EA048767B1 - PROBIOTIC COMPOSITION BASED ON PLANT RAW MATERIALS AND METHOD OF ITS PRODUCTION - Google Patents

Description

Field of technology to which the invention relates

The invention relates to the food industry, and more specifically to functional food products, in particular, to probiotic food products. In addition, the invention is applicable to obtaining products with a high content of essential nutrients enriched with protein, to biopreservatives, to the field of dietetics and medicine. The invention can also be used in the composition of medical preparations and in the field of cosmetics, in a feed additive in the field of animal husbandry, and also as a fertilizer.

State of the art

The modern human diet is dominated by non-functional food products, in which the balance of nutrients is strongly shifted towards the deficiency of some and excess of other food components. In the modern world, the consumption of refined food, inadequate diets, unfavorable environmental factors, stress, which lead to problems of microbiota imbalance, as evidenced by the increasing number and variety of dietary supplements and drugs produced to stimulate friendly gastrointestinal microbiota, predominate.

Currently, the food industry is actively developing the production of so-called functional food products, designed to reduce the risks of various diseases or have some additional benefits in terms of influencing the physiology of consumers compared to conventional products.

GOST R 52349-2005 introduces and defines the concept of a functional food product as a special food product intended for systematic consumption as part of diets by all age groups of the healthy population, possessing scientifically substantiated and confirmed properties, reducing the risk of developing diseases associated with nutrition, preventing a deficiency or replenishing an existing deficiency of nutrients in the human body, maintaining and improving health due to the presence of functional food ingredients in its composition.

Further, according to GOST R 52349-2005, a functional food ingredient (functional ingredient; physiologically functional ingredient; functional component; physiologically functional component; physiologically functional food component) is a living microorganism, a substance or a complex of substances of animal, plant, microbiological, mineral origin or identical to natural ones, included in the composition of a functional food product in an amount of at least 15% of the daily physiological requirement, calculated per one serving of the product, possessing the ability to exert a scientifically substantiated and confirmed effect on one or more physiological functions, metabolic processes in the human body with systematic use of a functional food product containing them.

Functional food ingredients include physiologically active, valuable and safe for health ingredients with known physical and chemical characteristics, for which health-promoting properties have been identified and scientifically substantiated, and a daily physiological requirement has been established. In particular, functional food ingredients currently include:

soluble and insoluble dietary fiber (pectins, etc.);

vitamins (vitamin E, tocotrienols, folic acid, etc.);

minerals (calcium, magnesium, iron, selenium, etc.);

fats and substances associated with fats (polyunsaturated fatty acids, plant sterols, conjugated isomers of linoleic acid, structured lipids, sphingolipids, etc.);

polysaccharides, secondary plant compounds (flavonoids/polyphenols, carotenoids, lycopene, etc.), probiotics, prebiotics and synbiotics.

It should be noted that the definition given in GOST R 52349-2005 does not disclose the possibility of using food raw materials from mushrooms and algae.

Functional (processed) products include:

fortified products to which vitamins, microelements, and dietary fiber have been added;

products from which certain substances not recommended for medical reasons have been removed (microelements, amino acids, lactose, etc.);

products in which the removed substances are replaced by other components.

GOST R 52349-2005 also introduces and defines the concept of a probiotic food product as a functional food product containing, as a physiologically functional food ingredient, specially isolated strains of useful for humans (non-pathogenic and non-toxicogenic) live microorganisms that have a beneficial effect on the human body through normalization of the microflora of the digestive tract. From this definition it follows that a probiotic food product is a special subclass of functional food products with one mandatory component (food strains of live microbes). The presence of functional food ingredients of non-microbial origin in this subclass of products is not specifically stipulated.

In essence, functional food ingredients of probiotic foods can be

- 1 048767 can include any functional food ingredients from plant, animal, microbial raw materials, as well as from fungi and algae. It should be especially noted that any products of this subclass (fermented milk products; tea and rice fungus of microorganism origin; fermented oat yoghurts or fermented vegetable and plant juices) are processed functional food products due to the high fermentation, synthetic and sorption activities of food strains of microorganisms.

The properties of functional products are described in GOST R 52349-2005 from the point of view of supplementing food with essential nutrients. The problem is that the GOST metrics used and the state of the art, which will be discussed below, do not address the impact of functional products on the known and important functions of the gastrointestinal tract for maintaining and improving health. Both the standard discussed above and the known solutions from the state of the art do not address either the digestive functions (digestion, absorption, substrate binding, motor-evacuation, secretory) or non-digestive functions of the gastrointestinal tract (metabolic, barrier, excretory, reutilization of endogenous substances, detoxification, entero-endogenous recirculation, regulation of microbiota, antioxidant). At the same time, the functions of the gastrointestinal tract critically depend on the balance of microorganisms of the normal flora (microbiota), which is often shifted towards pathogenic and opportunistic microorganisms in people suffering from diseases and/or disorders of the gastrointestinal tract and metabolism.

The fermented milk fat-free bioproduct Narilak Forte-V produced by ZAO Vector-Bialgam (see http://www.narilak.ru/doc/instructions.pdf) is well-known. According to the manufacturer's statement, it has the following health benefits: it improves the functioning of the gastrointestinal tract, prevents the proliferation of pathogenic microbes in the intestines, has a stimulating effect on the immune system, promotes the absorption of vitamins and microelements, normalizes the level of hemoglobin in the blood, helps to normalize cholesterol levels, eliminates the negative side effects of taking antibiotics, and stimulates the growth of lacto- and bifidoflora in the intestines. The well-known bioproduct contains a complex of bifido- and lactobacteria according to TU 9222-012-14392386-12 (see http://www.bialgam.ru/assets/images/123/Декларация%20Нарилак%20форте%20B%20Декларация%20новαя %2024.01.19.pdf).

The well-known bioproduct contains a multi-species consortium of lacto- and bifidobacteria in a live, biologically active state (1 ml of the product contains at least 1 billion lactobacilli L. acidophilus and at least 100 million bifidobacteria B. bifidum and B. longum, as well as their metabolites (vitamins B, PP, C, volatile fatty acids, enzymes, essential amino acids)). The composition of the well-known bioproduct includes: skim milk, lactobacilli starter, live culture of bifidobacteria. The manufacturer recommends the following method of use: 2-3 times a day 20-30 minutes before meals, with a small amount of liquid. The dosage recommended by the manufacturer: adults - 2 tablespoons; children 1-7 years old - 1-3 teaspoons, children 7-12 years old - 2 dessert spoons. The duration of admission is not less than 14 calendar days, with a recommendation for longer intake of the product. Courses of admission 2 times a year according to the recommendations indicated on the product website. The shelf life of an unopened package of the bioproduct is 3 months at a storage temperature of 4±2°C.

The well-known bioproduct has a number of disadvantages. In particular, it is not suitable for consumption by people suffering from lactose and dairy intolerance, as well as people following a strict vegan diet due to the content of skim milk. It lacks prebiotic dietary fiber, which helps maintain normal intestinal motility and is necessary for the normal functioning of the gastrointestinal tract's own microbiota. It contains a narrow range of bacterial digestive enzymes (ferments), which is due to the composition of the nutrient medium (milk). This medium does not contain complex carbohydrates, and, accordingly, there are no enzymes to digest them. In particular, when adding the well-known bioproduct to food with starches, there will be no bacterial enzymes to digest them; in addition, the enzymes from the well-known bioproduct will not participate in the digestion of such food.

The known bioproduct does not contain polyphenols, which serve as a nutrient medium for the friendly microflora of the gastrointestinal tract. In addition, the liquid medium of the bioproduct does not contain a full range of microelements. The known bioproduct is obviously not resistant to multiple freeze/thaw cycles due to the precipitation of milk proteins, which creates additional difficulties during transportation and storage. High requirements are imposed on milk raw materials for microbiological production, as well as on the production cycle of the bioproduct (sterile conditions due to the fact that milk-based media are not resistant to the germination of harmful microorganisms). The known bioproduct has a relatively high cost due to the high cost of the medium and high requirements for the production cycle. In addition, it has a short shelf life in unopened packaging (the shelf life of an opened package is not declared), and the temperature storage conditions of the known bioproduct do not allow it to be kept warm (at room temperature and during transportation). As a result, the significant disadvantages of the well-known bioproduct include the presence of animal-based components in its composition, low prebiotic and

- 2 048767 functional properties of the product.

Synbiotic products containing prebiotics and probiotics are known. In particular, the series of preparations of biologically active substances Vetom 1.23, 2.25, 2.26, 3.22, 4.24 contains corn extract fermented by microorganisms of the genus Bacillus, sodium chloride, distilled water. The products differ from each other in the species composition of strains aimed at suppressing one or another pathogenic flora of the gastrointestinal tract. The product is intended for oral administration.

The disadvantages of these well-known products include the absence of probiotic bacteria that complement the resident flora of the gastrointestinal tract and stimulate friendly resident microflora. The enzymatic digestive activity of the well-known products is limited, which is due to the environment (corn extract) and dosages (4-10 drops per day). The cost of this type of well-known products for regular use is high. The products are not suitable for fermentation of plant materials and molecular cuisine.

Also known is Komarov's powder from the same manufacturer, which is related to synbiotics, which contains Bacillus subtilis bacteria, wheat bran, blue-green algae (spirulina), used as a source of protein, essential amino acids, omega products and minerals, as well as citric acid. The well-known product is an additional source of essential nutrients, dietary fiber, as well as a beneficial strain of the transient bacterium Bacillus subtilis.

The disadvantage of this well-known product is the content of wheat bran, the use of which can cause unpleasant and painful sensations in people with lesions of the gastrointestinal mucosa. Another disadvantage of Komarov's powder is the lack of probiotic bacteria. This well-known product is not suitable for fermentation of plant materials and molecular cuisine.

A method for preparing a biologically active food product is known, disclosed in Russian patent No. RU 2558191 C1, wherein the known method includes drying the plant medium (oats, rice, millet with the optional addition of oat bran) in a dry-heat oven at 200°C for 5 minutes, grinding it, fermenting with types of lactic acid bacteria (residents of the gastrointestinal tract) until pH 3.5-4.5 is achieved, separating the supernatant liquid (postbiotics, including lactic acid and other water-soluble substances synthesized by bacteria), with the production of a sediment containing 39.78% protein in terms of dry matter. The final products are a sediment in the form of a creamy mass (1/3 of the original volume) and a supernatant liquid.

The resulting creamy mass can be used as a salad dressing, vinaigrette, and as a flavoring additive to sweet dishes. The supernatant liquid can be used as an additive to drinks or as an independent drink like kvass.

Source RU 2558191 C1 states that the product has no contraindications for use, is safe, contains live lactobacilli in high concentrations, easily digestible vegetable protein, a number of essential amino acids, and can be recommended for dietary nutrition, in particular for dietary nutrition of patients with gastrointestinal pathology.

The disadvantages of the product obtained by the method according to RU 2558191 C1 include:

the presence of coarse bran particles in the final product, which is contraindicated in various gastrointestinal diseases. This limits the use of the product for health purposes, which contradicts the safety and absence of contraindications indicated in this source;

drying at 200°C destroys polyunsaturated fats and vitamins in the original plant material; lack of microelements necessary, in particular, for the synthesis of vitamins and metalloproteins;

low concentration of accumulated postbiotics due to the absence of a product maturation stage;

a small amount of biopreservatives (lactic acid) in the creamy product, which reduces the shelf life of the known product.

