SU1604847A1 - Method of growing bifidobacteria - Google Patents

Abstract

This invention relates to biotechnology and can be used in the manufacture of fermented dairy products. In order to increase the viability of bifidobacteria (BB), their cultivation in milk is carried out in conjunction with acetic acid bacteria (UCB) of ACETOBACTER ACETI and ACETOBACTER PASTEURIANUM species selected for proteolytic activity against milk proteins. The BB and UCB strains are introduced into milk in a ratio of 3: 1 with a total amount of inoculum of 4-5%, and the cultivation is carried out at 30-37 ° C for 12-16 hours. 3 tab.

Description

This invention relates to biotechnology and can be used in the production of fermented dairy products.

The purpose of the invention is to increase the viability of bifidobacteria.

Cultivation of bifidobacteria in milk is carried out by co-cultivation with microorganisms that stimulate their growth, and acetic acid bacteria of the Acetobacter aceti and Acetobacter pasteuriamim species selected by the size of proteolytic activity, exceeding 7.5 mg% in relation to molars, are used as growth stimulants. proteins, while bifidobacteria and acetic acid bacteria are introduced in a ratio of 3: 1,

The method involves the use as microorganisms activating the development of bifidobacteria, acetic acid bacteria of species A.aceti and A. pasteurianum. Any strains of acetic-acid bacteria isolated from milk, x products can be used for the indicated species. In contrast to the acetic acid bakt.ieri, common in. other natural sources, they are endowed with proteolytic activity against milk proteins. These microorganisms are characterized by a strictly aerobic growth pattern, the ability to develop at low pH values, use organic acids as the sole carbon source, and are distinguished by their vitamin synthesis ability.

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The characteristics of acetic acid boilers of A.aceti and A.pasteurianum species are listed in Table. one.

The peculiarity of the metabolism of bifidobacteria consists in the anaerobic digestion of carbohydrates with the formation of organic acids, mainly lactic and vinegar, they easily enter into a symbiotic interaction with the acetic acid bacteria of A.aceti and A.pasteurianum species. In the process of cultivation in milk, these bacteria provide the biosynthesis of proteolysis products of proteins and vitamins of group B, which, being ± growth activators, promote the more rapid development of bifidobacteria. In addition, acetic acid bacteria during their development actively oxidize organic acids, accumulated by bifidus bacteria, and prevent the latter from inhibiting the final metabolites of their viability in the development of α-cells. 25 also obligate aerobiosis of acetic acid bacteria is important. Intensively absorbing oxygen contained in the culture medium, acetic acid bacteria create anaerobic conditions favorable for the development of bifidobacteria. The mutual benefit obtained by acetic acid bacteria in developing them together with bifidobacteria consists in feeding all available milk for acetic acid bacteria to carbon dioxide in the form of organic acids, among which the main role is taken by milk and acetic acid most easily assimilated by acetic acid bacteria At the same time, the anaerobic nature of bifidobacteria allows acetic acid bacteria to use all the oxygen — 1 s in the nutrient medium, in the absence of which they cannot nktsionirovat.

The activation of the development of bifidobacteria is most pronounced when they are combined with acetic acid bacteria in a 3: 1 ratio.

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A change in the ratio between microorganisms leads to a decrease in the cell content of both bifidobacteria and acetic acid bacteria in the combinations obtained by the proposed method, and consequently, a decrease in the milk-clotting activity of the latter (Table 2).

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The method is carried out as follows.

One of the production strains Bif1-1dobacteria and the strain of acetic acid bacteria are introduced into sterile milk in a ratio of 3: 1 with a total amount of inoculum of 4-5%. The cultivation is carried out at 30-37 C for 1216 hours.

The resulting combination is intertwined; 1015 times in sterile milk at intervals of 3-5 days. Between intersections, the combinations are stored at (6 + 2) ° C. After each replanting, the tissue is examined for a microscopic preparation, the number of viable cells of bifidobacteria and acetic acid bacteria, for milk-clotting activity and activity of acid accumulation.

Proven symbiosis is used as a separate starter or in combination with lactic acid bacteria in the production of fermented milk products.

