Strain enterococcus hirae, used to make cultured milk foods

FIELD: biotechnologies.

SUBSTANCE: strain Enterococcus hirae BC - 37 having high antagonistic activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under the registration number VKPMV-10090 and may be used, for instance, in production of such cultured milk foods as kefir, cottage cheese or ryazhenka.

EFFECT: invention makes it possible to produce a strain having high antagonistic activity and high speed of milk turning sour.

3 ex

 

The invention relates to biotechnology and the dairy industry represents a new mesophilic strain of Streptococcus, intended for the production of fermented milk products and bacterial agents.

Known strains of Streptococcus citrovorus, Streptococcus paracitrovorus, Streptococcus diacetilactis, highly resistant to growth inhibition, the ability to stably form a viscous milk clot, good compatibility with other strains of lactobacilli and pronounced physiological and biochemical properties.

The disadvantage of these strains are lower ultimate pH and the rate of fermentation of milk.

Known strain of Enterococcus durans RSH (VKPM B-8717) patent No. 2273662. E. durans since 1982 is applied in the dairy industry as a component of starter cultures, as well as a probiotic in medicine (Rocha, J.M., Malcata F.X., On the microbiological profile of traditional Portuguese sourdough. J. Food Prot. 62(12): 1416-29, 1999).

Also it is shown that E. durans on a number of characteristics (resistance to bile and acid shock in vitro assimilation of cholesterol) than the known probiotic microorganisms (Dora I.A. Pereira and Glenn R., Gibson Cholesterol Assimilation by Lactic Acid Bacteria and Bifidobacteria Isolated from the Humman Gut // Applied and Environmental Microbiology, vol.68, No. 9, p.4689-4693, 2002).

The disadvantage of this strain is the lower limit of acidity and speed of ripening milk.

Objective of the claimed invention, t is aetsa the creation of a fermented milk product with the use of lactobacilli on available nutrient media, with high antagonistic activity.

The task is solved by the use of nutrient dense environments and liquid consistency: sterile skimmed milk, gidrolizovannogo milk (pH of 6.8 to 7.0).

Local strain of Enterococcus hirae VKPM B-10090 allocated in the natural conditions of the microflora of kefir fungus.

Identification of the strain was performed according to the methodology described in the book Bannikova L.A. "Selection of lactic acid bacteria and their application in dairy industry. - M, "Food industry", 1975.

The strain is deposited in Russian national Collection of Industrial Microorganisms and has a number VKPM B-10090.

Local strain of Enterococcus hirae selected in vivo from the surface of the alfalfa plants (flower).

Identification of the strain was performed according to the methodology described in the book of Bannikova L.A. (1975). Strain Enterococcus hirae deposited in PMBC number In-10090.

Strain Enterococcus hirae VKPM B-10090 has the following characteristics:

1. Cultural-morphological:

- optional gone anaerobic;

- gram-positive cocci;

- diameter of 1.0±0.02 mm;

- fixed;

- located pair or short chains.

- endospore do not form.

On the surface of agar medium (gidrolizovannogo milk) after 24 hours at 37°C form:

- the colonies are round in shape,

- size 2.2 mm (average);

- har is cter contour edge smooth;

- relief - convex;

- the surface is smooth, shiny;

- color - white;

the structure is homogeneous;

- consistency is pasty.

The study of cultural or morphological properties of selected strains of lactic acid microorganisms were performed on dense nutrient - agar milk.

2. Physiological-biochemical:

- the speed of ripening milk - 9 hours;

- limit accumulation of acid - 201°T;

a strain sprayway glucose, galactose, lactose, maltose, dextrin;

- grow in milk at a temperature of 40-45°C;

- grows in gidrolizovannogo milk containing NaCl(2%, 3%, 4%, 6,5%);

- grows in meat-peptone broth (pH 9,2; 9,6);

- grows in gidrolizovannogo milk content of bile(20%, 30%, 40%);

grows in milk containing methylene blue(0,0005%, 0,005%, 0,01%, 0,1%);

- withstands heating for 30 minutes at a temperature of 55°C., 60°C.

The product synthesized by the strain - lactic acid.

3. Antibiotic activity:

the diameter of the zone of inhibition of growth relative to the representative of pathogenic Staphylococcus aureus was 23 mm;

the diameter of the zone of inhibition of growth relative to the representative of conditionally pathogenic Escherichia coli was 20 mm.

Determination of antibiotic activity of the strain Enterococcus hirae VKPM B-10090 conducted on solid nutrient medium is e (MPA) by the method of diffusion in agar.

The method, conditions and medium composition for long-term storage of strains of lyophilization; at 4°C; environment MPG; Rogosa; milk with chalk; the milk with yeast autolysate, glucose, litmus paper, liver extract and CaCO3; agar milk cheese whey.

The method, conditions and medium composition for growth of strain at a temperature of 30-37°C, medium: agar milk cheese whey; milk, unsalted cheese whey containing not less than 4.0% lactose.

Optimal conditions and the composition of the medium for fermentation is temperature - 37°C; medium: cow's milk, unsalted cheese whey containing not less than 4.0% lactose, skimmed cow's milk.

The proposed new strain of Enterococcus hirae VKPM B-10090, isolated from the surface of the alfalfa plants (flower), well adapted to local conditions, easy to grow and store, can be used in the production of dairy products, is not a zoo - and phytopathogenic not constitute a hazard for other reasons.

Due to their pronounced biological and antibiotic properties (ability to form viscous milk clot, the diameter of the zone of inhibition of growth of Staphylococcus aureus - 23 mm, Escherichia coli - 20 mm) strain represents the production value for the dairy industry.