Postbiotic products (also abbreviated as postbiotics) are known, prepared on plant raw materials, which contain metabolites of strains of probiotic bacteria, as well as components of their walls and enzymes of lysed cells. Recently, postbiotics have become widespread after repeated confirmation of their health effect on the human body, mainly due to the high stability and long shelf life of such preparations due to the absence of living microorganisms in them. Among the metabiotics, the following preparations are known: LACTIS, MultiLiz and a number of analogues.

MultiLiz (manufacturer Artlife LLC), state registration certificate KZ.16.01.98.003.E.000642.07.19 dated July 26, 2019, was chosen as the closest analogue of this group of products to the claimed invention. This well-known product is a product of pumpkin and kombucha fermentation by specialized strains of probiotic microorganisms. The product contains in liquid form: fermented pumpkin and kombucha puree, prebiotics (pumpkin and kombucha polysaccharides), metabolites of bacteria Lactobacillus casei, L. rhamnosus, L. salivarius, Bifidobacterium bifidum and their ultralysates (peptide fragments of the cell wall of microorganisms), lactulose, xanthan gum, guar gum, pyridoxine hydrochloride (Vitamin B6), cyanocobalamin (Vitamin B12), glucose. The scope of application of this well-known product, declared by

- 3 048767 by the manufacturer - as a biologically active food supplement - an additional source of vitamins B6 and B12. The method of using the well-known product, declared by its manufacturer: 1 sachet (5 g) per day, previously diluted in 50 ml of drinking water, consume for a month.

The disadvantages of this well-known product include:

the absence of live microorganisms in its composition, which have a wider range of positive effects on food and the human gastrointestinal tract than the combination of bacterial metabolites and microorganism ultralysate. Thus, unlike postbiotic preparations, live probiotic bacteria are able to flexibly adapt to the conditions of the gastrointestinal tract and the composition of food, actively participating in its digestion and detoxification, as well as in the neutralization of endotoxins. In addition, live probiotic bacteria have their own extremely high antioxidant activity in the gastrointestinal tract, which is associated with their low tolerance of free oxygen;

lack of beta-glucans;

deficiency of conjugates of polyunsaturated fats (omega 3);

incomplete spectrum of microelements in the fermentation medium; technological complexity of product production;

high cost of a single dose.

Unlike the claimed invention, which uses exclusively plant raw materials for fermentation, Multilys contains a fermentate of mixed plant and mushroom raw materials (pumpkin and kombucha), supplemented with thickeners xanthan and guar gum. The common thing between the claimed invention and this known product is the content (but not the spectrum) of prebiotics, polysaccharides, metabolites of probiotic bacteria, including digestive enzymes, lysates of probiotic microorganisms, and vitamin composition. However, the absence of active microorganisms, deficiency of microelements, polyphenols, omega 3 products in the composition of the known product does not allow using Multilys and its analogues as a prototype for the claimed invention.

A method for preparing raw materials for functional food products from a fermented plant medium is known, described in US Patent No. US 9161495 B2 (published 20.10.2015). The plant raw materials in the known method are grains of winter varieties of barley and wheat with the addition of cereal grain bran, if necessary. According to this well-known solution, vernalization is used to activate endogenous enzymes of sprouted grains, saccharify them with endogenous alpha-amylase and maltose or additionally with external alpha-amylase and maltose, inactivate the enzymes with high temperature, dry and grind if necessary, supplement them with organic materials and/or medicinal herbs, and/or medicinal mushrooms, and/or vegetables and their extracts, ferment them in an aqueous suspension with beneficial lactic acid bacteria (Streptococcus sp., Pediococcus sp., Leuconostoc sp., Lactobacillus sp., Bifidobacterium sp.) and/or food yeast (Saccharomyces sp.). The resulting fermented material is used, if necessary, in a processed lyophilized form as a functional food ingredient with antioxidant and antidiabetic activities, anti-obesity activity, proven in experiments on laboratory animals, and is used in functional food compositions in the form of powder, juice concentrate, drink, tablet, suspension, granules, emulsion, capsules and syrup. If necessary, the fermented product may be supplemented with sweeteners (including honey), flavorings, physiologically active ingredients such as catechins (catechin, epicatechin and gallocatechin) and vitamins (retinol, ascorbic acid, tocopherol, calciferol, thiamine and riboflavin), minerals (such as calcium, magnesium, chromium, cobalt, copper, fluorides, germanium, iodine, iron, lithium, magnesium, manganese, molybdenum, phosphorus, potassium, selenium, silicon, sodium, sulfur, vanadium and zinc), preservatives, emulsifiers (such as acacia gum, carboxymethylcellulose, xanthan gum and pectin), acidifiers and thickeners.

What this known solution has in common with the claimed invention is the use of plant raw materials, the stage of saccharification of such raw materials using endogenous or, as an addition, external enzymes, the stage of temperature inactivation of saccharification enzymes, the stage of blending the saccharified plant raw materials with extracts of medicinal herbs and mushrooms, fruits or vegetables, the stage of fermentation in an aqueous suspension of crushed blended saccharified plant raw materials using lactic acid bacteria.

However, the disadvantages of this well-known solution include:

a long minimum technological cycle for obtaining final products (according to the conditions disclosed in this well-known decision - 35-60 days) and the relative complexity of the method of preparing the product;

restrictions on the plant materials used (only viable grains of winter crops, primarily winter varieties of cultivated cereals such as barley and wheat, as well as rye, which meets these requirements but is not mentioned in this source);

a functional food ingredient is not intended to be used in its pure form, but only as a raw material for functional food products;

deficiency of protein monomers in the fermentation medium;

- 4 048767 use of products of animal origin; deficiency of biopreservatives (lactic acid); absence of live microorganisms in an active state in the product;

restriction on the use of the product by persons with increased sensitivity to optional additives of highly active medicinal products of natural origin;

lack of trace elements in bioavailable form.

A method is known, described in the patent of the Russian Federation No. RU 2332113 C1 for a bio-oat food and drink product based on oat raw materials, characterized by a standardized content of soluble oat dietary fiber - 1,3-1,4-βD-glucan, which have undergone a deep processing process using two-stage fermentation, and containing a standardized amount of high-molecular 1,3-1,4-βϋglucan, which has maximum bioavailability, to impart non-specific biological properties to it. As in the claimed invention, in the known method, plant raw materials are used (oat medium is a special case of plant raw materials), deep fermentation of raw materials with probiotic microorganisms is carried out, and prebiotics and probiotics are also contained.

However, the disadvantages of the product obtained by this known method include: deficiency of polyphenols, deficiency of postbiotics associated with a short period of post-fermentative accumulation of postbiotics, deficiency of the composition of microelements, long periods of preparation of plant raw materials for fermentation, limitations of the spectrum of plant raw materials, a decrease in the number of microorganisms in the product during the shelf life (from 1013 to 106-7), a long and excessively high-tech production method, and the unreasonableness of the declared quality criteria of the known product (absence of stratification) from the point of view of modern ideas about the functionality of the product.

The prior art discloses a fermented hydrolyzed plant material (see US Patent Application No. US 20180327792 A1), as well as a corresponding method and composition obtained on the basis of said material and method, wherein the known method consists of three stages. The first stage of the known method includes hydrolysis of plant material to obtain hydrolyzed plant material. At the second stage, a starting material for fermentation is obtained, containing a hydrolysate of plant material. The third stage includes fermentation of the starting fermentation material to obtain compositions of fermented plant material.

According to this known solution, the plant material is hydrolyzed while maintaining the original dietary fiber content and beta-glucan concentration, and then fermented, while individual desirable properties of the original plant material can be maintained, such as nutrients, whole grain condition, fiber content and beta-glucan content, and some organoleptic properties can be varied.

The potential benefit of the compositions described in this prior art solution may be their use as a prebiotic, a glycemic index reducer, an immune booster, a fiber source, a soluble fiber source, a nutritional supplement, a texture modifier, a viscosity modifier, or a combination thereof.

However, this known solution has a number of disadvantages, consisting, in particular, of a deficiency of postbiotics due to the absence of a special stage of their accumulation, a lack of metalloproteins and vitamins of bacterial synthesis due to the absence of a source of microelements at the stage of fermentate enrichment, as well as the absence of a stage of separation of the coarse fraction of the raw material.

A fermented probiotic drink with an increased content of bioactive (functional) ingredients is known, described in the article Development of a Dairy-Free Fermented Oat-Based Beverage With Enhanced Probiotic and Bioactive Properties. Front. Microbiol., December 3, 2020. The fermented probiotic drink is obtained on the basis of plant raw materials (whole grain oat flour) with the addition of natural honey by preliminary sterilization of the raw materials, followed by the addition of honey dissolved in water and bacteria of the Lactobacillus fermentum (PC1) strain to a count of 107 (fermentation for 72 hours), with incubation at 4 ° C for 10-14 days to accumulate metabolites and postbiotics. After this period, the number of microorganisms in the drink does not decrease, and the amount of postbiotics and metabolites (phenolic acids: gallic, vanillic, coffee, rycumaric, ferulic, as well as polyphenols catechin and quercetin) increases by more than 50%, while the level of glucose and fructose decreases, there is an increase in the content of antioxidants and short-chain fatty acids (acetic and lactic acid), and the content of the prebiotic beta-glucan does not change.

Honey is an additional source of antioxidants (phenolic acids and polyphenols) and monosaccharides. It has been shown that after fermentation with lactic acid bacteria, the bioavailability of phenolic acids in oats and barley increases by ~20-25 times (see Hole AS, Rud I, Grimmer S, Sigl S, Narvhus J, Sahlstrom S. Improved bioavailability of dietary phenolic acids in whole grain barley and oat groat following fermentation with probiotic Lactobacillus acidophilus, Lactobacillus johnsonii, and Lactobacillus reuteri. J Agric Food Chem. 2012 Jun 27;60(25):6369-75. doi: 10.1021/jf300410h. Epub 2012 Jun 15. PMID: 22676388. And Conway, P. L, Stern, R., and Tran, L. (2010). The value-adding potential of prebiotic components of Austra

- 5 048767 lian honey. Rural Industries Research and Development Corporation).

The disadvantages of the well-known fermented probiotic drink include: a deficiency of oligopeptides and amino acids, the absence of metalloproteins and bioavailable microminerals, a deficiency of the spectrum of accumulated metabolites and postbiotics due to a poorer environment. In essence, the disadvantages of this product are also low probiotic and functional properties of the product, a lack of metalloproteins and minerals.

A method is known, disclosed in the publication of international application No. WO 1991017672 A1, according to which a product is produced on the basis of grain flour or vegetables and fruits, or oilcake and meal of plant origin with a high content of dietary fiber, which is soaked in water, then gelatinized and pasteurized, fermented with bifido- and propionic bacteria, if necessary in combination with lactobacilli, if necessary supplemented with berries/fruits and/or sugars, kept at 5-8°C for 7-10 days, after which it is packaged and stored at the same temperature.

The product obtained by this known method can be modified in terms of functional characteristics and taste qualities by adding special ingredients (such as raw plant components, milk, milk powder or other milk derivatives, soy flour, starch, sugar, natural honey, berries, fruits, vegetables or jams, juices, coffee, cocoa, or food additives in the form of salt, aromatic or coloring substances, stabilizers).

