RU2425576C1 - Dry probiotically active biopreparation and method for production of fermented milk drink based on it - Google Patents

Abstract

FIELD: food industry. ^ SUBSTANCE: invention relates to biotechnology and may be applied in food industry, medicine, veterinary and allied industries. The probiotically active biopreparation contains wt %: lyophilised milk powder - 98-99.5; Tween 80 - 0.1-1.0; the rest is "Vitaflor" starter. The method for production of a fermented milk product based on the biopreparation consists in preliminary introduction of Tween 80 into fresh milk; the milk undergoes sublimation drying, milling into particles sized 20-150 mcm and mixing with dry "Vitaflor" starter at a preset ratio. The produced mixture is rehydrated with water addition at a preset concentration and placed into a thermostat at a temperature of 372C for 18-20 hours. ^ EFFECT: invention allows to produce a high-quality fermented milk product which has adaptogenic and immunostimulating effect under the conditions of milk deficit or agalaxy. ^ 3 cl, 1 dwg, 2 tbl, 4 ex

Description

The invention relates to the field of biotechnology, in particular to dry microbiological preparations and a technology for producing a fermented milk product with high probiotic potential in extreme conditions, in particular on board a spaceship, in conditions of prolonged scuba diving, etc. The invention can also be used in the food industry, medicine, veterinary medicine and related industries.

Dry biologics based on lactic acid microorganisms are known (EP 0097484, 1984, CL A23C 1/05; EP 0043962, 1980, CL A23C 19/086; US 4289888, 1979, CL A23C 9/123). Such preparations contain bacterial cells, residues of culture fluid (QOL), fats, proteins, lactose, gum and other additives. The technology for their preparation includes, as a rule, the preparation of a solution or suspension of the biomass of microorganisms, the introduction of special additives into it, freeze-drying, and the preparation of a finished dry product (sourdough), which is subsequently used as a seed for the production of a fermented milk product.

Known methods for producing dry preparations based on acidophilic microorganisms (RU 2064269, 1994, class A23C 9/12; SU 1227145, 1983, class A23C 9/123), including the cultivation of microorganisms on a nutrient medium, the introduction into the culture fluid (QOL ) or its concentrate protective environment and drying. The drug, as a rule, contains bacterial cells and residues of QOL with some additives that improve its operational characteristics.

The disadvantage of the above methods is the need for additional preparation of a special nutrient medium to obtain the final fermented milk product containing the above cells of microorganisms, as well as the relatively low biological activity of the seed.

The closest in technical essence to the claimed group of the invention is the preparation "VITAFLOR" ® and the technology for its preparation described in patent RU 2160992, 2000. Lyophilized dry sourdough of this preparation contains 2-100 billion viable cells of 2 strains of Lactobacillus acidophilus H-1 and N-2 (when depositing strains A-91 and H-91 or DN 75 and DN 76, respectively, see certificates of deposit), 0.5-2.5 wt.% Gelatin, 5-20 wt.% Sucrose, 3-5 wt. % skim milk, 2-8 wt.% water and the remains of the culture fluid. To obtain the final fermented milk product, the leaven is placed in milk or milk mixture and kept at elevated temperature for 16-18 hours. The resulting preparation has established itself as an effective antibacterial agent for the prevention and treatment of intestinal infections, as well as a promising immunostimulant and adaptogen (RU2371190, 2009 ; RU 342174, 2008; RU 2339389, 2008; RU 2278682, 2006), which, along with complete safety - the drug is approved for feeding children older than 3 months - makes it promising for use in extreme conditions, where a person is deprived of the opportunity to use inpatient treatment (space flight conditions or staying in a long voyage). The method of obtaining it involves obtaining a starter culture of bacteria Lactobacillus acidophilus in a hydrolyzate-milk nutrient medium containing 0.3-0.5 wt.% Calcium carbonate, the introduction of a protective medium containing gelatin, sucrose, skim milk, freezing at a temperature of -25-30 ° C, sublimation drying at a temperature of no higher than minus 30 ° C and a pressure of 13-15 Pa for 45-48 hours. To obtain a fermented milk drink, the yeast is dissolved in water, milk is poured at a temperature of 35-45 ° C and incubated for 12-18 hours.

