RU2216588C1 - Strain bifidobacterium longum ya2 used for preparing fermented-milk, fermented and nonfermented foodstuffs, ferments, hygienic and cosmetic agents, biologically active supplements and bacterial preparations - Google Patents

Abstract

FIELD: food, cosmetic and medicinal industry, biotechnology, microbiology. SUBSTANCE: strain Bifidobacterium longum YA2 is isolated from intestine content of health adult man and multiplies actively in nutrient medium with accumulation of industrial biomass for short (12-24 h) culturing period and with high concentration of bifidobacteria. Strain shows acid-producing and inhibitory properties with respect to pathogenic and putrefactive microflora. This allows preparing fermented-milk, fermented and nonfermented foodstuffs, ferments, hygienic and cosmetic agents, biologically active supplements and bacterial preparations providing normalization of microbiocenosis of human body, among them, digestive and urogenital tracts, cutaneous and mucous tissues. EFFECT: valuable biological and medicinal properties of strain, expanded assortment of analogous agents. 2 tbl, 5 ex

Description

 The invention relates to the food, cosmetic, biotechnological and medical industries, in particular, it is used for the preparation of sour-milk, fermented and non-fermented foods, hygiene and cosmetics, biologically active additives and bacterial preparations.

 Due to adverse environmental conditions, stress, prolonged antibacterial, radiation and chemotherapy, dysbiosis resulting from past diseases, the number of people suffering from impaired normal microflora of the body increases. For the treatment and prevention of these disorders, increasing attention is being paid to both drugs with a probiotic effect and food products containing bifidobacteria.

 Five types of bifidobacteria, B. bifidum, B. longum, B. adolescentis, B.breve and B.infantis, are found in the human intestines. The species B.bifidum, B.longum and B.adolescentis (39-75%) are found with the greatest frequency in the biocenosis, the latter two being predominant in adults, and the species B.breve and B.infantis predominantly in infants (17 -36%).

 Foreign mono-and complex preparations-eubiotics and sour-milk products are known, including bifidobacteria of the species B. bifidum, B. lonqum, B. adolescentis, B.breve (patent 2336886, patent 2411008 France; patent 637297 Switzerland; patent 2134179 , patent 2421066 Germany; patent 05227946, patent 0430642 Japan; patent 4645667 USA).

 In our country, domestic strains of bifidobacteria species of B. bifidum 1,791 are used to prepare bacterial preparations and dietetic food products; VKPM S-1257, Ac-1578; B.lonqum B379M; B.adolescentis MC-42 (copyright certificate 343588, patent RF 1347221, patent 849779, patent RF 2108383, patent RF 2152993, patent 863639).

 The quality of medicinal products and preparations is largely due to the properties of the producer strain - its specific activity and manufacturability. A large assortment of biologically active and technologically advanced strains of bifidobacteria makes it possible to develop new mono-and complex medical nutrition products and drugs that can provide high colonization resistance of the human intestine, restore its normal microflora and reduce the incidence of various intestinal infections.

 The objective of the present invention is to obtain a strain of microorganisms of the species B.longum with high biological activity, resistance to environmental conditions, with stabilizing acid formation in the entire temperature range of cell development, fermenting milk well and rapidly multiplying in artificial nutrient media.

 The problem is solved by using the strain of Bifidobacterium longum H2 for the production of sour-milk, fermented and non-fermented foods, starter cultures, hygienic and cosmetic products, dietary supplements and bacterial preparations.

 The strain Bifidobacterium longum I2 was isolated from the intestinal contents of a healthy adult and deposited in the Museum of Microorganisms of GU MNIIEM them. G.N.Gabrichevsky under 170.

 The inventive strain can be used in the production of fermented milk products, fermented foods, for example: kvass, beer, etc., unfermented foods, for example: juices, drinks, etc., starter cultures for the preparation of fermented milk products and dietary supplements. Also, the Bifidobacterium longum I2 strain can be widely used for the production of hygiene and cosmetics, for example: creams, lotions, napkins, soaps, powders, tampons, nourishing masks, gels, women's pads, cosmetic milk, various powders, etc.

 In addition, the strain of Bifidobacterium longum I2 can be used in the production of bacterial preparations, in the form of suppositories, oral and other drugs.

 The technical result of the claimed invention is the production of sour-milk, fermented and non-fermented foods, starter cultures, hygienic and cosmetics, dietary supplements and bacterial preparations, which as an active principle contain a strain of Bifidobacterium longum H2, which has acid-forming and antagonistic activity against pathogenic and conditionally pathogenic microflora, which ensures the normalization of the microbiocenoses of the human body, including the gastrointestinal and urinary tract of the tracts, skin and mucous membranes, as well as an expansion of the arsenal of similar drugs.

 Cultural and morphological properties of the strain.

 Cells are gr + polymorphic bacilli with bifurcation or thickening at the 1-2 ends, in liquid nutrient media they grow along the entire height of the volume, except for the aerobiosis zone. Separately grown colonies on the hepatic nutrient medium have a spherical, crumbly form of colonies. The logarithmic growth phase ends at 12-24 hours.

 Physico-biochemical properties of the strain.

 Indisputable, motionless anaerobic. The optimum temperature of cultivation is 37-39 degrees Celsius. Catalase does not form, gelatin does not dilute. It ferments glucose with the formation of acetic and L + lactic acids, without the formation of gas, acidifying the growth medium to a pH of 4.1-3.9. It ferments lactose, arabinose, raffinose, galactose, variable: cellobiose, mannose, xylose, sorbitol. The strain fermented milk by 12-24 hours with the formation of a clot.

 The strain actively increases microbial mass during growth on hydrolyzate-dairy medium (GM), Bifidum medium (BP), producing acids (Table 1).