This well-known product can be used independently or as a functional ingredient for any food of both plant and animal origin. The declared shelf life is up to 1 week at a temperature of 5°C. Exceeding this period leads to a deterioration in the taste of the product due to its souring. This well-known solution emphasizes the presence of dietary fiber, probiotic bacteria, lactic acid in the product, and also declares high taste qualities and reduced energy value. The functional properties of the well-known product help to reduce blood cholesterol levels, bind bile acids, and prevent the risk of developing oncological diseases (colon or breast cancer).

However, the known product disclosed in the above source has disadvantages, including:

the presence of coarse bran particles in the final product, contraindicated in various gastrointestinal diseases, which limits the use of the product for health purposes, a short declared shelf life of the product (from 1 to 2 weeks at 5°C), storage and transportation at a temperature not exceeding 5°C, a lack of oligopeptides and amino acids, a deficiency of a spectrum of microelements during fermentation, a lack of vitamins, the synthesis of which depends on the presence of microelements in the fermentation medium.

The method and product disclosed in WO 1991017672 A1 can be considered as the closest analogue (prototype) in relation to the claimed invention.

Disclosure of invention

Taking into account the level of technology discussed above, the technical problem solved by the present invention consists in the need to improve the effect of functional products on the known and important functions of the gastrointestinal tract for maintaining and improving health, with an emphasis on normalizing microflora and regulating metabolism.

The problem, which the invention is aimed at solving, consists in creating a probiotic composition for a wide range of purposes with no contraindications for use, using exclusively plant-based raw materials, containing a wide range of functional ingredients that help to increase the digestibility of food, increase the nutritional value and enrich the diet with essential nutrients, microelements in a bioavailable form, normalize and maintain the functions of the gastrointestinal tract (including detoxification of the body), normalize and maintain friendly resident microbiota, increase the shelf life of food products and food raw materials, expand the arsenal of functional bioproducts available to the population, with the possibility of local (external) and internal (oral, rectal, intranasal) use for health purposes.

The technical result achieved by using the invention consists in increasing the probiotic and functional properties of the probiotic composition with an increase in the level of functionality and digestibility of food (active participation in digestion). In addition, the said technical result is accompanied by such useful effects as normalization and maintenance of regulatory processes in the body, effective stimulation of friendly resident microbiota, elimination of contraindications for the use of the final product, expansion of the areas of application of the probiotic composition, maintenance of the stability of the properties of the elements of the probiotic composition over time (i.e. long shelf life without loss of properties and with maintenance of the balance of elements), simplification of the process of production of the probiotic composition.

- 6 048767

To solve the above-mentioned problem with achieving the mentioned technical result, an invention is proposed, with an inventive concept, containing both probiotic compositions on plant raw materials and methods for their production.

In the first aspect of the present invention, the said problem is solved by a method for producing a probiotic composition based on plant raw materials, according to which natural honey or material of plant origin, which is cereals, legumes, vegetables, fruits, green mass of edible plants, including wild plants, and their by-products (crushed grain, crushed seeds, flakes, granules, flour, juices, solid/semi-liquid/liquid/dry concentrates, pastes and cake), taken separately or in the form of blends of different compositions, are at least once enriched with a composition of microelements, converted into a liquid form of hydrolyzate with a dry matter concentration of 0.2 to 80%, pasteurized, a mesophilic and/or thermophilic starter containing strains of food microorganisms is introduced, fermented at a temperature optimal for the reproduction of microorganisms until they reach the stationary growth phase, transferred to the fermentation stage at suboptimal low temperature until the specified characteristics are achieved, filtered from coarse bran particles at least once after hydrolysis or one of the fermentation stages.

The micronutrient composition may be selected from a group comprising at least one of pink (Himalayan) salt, sea salt, seaweed powder, food ingredients or compositions containing a broad spectrum of micronutrients, or any combination thereof.

The above-mentioned mesophilic and/or thermophilic starter may be represented in embodiments of the invention by at least one of the strains suitable for use in the food industry, from among the species of lacto/bifido/propionic acid bacteria of the following genera: Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Propionibacterium, Sporolactobacillus, Streptococcus, Tetragenococcus, Vagococcus, Weissella, and representatives of other genera of lactobacilli that are not related to species and strains of microorganisms pathogenic for humans, both individually and in any possible combination with each other.

At any stage of the proposed method, additional introduction of at least one of the components selected from the group containing: minerals, polyphenols and phenolic acids individually or as a mixture, sources of omega 3 and 6, dietary fiber, herbal extracts, natural honey, molasses, natural sugars, natural thickeners, natural antioxidants and flavorings in the form of aqueous solutions, powder, granules, aqueous or oil extracts, pastes, crushed plant materials, plant parts, crystals, or any combination thereof may be provided.

If necessary, food grade alkalis and/or antifoams are added at the hydrolysis and fermentation stages.

In embodiments of the claimed method, the conversion into a hydrolyzate may be accompanied by its saccharification, or it may be converted into a hydrolyzate using food grade alkalis.

At the fermentation stage or after filtration, if necessary, the composition is diluted with water, juice, saline solution, herbal and plant solution, syrups or other drinks, edible oils.

The above-mentioned specified characteristics of completion of the fermentation stage at a suboptimal temperature are selected from a group comprising: the level of enzyme activity, the composition and quantitative content of vitamins, the content of monomers and/or oligomers of proteins, carbohydrates and fats, the content and composition of short-chain fatty acids, exopolysaccharides, bacteriocins, polyphenols and phenolic acids, natural antioxidants, minerals in bioavailable form, dietary fiber, the amount of postbiotic and/or metabolic layer, the pH level, the fermentation time at a given temperature, organoleptic properties, or any combination thereof.

In the second aspect, the present invention relates to a probiotic composition on plant raw materials, obtained by the method according to the above-mentioned first aspect, which is characterized by a pH level from 1.5 to 6.5, the absence of coarse bran particles, the content of a composition of microelements, natural antioxidants of plant origin, polyphenols and phenolic acids of plant and bacterial origin, dietary fiber, peptides, beta-glucans, mono- and oligomers of carbohydrates and proteins, polyunsaturated fatty acids, active microorganisms, accumulated metabolites and postbiotics of microorganisms, minerals in bioavailable form, hydrolytic enzymes and detoxification enzymes, bacteriocins, exopolysaccharides, B vitamins, short-chain fatty acids, neurotransmitters and their precursors, heme-containing proteins and other metalloproteins; and a mass fraction of dry matter from 0.2 to 80 wt.%.

In other aspects, the present invention relates to the use of a probiotic composition according to the above-mentioned second aspect as a means for health purposes for external and internal use, wherein internal use is selected from the group comprising oral, intranasal and rectal use, as well as at least one of a feed additive for animals, a food additive for a human diet, a cosmetic paste or lotion, a fertilizer. In embodiments of the invention, the probiotic composition is used with dilution with physiological or aqueous solutions, herbal solutions and other natural medicines. The present invention also relates to the use of a probiotic composition as a functional food ingredient in at least one of solid food, drinks and ready-made sauces and drinks, as at least one of a detoxifying agent, an ingredient of medical products, a component having effects directed against at least one of obesity, symptoms of type II diabetes, proinflammatory and toxicogenic glycation of biomolecules with elevated levels of post-lunch blood sugar, as a concentrate for the preparation of functional food products and/or an additional source of digestive enzymes.

In another aspect, the invention provides a probiotic product containing a retentate of coarse bran particles obtained after one of the fermentation stages in the method according to the above-mentioned first aspect, made with the possibility of use in wet and/or pressed form as at least one of a feed additive for animals, a food additive for a human diet, a cosmetic paste or lotion, a fertilizer.

It should be noted that the inventive concept is not limited to the above aspects, and based on the information characterizing the invention in the description, it will be obvious to those skilled in the art that the invention may also take the form of other products and/or methods different from those listed above. The scope of legal protection of the invention is determined solely by the attached claims and is not limited by any specific details provided in the following description of the invention.

Implementation of the invention

Based on the state of the art, presented in particular by the known solutions described above, the authors of the claimed invention came to the conclusion about the possibility and necessity of supplementing the metrics of functional products described in GOST R 52349-2005 with metrics of the effect on the functions of the gastrointestinal tract, with an emphasis on the normalization of microflora and the regulation of metabolism by functional ingredients from probiotic food products prepared on pre-processed and fermented plant raw materials.

The proposed invention, as will be described in more detail below, is characterized, in particular, by an increase in the concentration of postbiotics against the background of an extremely low concentration of monosaccharides and the absence of hidden sugars, the content of a significant amount of oligomers of proteins, carbohydrates and fats, as well as oligopeptides, oligosaccharides, conjugates of polyunsaturated fatty acids (omega 3) and other essential nutrients, prebiotics and dietary fibers. The proposed composition contains a wide range of digestive enzymes of bacterial origin, a wide range of microminerals in a bioavailable form. At the same time, it does not contain coarse fragments of bran, since they are filtered during the preparation of the composition, and is characterized by a long shelf life. In addition, in accordance with literature data (Corcoran BM, Stanton C, Fitzgerald GF, Ross RP. Survival of probiotic lactobacilli in acidic environments is enhanced in the presence of metabolizable sugars. Appl Environ Microbiol. 2005 Jun;71(6):3060-7. doi: 10.1128/AEM.71.6.3060-3067.2005. PMID: 15933002; PMCID: PMC1151822.), probiotics are better preserved in an acidified fermented medium with exopolysaccharides, comparable to the compositions proposed by the applicant, and more easily tolerate the aggressive environment of the gastrointestinal tract.

The inventive concept is based on the idea that normalization of microflora is not the only useful characteristic of probiotic food products of any origin. No less important is the role of functional food ingredients that regulate metabolism in the human body and, in particular, numerous digestive and non-digestive functions of the gastrointestinal tract and liver.

Increased digestibility of food is achieved thanks to the digestive activity of bacterial enzymes, both accumulated in the claimed probiotic composition and newly synthesized by active microorganisms when added to food.

Enrichment of food with essential nutrients in the probiotic composition according to the invention is achieved due to the presence of vitamins synthesized by bacteria, vitamins of the original plant raw material, oligomers and monomers of plant proteins and carbohydrates obtained as a result of fermentation, polyunsaturated fats from the composition of the raw material and their bacterial conjugates (omega 3), microelements in bioavailable form and metalloproteins.

Normalization of the digestion functions of the human gastrointestinal tract when using the probiotic composition according to the invention is achieved due to bacterial enzymes both accumulated in the product and newly synthesized by active microorganisms in the food. Normalization of the absorption function of the human gastrointestinal tract when using the probiotic composition according to the invention is achieved due to the presence of oligomers and monomers of plant proteins and carbohydrates obtained as a result of hydrolysis and fermentation.

Normalization of the motor-evacuation function of the human gastrointestinal tract when using a probiotic composition according to the invention is achieved due to the presence of fermented dietary fibers in the composition of probiotic food compositions.

Normalization of the secretory function of the human gastrointestinal tract when using a probiotic composition

- 8 048767 according to the invention is achieved due to the presence of neurotransmitters and their precursors of bacterial origin present among postbiotics in the composition of probiotic food compositions.

Normalization of the metabolic function of the human gastrointestinal tract when using a probiotic composition according to the invention is achieved due to the presence of detoxifying enzymes of bacterial origin in the composition of probiotic food compositions.

