RU2449011C2 - Lactobacillus gallinarum vkpm v - 10131 strain used to produce fermented milk products - Google Patents

Abstract

FIELD: chemistry.

SUBSTANCE: Lactobacillus gallinarum strain is extracted from kefir fungus microflora in natural conditions. The strain is deposited in the Russian National Collection of Industrial Microorganisms (VKPM) under registration number VKPM V - 10131. The strain is cultured on the following culture media: agarised dairy cheese whey, skimmed milk, unsalted milk whey.

EFFECT: invention enables to obtain fermented milk products - kefir, acidophilus milk, acidophilus paste, acidophilin and kumis.

6 ex

Description

The invention relates to biotechnology and the dairy industry and is a new strain of mesophilic bacillus, intended for the production of fermented milk products and bacterial preparations.

Known strains of Lactobacterium acidophilum, Lactobacterium lactis, Lactobacterium plantatum, Lactobacterium fermenti, which are highly resistant to growth inhibitors, the ability to stably form a viscous milk clot, have good compatibility with other strains of lactobacilli and pronounced physiological and biochemical properties.

The disadvantage of these strains is lower antibiotic activity.

The objective of the invention is the creation of a fermented milk product using lactobacilli on accessible nutrient media with high antagonistic activity.

The problem is solved by using nutrient media of dense and liquid consistency: sterile skim milk, hydrolyzed milk (pH 6.8-7.0).

The local strain of Lactobacillus gallinarum is isolated in vivo from the microflora of kefir fungus.

The identification of the strain was carried out according to the method described in the book by L.A. Bannikova (1975).

The strain is deposited in the All-Russian collection of industrial microorganisms and has the number VKPM B-10131.

The strain of Lactobacillus gallinarum VKPM B-10131 has the following characteristics:

1. Cultural and morphological:

- optional anaerobic;

- gram-positive sticks;

- length 5 ± 0.03, width 0.7 ± 0.03;

- motionless;

- located singly and in chains;

- do not form endospores;

On the surface of the agar medium (hydrolyzed milk) after 24 hours at a temperature of 37-42 ° C form:

- round colonies,

- size 1-1.5 mm (spot);

- the nature of the contour of the edge is vague;

- the relief is flat;

- surface - smooth, matte;

- White colour;

- structure - granular;

- the texture is soft.

The study of the cultural or morphological properties of the selected strain of lactic acid microorganisms was carried out on a dense nutrient medium - agarized milk.

2. Physiological and biochemical:

- milk fermentation rate - 6 hours.

- the limit of acid formation is 300 ° T.

- the strain ferments glucose, lactose, maltose.

- grows at a temperature of 45 ° C -48 ° C;

- grows in hydrolyzed milk containing 2% and 4% NaCl;

- grows in hydrolyzed milk containing 20% bile;

- grows in meat and peptone broth at pH 8.3;

- grows in milk with 0.4% phenol.

- the strain withstands heating at a temperature of 60 ° C for 90 minutes;

- the strain withstands heating at a temperature of 65 ° C for 30 minutes.

3. Antibiotic activity:

- the diameter of the zone of inhibition of growth in relation to the representative of the pathogenic microflora Staphylococcus aureus was 25 mm;

- the diameter of the zone of inhibition of growth in relation to the representative of opportunistic microflora of Escherichia coli was 25 mm.

The antibiotic activity of the strain of Lactobacillus gallinarum VKPM-B 10131 was determined on a solid nutrient medium (MPA) by agar diffusion.

The method, conditions and composition of the media for long-term storage of the strain - lyophilization; at a temperature of 4 ° C; MPG environments Rogosa e1a1 .; milk with chalk; milk with yeast autolysate, glucose, litmus, liver extract and CaCO ₃ ; agarized milk cheese whey.

The method, conditions and composition of the media for the propagation of the strain at a temperature of 37 ° C; environments: agarized milk cheese whey; skim milk, unsalted cheese whey with a content of at least 4.0% lactose.

Optimum conditions and composition of the medium for fermentation - temperature 37 ° C; environments: cow's milk; unsalted cheese whey with a content of at least 4.0% lactose, skimmed cow's milk.

The proposed new strain isolated from the microflora of kefir fungus is well adapted to local conditions, easy to cultivate and store. In addition, the strain has high productivity and atibiotic activity in relation to opportunistic and pathogenic microflora.

Due to its pronounced biological and antibiotic properties (the ability to form a viscous milk clot, the diameter of the zone of inhibition of growth of Staphylococcus aureus is 25 mm, Escherichia coli is 25 mm), the strain is of industrial value for the dairy industry.

Example 1. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of kefir

Milk is weighed, sorted and filtered, subjected to primary processing. It must be fresh, natural and meet the requirements of the standard.

In the production of kefir, milk is cooled to a fermentation temperature of 20-25 ° C. Already pasteurized milk is poured into the fermenter tank and pre-prepared yeast is added - 5-6% of the milk mass. The starter culture is also enriched with a strain of Lactobacillus gallinarum VKPM-B 10131. After thorough mixing, the contents of the tank are left alone, ensuring a constant temperature with a water jacket. The milk is fermented to obtain a dense clot with an acidity of 90-100 ° T. The duration of ripening is 10-12 hours at a temperature of at least 20 ° C. Upon reaching the necessary acidity and viscosity of the clot, the circulation of water in the jacket is stopped, ice water with a temperature of 1-2 ° C is supplied to the interstitial space and a stirrer is included to stir the clot and more quickly cool it to the ripening temperature. After reaching a clot temperature of 12-16 ° C, the supply of cooling water is stopped and it is left alone for 4-6 hours for the maturation and development of yeast. Then the contents of the tank are further cooled to a temperature of 8-10 ° C to complete ripening.