The disadvantages of this drug and the technology for its preparation include the fact that in order to obtain a final fermented milk product with the required probiotic properties, significant quantities of fresh milk are necessary, which is difficult to do in conditions of prolonged separation from sources of supply. However, the use of standard powdered milk obtained by spray drying does not allow to obtain a high-quality drug with a high titer of living microorganisms.

The problem solved by the authors was the creation of a technology for producing a dry preparation based on “Vitaflor” sourdough, which, after long-term storage, makes it possible to obtain a high-quality fermented milk drink that has adaptogenic and immunomodulating effects in the absence or absence of ingredients, in particular milk.

The technical result was achieved by the creation of “Vitaflor-K”, which, along with “Vitaflor” sourdough, contained lyophilized milk powder (LMP) with the addition of Tween-80 surface-active substance (surfactant) in the following ratio of ingredients (wt.%):

lyophilized milk powder 98-99.5;

Twin-80 - 1.0-0.1;

“Vitaflor” sourdough - the rest.

Along with Tween 80, other non-ionic surfactants approved for pharmaceutical use can also be used.

A method of obtaining dry seed (sourdough) and a fermented milk product based on it is that Tween-80 is added to fresh milk in a ratio of 1.0 g / l and subjected to freezing on a shelf of a sublimation unit at a temperature of minus 25-30 ° С for 1 , 5-2.0 hours, and then freeze-drying at a temperature not exceeding minus 20 ° C and a pressure of 13-15 Pa for at least 48 hours. The drying thermogram is shown in the drawing. Then, the dried milk is subjected to grinding to obtain a polydispersed powder with a particle size of from 20 to 150 microns, mixed with freeze-dried and chopped Vitaflor sourdough under similar conditions and, if necessary, packaged in hermetically sealed containers. To obtain the final fermented milk product, the drug is diluted with water in a predetermined concentration (usually in a ratio of about 1:10) and placed for 18-20 hours in a thermostat at 37 ± 2 ° C.

A feature of the obtained fermented milk product is its increased stability during subsequent storage. This is a consequence of the fact that the cells of microorganisms in this case are “packed in capsules” of milk casein, which, when dissolved in water, provide the bacteria with an optimal accessible environment for further development. The invention is illustrated by the following examples.

Example 1. Obtaining starter culture. In ampoules with lyophilized standards of production strains of Lactobacillus acidophilus A-91 and H-91, 1 ml of distilled water was added, after which they were thoroughly mixed, placed separately in test tubes with skim milk and kept at 37 ° C for 16-20 hours. The seed was introduced into flasks containing 100 ml of milk hydrolyzate medium containing 0.2 g peptone, 0.06 g sodium chloride, 0.002 g cysteine and 0.1 g lactose in a pancreatic milk hydrolyzate, and kept at 37 ° C for 16-20 hours. Ready seed was obtained by mixing separately grown cultures so that the cell ratio of individual strains was close to 1: 1 according to optical density at a wavelength of 600 nm.

The native culture was grown on a medium containing 1 g of pancreatic hydrolyzate of milk 2 g of peptone, 0.6 g of sodium chloride, 0.02 g of cysteine, 4 g of calcium carbonate and 1 g of lactose at 37 ° C for 16-20 hours. A 3-component medium consisting of sucrose, gelatin and skimmed milk powder was added to the culture liquid in a ratio of 10%: 2%: 4% based on the dry matter of the final product. The resulting product was poured onto aluminum trays of 250 ml each, which is 3-4 mm thick of the dried layer, and frozen in a chamber of a TG-5 sublimation unit (Germany) at minus 25-30 ° C, after which it was subjected to freeze-drying at a temperature condenser-freezer not higher than minus 50 ° C and a pressure in the sublimation chamber 13-15 Pa. The time to reach the working vacuum was 25 min; the total duration of the drying process was 48 hours.

The preparation obtained after drying had the appearance of a uniform color and height mass. After drying, the preparation was subjected to grinding on a KEM-36 brand screw shredder manufactured by Belwar company. The resulting polydisperse powder consisted of particles ranging in size from 20 to 150 microns. The color of the powder is from light to dark beige, humidity is usually about 4% (humidity up to 7% is allowed). The biological activity of the powder was 2 × 10 ⁹ CFU / g of product.

The resulting powder can be stored in a hermetically sealed container at a temperature of 5-6 ° C, the warranty shelf life of the powder is 2 years.