Normalization of the barrier function of the gastrointestinal tract when using the probiotic composition according to the invention is achieved due to the presence of bacterial exopolysaccharides, both free and associated with bacterial cells, in the composition of probiotic food compositions.

Normalization of the detoxification function of the human gastrointestinal tract when using the probiotic composition according to the invention is achieved due to the presence of bacterial detoxification enzymes, sorption of toxins and endotoxins by fermented dietary fibers and bacterial lysates (bacterial membrane proteoglycans), as well as due to the antioxidant activity of live bacteria, the presence of short-chain fatty acids, polyphenols and phenolic acids.

Normalization of microbiota and effective maintenance of friendly gastrointestinal microflora when using the probiotic composition according to the invention is achieved due to the presence of exopolysaccharides (free and associated with bacterial walls), the presence in the product of probiotics that stimulate the resident gastrointestinal microflora, and prebiotics (fermented dietary fibers, beta-glucans of plant and bacterial origin, bacterial exopolysaccharides), as well as the presence of bacteriocins that suppress pathogenic and opportunistic gastrointestinal flora, bacterial lysates, and minerals in bioavailable form.

Normalization of the antioxidant function of the human gastrointestinal tract when using the probiotic composition according to the invention is achieved due to the presence of antioxidants of natural origin in the composition of plant raw materials and bacteria, additional enrichment with plant polyphenols, and the inherent antioxidant activity of living microorganisms in the composition of probiotic food compositions.

Improvement of interaction between the intestine and the liver when using the probiotic composition according to the invention is achieved due to the presence in the product of short-chain fatty acids of bacterial origin, in particular lactic acid.

Improvement of interaction between the intestine and the brain is achieved due to the presence in the probiotic composition according to the invention of neurotransmitters and their precursors of bacterial origin, as well as the destruction or binding of harmful metabolites of gastrointestinal microorganisms, in particular, proinflammatory lipopolysaccharides of gram-negative bacteria.

The adjustment of the functions of the human gastrointestinal tract and the metabolic activity of the microbiota when using the probiotic composition according to the invention is achieved by normalizing the regulatory processes in the human body.

The complex effect of the functional ingredients of the probiotic composition according to the invention regulates metabolic processes (regulatory processes) in the human body, improves the functional properties of any food and increases the shelf life of products.

The absence of coarse dietary fiber and medicinal compounds that have individual contraindications eliminates contraindications for the use of the claimed probiotic food compositions. A long shelf life of the probiotic composition according to the invention is achieved without changing the number of microorganisms due to the presence in the medium of available nutritional nutrients (such as peptides, oligopeptides, amino acids, oligosaccharides, etc.) obtained at the stage of hydrolysis of plant raw materials in the method for producing a probiotic composition on plant raw materials in accordance with the proposed invention.

Method for producing a probiotic composition.

First, a method for producing a probiotic composition based on plant material according to the first aspect of the present invention will be described. In general, the method according to the first aspect of the invention comprises the steps of:

natural honey or material of plant origin, which is cereals, legumes, vegetables, fruits, green mass of edible plants, including wild plants, and their by-products (crushed grain, crushed seeds, flakes, granules, flour, juices, solid/semi-liquid/liquid/dry concentrates, pastes and cake), taken separately or in the form of blends of different compositions, are at least once enriched with a composition of microelements, converted into a liquid hydrolyzate with a dry matter concentration of 0.2 to 80%, pasteurized, a mesophilic and/or thermophilic starter containing strains of food microorganisms is introduced, fermented at a temperature optimal for the reproduction of microorganisms until they reach a stationary growth phase, transferred to the fermentation stage at a suboptimal low temperature until they reach

- 9 048767 of these characteristics, filter from coarse bran particles at least once after hydrolysis or one of the fermentation stages.

To obtain the probiotic composition according to the invention, such modes of hydrolysis with food enzymes and pasteurization are used, the combined effect of which does not lead to the destruction of prebiotic and bioactive components of the original plant material. Food hydrolytic enzymes are used to liquefy the plant material, including starch and other polysaccharides, which are decomposed into a mixture of oligo- and monosaccharides. This (saccharification and liquefaction) makes it possible to obtain a more concentrated environment favorable for the reproduction and active existence of probiotic bacteria.

The method according to the invention imposes relatively low requirements for sterility, which is due to the fact that the hydrolysate of the original plant material, containing more than 2.5% of sea or Himalayan salt, prevents the proliferation of harmful microorganisms. The fermentation medium after fermentation of such a hydrolysate becomes even less favorable for harmful microorganisms, due to its enrichment with bacteriocins and lactic acid from the composition of the starter.

The enrichment of the product with a multi-component mineral composition according to the claimed method is carried out before the production of a hydrolyzate of plant-based material. This ensures the optimal ionic composition of the reaction mixture for the implementation of enzymatic hydrolysis and eliminates the possible surplus of metal ions necessary for the effective operation of hydrolysis enzymes.

At the first stage of fermentation of plant material under stationary conditions (without maintaining the composition of the nutrient medium) in a saccharified medium and at an optimal temperature, a logarithmic increase in the number of microorganisms occurs until the stationary phase is reached.

If there are fast sugars in the environment, other components available for feeding microorganisms are used only for biosynthesis purposes. The transition to their use as an energy source occurs after the pool of fast sugars is exhausted.

A distinctive feature of the claimed method for producing the probiotic composition is the second fermentation stage (at a low temperature, suboptimal for the reproduction of microorganisms). Under these conditions, the consumption of low-energy compounds present in large quantities in the fermentation medium is sufficient both to maintain the number of microorganisms and to accumulate useful metabolites and postbiotics in the fermentolysate. The low fermentation temperature also contributes to an increase in the lifespan of bacterial metabolites and postbiotics, in particular, digestive enzymes accumulated at the fermentation stage of fast sugars. The qualitative composition of metabolites and postbiotics at the second fermentation stage is richer than at the first fermentation stage. Such enrichment is the result of stepwise hydrolysis of specific nutritional components of the fermentation medium (polysaccharides and their fragments, including oligosaccharides, peptides, oligopeptides and amino acids, oligolipids and fragments of the original fat molecules).

Thus, at the second stage of fermentation, the number of microorganisms achieved at the first stage of fermentation is maintained, and the quantity and spectrum of essential nutrients, prebiotics, postbiotics and beneficial metabolites of microorganisms in the probiotic composition are increased and enriched.

In the absence of the second stage of fermentation, the number of microorganisms gradually decreases by 1-3 orders of magnitude, and the accumulated volume of essential nutrients, prebiotics, postbiotics and beneficial metabolites of bacterial origin becomes unstable both quantitatively and qualitatively.

The authors of the invention have established that the addition of fast sugars at the second stage of fermentation increases the number of microorganisms in the environment by 1-2 orders of magnitude (up to 108-1014 CFU in 1 ml) while maintaining the composition of the accumulated metabolites and postbiotics that are beneficial to humans.

The said second stage of fermentation at a reduced temperature is completed upon reaching the characteristics specified by the functional orientation of a specific probiotic composition. Such characteristics can be specified in relation to both individually and in combination of enzymatic activity, spectrum and quantitative content of vitamins, mono- and oligosaccharides, oligopeptides and amino acids, short-chain fatty acids, exopolysaccharides, bacteriocins, polyphenols and phenolic acids, minerals in bioavailable form.

For fermentation in the method according to the invention, the following cultures of probiotic microorganisms are used, which in the final product are in a functionally active state and can be represented by at least one of the strains suitable for use in the food industry, from among lacto/bifido/propionic acid bacteria of the genera: Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Propionibacterium, Sporolactobacillus, Streptococcus, Tetragenococcus, Vagococcus, Weissella, as well as representatives of other genera of lactobacilli that are not pathogenic for humans.

- 10,048,767 languages, both individually and in any acceptable combination with each other.

The concentration of microorganisms in the finished product is set in advance in the range from 106 to 1013 CFU/ml depending on the functional orientation of a specific probiotic composition and the probiotic strains used in the technological process. The stages of fermentation and accumulation of postbiotics are carried out at a given temperature (optimal and suboptimal for specific microorganism strains, respectively) until they reach the stationary growth phase.

The microorganisms used and permitted for use in the composition can function in the temperature range from 0 to 50°C. Transportation at elevated temperatures potentially reduces the shelf life of the composition, but does not affect the beneficial properties of the product. At elevated temperatures, the concentration of metabolites and postbiotics of probiotic bacteria in the composition increases intensively, and the amount of food (mainly dietary fiber of plant origin) decreases proportionally. At slightly negative temperatures (-1°C), the product does not freeze.

The pH level, the amount of postbiotic and/or metabolic layer, organoleptic properties, including odor, and the known time to achieve such characteristics at the temperature used during the second fermentation stage can be used as indirect signs determining the achievement of the required characteristics of the composition and the end of the fermentation stage.

The introduction of food alkalis, carried out if necessary in some embodiments of the invention, at the fermentation stage at an optimal or reduced temperature reduces the concentration of lactic acid, which stimulates further growth in the number of microorganisms, leads to the appearance of biopreservatives (such as sodium and/or potassium lactates), and also changes the organoleptic properties of the product.

When enriching a product with fat-based additives (in particular, olive oil, fish oil, etc.), emulsification and conjugation of polyunsaturated fatty acids (omega 3 and 6) in the product occurs due to lactic acid.

In accordance with the claimed method, the plant raw materials are cereal grains, legumes, vegetables, fruits, green mass of edible plants, including wild plants, taken separately or in the form of blends of different compositions. The variety of raw materials allows you to select an environment with a given enrichment option with certain functional ingredients, which allows you to adapt production to available local plant raw materials.

Examples

Further, the method according to the invention will be illustrated by specific examples of the implementation of the invention, which serve solely to confirm the possibility of implementing the purpose of the claimed invention with the achievement of the declared technical result, but not to limit the scope of legal protection of the present invention by any specific details.

Example 1. Concentrate of probiotic composition on oat medium with minerals and polyphenols (peppermint).

In this example, the method according to the invention is carried out to obtain a probiotic composition in the form of a concentrate on an oat medium with minerals and polyphenols. The process in accordance with the method takes place in the form of the following sequence of stages:

1. Soak dry oat flakes in water at a temperature of 76°C while stirring, at a rate of 600 g of flakes per 1 liter of oat suspension.

2. A composition of macro- and microelements is introduced - pink Himalayan salt - to a final concentration of 2.2% by weight.

3. Add the hydrolytic enzyme Amylosubtilin at a rate of 0.75 g per 1 kg of dry oat flakes.

4. Hydrolyze the hydrated oats with additives in the grinding mode at 76°C for 15 minutes until a homogeneous suspension with a semi-liquid consistency is formed, which does not change to liquid in the last 3-5 minutes of hydrolysis.

5. Cool the semi-liquid oat suspension while stirring to 58°C.

6. Add the hydrolytic enzyme Protosubtilin at a rate of 3 g per 1 kg of dry oat flakes.

7. Hydrolyze the semi-liquid suspension with additives in grinding mode at 58°C for 30 minutes until a finely dispersed liquid suspension is formed, the consistency of which does not change noticeably during the last 5-10 minutes of hydrolysis.

8. Pasteurize the hydrolyzate in stirring mode for 10 minutes at a temperature of 92°C, which simultaneously leads to the inactivation of external hydrolysis enzymes.

