Bifidobacterium adolescentis pbb-3 strain used to obtain bifido-containing product

FIELD: chemistry.

SUBSTANCE: Bifidobacterium adolescentis PBB-3 strain is isolated from the intestinal contents of a healthy adult person and is deposited in the National Collection of Normal Microflora of the G.N.Gabrichevsky Moscow Research Institute of Epidemiology and Microbiology under registration number 218. The strain propagates in different culture media with accumulation of production biomass with high concentration of bifidobacteria.

EFFECT: strain has acid-forming activity, antagonistic activity on pathogenic and opportunistic microorganisms and ferments milk, which enables its use to obtain a bifido-containing product.

2 tbl, 5 ex

 

The invention relates to the food, cosmetic, biotech and medical industries, in particular, is used to prepare fermented milk, fermented and non-fermented food, hygiene and cosmetics, dietary supplements and bacterial preparations.

Due to unfavorable environmental conditions, stress, prolonged antibiotic, radiotherapy and chemotherapy, dysbiosis resulting from illness, an increasing number of people suffering from disorders of the normal microflora. For the treatment and prevention of these disorders is gaining attention as drugs, with probiotic action, and food products containing bifidobacteria.

In the human intestine, basically, there are 5 species of bifidobacteria - B.bifidum, B.longum, B.adolescentis, B.breve and B.infantis. Of them with the greatest frequency in the biocenosis detected species B.bifidum and B.longum. View B.adolescentis more common in adults, and species B.breve and B.infantis - predominantly in infants.

Well-known foreign mono - and combination preparations-eubiotics and dairy products, including bifidobacteria species B.bifidum, B.longum, B.adolescentis, B.breve (patent 2336886, patent 2411008, France; patent 637297, Switzerland; patent 2134179, patent 242066, Germany; patent 05227946, patent 0430642, Japan; patent 4645667, U.S. patent 6063935, Japan, the patent 5922375, U.S. patent 5902743, U.S. patent 6746672, U.S. patent 6548057, USA). In our country for the preparation of bacterial preparations, dietary supplements, medicinal products-dietary use of domestic strains of bifidobacteria species B.bifidum 1,791,792; B.longum VM, I-2, I-4; B.adolescentis MC-42, G 7513; .breve 79-119; B.infantis 79-43 (copyright certificate №343588, RF patent 1347221, RF patent 849779, RF patent 2108383, RF patent 2152993, patent 863639, RF patent 2120991, RF patent 2078814, RF patent 2111248).

The quality of medicinal products and drugs is largely due to the properties of the producer strain - specific activity and adaptability. A large range of biologically active and technologically advanced strains of bifidobacteria makes possible the development of new mono - and integrated products for medical nutrition and bakpreparatov, capable of providing high colonization resistance of the human intestine, to restore the normal microflora and reduce the incidence of various intestinal infections.

The present invention is to obtain a strain of microorganism of the species .adolescentis PBB-3 with high biological activity, resistance to environmental conditions, with a stabilizing kislotoobrazovanie in the whole temperature range the development of cells, squarepusher milk and breeding on artificial nutrient media of different composition.

Strain Bifidobacterium adolescentis PBB-3 isolated from the contents of the intestines healthy adult and deposited in the Museum of microorganisms fsri mniam them. Hingamisega of the CPS No. 218.

The inventive strain can be used in the production of dairy products, fermented foods, such as kvass, beer, etc., non-fermented foods, such as juices, drinks, etc., starter cultures for the preparation of dairy products and biologically active additives to food. Strain Bifidobacterium adolescentis PBB-3 can also be used for the production of hygienic and cosmetic products such as creams, lotions, wipes, Soaps, powders, tampons, nourishing masks, gels, feminine sanitary pads, skin milk, various powders, etc.

In addition, the strain of Bifidobacterium adolescentis PBB-3 can be used in the production of bacterial preparations, in the form of candles, oral, and other tools.