 The strain maximally accumulates microbial mass by 12-24 hours, reaching 1-9 lg of 8-9 microbial cells. The strain exhibits an antagonistic effect against pathogenic and conditionally pathogenic microflora (Table 2).

 Thus, the proposed strain B.longum H2 is technologically advanced, since it actively propagates in nutrient media with the accumulation of production biomass in short, 12-24 hours, cultivation periods with a high concentration of bifidobacteria; It has acid-forming and inhibitory properties against pathogenic and putrefactive microflora.

 Sour-milk, fermented, non-fermented foods, hygienic, cosmetics, dietary supplements and bacterial preparations containing B.longum H2 strain can be widely used in the food, medical, biotechnological and cosmetic industries.

 The ability of the strain B.longum H2 to increase biomass in a nutrient medium in a short cultivation period, its antagonistic, acid-forming activity, as well as other properties useful to the human body, make it possible to use the strain as part of various corrective agents for therapeutic and prophylactic purposes.

 Example 1. The use of strain B.longum H2 in the composition of biologically active additives.

1 passage: lyophilized B.longum H2 in the amount of 0.1 g of dry biomass is introduced into test tubes with a nutrient medium and titrated by the method of ten-fold dilutions to 10 ^-9 . Incubated at a temperature of 37-38 degrees Celsius until the formation of visible growth in the last dilutions;
2 passage: the grown culture from 10 ^-7-9 dilutions in a volume of up to 10% is introduced into the nutrient medium and cultivated at a temperature of 37-38 degrees Celsius for 12-24 hours until a dense growth with a visible aerobiosis zone is formed;
3 passage: the resulting biomass in an amount of up to 10% of the volume is introduced into the accumulation medium and cultivated for 12-24 hours at 37-38 degrees Celsius to obtain biomass with a content of live bifidobacteria of at least 10 ⁸ / ml.

The result is a biologically active additive, which is a liquid concentrate with a high, at least 10 ⁸ / ml, content of bifidobacteria.

 Example 2. The use of strain B. longum H2 for the preparation of unfermented food products.

1 passage: lyophilized B.longum H2 in the amount of 0.1 g of dry biomass is introduced into test tubes with a nutrient medium and titrated by the method of ten-fold dilutions to 10 ^-9 . Incubated at 37-38 degrees Celsius until the formation of visible growth in the last dilutions;
2 passage: the grown culture from 10 ^-7-9 dilutions in a volume of up to 10% is introduced into the nutrient medium and cultivated at a temperature of 37-38 degrees Celsius for 12-24 hours until a dense growth with a well-visible zone of aerobiosis is formed;
3 passage: the resulting biomass in an amount of up to 10% of the volume is introduced into the accumulation medium and cultivated for 12-24 hours at 37-38 degrees Celsius to obtain biomass with a content of live bifidobacteria of at least 10 ⁸ / ml.

Obtaining an unfermented food product: the grown biomass is introduced into food raw materials (milk, juices, etc.), mixed and poured into containers. The result is a product with a concentration of live bifidobacteria of at least 10 ⁶ / ml.

 Example 3. The use of strain B. longum I2 for the preparation of fermented milk product.

1 passage: lyophilized strain is introduced into the culture medium and titrated by ten-fold dilution to 10 ^-9 . Incubated at 37-38 degrees Celsius until the formation of visible growth in the last dilutions. Or dry lyophilized biomass of bifidobacteria is added in an amount of 1-2 doses in a test tube with 30 ml of culture medium. Cultivate at a temperature of 37-38 degrees Celsius to obtain biomass with a content of live bifidobacteria not less than 10 ⁷ / ml;
2nd passage: a grown culture of B. longum H2 from the last dilutions or from a test tube with grown biomass is introduced into the accumulation medium with a volume of up to 10% and cultivated at a temperature of 37-38 degrees Celsius to obtain biomass with a content of bifidobacteria of at least 10 ⁸ / ml;
3 passage: the resulting biomass of B. longum H2 in a volume of up to 10% is introduced into sterile milk, cultivated until a clot forms (fermented milk product). The number of passages 2 can be increased depending on the volume of production.

 Example 4. The use of B. longum H2 to obtain bacterial preparations.

The culture of the strain of the 1st or 2nd passage (Example 1) is introduced into the nutrient medium, cultivated at a temperature of 37-38 degrees Celsius until a biomass of bifidobacteria is obtained with a living cell content of at least 10 ⁸ in 1 ml. Protective drying media (sucrose-gelatin medium, skim milk) are added to the resulting biomass. The resulting suspension is poured into ampoules, vials and other containers, frozen at a temperature of minus 35-45 degrees Celsius and lyophilized from a frozen state in a vacuum dryer. The resulting preparation contains at least 10 ⁷ living bifidobacteria cells. This content of bifidobacteria is consistent with the scientific and technical documentation for a similar bifidobacterium preparation - dry bifidumbacterin (FS 42 132 BC-94) and is sufficient to use the resulting agent as a drug - eubiotics.

 Example 5. The use of B. longum H2 for hygiene and cosmetics.

The culture of the strain of the 1st or 2nd passage (Example 1) is introduced into the nutrient medium, cultivated at a temperature of 37-38 degrees Celsius until a biomass of bifidobacteria is obtained with a living cell content of at least 10 ⁸ in 1 ml.

 The resulting biomass of bifidobacteria is introduced into the composition of a hygienic or cosmetic product.

Claims (1)

 The strain Bifidobacterium longum Y2 (collection 170 of the State collection of human microflora GKNM GU MNIIEM Ministry of Health of the Russian Federation named after G.N. Gabrichevsky) used to produce fermented milk, fermented and non-fermented foods, starter cultures, hygienic and cosmetic products, dietary supplements and bacterial preparations.

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