9. Add a composition of polyphenols (a product of hot water extraction of crushed dry peppermint, at a rate of 10 g of mint per 1 liter of extract) in an amount of 50 ml of extract per 1 liter of pasteurized hydrolysate.

10. Cool the hydrolyzate to 42°C.

11. Introduce a mesophilic starter (any non-pathogenic strains selected from the group containing Lactobacillus acidophilus, L. casei, L. pediococcus, L. plantarum, Lrhamnosus, L. bulgaricus, L. helveticus, L. salivarius, L. fermentum, L. reuteri, Lactococcus lactis, Bifidum bifidum, B. breve, B. longum, B. infantis,

- 11 048767

Streptococcus thermophilus (singly or in any combination).

12. Ferment the suspension in stirring mode for 10-15 hours at a constant temperature of 35-40°C until a pH of 3.8-4.8 is reached.

13. Filter out coarse bran particles using sieves with 80-100 mesh cells.

14. Ferment at 12°C until the desired amount of metabolites and postbiotics is achieved.

The resulting probiotic composition is characterized by an extremely low content of monosaccharides (<0.9% mass fraction according to GOST R 54607.6-2015) and contains the following bioactive components:

probiotic microorganisms at a concentration of 109-1014 CFU/ml with high intrinsic antioxidant activity (the content is assessed, for example, according to GOST 10444.11-2013, MUK 4.2.577-96, the methods described in Aline Moser, Helene Berthoud, Elisabeth Eugster, Leo Meile, Stefan Irmler, Detection and enumeration of Lactobacillus helveticus in dairy products, International Dairy Journal, Volume 68, 2017, Pages 52-59, ISSN 0958-6946, https://doi.org/10.1016/j.idairyj.2016.12.007, The spiral plate count (SPLC) method, and other methods that determine the number of microorganisms);

digestive enzymes of lactic acid bacteria (proteolytic activity (the content is estimated, for example, according to GOST 34430-2018, as well as according to the methods described in the sources CHOW, B. F., & PETICOLAS, M. A. (1948). A rapid method for the determination of proteolytic activities of enzyme preparations. The Journal of general physiology, 32(1), 17-24. https://doi.org/10.1085/jgp.32.1.17; S CASTRO and A M B CANTERA, A rapid and inexpensive procedure for the determination of proteolytic activity - 1995 Biochemical Education - Wiley Online Library https://iubmb.onlinelibrary.wiley.com/doi/pdf/10.1016/03074412%2894%2900104-W);

short-chain fatty acids (the content is determined, for example, by gas chromatography);

prebiotics (dietary fiber of oats and probiotic bacteria) (the content of soluble and insoluble dietary fiber is determined, for example, by the enzymatic-gravimetric method according to GOST R 54014-2010 or by the method described in the source Li BW. Determination of total dietary fiber in foods and food products by using a single-enzyme, enzymatic-gravimetric method: interlaboratory study. J AOAC Int. 1995 Nov-Dec;78(6):1440-4. PMID: 8664579);

bacteriocins (the content is determined, for example, by the method described in the source Catherine B. Lewus, Thomas J. Montville, Detection of bacteriocins produced by lactic acid bacteria, Journal of Microbiological Methods, Volume 13, Issue 2, 1991, Pages 145-150, ISSN 0167-7012, https://doi.org/10.1016/01677012(91)90014-H.);

exopolysaccharides of probiotic bacteria (the content is determined, for example, by the method described in Ruhmann B, Schmid J, Sieber V. Methods to identify the unexplored diversity of microbial exopolysaccharides. Front Microbiol. 2015 Jun 9;6:565. doi: 10.3389/fmicb.2015.00565. PMID: 26106372; PMCID: PMC4460557);

conjugates of polyunsaturated fatty acids (omega 3 in bioavailable form);

microminerals in bioavailable form (for example, the presence of selenium proteins and selenium-cysteine as a bioavailable form of micromineral);

antioxidants of oats and probiotic bacteria (polyphenols and phenolic acids) (determined by spectrophotometric method);

essential nutrients (free amino acids, oligopeptides, short lipids, B vitamins, etc.);

enterosorbents of oats and lactic acid bacteria;

neurotransmitters and their precursors of bacterial origin (serotonin, gamma-aminobutyric acid, etc.) (determination based on the methods disclosed in the article Briguglio M, Dell'Osso B, Panzica G, Malgaroli A, Banfi G, Zanaboni Dina C, Galentino R, Porta M. Dietary Neurotransmitters: A Narrative Review on Current Knowledge. Nutrients. 2018 May 10;10(5):591. doi: 10.3390/nu10050591. PMID: 29748506; PMCID: PMC5986471).

The probiotic composition obtained in this example can be used as a probiotic food concentrate, a beverage concentrate, a functional food additive, an animal feed additive, a fertilizer.

Example 2. Concentrate of probiotic composition on buckwheat medium treated with food alkali and supplemented with minerals, enriched with omega 3 (olive oil) and polyphenols (peppermint).

In this example, the method according to the invention is carried out to obtain a probiotic composition in the form of a concentrate on a buckwheat medium treated with food alkali and supplemented with minerals. The process in accordance with the method takes place in the form of the following sequence of stages:

1. Soak buckwheat groats in water at a temperature of 60°C at a rate of 500 g per 1 liter of buckwheat suspension for 30 minutes.

2. Grind and mix until a homogeneous suspension with a semi-liquid consistency is formed.

3. Introduce in stirring mode a composition of macro- and microelements - pink Himalayan

- 12 048767 salt - to a final concentration of 1.0 wt%.

4. Add food grade alkali in the same mode at a rate of 15-20 g per 1 liter of suspension.

5. Olive oil is added as an antifoaming agent and a source of omega 3 at a rate of 50 ml per 1 liter of suspension.

6. Hydrolyze the semi-liquid suspension with additives for 30 minutes in grinding mode with a gradual increase in temperature to 92°C until a finely dispersed liquid suspension is formed, the consistency of which does not change noticeably during the last 5-10 minutes of hydrolysis.

7. Pasteurize the hydrolyzate in stirring mode for 10 minutes at a temperature of 92°C.

8. Add a composition of polyphenols (a product of hot water extraction of crushed dry peppermint, at a rate of 10 g of mint per 1 l of extract) in an amount of 50 ml of extract per 1 l of pasteurized hydrolysate.

9. Cool the hydrolyzate to 37°C.

10. Mesophilic starter culture of lactic acid bacteria (strains Lactobacillus acidophilus and Streptococcus thermophilus) is introduced.

11. The suspension is fermented in stirring mode for 15-17 hours at a constant temperature of 37°C.

12. Filter out coarse bran particles using sieves with 80-100 mesh cells.

13. Ferment at 12°C until pH 4.0-4.8 and a specified amount of metabolites and postbiotics are achieved.

The probiotic composition obtained in this example can be used as a probiotic food concentrate, a beverage concentrate, a functional food additive.

Example 3. Concentrate of probiotic composition on a medium of wild plants with minerals.

In this example, the method according to the invention is carried out to obtain a probiotic composition in the form of a concentrate on a medium of wild plants (green mass of goutweed and dandelion root) with minerals. The process in accordance with the method takes place in the form of the following sequence of stages:

1. Obtain an aqueous suspension of a mixture of frozen (or raw) wild plants (green mass of common goutweed and dandelion root with a mass ratio of 3 to 1) or powders prepared from them, taken in the same ratio, in a grinding or mixing mode, at a rate of 600 g of moistened wild plants per 1 liter of aqueous suspension or 150 g of a mixture of powders per 1 liter of suspension.

2. Add a microelement-enriched composition of an aqueous extract of crushed fucus powder (a product of hot aqueous extraction of crushed fucus powder, at the rate of 30 g of fucus in 1 l of extract) to a final concentration of 0.9 wt%.

3. In grinding or mixing mode, heat the mixture to 59°C.

4. Add a mixture of hydrolytic enzymes (amylosubtilin and cellolux) in a 1 to 1 ratio while stirring, at a rate of 2 g of enzyme mixture per 1 kg of powder or 1 kg of dry weight of the wild plant mixture.

5. Hydrolyze for 30 minutes at 58°C until a homogeneous suspension with a stable semi-liquid consistency is formed.

6. Heat the hydrolyzate in stirring mode to 65°C.

7. Add the proteolytic enzyme papain at a rate of 5 g of enzyme per 1 kg of powder or dry weight of the wild plant mixture.

8. Hydrolyze the suspension in stirring mode at 65°C for 15 minutes until a homogeneous suspension of liquid consistency is formed.

9. Pasteurize the hydrolyzate in stirring mode for 10 minutes at a temperature of 92°C, which simultaneously inactivates all external hydrolysis enzymes.

10. Cool the hydrolyzate in the same mode to 45-50°C.

11. Thermophilic starter is introduced (strains of Streptococcus salivarius ssp. thermophilus, Lactococcus lactis ssp. cremoris, Lactococcus lactis ssp. lactis, Lactobacillus casei, Leuconostoc mesenteroides ssp. mesenteroides individually or in any combination).

12. Ferment in stirring mode for 15-18 hours at a constant temperature of 4550°C until pH 4.0-4.8 is reached.

13. Filter out coarse bran particles using sieves with 80-100 mesh cells.

14. Add food grade alkali (Na or K bicarbonate, or a mixture of them in a 1 to 1 ratio) to the fermentate while stirring at a rate of 15 g per 1 l of suspension.

15. Olive oil is added as an antifoaming agent and a source of omega 3 conjugates.

16. Ferment at 20°C until pH 4.0-4.8 and the specified amount of metabolites and postbiotics are achieved.

The probiotic composition obtained by the method according to this example, in addition to bioactive substances of bacterial origin, includes healthy components of wild plants and brown algae fucus;

prebiotics (dietary fiber from wild plants);

natural thickeners (inulin from dandelion roots, agar-agar from fucus);

- 13 048767 polyphenols and phenolic acids (wild plants, fucus);

vitamins and microelements in bioavailable form (wild plants, fucus);

essential nutrients (wild plants, fucus).

The probiotic composition obtained in this example can be used as a probiotic food concentrate, a beverage concentrate, a functional food additive, and an animal feed additive.

Example 4. Probiotic honey drink concentrate with added prebiotic (dietary fiber).

In this example, the method according to the invention is carried out to obtain a probiotic composition in the form of a probiotic honey drink concentrate with the addition of a prebiotic (dietary fiber). The process in accordance with the method takes place in the following sequence of stages:

1. Dissolve natural honey in water at 55°C while stirring at a rate of 600 g of honey per 1 liter of water.

2. A composition of macro- and microelements is introduced - pink Himalayan salt - to a final concentration of 3.4 weight%.

3. Cool the solution to 42°C.

4. Add a concentrated solution of red grape polyphenols in the amount of 10 ml per 1 liter of suspension while stirring.

5. Mesophilic starter culture is introduced (strains of Lactobacillus acidophilus, L. casei, L. pediococcus, L. plantarum, L. rhamnosus, L. bulgaricus, L. helveticus, L. salivarius, L. fermentum, Bifidobacterium bifidum, B. breve, B. Longum, B. infantis, Lactococcus lactis, Streptococcus thermophilus, individually or in any combination).

6. Ferment the suspension in stirring mode for 10-15 hours at a constant temperature of 35-40°C until a pH of 4.0-4.8 is reached.