The technical result of the claimed invention to provide a strain Bifidobacterium adolescentis PBB-3, which can be used as biologically active substances in the composition of fermented milk, fermented and non-fermented food starters, cosmetics, pharmacy hygiene and the logical devices, biologically active additives and bacterial preparations. Strain Bifidobacterium adolescentis PBB-3 has biologically and technologically important properties, such as acid-forming activity, the ability to increase microbial biomass in different nutrient media. Strain Bifidobacterium adolescentis PBB-3 antagonistically active against pathogenic and conditionally pathogenic microorganisms. This will allow you to use it as a biologically active component in the composition of different probiotic means of ensuring the normalization of microbiocenosis of the human body, including the gastrointestinal and urogenital tracts, skin and mucous membranes of the skin, and broaden the Arsenal of similar means.

Cultural and morphological properties of the strain

Cells are gr + polymorphic sticks with bifurcation or thickening 1-2 ends, in a liquid nutrient medium to grow over the entire height of the volume, except for the zone of aerobiosis. Separately grown colonies on hepatic nutrient medium have cometabolism form colonies. Logarithmic growth phase ends for 16-24 hours of cultivation.

Physical and biochemical properties of the strain

Non-sporulating still gone anaerobic. The optimum temperature for growing 37-39 degrees Celsius. Catalase does not form, gelatine does not dilute. Sprayway glucose with the formation of the UKS the red and L + lactic acid without the formation of gas, sakissa environment growing to a pH from 4.3 to 3.9. Ferments lactose, arabinose, xylose, cellobiose, inulin, melibiose, melezitose, raffinose, salicin, not fermented mannitol, mannose, sorbitol, Inositol. Strain skachivat milk for 12-24 h of cultivation.

Strain Bifidobacterium adolescentis PBB-3 has an antagonistic activity against pathogenic and conditionally pathogenic microflora (table 1).

Table 1
Antagonistic activity of strain .adolescentis PBB-3
StrainThe zone of growth inhibition of the test cultures, mm/ M±M
Sh.sonnetSh.flexneriE.coliS.aureusPr.vulgaris
.adolescentis PBB-320±1,218±1.822±1,610±1,418±1,6

The strain is expanding microbial mass and increase hydrolysate-dairy nutrient medium (GM), bifidum environment (BF), thus producing acid (table 2).

Table 2
The accumulation of microbial biomass (CFU /ml 10 lg) and the activity of acid generating (in degrees Turner) strain .adolescentis PBB-3 in the dynamics of development
NUTRIENT MEDIUMTIME of CULTIVATION
BIOLOGICAL ACTIVITY
12 hours24 hours48 h
SOMESOMESOME
GM8,41088,81187.8164
BF8,2888,6987,4142

The maximum strain accumulates microbial mass to 12-24 hours of cultivation, reaching in 1 ml lg 8-9 microbial cells producing acid.

So about what atom, the proposed strain .adolescentis RVV-3 technology, as propagated in various nutrient media with the accumulation of production of biomass with a high concentration of bifidobacteria; has acid-forming activity and antagonistic activity against pathogenic and conditionally pathogenic microorganisms.

Fermented milk, fermented, unfermented food, hygiene, cosmetics, dietary supplements and bacterial preparations containing strain .adolescentis RVV-3, can find application in the food, medical, biotechnology and cosmetic industries.

The ability of strain .adolescentis RVV-3 to increase biomass in different nutrient media, acid-forming activity, antagonistic activity against pathogenic and conditionally pathogenic microflora, as well as other useful for the human body properties, typical for the species .adolescentis, make possible the use of the strain in the various corrective means of therapeutic and prophylactic purposes.

Example 1. The use of strain .adolescentis RVV-3 in the composition of dietary supplements.

1 passage: lyophilized strain .adolescentis RVV-3 in an amount of 0.1 g dry biomass contribute in tubes with nutrient medium and titrated by the method of tenfold intelligence is s to 10 -9. Incubated at a temperature of 37-38 degrees Celsius before the formation of visible growth in the last dilutions.

2 passage: a grown culture of 10-7-9dilutions up to 10% contribute to the nutrient medium and cultured at a temperature of 37-38 degrees Celsius within 12-24 h before formation of the dense growth of the visible area of aerobiosis.