Example 2. Obtaining funds "Vitaflor-K". Tween-80 (0.5 g / l) was added to drinking milk, poured into aluminum trays of 300 ml each and subjected to freezing on a shelf of a sublimation unit at a temperature of minus 25-30 ° C for 1.5-2.0 hours and then freeze-drying at a temperature not exceeding minus 20 ° C and a pressure of 13-15 Pa for at least 48 hours. A drying thermogram is also shown in the drawing.

In the preparation of the Vitaflor-K product, the following ratio of components was used: sublimated milk powder dried with Tween-80 - 50 g; freeze-dried crushed sourdough containing 2 strains of lactobacilli, obtained under the conditions of example 1, 0.260 g. The obtained samples were mixed and packaged in airtight containers.

To obtain a fermented milk product, 50 g of Vitaflor-K was placed in a sterile container into which 500 ml of distilled water was added. The container with rehydrated material was closed and placed in a thermostat heated to 37 ± 2 ° C for 18 hours. Upon reaching the above exposure, the obtained fermented milk product was removed from the thermostat and checked for probiotic properties: biological activity, antibiotic resistance, antagonism with respect to pathogenic and conditionally pathogenic microorganisms and acid formation. The biological activity of the obtained fermented milk product was 2 × 10 ⁸ CFU / ml, the pH of the obtained product was 3.6. Antibiotic resistance and antagonism were in line with the norm declared for the Vitaflor probiotic.

Example 3. The effect of the composition of the drug on its properties after rehydration. In the conditions of example 2 received the tool "Vitaflor-K" with a different ratio of ingredients. Using a similar technology, a mixture was prepared in which, for comparison, in the composition of the preparation, LMP was replaced with milk powder, which was obtained industrially by spray drying in a stream of hot air. The results are shown in table 1.

Table 1 The effect of the composition on the biological activity of the drug "Vitaflor-K" The composition, wt.% Biological activity, CFU / ml Sourdough Vitaflor Dairy Product (MK) MK concentration Twin 80 0.5 LMP 99.0 0.5 2 × 10 ⁸ 0.5 Powdered milk 99.0 0.5 1 × 10 ⁵ 1.0 LMP 98.0 1.0 6 × 10 ⁷ 0.5 LMP 99.0 0.5 2.5 × 10 ⁸ 0.4 LMP 99.5 0.1 2 × 10 ⁸

Example 4. The composition "Vitaflor-K" obtained in the conditions of example 3 was subjected to storage at room temperature for 1 month, after which they were rehydrated, cultured in an incubator at 37 ° C and determined the activity of the obtained fermented milk product. The test results are shown in table 2.

table 2 The effect of the composition on the properties of the rehydrated fermented milk product. The composition, wt.% Biological activity, CFU / ml, Sourdough Vitaflor Dairy Product (MK) MK concentration Twin 80 0.5 LMP 99.0 0.5 2 × 10 ⁸ 0.5 Powdered milk 99.0 0.5 1 × 10 ⁴ 1.0 LMP 98.0 1.0 6 × 10 ⁷ 0.5 LMP 99.0 0.5 2.5 × 10 ⁸ 0.4 LMP 99.5 0.1 2 × 10 ⁸

The results obtained showed that the claimed technology allows to obtain a high-quality sour-milk product with desired properties using a simpler production technology suitable for use in space flight and long voyages.

Claims (3)

1. A probiotic active biological product based on “Vitaflor” sourdough and a milk-containing component, characterized in that it additionally contains Tween 80, and as a milk-containing component, lyophilized milk powder in the following ratio of ingredients, wt.%:
Lyophilized milk powder 98-99.5 Twin 80 0.1-1.0 Sourdough Vitaflor rest 2. The method of producing a fermented milk product based on the dry probiotic active biological product according to claim 1, which consists in the fact that fresh milk after adding Tween 80 is lyophilized, crushed to a particle size of 20-150 microns, and then mixed with dry crushed to similar the size of the vitaflor starter, and then rehydrated before use by adding water to a predetermined concentration and placed for 18-20 hours in a thermostat at 37 ± 2 ° C. 3. The method according to claim 2, characterized in that during the freeze-drying, the milk is subjected to freezing on a shelf of a freeze-drying unit at a temperature of minus 25-30 ° C for 1.5-2.0 hours, and then freeze-drying at a temperature not higher than minus 20 ° C and a pressure of 13-15 Pa for at least 48 hours

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