7. Add Jerusalem artichoke powder as a prebiotic and thickener at a rate of 4 g per 1 liter of suspension.

8. Ferment at 12°C until the desired amount of metabolites and postbiotics is achieved.

Example 5. Concentrate of probiotic composition on rice medium with added minerals, green tea extract and natural honey.

In this example, the method according to the invention is carried out to obtain a probiotic composition in the form of a concentrate on a rice medium with the addition of minerals, green tea extract and natural honey, supplemented with turmeric (or ginger root extract). The process in accordance with the method takes place in the form of the following sequence of stages:

1. Soak the parboiled rice grains in water at a temperature of 76°C in a stirring mode at a rate of 210 g of rice per 1 liter of suspension.

2. A composition of macro- and microelements is introduced - pink Himalayan salt - to a final concentration of 1.1% by weight.

3. Add the hydrolytic enzyme Amylosubtilin at a rate of 0.75 grams per 1 kg of dry grains.

4. Hydrolyze the watered rice in grinding mode at 76°C for 15 minutes until a homogeneous suspension of semi-liquid consistency is formed.

5. Cool while stirring to 58°C.

6. Add the proteolytic enzyme bromelain in the same mode at a rate of 5 g per 1 kg of dry grains.

7. Hydrolyze the semi-liquid suspension at 58°C for 30 minutes in grinding mode until a finely dispersed liquid suspension is formed.

8. Pasteurize the hydrolyzate in stirring mode for 10 minutes at a temperature of 92°C, which simultaneously leads to the inactivation of external hydrolysis enzymes.

9. Cool in the same mode to 55°C.

10. Add an aqueous extract of green tea polyphenols (extraction of 10 g of green tea with 1 liter of boiling water) at a rate of 50 ml of tea extract per 1 liter of pasteurized hydrolysate.

11. Dissolve natural honey in the hydrolyzate at 55°C while stirring at a rate of 600 g of honey per 1 liter of water.

12. Cool the hydrolyzate to 42°C.

13. Mesophilic starter culture is introduced (strains of Lactobacillus acidophilus, L. casei, L. pediococcus, L. plantarum, L. rhamnosus, L. bulgaricus, L. helveticus, L. salivarius, L. fermentum, Bifidobacterium bifidum, B. breve, B. Longum, B. infantis, Lactococcus lactis, Streptococcus thermophilus individually or in any combination).

14. The suspension is fermented in stirring mode for 10-15 hours at a constant temperature of 35-40°C until a pH of 4.0-4.8 is reached.

15. Filter out coarse bran particles using sieves with 80-100 mesh cells.

16. Ferment at 12°C until the desired amount of metabolites and postbiotics is achieved.

17. Add turmeric powder in stirring mode at a rate of 2 g per 1 l of fermentate or cooled aqueous extract of ginger root (extraction of 50 g of crushed root in 1 l of boiling water) at a rate of

- 14,048,767 ml of extract per 1 liter of fermentate.

Example 6. Probiotic composition with increased content of microorganisms, postbiotics and metabolites of bacterial origin.

This example of implementing the method according to the invention essentially represents a modified version of the method described above in Example 1, and allows to obtain a probiotic composition with an additionally increased content of microorganisms, postbiotics and metabolites of bacterial origin. The process in accordance with the method essentially corresponds to the process described above in Example 1. However, it is supplemented by the fact that food alkali is introduced into the probiotic composition prepared by the method described in Example 1 at the fermentation stage at 12°C in an amount of 18 g per 1 l of fermentate.

The result of this additional stage is an increase in the number of microorganisms in the finished product by 1-2 orders of magnitude.

Example 7. Probiotic composition with propionic acid bacteria and propionic acid.

This example of implementing the method according to the invention essentially represents a modified version of the method described above in any of the examples 1-5, and allows to obtain a probiotic composition supplemented with propionic acid bacteria and propionic acid. The process in accordance with the method essentially corresponds to the process described above in any of the examples 1-5. However, it is supplemented in that a starter containing the strains Propionibacterium acidipropionici, P. freudenreichii, P. jensenii, P. microaerophilum individually or in any combination is added to the hydrolyzate prepared at the corresponding stage of implementing the method described in any of the above examples 1-5, after which a two-stage fermentation is carried out.

This allows the resulting probiotic composition to be supplemented with propionic acid bacteria and their metabolite propionic acid. Propionic acid and propionates have the following functions: energy supply to the epithelium, antibacterial effect, regulation of proliferation and differentiation of the epithelium, supply of gluconeogenesis substrates, blocking the adhesion of pathogens to the epithelium, maintaining ion exchange (see, for example, Ardatskaya M.D., Minushkin O.N. Modern principles of diagnostics and pharmacological correction // Gastroenterology, supplement to the journal Consilium Medicum. - 2006. - Vol. 8. - No. 2. Pp. 4-17., Hosseini E, Grootaert C, Verstraete W, Van de Wiele T. Propionate as a health-promoting microbial metabolite in the human gut. Nutr Rev. 2011 May;69(5): 245-58. doi: 10.1111/j.1753-4887.2011.00388.x. PMID: 21521227). Propionic acid is also involved in the synthesis of hormones, neurotransmitters (serotonin, endorphins) and has an antifungal effect and increases physical endurance of the body (Jonathan Scheiman, Sarah Lessard, Aleksandar D. Kostic, et al. Meta-omics analysis of elite athletes identifies a performance-enhancing microbe that functions via lactate metabolism. Nature Medicine, 2019).

These factors enhance the functionality of the final product.

Example 8. Preparation of an oat probiotic drink with aqueous peppermint extract.

This example of implementing the method according to the invention essentially represents a modified version of the method described above in Example 1, and allows to obtain a probiotic composition in the form of an oat probiotic drink with an aqueous extract of peppermint. The process according to the method essentially corresponds to the process described above in Example 1. However, it is supplemented in that the concentrate of the probiotic composition prepared by the method described in Example 1 is diluted with water in a volume ratio of 6 to 1. After this, a freshly prepared aqueous extract of peppermint powder is added in an amount of 50 ml of extract per 1 liter of drink.

In one or more embodiments of the invention, the use of aqueous extracts of peppermint, which have antispasmodic, analgesic, anti-inflammatory, antispasmodic, decongestant properties, which is known from biomedical research (see, in particular, Almeida RN, Hiruma CA, Barbosa-Filho JM: Analgesic effect of rotundefolone in rodents. Fitoterapia 1996, 67:334-338.; Delia Loggia R, Tubaro A, Lunder T: Evaluation of some pharmacological activities of a peppermint extract. Fitoterapia 1990, 61:15-221; Raya MD, Utrilla MP, Navarro MC, Jimenez J: CNS activity of Mentha rotundifolia and Mentha longifolia essential oil in mice and rats. Phytother Res 1990, 4:232-234; MimicaDukic N, Bozin B, Sokovic M, Mihajlovic B, Matavulj M: Antimicrobial and antioxidant activities of three Mentha species essential oils. PlantaMed 2003, 69:413-419). In addition, aqueous extracts of peppermint are also a natural source of polyphenols and antioxidants.

Example 9. Probiotic compositions with medicinal herbs.

This embodiment of the method according to the invention is essentially a modified version of the method described above in any of Examples 1-5, and allows to obtain a probiotic composition supplemented with medicinal herbs. The process according to the method essentially corresponds to the process described above in any of Examples 1-5. However, it is supplemented in that extracts of medicinal herbs are added to the probiotic composition prepared by the method according to any of Examples 1-5 after the fermentation step, such as, as a non-limiting example, an aqueous extract of chamomile (in an amount of crushed chamomile flowers based on 100 g of chamomile per 1 liter of hot water extraction) in an amount of 50 ml of extract per 1 liter of pro

- 15 048767 duct. This additionally provides anti-inflammatory, antispasmodic, regenerating and calming (sedative) effect of the probiotic composition on humans.

Example 10. Probiotic composition in the form of a paste.

This example of implementing the method according to the invention is essentially a modified version of the method described above in Example 3, and allows to obtain a probiotic composition in the form of a paste. The process in accordance with the method essentially corresponds to the process described above in Example 3. However, it is supplemented by the fact that Jerusalem artichoke root powder is added to the probiotic composition prepared in accordance with Example 3 until a paste-like consistency is achieved, which increases the prebiotic component of the product, since Jerusalem artichoke root contains a lot of inulin. The functional focus of such a product is for people with type 2 diabetes, obesity, and dysbiosis.

Example 11. Hypoallergenic probiotic composition.

This example of implementing the method according to the invention essentially represents a modified version of the method described above in any of the examples 1-5, and allows to obtain a hypoallergenic probiotic composition. The process according to the method essentially corresponds to the process described above in any of the examples 1-5. However, it is supplemented in that a starter containing the strain Lactobacillus acidophilus EP 317/402 (Narine) is added to the hydrolyzate prepared within the framework of implementing the method according to any of the examples 1-5, and the first fermentation stage is carried out at 37°C, the second fermentation stage - at 10-15°C.

Example 12. Probiotic composition with increased mineral content.

This example of implementing the method according to the invention essentially represents a modified version of the method described above in Example 1, and allows to obtain a probiotic composition in the form of a concentrate with an increased mineral content. The process according to the method essentially corresponds to the process described above in Example 1. However, it is supplemented by the fact that a 30% aqueous solution of macro- and microelements (pink Himalayan salt) is added to the probiotic composition prepared according to Example 1 to a final concentration of 4.0 wt.%. Pink (Himalayan) and sea salt are intended to further improve the organoleptic properties of the probiotic composition according to the invention.

It should be noted that the above-described examples of preparing probiotic compositions by the method according to the invention represent only some of the possible embodiments of the method according to the invention. The specific stages of the method and modes of their implementation described in various examples can be used in other embodiments of the method according to the invention, forming a wide range of other possible specific embodiments of the invention, despite the fact that their description is not provided explicitly. The examples are provided solely to explain possible embodiments and demonstrate the variability of technological processes within the framework of the inventive concept, as well as to confirm the possibility of implementing the purpose of the invention and achieving the technical result.

The choice of a specific method for preparing a probiotic composition according to the invention and its specific composition is determined by nutritionists or food technologists taking into account the required functional orientation of the product, industrial capabilities, availability of plant materials and available or intentionally used probiotic strains of microorganisms, examples of which are given above.

Probiotic composition.

In a second aspect, the invention relates to a probiotic composition on plant raw materials, obtainable by the method according to the first aspect of the invention described above. The probiotic composition according to the invention is generally characterized by the following:

pH level from 1.5 to 5.8, absence of coarse bran particles, content:

natural antioxidants of plant origin, polyphenols and phenolic acids of plant and bacterial origin, dietary fiber, peptides, beta-glucans, mono- and oligomers of carbohydrates and proteins, polyunsaturated fatty acids, active microorganisms, accumulated metabolites and postbiotics of microorganisms, minerals in bioavailable form, hydrolytic enzymes and detoxification enzymes, bacteriocins, exopolysaccharides,

- 16,048,767 B vitamins, short-chain fatty acids, neurotransmitters and their precursors, heme-containing proteins and other metalloproteins; and a dry matter content of 0.2 to 80 wt.%.

Below is a brief description and explanation of the functions of some key components of the probiotic composition included in the finished product and/or obtained during the implementation of the above-described method for producing the probiotic composition according to the invention.

