane capable of blocking small molecules leads to the formation of a product containing smaller peptides and more free amino acids.

On the penetrating side, a stream of water is created, passing along the membranes, corresponding to the flow on the penetrating side. The same pressure can be maintained on both sides of the membrane and permeate passes through the membrane under osmosis. Dissolution occurs in a fluid circulating on the penetrating side of the membrane.

Osmosis conditions are created in case of an increased concentration of amino acids and peptides on the retaining side of the membrane. This is achieved when the distillate from the concentration plant is a liquid circulating from the penetrating side of the membrane. In principle, this option can be characterized as reverse diafiltration occurring on the penetrating side when pure water is added.

The permeate concentration can be carried out using another membrane filter designed for reverse osmosis (RO). The function of the membrane filter remains the same.

A series of filters can be used to separate the various fractions, with regard to the maximum size of the peptides, but on subsequent filters the enzyme complex does not contribute to the release of the retaining side of the filter membrane from the filter cake. In this case, in order to prevent blocking, it is useful to carry out a one-stage filtering.

The following results were obtained in laboratory tests using a dialysis membrane standard hose type used in clinical conditions, manufactured by the manufacturer Spectrum Laboratories.

Spectra / Por 1 — Artificial Cellulose (RC) with a molecular weight cut-off (MWCO) of 6000-80000 daltons (6k-8k MWCO) was used in these tests. The initial flow through such membranes with a total content of DM (dry matter) on the holding side of 14.7% at a temperature of 48.7 ° C and a pH of 7.85 was 3.7 ml / cm ² / h. After 12 hours, the flow was 3.8 ml / cm ² / h and after 24 hours also 3.8 ml / cm ² / h. Within 60 hours of operation, no blocking of the system was observed. No molecules weighing more than 9,000 daltons were detected in the permeate when analyzing the size of the peptides before and after the concentration of 23 liters of liquid containing 36% DM formed over 60 hours. The largest peak in the spectrogram corresponded to the region 410-1350 daltons, which, without correction, amounted to 42% of the spectrogram area.

Claims (18)

1. The method of producing peptides / amino acids with a fat content of less than 0.5 wt.%, From raw materials containing protein, characterized in that it includes the following stages: a. grinding of raw materials; b. heating the crushed raw materials to a temperature in the range of 40-62 ° C, preferably 45-58 ° C; d. the addition of water having about the same or the same temperature as the raw materials, and a pH value of 7.0-8.5, established as a result of the addition of calcium; e. hydrolysis of raw materials using endogenous enzymes in order to obtain a hydrolyzate; g. removal from the hydrolyzate of solid particles and non-hydrolyzed proteins that may return to the hydrolysis stage; h. periodic or continuous separation of fat / oil in order to obtain a second oil product; j. the selection of the desired molecular weight fraction of peptides / amino acids by membrane filtration, preferably transfer type; k. returning portions of the hydrolyzate not penetrating through the membrane filter according to item j to the hydrolysis step e; l. the concentration and optional drying of the permeate in order to obtain peptides / amino acids; and m. full or partial return of the distillate from the concentration stage to the permeable side of the membrane filter. 2. The method according to claim 1, characterized in that it includes, before and / or after stage b from. the selection of oil / fat from raw materials in order to obtain the first oil product. 3. The method according to claim 1, characterized in that it further includes during the hydrolysis step e f. adding a pH adjuster to maintain the desired pH of 7.0-8.5 hydrolyzate. 4. The method according to claim 1, characterized in that it further includes after stage h i. treatment of the hydrolyzate, aimed at suppressing the growth of microorganisms, preferably using UV radiation. 5. The method according to claim 1, characterized in that it is carried out in a closed process. 6. The method according to claim 1, characterized in that the pH regulator in step f is nitrogen gas or bone meal. 7. The method according to claim 1, characterized in that it further comprises dividing the solid particles from step g into hydroxyapatite, protein residues, and other solid particles. 8. The method according to claim 1, characterized in that the second oil product regenerated in stage h is passed through a filter and heavy components (for example, stearic acid) are removed to obtain a cold-pressed, protein-free sterile oil. 9. The oil obtained by the method according to claim 8, characterized in that it does not contain allergens or traces of DNA. 10. Hydroxyapatite obtained by the method according to claim 7, characterized in that it does not contain allergens or traces of DNA. 11. The method of hydrolysis of one or more types of protein-containing raw materials and the selection of amino acids / peptides, characterized in that the hydrolysis is carried out using endogenous enzymes of the protein-containing material or materials; while the heated mixture is pumped into the hydrolysis tank, water is added at a temperature of 45-58 ° C in an amount of 10-40%, the mixture is constantly mixed and pumped, and during the hydrolysis, the pH is 7.6-8.2. 12. A method for removing peptides and amino acids from a hydrolysis mixture, characterized in that the hydrolysis mixture containing active enzymes, amino acids, peptides and unconverted proteins is passed through a membrane filter, where amino acids and peptides with a molecular weight of less than 10,000 D are filtered, and the active enzymes present destroy the proteins deposited on the membrane filter. 13. Amino acids / peptides obtained by the method according to claim 1, characterized in that they do not contain allergens and traces of DNA, and the content of free amino acids is 30-60 wt.%, The fat content is less than 0.5 wt.%, And the content salt is less than 1 wt.%. 14. Amino acids / peptides obtained by the method according to claim 1, characterized in that they do not contain allergens and traces of DNA, and the content of free amino acids is 5-100 wt.%, The fat content is less than 0.1 wt.%, And the content salt is less than 0.5 wt.%. 15. The use of amino acids / peptides according to item 13 or 14 to obtain a pharmaceutical product. 16. The use of amino acids / peptides according to item 13 or 14 to obtain a biotechnological product. 17. The use of amino acids / peptides according to item 13 or 14 to obtain a food product. 18. The use of amino acids / peptides according to item 13 or 14 to obtain a feed product.

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