EA017593B1 - Consortium of bacteria (streptococcus thermophiles a1, lactococcus lactis a1-1, lactobacillus plantarum sh2, lactobacillus fermentum pl3, lactobacillus acidophilus k4, bifidobacterium longum pl5, bifidobacterium bifidum ki 6) for yoghurt production - Google Patents

Abstract

The invention relates to medical microbiology and diary industry and represents a consortium of bacterial probiotic cultures which can be used for preparation of fermented milk foods and bacterial preparations. The invention is aimed at producing a novel consortium of bacterial cultures having good organoleptic, textural properties in milk fermentation expressed by probiotic, antioxidant and antigenotoxic effect. The consortium is created by a method of combined culturing of seven bacteria strains - Streptococcus thermophiles a1, Lactococcus lactis a1-1, Lactobacillus plantarum sh2, Lactobacillus fermentum pl3, Lactobacillus acidophilus k4, Bifidobacterium longum pl5, Bifidobacterium bifidum ki 6. The consortium possesses not only high antagonistic activity to opportunistic pathogenic microflora, but also to a high potential to reduce the action of free radicals expressed by a DNA- protective action, thus providing to use thereof for prophylaxis of colorectal cancer and for correction of age disturbances in persons of elder and old age.

Description

The invention relates to medical microbiology and the dairy industry and is a consortium of bacterial probiotic cultures, which can be used for the preparation of fermented milk products, bacterial preparations.

A consortium of bacterial cultures for the preparation of yogurt 81gsr1ososp§ 111sgtor1i1s§ a1. Bac1osossi§1aci§ a1-1. LB1Bb8B8B1A1BIT §12. Baobbbbgrrrrrr13. Baclofacialis aa6or1i§k4, Bbbacompop 1 pd. VgMobas1sgsht Shbbit k1 6 is used for the preparation of fermented milk products.

The inventive consortium has pronounced probiotic. antioxidant and antigenotoxic properties and is intended for fermenting milk in order to prepare a functional product. The resulting product can be used for the prevention of colorectal cancer due to antigenotoxic properties and is used for elderly people to correct age-related disorders.

The invention relates to medical microbiology and the dairy industry and is a consortium of bacterial probiotic cultures. which can be used for the preparation of dairy products. bacterial preparations.

Known strains of bacillus bacillus aa6orb1i§ are known. VGMoasShgsht Shbbit. VgMoaschscht 1opdit and 81gcr1osossi§1sgtorO1i§ to obtain the symbiotic fermented milk product Bifacil. The fermented milk product Bifacil has therapeutic and prophylactic properties for infectious diseases of the body due to the direct bacteriostatic suppression of pathogenic microorganisms of the digestive tract. and due to the activation and stimulation of immunocompetent organs - the thymus and spleen.

A consortium of bacterial cultures for the preparation of yogurt has not only a high antagonistic activity for opportunistic and pathogenic microflora. but also a high potential to reduce the effect of free radicals. pronounced DNA protective effect. which allows it to be used for the prevention of colorectal cancer and for the correction of age-related disorders in the elderly and senile.

The objective of the invention is to obtain a new consortium of bacterial cultures. possessing good organoleptic. textural properties when fermenting milk. pronounced probiotic. antioxidant and antigenotoxic effect. The consortium was obtained by co-cultivation of seven strains of bacteria: 81gcr1ossoss§111gtor1i1s§ a1. Bac1osossi§1aci§ a1-1. BASBASASpk p1ap! Agit §12. Habiton d'Article p13. Baclococcus aa6or1i§ k4. Bbbasbrst 1pd p15. WBWAWWWWWWWWWWWWWWWWHF 6.

The components of the consortium are isolated from dairy products: Ayran. koumiss. shubat and clinical samples.

A consortium of bacterial cultures for the preparation of yogurt 81er1ossoss§111gtor1n1s§ a1. Bac1osossi§1aci§ a1-1. Baclococci8 p1ap1 agit §12. Habiton d'Article p13. Baclococcus aa6or1i§ k4. Bbbasbrst 1pd p15. BhMoBac1crf bShbit k1 6 is used for fermenting milk in order to prepare a functional product.

In studies of ν ίΙΐΌ, the colonization potential of individual strains was also demonstrated in the consortium. antioxidant activity (OAA = 68.4%). antigenotoxic properties (PI = 0.6). a wide range of antagonistic properties (with respect to E. coli. Prigo sigma tigma. K1cbg1c11a. A safety assessment has been carried out.