The fermentolysate is a fermented plant medium obtained in the process of implementing the method for producing a probiotic composition, characterized by a high content of peptides and amino acids. The fermentolysate helps maintain the required number of microorganisms in the probiotic composition according to the invention at the stage of its storage.

The wide variability of plant materials and their extracts used in the production of the probiotic composition (pomegranate, turmeric, peppermint, red grapes, green tea, pepper and other spices, various berries) ensures high variability of the final composition of the composition in terms of polyphenols and phenolic acids. Polyphenols contained in the probiotic composition obtained by the proposed method are antioxidants, which has a positive effect on the shelf life of the composition.

Due to the above-described enzymatic activity of microorganisms, polyphenols in the probiotic composition obtained by the proposed method can be modified, which expands the spectrum of polyphenols in the gastrointestinal tract. Polyphenols and phenolic acids are common components of hydrolyzed products of plant origin, as well as metabolites of probiotic microorganisms obtained on media containing materials of plant origin. They are effective antioxidants and improvers of the organoleptic properties of the probiotic composition.

Postbiotics and metabolites of probiotic bacteria in the composition are represented by biomolecules useful for humans: short-chain fatty acids, essential nutrients, including vitamins and minerals in bioavailable form, exopolysaccharides, digestive enzymes, neurotransmitters and their precursors, bacteriocins, detoxifiers and activators of detoxification of various types of human cells and organs, food fibrils, including bacterial beta-glucans, growth factors and protection of human cells and friendly resident intestinal microflora, anti-inflammatory agents and other chemical compounds that help normalize metabolism in the human body.

Postbiotics in the composition accumulate during its maturation at a suboptimal low temperature and during its storage period at a temperature of 2-8°C. Postbiotics include both heat-stable (in the range from 0 to 50°C) products of bacterial synthesis with a long lifespan, and products of bacterial synthesis with reduced heat stability and/or with a medium and short lifespan. The concentration of short-lived and/or non-heat-stable postbiotics in the composition can be maintained at a constant level at a low temperature due to the constant metabolic activity of microorganisms. At the same time, the amount of heat-stable postbiotics increases during the periods of maturation and storage of the product at low temperatures.

Positive dynamics of the level of postbiotics in the product are a distinctive feature of the probiotic composition from known postbiotic preparations (metabiotics), in which the level of thermostable biomolecules is a relatively constant value, and the content of non-thermostable and/or short-lived biomolecules rapidly decreases during the shelf life.

Lactobacilli are known for their ability to convert microelements into a form accessible to human cells. An example is the bacterial transformation of selenium ions into nanoparticles or selenized amino acids, which allows for the rapid and effective elimination of the deficiency of this vital microelement and thus the prevention of cancer, aging and other chronic diseases associated with a deficiency of bioavailable selenium.

The functionality of food supplemented with the probiotic composition according to the invention is increased by bacterial protein (enzymes and peptides secreted by bacteria, as well as proteins in the bacteria, which make up at least 1/4-1/3 of their dry weight). Bacterial protein, unlike the reserve proteins of many plants, is well balanced in amino acid composition. Such protein contains all essential and conditionally essential amino acids for humans in sufficient quantities.

Bacteria in the proposed probiotic composition can be destroyed during natural death, under the action of peroxides released by them (for example, hydrogen peroxide), the enzyme lysozyme and third-party bacteriocins from bacteria of another species. In this case, the original composition of the probiotic composition is enriched with useful products of bacterial lysis (lytic enzymes, components

- 17 048767 cell wall, total protein and peptides, oligosaccharides, etc.).

Bacteriocins synthesized by probiotic bacteria accumulate in the probiotic composition produced by the method according to the invention. These agents disinfect and thereby prevent spoilage of the product obtained by this method, acting as biopreservatives. Bacteriocins normalize the resident microbiota of the large intestine by suppressing pathogenic and opportunistic microorganisms. Probiotic bacteria from the composition obtained by the proposed method and their components (cell wall fragments) entering the large intestine are stimulating factors for the growth and development of normal flora.

The initial plant material used to obtain the probiotic composition by the method according to the invention may have a deficiency in some microelements. Microelements are necessary for effective bacterial synthesis of some vitamins and metalloproteins. In particular, the synthesis of vitamin B12 requires the presence of cobalt ions. The wider the range of microelements in the medium, the higher the level of synthesis of vitamins and metalloproteins by probiotic bacteria in the probiotic composition and the normal flora of the human gastrointestinal tract. To compensate for the potential deficiency of microelements in the final product according to the method, at least one enrichment is carried out with a composition of microelements, which can be represented by either pink (Himalayan) salt, or sea salt, or seaweed powder, or other food ingredients or compositions containing a wide range of microelements.

Microelement enrichment can be achieved through food ingredients or compositions - natural sources of different types of microelements, such as, for example, mumiyo, extracts of chaga and varnished tinder fungus, apricot or nut seeds, some wild plants such as licorice root, rhubarb, dandelion or licorice, as well as pharmaceutical preparations with microelement additives (such as flaxseed oil with selenium) and fermented legume seeds (such as miso paste) alone or in combination.

The composition and quantitative content of microelements in the recommended daily dose of the probiotic composition according to the invention does not exceed half of the daily human requirement for them. Maximum permissible concentrations are determined for different categories of the population in relation to such sources of minerals as pink (Himalayan) and sea salt, as well as for fucus powder and other algae used in food. This makes it easy to calculate the maximum permissible daily consumption of the composition according to the invention for different age and sex groups of a person for microelements, both for conditionally healthy people and for people suffering from diseases that require corrections in the daily consumption of certain minerals.

The presence of microelements in the probiotic composition obtained by the method according to the invention increases the functionality of food consumed together with it, regardless of the diet.

An important role in the composition according to the invention is played by short-chain fatty acids, which are metabolites of probiotic microorganisms fermenting saccharified media. According to the claimed method, saccharification of plant-based materials occurs at the hydrolysis stage. Subsequent fermentation by bacteria is carried out up to the maximum possible level of desaccharification. A further increase in the concentration of short-chain fatty acids is possible, if necessary, by additionally enriching the raw materials with saccharified mixtures. Excess lactic acid inhibits bacterial proliferation, which can be avoided by introducing food grade alkali (Na or K bicarbonate, or a mixture thereof in a 1 to 1 ratio) into the fermentation medium. Short-chain fatty acids are detoxifying agents, bacteriostatics and nutrients involved in lipid metabolism.

The plant-based materials used to obtain the fermentation medium contain both water-soluble and insoluble dietary fibres (poly- and oligosaccharides), which are prebiotics available as food products for the resident microbiota.

Due to bacterial fermentation, the dietary fibers of the original raw material are fragmented into shorter sequences. This expands the spectrum of food prebiotics due to the diversity of food preferences of beneficial bacteria of the human normal flora, which completely eliminates the risks of damage to the protective mucous layer of the intestine.

Dietary fibers of different composition and origin have selectivity in relation to the sorption of toxicants of different types. This allows for the effective sorption of toxicants of various types, both in food and in the gastrointestinal tract.

One of the body cleansing mechanisms is the process of physiological filtration and reabsorption of fluid from the vascular bed into the intestinal lumen by its villi. Contacting with sorbents from the probiotic composition (dietary fiber, bacterial walls) and under the influence of bacterial detoxification enzymes, harmful substances are removed from the body. Thus, the composition is an effective remedy for poisoning, toxicosis of various etiologies, including endotoxicosis. The sorption properties of the composition improve the condition of the human body in dysbiosis.

Polyphenols in the composition serve as antioxidants, which has a positive effect on the preservation of the organoleptic and functional properties of the composition. In turn, bacte

- 18 048767 riocins block the reproduction of foreign microorganisms in the composition. These factors ensure a long shelf life of the composition. The applicants have experimentally established that the shelf life of the composition is at least one year at a storage temperature of 2-8°C.

Plant raw materials for the production of probiotic compositions include cereals, legumes, vegetables, fruits, green mass of edible plants, including wild plants, taken separately or in the form of blends of different compositions.

It is possible to supplement the original plant material with dietary fiber and natural thickeners (inulin or inulin-rich additives, resistant starch, xanthan or guar gum, pectin, etc.).

In addition, a significant place in the probiotic composition according to the invention is occupied by hydrolytic enzymes of bacteria. It is well known that hydrolytic enzymes of bacteria of normal flora participate in the digestion of food together with human digestive enzymes. In the large intestine of a person, digestion is carried out mainly by bacterial enzymes.

Bacteria secrete digestive enzymes depending on the composition of the nutrient medium. In the presence of mono- and disaccharides, probiotic bacteria primarily secrete enzymes for digestion and assimilation of these energetically advantageous compounds. After the saccharides are exhausted, the bacteria switch to other less energetically valuable compounds, secreting the corresponding hydrolytic enzymes. The synthesized enzymes accumulate in the fermentation medium.

The composition of the hydrolysates or alkaline lysates of plant material used in the composition includes various simple and complex sugars (oligo- and polysaccharides). This circumstance determines the heterogeneity of bacterial digestive enzymes accumulated in the probiotic composition. This allows the probiotic composition to be used as an effective means for reducing the glycemic index of food. As a result, the use of the probiotic composition has a positive effect on the dietary properties of the food consumed and the well-being of people suffering from type II diabetes.

When adding the probiotic composition to food as a functional food ingredient, clear signs of food digestion in the plate are recorded (liquefaction when stirred, enrichment of the organoleptic bouquet, noticeable desugaring) by enzymes accumulated in the probiotic composition. Food with the ingredient probiotic composition is designated by the rally of the present invention as probiotic food.

Consumed food usually contains various mono- and disaccharides and hidden sugars (starchy compounds), which increase its glycemic index. Bacterial digestive enzymes from the probiotic composition according to the invention hydrolyze them.

The wider the spectrum of sugar-destroying enzymes accumulated in the probiotic composition, and the greater the quantity of them accumulated in the probiotic composition, the faster and more effectively the desugaring and reduction of the glycemic index of the food consumed will occur after supplementing it with the composition. Food processed or consumed with the composition becomes more suitable for people suffering from obesity, as well as type II diabetes and high blood sugar levels.

In the absence of special preservatives, digestive enzymes are quickly destroyed. However, due to the technical features of the invention described above, the probiotic composition maintains a balance of acidity, antioxidants, minerals and natural preservatives (sodium lactate and lactic acid), which prevent the rapid destruction of the digestive enzymes present and a number of other functional ingredients of the composition.

In addition, in the probiotic composition according to the invention, due to the synthetic activity of probiotic bacteria, both serotonin precursors and serotonin itself are formed. For people suffering from gastritis and ulcerative diseases of the gastrointestinal tract, heartburn, increased acidity, the presence of serotonin in the composition is a significant factor in normalizing the functions of the gastrointestinal tract. This allows the composition to be used both independently and as a food ingredient for the prevention and relief of symptoms of the listed gastrointestinal diseases.

Use of probiotic composition.

In other aspects, the present invention relates to the use of the probiotic composition obtained by the method according to the first aspect described above, for various purposes, non-limiting examples of which will be described below. It should be noted that, based on the above information about the product (probiotic composition) and the method for its production, other purposes for which the probiotic composition according to the invention can also be used with the implementation of the said purpose and the achievement of the said technical result may be obvious to specialists in this field of technology, wherein all such purposes are also considered to be included in the scope of legal protection of the claimed invention.