Example 1. Ispolzovaniem Enterococcus hirae VKPM B-10090 in the production of yogurt

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

In the production of kefir milk is cooled to the fermentation temperature of 20-25°C. Already pasteurized milk is poured into the reservoir-zakuani and make a pre-prepared yeast - 5-6% of the mass of milk. Leaven also enrich strain of Enterococcus hirae VKPM B-10090. After thorough mixing of the contents of the tank left alone, providing you with the water jacket a constant temperature. Milk sours to produce a dense clot acidity 90-100°So the duration of the fermentation is 10-12 hours at a temperature below 20°C. Upon reaching the desired acidity and viscosity of the bunch, the water circulation in the jacket to cease, in the annular space serves ice water at a temperature of 1-2°C and include a mixer for mixing the bunch, and more rapid cooling to a temperature of ripening. After reaching the temperature of a bunch of 12-16°C the cooling water stop and leave it alone for 4-6 h for maturation and development of yeast. Then the contents of the tank Doklady to a temperature of 8-10°C. to complete maturation.

Example 2. The use of a strain of Enterococcus hirae VKPM B-10090 in the production of cheese

Production Toro is rather traditional way. The technological process consists of the following operations: receiving and preparation, separation of milk, standardization, pasteurization, cooling, fermentation and ripening of normalized milk, cutting the bunch, the separation of serum and bottling clot, samopostovanje and pressing clot, cooling, filling, packaging, storage and transportation of cheese.

Raw milk intended for the production of cheese, clean the cages-molotovcoketail or filtered through three layers of gauze or other filtration fabric. Purified milk is heated to 37±2°C and separated by separators-livaudais. In the manufacture of cottage cheese fat, bold and peasant milk normalize for fat considering the mass fraction of protein in whole milk to get the finished product with the specified fat content and moisture. Skim or normalized milk pasteurized at a temperature of 78±2°C with a holding time of 15-20 C in plate or tubular pasteurization and cooling installations or capacitive devices. After pasteurization, the milk is cooled to the fermentation temperature.

Leaven is prepared with pure cultures of mesophilic lactic streptococci, including making the culture of Enterococcus hirae VKPM B-10090. Before you make the milk surface layer ferment gently remove clean proteinthis is included in the bucket and remove. Then the starter mix to a uniform consistency with clean whisk (when cooking in the starter tubs) or with a mixer and pour into the prepared milk in an amount of 5% of the total mass. The duration of ripening milk 6 o'clock

An aqueous solution of calcium chloride (mass fraction of calcium chloride 30-40%) add in the milk after fermentation: 400 g per 1000 kg of the fermented milk. Training and preparation of a solution of calcium chloride produced in accordance with the regulations on technical and chemical control at the enterprises of the dairy industry. After making the solution of salt in fermented milk impose a 1%solution of enzyme per 1 g of the drug activity ME 100000 1000 kg of milk.

Rennet powder or pepsin make milk in a 1%aqueous solution, prepared by boiling and cooled to 36±3°C water. A solution of the enzyme contribute to the milk with constant stirring. After 10-15 minutes after making the solution of the enzyme finish mixing and leave the milk alone to form a tight bunch acidity 61±5°T for cottage cheese 9%and 18%fat, 65±5°T for peasant and 71±5°T for low-fat cottage cheese.

For processing clot using hand lyre, in which the knife is stretched thin stainless steel wire. Such wire knives curd is cut into cubes RA which measures 2×2×2 cm The clot is first cut along the length of the bath in horizontal layers, and then the length and width on the vertical. After this treatment, the clot is left for 40-60 min to separate the serum and increasing acidity. The separated whey is drained from the bath. Clot after draining the whey is poured into basebya or Mylar bags with dimensions of 40×80 cm Bags fill about 70%, which is 7-9 kg of cheese. Then the bags are tied and stacked one upon the other in the tub for the pressing itself, press the cart or the installation of UPT for pressing and cooling of the curd.

To speed up the separation of serum, or in poor allocation of serum clot is heated by filing in the space between the walls of cottage cheese tubs, steam or hot water. For uniform heating of the upper layers of the bunch move a wooden or metal plate (shovel) from one wall of the bath to the other. Clot heated to 40±2°C for 30-40 min for cottage cheese 9%and 18%fat, 35±2°C for 20-40 min for peasant and 36±2°C for 15-20 min for low-fat cheese.

Samopostovanje of cheese is not less than 1 hour continue Pressing until you get a cheese with a mass fraction of moisture, provided for legal documentation. For cottage cheese 18%fat she is 65%; 9%fat - 73; peasant - 74,5; table - 76; not the IRNA - 80; for a diet of fruit 11%fat) - 64, 9%fat - 66, 4%fat - 77 and lean - 79% moisture. When formulating low-fat cottage cheese dehydration clot can be curd separator. After separation and pressing the curd is cooled. The curd is cooled in bags placed on the shelves of the refrigerating chamber or placed in a truck or tubs. The cheese is cooled to a temperature of 12±2°C and sent for packing and marking.

Example 3. The use of a strain of Enterococcus hirae VKPM B-10090 in the manufacture of this product

Normalized milk fat content of 4% is subjected to heat treatment at 100°C without exposure, cooled to 38°C, make a starter for this product at 2%, activated biomass of bifidobacteria in milk in an amount of 0.1% and a strain of Enterococcus hirae VKPM B-10090 sterile milk in the amount of 0.5%. The fermentation is carried out at a temperature of 37°C to achieve titratable acidity 65°So Then the product is slowly cooled down to 6°C, followed by bottling.

Strain Enterococcus hirae VKPM B-10090 used in the production of dairy products.



 

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