RU2567149C1 - Lactobacillus plantarum STRAIN USED FOR OBTAINING OF FERMENTED MILK PRODUCTS AND PROBIOTIC PREPARATIONS - Google Patents

Abstract

FIELD: biotechnologies.

SUBSTANCE: strain Lactobacillus plantarum B2-II with high antagonistic activity is deposited in the All-Russian Collection of Industrial Microorganisms under the registration number VKPM V-10816 and can be used for production of fermented milk products and pro-biotic preparations.

EFFECT: invention allows to obtain more viscous milk coagulum.

1 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: Culture LT-7 is assigned to Lactobacillus plantarum on the basis of phylogenetic data, as well as a number of physiological and biochemical characteristics that distinguish it from other related species of lactobacilli: the ability to grow at + 15 ° C, ferments ribose, but does not ferment glycerol and xylose ; 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 dairy product using lactobacilli on accessible nutrient media with high biological and 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), unsalted cheese whey with a content of at least 4.0% sugar.

The local strain of Lactobacillus plantarum was isolated from the surface of the plant, the large-flowered initial.

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

The strain is deposited in the All-Russian collection of industrial microorganisms and has a number VKPM B-10816.

The strain of Lactobacillus plantarum VKPM B-10816 has the following characteristics:

1. Cultural and morphological:

- optional anaerobic;

- gram-positive sticks;

- 5-6 microns long, 0.4-0.5 microns wide;

- motionless;

- located in the form of long interwoven chains of large sticks;

- do not form endospores.

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

- rounded colonies;

- the size is small;

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

- profile - convex;

- White colour;

- structure - fine-grained;

- the consistency 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 and MRS agar.

2. Physiological and biochemical:

- milk fermentation rate - 6 hours;

- the limit of acid formation is 259 ° T;

- the strain ferments glucose, lactose, beckons, sucrose, fructose, galactose;

- grows at a temperature of 37 ° C - 45 ° C;

- grows in hydrolyzed milk containing 2%, 4% and 6.5% NaCl;

- grows in hydrolyzed milk containing 20%, 30% bile;

- grows in milk containing methylene blue (0.01% and 0.1%);

- grows in meat and peptone broth at pH 9.2;

- 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.

The product synthesized by the strain is lactic acid, antibiotic substances.

3. Antibiotic activity:

- the diameter of the zone of inhibition of growth in relation to the representative of the pathogenic microflora Staphylococcus aureus was 20 mm;

- the diameter of the zone of growth inhibition in relation to the representative of opportunistic microflora of Escherichia coli was 20 mm.

The antibiotic activity of the strain of Lactobacillus plantarum VKPM B-10816 was determined on solid nutrient medium (MPA) by agar diffusion.

The method, conditions and composition of the media for long-term storage of the strain - lyophilization; storage at 4 ° C; MPS environments: Rogosa et. al .; 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: MPS; skimmed milk.

Optimum conditions and composition of the medium for fermentation - temperature 37 ° C; environment: unsalted cheese whey with a content of at least 4.0% sugar; milk of farm animals.

The proposed new strain of Lactobacillus plantarum VKPM B-10816. isolated from the surface of the plant - large-flowered initial letters, well adapted to local conditions. Easy to cultivate and store, has a high marginal acidity and clotting speed of milk, which allows to increase economic efficiency in production.

The strain exhibits a high antagonistic ability in relation to opportunistic and pathogenic microflora.

The strain Lactobacillus plantarum VKPM B-10816 has a high biological activity.

Example 1. The use of a strain of Lactobacillus gallinarum VKPM - B 10132 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 10132. 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.

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 20 mm, Escherichia coli is 20 mm), the strain is of industrial value for the dairy industry.

Claims (1)

The strain Lactobacillus plantarum VKPM B-10816 used in the production of dairy products and probiotic preparations.