Multicomponent probiotics based on two or more types and genera of microorganisms have several advantages over monocomponent preparations: they have a wider spectrum of antagonistic activity against pathogenic and conditionally pathogenic microflora. more technological in practice. since they have a greater number of transformable substances.

Before conducting joint cultivation of bacterial strains, they were controlled for the manifestation of antagonistic properties in relation to each other. for these experiments, the method of delayed antagonism was preferred. Zones of growth inhibition in the cultures of the obtained consortium were not found. As a result, they started an experiment with a simultaneous increase in the cultures of the consortium. Combined starter cultures are usually less susceptible to contamination from extraneous microflora and have a low sensitivity to phage effects.

Strains 81er1ossoss§ 111sgtor1n1s§ a1. Bas1ososg§ 1asB§ a1-1. Bas1basbnp1 p1ap1agit §12. Habiton d'Article p13. Baslobaspida aa6or1i§ k4. Bbbasbrst 1pd p15. VGMoasShgsht Ybbpt k1 6. members of the consortium. demonstrated resistance to high concentrations of hydrochloric acid. bile. gastric juice. phenol resistance (table. 1). Those. these cultures are characterized by high survival in adverse conditions of the gastrointestinal tract. which ensures their viability and functioning.

The consortium has a high potential to reduce the effect of free radicals. the total antioxidant activity of the consortium was 68.4%. It is known. that the level of antioxidant activity of food components in the diet of people of old age from long-lived populations is somewhat

- 1 017593 higher than officially recommended values.

The strains included in the consortium have a high antagonistic activity against such representatives of the conditionally pathogenic microflora as Vasik Shikkuh, Ooohpuh, reideideik, Rkoebotoyak aegidshoka, Excepsois soya, Prgo! 8a1toie11a [ur. Zones of growth inhibition were recorded in the range of 15-30 mm.

One of the key indicators of the functional effectiveness of the consortium is the DNA protective effect. The calculation of the damage index was 0.6, which indicates a pronounced DNA protective effect of the consortium.

Organoleptic, physico-chemical and microbiological indicators of the finished product largely depend not only on the types of microorganisms used, the properties of the strains, but also on their ratio in bio-starter culture. Therefore, it was necessary to select the optimal ratio of cultures in the combined multiprobiotic complex.

In this regard, at this stage of the work, the properties of the created associations were studied, consisting of cultures of lactobacilli and bifidobacteria in various ratios. The temperature regime of fermentation is 37 ± 1 ° C. In the fermentation process, indicators such as acid formation activity, aroma and taste of the product, viscosity and density were studied.

The ratio of cultures in the combined starter culture was chosen taking into account balanced growth, product viscosity and organoleptic characteristics. It was revealed that the ratio 1: 1 (culture of consortia: bifidobacteria) is the most optimal.

Organoleptic indicators taken from products obtained by consortium fermenting milk completely satisfied the needs of the required fermented milk products. Fermentation of milk was carried out in no more than 6 hours, while the finished product acquired a thick consistency with a content of at least 10 ⁸ live bacteria in 1 ml, high density, satisfactory ductility; the color is milky white, the taste is sour, yogurt, the smell of sour milk; pH 6.0; the energy of acid formation 70 ° T (table. 2).

The strains included in the consortium are deposited with the National Securities Depository under registration number B-372 and have the following morphological and physiological characteristics.

Strain 81er1ossossi5 1egtorO1i8 a1 isolated from ayran. The strain has the following characteristics.

Oval gram-positive cells are assembled in pairs and short chains, motionless. On nutrient agar form round smooth small colonies. The optimum development temperature is 40-45 ° C. Ferment maltose, raffinose.

The strain Bac! Ossossi 8 1ac115 a1-1 is isolated from ayran and has the following characteristics.

Oval gram-positive cells are arranged in the form of short chains of spores, they do not form capsules. On an agar medium, the culture forms beige, shiny, small colonies, and deep boat-shaped colonies. The temperature of the cultivation is 30-35 ° C, the optimum acidity of the medium is 6.0. Does not ferment ramnose, sucrose, raffinose.

Strain Lactobacillus r1ai! Asht 5112 isolated from shubat has the following characteristics.

Heterofermentative. Gram-positive sticks, medium length, single location, short chains. Stationary, do not form a dispute. Colonies on agarized culture media are whitish, smooth, convex, and the edges are even. In liquid media, moderate turbidity is formed, the sediment is dense. Optional anaerobic. The optimum development temperature is 30 ± 1 ° C; the optimum acidity of the medium is 6.0. Catalase-negative. It ferments lactose, cellobiose, galactose, fructose, mannose, sucrose, ribose, mannitol, arabinose, maltose, raffinose.