Example., 10 ml of sterile milk is made of 0.3 ml of the strain Bifido-bacter longum 415 and 0.1 MIT of the acetic acid bacteria strain A.aceti K-92, VKPM B-2847. The mixture is thoroughly mixed and left at 30 ° C for 16 hours. Then the combination is transferred 15 times into sterile milk every 3 days. After each /; second reseeding, the combination is examined for production-valuable properties.

PRI mme R 2. 0.75 ml of B. longum 447 and 0.25 ml of the strain: acetic acid bacteria of A. pasteur i.anum M-67 VKPM-3395 is added to 20 ml of sterile milk. Blend thoroughly. mix and incubate at 37 ° C for 12 hours. Then the combination is changed 10 times into sterile milk at intervals of 5 days. After each subculture, the combination is examined for valuable production properties.

PRI me R 3-. 0.3 ml of B. longum 447 strain and 0.1 ml of A. aceti U-16 strain of acetic acid bacteria are added to 10 ml of sterile milk. The mixture is stirred and incubated for 14 hours at 32 ° C, then the combination is transferred 12 times into sterile milk at intervals of 4 days. After each subculture, the combination will be discussed in production} of the valuable properties.

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EXAMPLE 4: In 10 P1 sterile milk, 0.3 ml of the B.Iongum strain and 0.1 ml of the A.pasteurianum strain are introduced.

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143, VKPM B-2751. The mixture is unmixed and left for 14 hours at 35 C. Then the combinations are transferred 12 times into sterile milk at intervals of 4 days. After each subculture, the combination is examined for production-valuable properties. In tab. 2 shows a comparative characteristic of combinations, the floor of the snake offered and known methods. From the above data, it follows that with the co-cultivation of bifidobacteria and acetic acid bacteria of A.aceti and A.pasteurianum species, taken in a 3: 1 ratio, the content of bifidobacteria cells in milk increases approximately 100 times as compared with pure cultures and . approximately 10 times compared with the known. The combinations resulting from the implementation of the method have a higher milk-clotting and proteolytic activity, and are characterized by stability of the composition and properties.

The use of these combinations in the production of fermented dairy products will increase the content of viable cells of bifidobacteria and protein proteolysis products in them.

PRI me R 5. Semi-tion of fermented milk drink.

First, a combination of bifidobacteria with acetic acid is prepared}. bacteria. For this purpose, 0.3 MP of B.Iongum strain 415 and 0.1 MP of A.aceti strain K-92 (VKPM B-2847) are introduced into 10 ml of sterile milk. The mixture is thoroughly mixed and kept at 30 ° C for 16 hours. The resulting mixture is transferred to fresh milk every 3 days under the same conditions.

The combination is characterized by the following features: the bifidobacteria cell content is 2.2 billion in 1 ml, the number of acetic acid bacteria cells is 0.7 billion in 1 ml, the combination rolls up the milk at a dose of 3% inoculum for 10 hours, increases the content of nitrogen. in milk at 5.2 mg%, thin long sticks and small moving sticks are visible in the field of vision of the microspot.

The combination is used in the composition of the starter culture for the production of a fermented milk drink.

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 To obtain a laboratory ferment, about 1 liter of sterile skim milk was used to add 4 ml of a combination of bifibacteria with acetic acid bacteria and 1 ml of the lactic acid bacteria strain Lactobacillus acidophilus 5 Ds. The mixture of milk with sourdough is stirred and held at 37 ° C for 10 hours.

To obtain a production ferment in 3 kg of sterile skimmed milk, 0.1 kg of laboratory ferment is added. The mixture is stirred up and left for 7 hours. To obtain a fermented milk drink, 100 kg of milk are pasteurized at 92 ° C. After 5 minutes, cooled to 37 ° C and added to it 3 kg of production ferment. Squamification is carried out for 6 hours before acidification of the clot.

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Characteristics of the product are given in table. 3

Cottage cheese preparation is carried out in the following way.

To obtain a combination of bifidobacteria with acetic bacteria in 10 ml of sterile milk, 0.3 ml of Bifidobacterium longum 447 and 0, 1 ml of A. aceti U-16 strain (VKPM B-3373) are introduced. The mixture after mixing 1-vania is held at 32 ° C for 35 1 i h, and then transferred 10 times into sterile milk with a periodicity of 4 days.

