SU1004471A1 - Method for preparing leaven of acidophilic cultures - Google Patents

Description

This invention relates to technical microbiology, in particular to the preparation of bacterial starters for the preparation of a preparation with high antagonistic properties.

In recent years, the attention of researchers to the study of normal intestinal microflora has increased, since long-term treatment with antibiotics disrupts its normal composition, and this often leads to severe intestinal disorders. In connection with this, it is of great interest to use the starter with Lactobacte iumac idoph i1 urn for the manufacture of dietary and therapeutic products.

Work was carried out on the selection of acidophilic lactic acid bacteria strains of the most valuable in activity, organoleptic characteristics, antibiotic activity and phenol resistance, which have the highest yields at an optimum cultivation temperature of 37-4 ° C and a high reducing ability 1.

The closest to the invention to the technical essence and the achieved positive effect is the method of preparation of the acidophilus starter culture, used for the production of fermented milk products such as acidophilus milk, acidophilus, acidophilic paste. According to a known method, Lbra acidophilus bacillus strains are used. acidophilum or their combination with the average activity of acid formation, coagulating pasteurized milk for 3.5-4.0 hours and possessing antibiotic activity. For the preparation of the starter culture by a known method, 0.5 sterilized milk of the Lbm culture is introduced into 15 sterilized milk. asidophi 1um or a combination of both and cultivate for 4.5-5.5 hours at 37-40 ° C; The ferment obtained at this has

20 pure sour milk taste with a specific flavor. Its acidity is 100-130 ° T 2.

However, leaven; obtained by a known method, not suitable for

25 production of concentrated preparations with high antagonistic properties ..

The purpose of the invention is to improve the biochemical properties of the starter by

30 increase the resistance of its microflora to antibiotics, drying, and storage.

The goal is achieved in that in a method involving the heat treatment of milk, cooling to a temperature of fermentation, making strains of lactic acid sticks and subsequent cultivation, according to the invention, for preparing a yeast, strains of lactic acid sticks are used that are selected according to the sum of indicators of such properties as high antagonistic activity. pathogenic microflora in the range of 98.1-100%, proteolytic activity in the range of 56.0-58.8 mg% based on the sum of three free amino acids in peres on tyrosine, resistance to 5-L types of antibiotics, resistance to drying within 45-58% of living cells during freeze-drying, storage stability within 80-90% of living cells every month when stored dry at room temperature, combination strains are selected with the condition that two of them possess high antagonistic, and one high proteolytic activity, with this strains contributing in an amount of 0.2-0.4% by weight of milk, and cultivation is carried out 18-24 hours at which check the antagonistic properties of zac Vaska.

In this case, it is advisable to take two strains with high antagonistic properties of Lbm. asidophilum NK-i and Ka, I2.4, and one with a high proteotic activity (100 A), at a ratio of 2: 2: 1.

Strains are selected according to the following signs.

Antagonistic activity in relation to pathogenic microflora in the range of 98-100% in relation to Shigella Sonne, Flexner, Salmonella, Protea High proteolytic activity. For this, the quantitative content of some amino acids in terms of tyrosine is determined, in this case, it is 56-58.8 mg% based on the sum of three free acids in terms of tyrosine. Antibiotic resistance (each culture is resistant to 5-7 antibiotics). Resistant to drying. In sublimation-dried cultures, 58-45% of living cells are conserved. Durability. When stored dry. During storage at room temperature, 80-90% of living cells are stored monthly. Compatibility With the joint cultivation of the properties of the strains of poyiazots.

The proposed method is as follows.

From the strains selected for the indicated indicators, the starter is prepared. Two

from the selected strains, they have high antagonistic, and one has high proteolytic properties (about 100% of the pathogenic microflura die, the proteolytic activity is 58.8 mg% in terms of tyrosine),

They are cultivated in skimmed milk at 35-37 C (each separately) for 16 hours.

Then, 0.2-0.4% of the combination of these three strains is added to the milk by weight of the milk — out of the calculation: ta-0.4% with high antagonistic and 0.2% each with high proteolytic activity. Cultivation lead 1824 h at 30-34 ° C.

The leaven prepared in this way has a pure sour-milk cocoa with a specific flavor of acidophilic cultures. Its acidity is 140-210 ° T. It is a homogeneous mass with a slightly mucous consistency.

Leaven can be used both in liquid form and dry when preparing bacterial preparations with high antagonistic properties.

The acidobacterial strains used in the preparation of the starter are characterized by the following signs.

Strain; 1K.

Morphological and cultural characteristics. The size of the cells is 5-10 μm, the cells are uniformly red, single or arranged in a chain of 34 segments. On agar with hydrolyzed milk forms colonies - surface forms, dark, curved, deep in the form of spiders. The diameter of the colonies is 1,0-.1,5 tiM.

Physiological signs. Optimum temperature of development 34-40 0 Sprays lactose, glucose, maltose, does not ferment sucrose.

Milk coagulates with the addition of 1% ferment and the optimum temperature develops 3ia 6-7 hours. Reduces the pH of milk to 3.8-3.9 over 10-11 hours. It develops in milk with a phenol content of 0.4-0.6. The proteolytic activity is 35.4 mg%, the resistance to drying is 48-51%, the storage stability is 88-90% of living cells per month. It has a pronounced antagonistic activity against pathogenic bacteria. The percentage of dead cells after 48 hours of co-culture was not, respectively, at Shigella Sonne 99.8} Flexner 100, Salmonella 98.1 and Proteae 99.4.

Strain .

Morphological and cultural characteristics. Rod-shaped cells of various lengths, 3-14x1,01, 5 microns in size. They are located as single cells or in chains of 2-3 cells each, gram-positive. Proteolytic activity is 36.5 mg%, resistance to drying is 45-52%, storage stability is 88-85% of living cells.