Strain Bac! Bacillus8 Gigenshit p13 isolated from clinical material, has the following characteristics.

Heterofermentative. Optional anaerobic. The bacterial cells are rod-shaped, regular in shape, gram-positive, arranged in short chains. They do not form a dispute. On solid nutrient media, the colonies are round, beige, with smooth edges. When grown on MPC broth for 24 hours at a temperature of 37 ° C, uniform turbidity and sediment are formed at the bottom. Ferments lactose, glucose, trehalose, galactose, fructose, mannose, sucrose, maltose, raffinose.

The strain Basfasfik asyoryik k4 isolated from koumiss has the following characteristics.

Homoenzymatic. Rods of different lengths, 3-5 microns in size, are arranged in short chains, singly, gram-positive, motionless, spore, does not form capsules, optional anaerobic. On an agarized medium MPC culture forms white, round colonies, the edges are even. The temperature of the cultivation is 37-43 ° C, the optimum acidity of the medium is 6.0. It ferments lactose, glucose, cellobiose, trehalose, galactose, fructose, maltose, mannose, sucrose.

Strain HbObas! Egst 1 poiditis p15 isolated from clinical material, has the following characteristics.

Cells are gram-positive polymorphic sticks with club-shaped thickenings, single, double, in the form of chains. In liquid culture media, the culture grows along the entire height of the volume, except for the aerobiosis zone. Fixed, undisputed anaerobes. Optimum temperature

- 2 017593 cultivation of 37.5-38 ° C, the pH of the nutrient medium 7.2. Catalase is not formed, gelatin is not liquefied. Ferments lactose, glucose, trehalose, galactose, fructose, maltose, mannose, sucrose, ribose, melibiosis, arabinose, xylose.

Strain B1yyobasepit Iyyit k1 6 isolated from clinical material, has the following characteristics.

Cells are gram-positive polymorphic rods with a thickening at the ends, form clusters. In liquid nutrient media, they grow along the entire height of the volume, except for the aerobiosis zone. Fixed, undisputed anaerobes. The optimum temperature of cultivation is 37.5-38 ° C, the pH of the nutrient medium is 7.2. Catalase is not formed, gelatin is not liquefied. Ferments glucose, lactose, fructose, arabinose, xylose.

The resulting consortium can be recommended for use as a starting culture for the preparation of fermented milk products, other fermented milk therapeutic and dietary products, unfermented food products and bacterial preparations.

Table 1. Data on the resistance of bacterial strains

The name of the strains Phenol Hydrochloric acid (pH) Bile Gastric juice 5 4 2 twenty% thirty% 40% twenty% thirty% 40% £. ρίατιίαηιιη 4- + + ± ± ± ± + + B. / eGetShit - + + + + + 4- + 4- 4- B. AsShorkIp ^ + + 4- - + ± ± 4- 4- 4- 5.1 kegtorkIiz + + 4- - 4- + 4- 4- + B. 1acNz + + - - ± ± + + 4- ± B. 1op% um + + 4- - 4- 4- 4- 4- 4- B. Βΐβάιιηι - + + - 4- + - + 4- 4-

Table 2. Characteristics of the consortium

Organoleptic: Colour White, uniform throughout the mass Taste and smell Pleasant, sour-milk, without odors and odors Appearance and consistency: Homogeneous, in the form of a dense, uniform clot Physicochemical: Mass fraction of solids,% 16.1 Mass fraction of fat,% 2,5 Mass fraction of protein,% 3.8 Mass fraction of sucrose in terms of invert sugar,% 3,5 Acidity, ° T 86.0

The proposed invention ferments milk with the formation of a clot with good taste and organoleptic characteristics, has pronounced probiotic, antioxidant and antigenotoxic properties, which allows it to be used for the preparation of a fermented milk product for the correction of functional intestinal disorders.

Claims (1)

CLAIM A consortium of bacteria (81ger1osossiz 1yegtorY1ez a1 a1-1 Eas1osossiz 1asyz, BaLoasShiz r1ap1agit §112, BaLoasShiz £ egtep1it p13, BaLoasShiz aszyorPiz k4, V1yyoas1eg1it 1opdit p15, k1 V1yyoas1epit Yyyit 6) deposited in the Collection of Industrial Microorganisms Depository AO KazAgroInnovation under the number B-372 for making yogurt.