The resulting combination contains in 1 ml of 1.3 billion cells of bifidobacteria 40 and 0.5 billion cells of acetic acid bacteria, coagulates milk for 8 hours, increases the content of amino nitrogen by 4.6 mg%, the microscopic preparation contains 45 long granular sticks and small mobile sticks.

To obtain a laboratory ferment in 0.1 l of sterile skim milk, 1.5 ml of a combination of bifidobacteria with acetic acid bacteria and 50 h of cottage cheese sourdough consisting of mesophilic and thermophilic milk bacteria are added. The mixture is stirred and maintained at for 10 hours

To obtain a production ferment, 0.1 kg of laboratory ferment is added to 3 kg of sterile milk. The mixture is stirred and held at 32 ° C for 6 hours;

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To obtain cottage cheese, 100 kg of mol.ok are pasteurized, cooled to 32 ° C, 3 kg of production is fermented, the vegetable ferment is left for 6 hours until the acidity is reached, 75 T, the whey is separated and pressed.

Characteristics of cottage cheese is given

in table 3.

Claims (1)

Invention Formula The method of pulling bifidobacteria, involving the co-cultivation in milk of bifidobacteria and microorganisms, stimulating their growth, characterized in that, in order to increase the viability of bifidobacteria, acetic acid bacteria of the species Acetobacter aceti and Acetobacter pasteurianum selected by the magnitude of proteolytic activity exceeding 7.5 mg% are used as growth stimulants against milk proteins, with bifidobacteria and acetic acid bacteria combined in a 3: 1 ratio. Table 1 Morphological features Relation to oxygen Temperature development, G: optimal and maximum is minimal. Optical acidity, units pH Using organic acids as the sole carbon source milky vinegar propiona amber lemon Dioxyacetone formation from glycerol Proteolytic activity against casein, mg% amino nitrogen. (With a control of 8, 0 mg%) Vitamin and synthesizing ability: AT" AT g PP Short sticks arranged in pairs or in chains Obligatory aerobes 28-30 37-45 4-6 3.7-5.0 + + + + + 20-35 20-35 + + + 4- + + Microscopic preplt Grainy thin sticks amount . ZHIZNRSHYUSOS NGH K. ifto current of bifil-a-cythria in I ml of milk The number of cells of acetic acid bacteria in t MJi of milk The increase in acyclic acidity for 26 hours ate. Molokospertyvaya activity, h Prprolitcheska ka LKKRYOST, MrZ (at the control of knockdowns of 7.3 mg.7) The number of pee-pev with preservation of st Runcti composition and properties Granular thin sticks of bnfi-lobakterij and cocci or chopchi lactic acid bacteria Grainy thin sticks, small movable sticks Dead thin p-pads, small mobile rods Serrated Grain thin pa-thin breaking, sticks, small small mobile movable sticks of a stick Grainy; Fine granules - thin sticks, palsukn, small small mobile movable rods (2-4). u (0,2-IJ) -IO 2,, 5 - (, 3-10 1, B-10 Z., 0 1.0-1.5 2.5-J, n3.2 26-7212-16 7.0-7.2 7.0-7.5 2-6 12.5 3.3 8 13.0 11.9 12.8 11.7 Periolic reseeding within two years of IR was attributed to the composition and characteristics of combinations ten , 5 10 3.7-10 4.3-10 5.0., 7.10 2.2 .tO “i 3.13,33,02,9 8911II 12.0 Indicators rrganoleptic assessment The content of cells 6yfidobakterii in 1 g of the product The nitrogen content of free amino groups. 1604847 ten table 2 Serrated Grain thin pa-thin breaking, sticks, small small mobile movable sticks of a stick Grainy; Granular thin - thin sticks, palsukn, small small movable movable sticks ten 11.9 12.8 11.7 3.13,33,02,9 8911II 12.0 Sour milk drink Pure sour-milk taste, homogeneous on the basis of moisture 6 -10 1.7-10 at