3 passage: the biomass in an amount up to 10% of the volume contribute to the accumulation and cultivate 12-24 h at 37-38 degrees Celsius to obtain a biomass content of live bifidobacteria not less than 108/ml.

The result is biologically active additive, which is a liquid concentrate with a high, not less than 108/ml, content of bifidobacteria.

Example 2. The use of strain .adolescents PBB-3 for the preparation of fermented and non-fermented foods.

1 passage: lyophilized strain .adolescentis PBB-3 in an amount of 0.1 g dry biomass contribute in tubes with nutrient medium and titrated by the method of ten-fold dilutions up to 10-9. Incubated at 37-38 degrees Celsius before the formation of visible growth in the last dilutions.

2 passage: a grown culture of 10-7-9dilutions up to 10% contribute to the nutrient medium and cultured at a temperature of 37-38 degrees Celsius within 12-24 h before formation of dense growth shorosho visible area of aerobiosis.

3 passage: the biomass in an amount up to 10% of the volume contribute to the accumulation and cultivate 12-24 h at 37-38 degrees Celsius to obtain a biomass content of live bifidobacteria not less than 108/ml.

Obtaining a fermented or non-fermented food: grown biomass bring in food raw materials (milk, juices, beer, kvass, etc. up to 10% of the volume, mix and pour into containers. The result is a product with a concentration of live bifidobacteria not less than 106/ml.

Example 3. The use of strain .adolescentis PBB-3 for the preparation of a fermented milk product.

1 passage: lyophilized strain introduced into the environment of cultivation and titrated by the method of ten-fold dilutions up to 10-9. Incubated at 37-38 degrees Celsius before the formation of visible growth in the last dilutions. Or dry lyophilized biomass of bifidobacteria contribute in 1-2 doses in the tube with 30 ml of culture medium. Cultivated at a temperature of 37-38 degrees Celsius to obtain a biomass content of live bifidobacteria not less than 107/ml.

2 passage: a grown culture of strain .adolescentis PBB-3 of the last dilution or test-tube with the increased biomass contribute to the accumulation of up to 10% and cultured at a temperature of 37-38 degrees Celsius to obtain the biomass with the holding of bifidobacteria not less than 10 8/ml.

3 passage: the biomass of the strain .adolescentis PBB-3 in the amount of up to 10% bring in sterile milk, cultured to the formation of a clot (milk product). The number of passages No. 2 may be increased depending on the volume of production.

Example 4. The use of strain .adolescentis PBB-3 to obtain the bacterial preparations.

The culture of the strain of the 1st or 2nd passage (example 1) contribute to the nutrient medium, cultured at a temperature of 37-38 degrees Celsius to obtain the biomass of bifidobacteria with the contents of living cells is not less than 108in 1 ml. To the obtained biomass add a protective environment drying (saharso-gelatin medium, skim milk). The resulting suspension is poured into ampoules, vials and other containers, frozen at a temperature of minus 35 to 45 degrees Celsius and lyophilizers from the frozen state in a vacuum drying apparatus. The resulting preparation contains not less than 107living cells of bifidobacteria. The content of bifidobacteria is consistent with the scientific and technical documentation on similar bifidobakterii drug "Bifidumbacterin dry" and is sufficient to use the funds received as a drug - eubiotics.

Example 5. The use of Bifidobacterium adolescentis PBB-3 to obtain hygienic and cosmetic products.

The cult is the strain of the 1st or 2nd passage (example 1) contribute to the nutrient medium, cultivated at a temperature of 37-38 degrees Celsius to obtain the biomass of bifidobacteria with the contents of living cells is not less than 1081 ml, dried by spray or freeze drying.

The obtained dry biomass of bifidobacteria strain Bifidobacterium adolescentis PBB-3 is introduced into the composition of the nutrient cream in an amount of not more than 1% by volume.

Safety: with the introduction of the five white mice dose per os microbial culture strain of bifidobacteria in an amount of 5×107SOME of the animal's death mice is not marked.

Strain Bifidobacterium adolescentis PBB-3 used to obtain bifidobacteria products deposited in the Public collections of microorganisms of the normal microflora (HCNM) fgun "mniam them. Hingamisega Rospotrebnadzor", registration No. 218.



 

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