First of all, it should be noted that the characteristics of the product (probiotic composition) obtained by the method according to the invention allow its internal (as an example, oral,

- 19 048767 intranasal and rectal) and external use for health purposes, which is due, in particular, to the composition of the probiotic composition and its physical form (finely dispersed suspension).

Tests of the probiotic composition according to the invention when used in the above-mentioned ways demonstrated the beneficial effects of the probiotic composition based on the following mechanisms of action of the probiotic composition.

It has been shown that the action of the probiotic composition according to the invention when administered intranasally and orally (in the latter case, as a mouthwash) is similar in its mechanism of action to such preparations with probiotics, bactericides, enzymes, and metabiotics, such as, for example, the anti-cold drug Nozdrin, manufactured by VETOM (composition: Bacillus amyloliquefaciens bacteria strain VKPM B-10642 (DSM 24614), their metabolic products, and filler), AVSYSTEMS (active ingredient: vitamins, microelements, and metabolites of symbiotic bacteria Bifidobacterium sp, Lactobacillus delbrueckii subsp. Bulgaricus, Faecalibacterium prausnitzii, Lactobacillus acidophilus, see https://a-v-systems.com/), LiviaOne Topical Spray Probiotics (active ingredient: a mixture of probiotics on an enzyme-enriched substrate, water, and and a 3-herb blend, see https://store.liviaglobal.com/products/topical-probiotic-spray-4oz).

The effect of the probiotic composition according to the invention upon rectal administration was assessed according to the recommendations given in the sources Perlmutter, D., Loberg, K. The Gut and the Brain. How Gut Bacteria Heal and Protect Your Brain 2020 ISBN 978-5-00146-890-5, Perlmutter, David (2019). The Microbiome and the Brain. CRC Press. ISBN 978-0815376729.

The effect of the probiotic composition according to the invention upon external application was assessed using the example of cosmetic products with metabiotics/postbiotics, enzymes, vitamins and probiotics (such as, for example, USDA Organic Topical (Skin) Probiotic Spray by MaryRuth's https://www.maryruthorganics.com/products/skin-usda-organic-topical-probiotic-spray-vegan-non-gmo-4oz).

The variability of the plant raw materials used creates rich technological possibilities for the production of a wide range of probiotic food compositions with different regulatory effects on the human body. This allows taking into account individual differences between people and selecting the optimal product that is most effective for normalizing the gastrointestinal microflora of a particular person.

External use is effective for:

itching (due to the complex action of short-chain fatty acids, antioxidants and anti-inflammatory agents from the probiotic composition);

wounds and ulcers of the epithelial integuments, skin rashes (due to the complex action of bacteriocins, antioxidants, anti-inflammatory agents from the probiotic composition);

in skin and hair care (due to essential nutrients, minerals in bioavailable form, exopolysaccharides and other bioactive metabolites of microorganisms).

Rectal use is effective for:

dysbacteriosis, impaired peristalsis, constipation and diarrhea;

diseases associated with the accumulation of proinflammatory cytokines in human organs (type II diabetes mellitus, obesity, local inflammation of the mucous membranes, etc.).

Intranasal use is effective for:

rhinitis associated with disruption of the nasal microbiota;

prevention of viral and bacterial infections of the nasopharynx;

difficulties with nasal breathing.

Rinsing and spraying the oral cavity is effective:

for inflammation of the oral cavity, salivary glands and tonsils, as well as for the prevention of viral and bacterial infections;

to reduce plaque;

to improve the condition of the oral mucosa and the surface of the teeth.

The properties and composition of the composition allow the invention to be used:

in military and expeditionary conditions due to the use of fermented wild plants during their growing season, preservation of meat products of local origin;

allows relatively healthy people to diversify their diet and strengthen their immune system through the use of fermented foods enriched with vitamins, natural antiseptics, antioxidants and immunomodulators;

in areas with food shortages, to obtain products suitable for adequate nutrition from low-nutritious and difficult-to-digest plant materials;

to restore and maintain gastrointestinal functions and normal flora;

in the food industry to reduce the cost of food production and increase the shelf life of products, fermentation and marinating of food products of animal origin (meat, fish, seafood);

as part of health and rehabilitation courses;

- 20 048767 for individuals with type II diabetes and/or obesity, to reduce levels of postprandial sugar and proinflammatory cytokines in affected organs and tissues, as well as partial blockade of the hunger center in the brain using the YY peptide of bacterial origin;

for women during pregnancy and after childbirth to relieve symptoms of toxicosis and enrich the diet;

for nursing women to enrich their diet with essential nutrients and minerals in bioavailable form;

in the penitentiary system as an accessible means of providing a nutritious diet;

as a functional ingredient and flavor enhancer for beverages and other food products;

to relieve intoxication during treatment for oncological diseases;

in the rehabilitation of coronavirus infection and during the course of the disease - as a means of eliminating the cytokine storm, restoring the microbiota and strengthening the immune system, thanks to a large number of anti-inflammatory compounds of bacterial origin and detoxifying agents of plant and bacterial origin.

The use of the probiotic composition according to the invention in food helps to normalize weight and reduce post-lunch blood sugar levels, due to the partial destruction of sugars by bacterial enzymes and slowing down the transport of monosaccharides through the intestinal wall, increasing the nutritional value of food, which allows to reduce the single volume of food consumed.

In addition to the above applications, it is also possible to use the probiotic composition according to the invention as feed and feed additives for animals, as well as a fertilizer for agriculture and a plant treatment agent.

Retentate of coarse bran particles.

In another aspect, the invention relates to a probiotic product containing a retentate of coarse bran particles obtained after one of the fermentation stages in the method for producing a probiotic composition based on plant material according to the above-described first aspect of the present invention.

In particular, the retentate of coarse bran particles is obtained by implementing the method according to the first aspect of the present invention at the stage of filtering the probiotic composition from coarse bran particles, performed at least once after hydrolysis or one of the fermentation stages. Filtered coarse bran particles are used to prepare the product according to this aspect of the invention. The product, which is a retentate of coarse bran particles, can be used in wet and/or pressed form as at least one of a feed additive for animals, a food additive for the human diet, a cosmetic paste or lotion, a fertilizer. The retentate of coarse bran particles according to the invention is a product ready for consumption or use with a long shelf life in a vacuum-packed form or after lyophilization, which, in turn, allows for an additional extension of the shelf life of the product.

Claims (18)

1. A method for producing a probiotic composition based on plant materials, comprising the following stages: natural honey or material of plant origin, which is cereals, legumes, vegetables, fruits, green mass of edible plants, including wild plants, and their by-products, taken separately or in the form of blends of different compositions, enriched with a composition of microelements, converted into a liquid form of hydrolyzate with a dry matter concentration of 0.2 to 80%, pasteurized, mesophilic and/or thermophilic starter containing strains of food microorganisms is introduced, fermented at a temperature optimal for the reproduction of microorganisms until they reach a stationary growth phase, transferred to the fermentation stage at a suboptimal low temperature, filtered from coarse bran particles after hydrolysis or one of the fermentation stages. 2. The method according to claim 1, wherein said by-products are selected from the group consisting of crushed grain, ground seeds, flakes, granules, flour, juices, solid/semi-liquid/liquid/dry concentrates, pastes and cake. 3. The method of claim 1, wherein the micronutrient composition is selected from the group comprising at least one of pink (Himalayan) salt, sea salt, seaweed powder, food ingredients or compositions containing a broad spectrum of micronutrients, or any combination thereof. 4. The method according to claim 1, wherein the mesophilic and/or thermophilic starter is represented by at least one of the strains suitable for use in the food industry, from among the species of lacto/bifido/propionic acid bacteria of the following genera: Aerococcus, Bifidobacterium, Carnobacterium, Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Oenococcus, Pediococcus, Propionibacterium, Sporo - 21 048767 lactobacillus, Streptococcus, Tetragenococcus, Vagococcus, Weissella, both individually and in any combination with each other. 5. The method according to claim 1, further comprising the step of introducing at least one of the components selected from the group consisting of: minerals, polyphenols and phenolic acids individually or as a mixture, sources of omega 3 and 6, dietary fiber, herbal extracts, natural honey, molasses, natural sugars, natural thickeners, natural antioxidants and flavors in the form of aqueous solutions, powder, granules, aqueous or oil extracts, pastes, crushed plant materials, plant parts, crystals, or any combination thereof. 6. The method according to item 1, wherein food grade alkalis and/or antifoaming agents are introduced at the hydrolysis and fermentation stages. 7. The method according to claim 1, wherein converting the hydrolyzate into liquid form additionally includes saccharification of the hydrolyzate. 8. The method according to item 1, in which the conversion of the hydrolyzate into liquid form is carried out using food grade alkalis. 9. The method according to claim 1, wherein at the fermentation stage or after filtration the composition is diluted with water, juice, saline solution, herbal and plant solution, syrups or other beverages, edible oils. 10. The method according to claim 1, wherein the fermentation step at a suboptimal temperature is completed upon reaching specified characteristics selected from the group consisting of: level of enzyme activity, composition and quantitative content of vitamins, content of monomers and/or oligomers of proteins, carbohydrates and fats, content and composition of short-chain fatty acids, exopolysaccharides, bacteriocins, polyphenols and phenolic acids, natural antioxidants, minerals in bioavailable form, dietary fiber, amount of postbiotic and/or metabolic layer, pH level, fermentation time at a given temperature, organoleptic properties, or any combination thereof. 11. A probiotic composition based on plant materials, obtained by the method according to paragraph 1, characterized by: pH level from 1.5 to 6.5, absence of coarse bran particles, content: compositions of microelements, natural antioxidants of plant origin, polyphenols and phenolic acids of plant and bacterial origin, dietary fiber, including fermented dietary fiber, peptides, beta-glucans, mono- and oligomers of carbohydrates and proteins, polyunsaturated fatty acids, active food microorganisms, accumulated metabolites and postbiotics of food microorganisms, minerals in bioavailable form, hydrolytic enzymes and detoxification enzymes, bacteriocins, exopolysaccharides, B vitamins, short-chain fatty acids, neurotransmitters and their precursors, heme-containing proteins and other metalloproteins; and a mass fraction of dry matter from 0.2 to 80 wt.%. 12. The probiotic composition of claim 11, wherein the micronutrient composition is selected from the group comprising at least one of pink (Himalayan) salt, sea salt, seaweed powder, food ingredients or compositions containing a broad spectrum of micronutrients, or any combination thereof. 13. Use of the probiotic composition according to item 11 as a probiotic health product for external and internal use, wherein internal use is selected from the group containing oral, intranasal and rectal use. 14. Use according to item 13, characterized in that the probiotic composition is used with dilution in physiological or aqueous solutions, herbal solutions and other natural medicines. - 22,048,767 funds. 15. Use of a probiotic composition according to claim 11 as a functional food ingredient in at least one of solid food, pastes, beverages and ready-made sauces. 16. Use of a probiotic composition according to claim 11 as a means selected from a group containing a detoxifying means, an ingredient of medical means, a component having effects directed against obesity, symptoms of type II diabetes, proinflammatory and toxicogenic glycation of biomolecules with elevated levels of postprandial blood sugar. 17. Use of a probiotic composition according to item 11 as a concentrate for the preparation of functional food products and an additional source of digestive enzymes. 18. Use of a probiotic composition according to claim 11 as a product selected from a group containing a feed additive for animals, a food additive for the human diet, a cosmetic paste or lotion, or a fertilizer.