Physiological signs. The optimum growth temperature is .37-4Z C. Acidity after 16. h of cultivation on sterile milk is 175-180 T. Extreme acidity is 400T. The rate of acidity is high. After 6 hours of cultivation in milk, with the addition of a 3% culture, the acidity changes to 125-140 ° T. Assimilates g to lucosis, arabinose, lactose. Does not assimilate raffinose, maltose, attracts. Antibiotic properties. It has a suppressive effect on the bacteria of the Escherichia coli group. The lack of growth of the test culture on a liquid nutrient medium in breeding 1/32, delay, growth in breeding 1/128. The zone of no-20 growth on a solid nutrient medium is up to 5 mm. It has antagonistic activity against pathogenic, microflorn Shigella Sonne, Flexner, Salmonella, Proteev (98-25

99.9%).

I.

It has a permanent resistance to spray-dried conditions. The yrotheolytic activity is 31.5 mg% in terms of tyrosine.

Strain 100 ASH.

Optional anaerobic. The minimum temperature of development, optimal 38-4O s, maximum 60b2 s. ; . 35

The milk coagulates to form a tight clot. Milk does not peptanize. Gelatin does not excite. Starch does not hydrolyze. Citrates. Does not restore. Ferments glucose, 40 galactose, lactose, sucrose and maltose. Forms carbonyl compounds in the amount of 8.5 mg% in terms of acetaldehyde, four-carbon compounds of diacetyl and acetone in 45 amount of 0.31 mg% .-,

It has antagonistic activity in relation to Shigella’s pathogenic microflora. Sonne, Flexner, Sal-- monell Proteus in the range of 90-92%. Proteolytic activity in the range of 58.8 mg% by the sum of three amino; acids in terms of tyrosine. Resistant to penicillin and other antibiotics, resistant to drying - 55 survival rate by drying by sublimation of 50-58% of cells, resistant to - 80-85% of living cells when stored at room temperature.

Example. Take 1 liter of sterile milk at 60% and put in 0.16% of NK culture; 0.16% culture. A and 0.080 culture 100 ASH. All ttstalno mixed and maintained at for 24 h.45

The resulting starter is characterized by antagonistic activity against Shigella Sonne. 99.9% 7 Flasner 100% ;. Salmonella 99.3% i Proteev 99., 9%. Proteolytic activity is 62.5 mg%. Sourdough resistant to monocycline, neomycin, kanamycin, polymyxin, penicillin, erythromycin, streptomycin and tetracycline ..

Thus, all the properties of the leaven are enhanced, the strains combine well with each other.

Example2. Take 1 liter of sterile milk at and bring 0.08% of the NK culture.; F, 0.08% of the K culture, Shi 0.04% of the culture 100 AU. .

All mixed well and kept for 18 hours. Antagonistic activity of the starter with respect to Shigella Sonne, 99.8% Flexner, 100%, Grinmonella, 99.9%. Proteolytic activity is 64 mg%. Sourdough resistant to monocycline, neomycin, penicillin, erythromycin, novrbytsill 1nu, streptomycin, tetracycline. 1 In 1 ml of the prepared ferment according to examples 1 2 (250-700 L x X V of viable cells. The acidity of the starters is 180 and 210, respectively. The cell survival during drying is 45 and 50%, the death: of cells during storage is 10-20%, respectively.

The obtained ferment of this type makes it possible to organize an industrial production of a dairy-based preparation with various fillers, which has dietary and preventive and medicinal properties.

The table shows the quality of the dry bacterial preparation of acidophilic rods, experimentally obtained from a leaven, prepared. prepared according to example 1 or 2.

The feasibility of organizing the production of a bacterial preparation based on the starter cultures of acidophilic cultures is necessary in connection with the fact that acidophilic lactic acid bacilli with respect to pathogenic and conditionally pathogenic microflora have high antagonistic activity. Such conditionally pathogenic microflora is the determining etiological structure of various forms of intestinal diseases, and the antagonistic activity of acidophilic lactic acid bacilli correlates with their pronounced sanitizing efficiency. In addition, the ability of acidophilic lactic acid bacilli to actively colonize the mucous membrane of the digestive tract and their positive effect on reparative processes in the intestine during dysentery has been established. This determines the feasibility of using bacterial preparations containing highly active, selective strains of acidophilic lactic acid bacillus as a dietary supplement.

and therapeutic foods that promote the release of pathogens and the normalization of intestinal microflora.

Claims (2)

Claim 1. A method of preparing acidophilic culture starter culture intended for the preparation of lactic acid products, comprising heat treatment of milk, cooling to a fermentation temperature, introducing strains of lactic acid bacilli and their subsequent cultivation, characterized in that, in order to improve the biochemical properties of the starter culture by increasing its microflora resistance to antibiotics, drying and storage, for the preparation of yeast using strains of lactic acid bacillus, selected by the sum of indicators properties, As high antagonistic activity with respect to pathogenic microflora. within 98.1-100%, proteolytic activity within 56.0 r8.8 mg% by the sum of three free amino acids in terms of tyrosine, resistance to 5-7 types of antibiotics, resistance to drying within 45-58% of living cells during freeze-drying, storage stability within 80-90% of living cells monthly when stored dry at room temperature, the combination of strains is selected with the condition that two of them have a high antagonistic , a high proteolytic activities, with strains introduced in an amount of 0.2-0.4% by weight of milk, culturing are 18-24 hours at 30-34 ° C, during which is checked 3Q antagonistic properties leaven. 2. The method according to π. 1, characterized in that the ratio between the strains having high antagonistic properties and odes ^E them having high proteolytic activity, is 2: 2: 1, wherein the strains used Lactobacterium acidophilum NK ^, KjUI ^ and 100ASH respectively.