Example 2. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of acidophilus milk

Pasteurized milk is fermented with a culture of Lactobacillus gallinarum VKPM-B 10131. 5% of the starter culture is introduced into pasteurized milk at a temperature of 42-45 ° C. The sliminess of the product helps to maintain a uniform consistency even after thoroughly mixing it.

Example 3. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of acidophilus-yeast milk

In pasteurized milk, cooled to 35 ° C, make starter culture prepared by co-cultivation of the strain of Lactobacillus gallinarum VKPM-B 10131 and milk yeast. Fermented milk is poured into bottles, corked and placed for fermentation in a thermostat, where it is kept at a temperature of 30 ° C until a clot forms. At the end of ripening, i.e. when a clot forms, the bottles with milk are transferred to a room with a temperature of 18 ° C, where they are kept for 12-18 hours.

In order to avoid too rapid an increase in acidity and souring of the product due to the enhanced development of the strain of Lactobacillus gallinarum VKPM-B 10131, the milk is fermented at a temperature of 30 ° C. Moreover, the development of the strain of Lactobacillus gallinarum VKPM-B 10131 is somewhat delayed, but yeast develops and a clot is penetrated by tiny carbon dioxide bubbles. After the formation of a clot, the product is not subjected to immediate cooling, but is kept in a warm chamber at 18 ° C for better development of yeast. After unloading from a warm chamber, the product is cooled to a temperature below 8 ° C.

Example 4. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of acidophilic paste

In sterilized or pasteurized milk, cooled to 45 ° C, make a starter culture containing pure cultures of mucous and non-mucous races of acidophilus influenzae, including the strain Lactobacillus gallinarum VKPM-B 10131.

The clot is cut into cubes 2 cm in size along the edge. The cut clot is poured into sterilized coarse calico bags, which are placed on the waste tables or in press trolleys for self-pressing and partial dehydration of the clot. The duration of self-pressing is 2-3 hours at 6-8 ° C. Ready acidophilus paste has a consistency of thick sour cream.

Acidophilic pastes are packaged in glass or ceramic jars and quickly cooled to 4-6 ° C.

Example 5. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of jasper

In sterilized or pasteurized milk, cooled to 45 ° C, make a starter culture containing pure cultures of mucous and non-mucous races of acidophilus influenzae, including the strain Lactobacillus gallinarum VKPM-B 10131.

The clot is cut into cubes 2 cm in size along the edge. The cut clot is poured into sterilized coarse calico bags, which are placed on the waste tables or in press trolleys for self-pressing and partial dehydration of the clot. The duration of self-pressing is 2-3 hours at 6-8 ° C. Ready acidophilus paste has a consistency of thick sour cream.

Then add fillers sugar, cocoa, candied fruit, raisins.

Acidophilic pastes are packaged in glass or ceramic jars and quickly cooled to 4-6 ° C.

Example 6. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of acidophilus

Pasteurized milk is fermented with a combination of starter cultures consisting of cultures of the strain Lactobacillus gallinarum VKPM-B 10131, lactic acid streptococcus and kefir starter culture. The total amount of yeast should be at least 5% of the amount of fermented milk.

With the reservoir method of acidophilus production, the mixing of milk with sourdough should be completed 20 minutes after the introduction of the last portions of sourdough. Then the mixer is turned off and the milk is left alone until the end of the fermentation, which is determined by the acidity and density of the clot. The fermentation of milk cannot be stopped until the acidity reaches 85 ° C, since after the spill of acidophilus into the bottles, whey may settle. The finished product can be cooled in a tank or in a stream.

With a thermostatic method of production, milk is mixed with sourdough until the end of filling, lasting no more than 30-40 minutes for each batch of fermented milk. Fermented milk bottled is sent to a thermostat, where at a temperature of 30-35 ° С it is fermented until a dense clot forms. For cooling, the product enters the refrigerator. In the implementation of acidophilus is produced at a temperature not exceeding 8 ° C.

Example 7. The use of strain Lactobacillus gallinarum VKPM-B 10131 in the production of koumiss

Mare's milk koumiss

In the production of koumiss, fresh mare’s milk with an acidity not exceeding 7 ° T, pure without extraneous smacks and smells, was used. Sugar was added to the milk and then fermented with cultures of Lactobacillus gallinarum VKPM-B 10131, as well as Bulgarian bacillus and yeast, fermenting lactose and having antibiotic properties. Ready koumiss is characterized by a sour-milk, spicy, slightly alcoholic taste and a liquid foamy consistency; it is not very different in density from cow's milk.

Koumiss from cow's milk

In the production of koumiss, cow's milk was used with an acidity not exceeding 7 ° T, pure without extraneous smacks and odors. Sugar was added to skim milk and then fermented with cultures of Lactobacillus gallinarum VKPM-B 10131, as well as Bulgarian bacillus and yeast, fermenting lactose and having antibiotic properties. The ripening process lasts 5 hours.

Ready koumiss is characterized by a sour-milk, spicy, slightly alcoholic taste and a liquid foamy consistency; it is not very different in density from cow's milk.

Claims (1)

The strain Lactobacillus gallinarum VKPM B-10131 used in the production of dairy products.