Strain of enterococcus hirae used for production of fermented milk products

FIELD: biotechnology.

SUBSTANCE: strain of Enterococcus hirae "И"-29 with high antagonistic activity, deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under registration number of RNCIM B-10088, and can be used in production, for example, of fermented milk products such as kefir, cottage cheese or fermented baked milk.

EFFECT: invention enables to obtain a strain with high antagonistic activity and a high rate of milk fermentation.

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, milky clot, have 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.

The task of the claimed image is the shadow - 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-10088 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 the dairy industry), "Food industry", 1975.

The strain is deposited in Russian national Collection of Industrial Microorganisms and has a number VKPM B-10088.

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

1. Cultural-morphological:

- optional gone anaerobic;

- gram-positive cocci;

- size of 0.3-0.02 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 of 1-1 .5 mm (point);

the character contour edge level;

- the terrain is flat;

- smooth surface, Matt;

- color - white;

the structure is fine - grained;

-consistency - soft.

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 - 100°T.

a strain sprayway glucose, galactose, lactose, sucrose, maltose, raffinose, glycerol, dextrin.

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

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

- 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 strains of lactic acid.

3. Antibiotic activity:

the diameter of the zone of inhibition of growth relative to the representative of pathogenic Staphylococcus aureus was 24 mm;

the diameter of the zone of inhibition of growth relative to the representative of conditionally pathogenic Escherichia coli was 26 mm

Determination of antibiotic activity of the strain Enterococcus hirae VKPM B-10088 conducted on solid nutrient medium (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; Molo is about 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 the temperature of 30-37°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-10088 isolated from the microflora of kefir fungus is 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 - 24 mm, Escherichia coli - 26 mm) strain represents the production value for the dairy industry.

Example 1. The use of a strain of Enterococcus hirae VKPM B-10088 in the production of yogurt.

The milk is weighed, sorted and filtered, subjected to primary processing. It should be fresh, natural and udovletvorat 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 contribute in a pre-prepared yeast - 5-6% of the mass of milk. Leaven also enrich strain of Enterococcus hirae VKPM B-10088. 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°T. 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 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 for complete maturation.

Example 2. The use of a strain of Enterococcus hirae VKPM B-10088 in the production of cheese.

Production of cheese in the traditional way. The technological process consists of the following operations: receiving and preparation, separation of milk, standardization, pasteurization, shall hladina, 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-10088. Before you make the milk surface layer ferment gently remove clean sanitized bucket and remove. Then the starter mix to a uniform consistency with clean whisk (when cooking in the starter tubs) is whether stirrer and pour into the prepared milk in an amount of 5% of the total mass. The duration of ripening milk - 6 hours

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 of size 2×2×2, see Clot first cut the length of the tub on the horizontal layers, and then the length and width on the vertical. After this treatment, the clot is left for 40-60 min to separate from the adjustable tap wrenches 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.

In order to accelerate the separation of the serum, as well as the bad allocation serum, clot 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; lean - 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 Sep'a is atore. 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-10088 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-10088 sterile milk in the amount of 0.5%. The fermentation is carried out at a temperature of 37°C to achieve titratable acidity 65°T. Then the product is slowly cooled down to 6°C, followed by bottling.

Strain Enterococcus hirae VKPM B-10088 used to prepare dairy products.



 

Same patents:

FIELD: chemistry.

SUBSTANCE: invention relates to field of biochemistry. Sewage water with increased content of phenol compounds 300 - 1200 mg/l, content of sulfide-ions to 64.1 mg/l and content of sulfate-ions to 401.4 mg/l and 1843.7 mg/l in case of sparging with air, is passed through column type reactor. Reactor contains adsorbent - petroleum coke, onto which cells of strain of aerobic bacteria Pseudomonas putida 131 All-Russian collection of industrial microorganisms B-10894 are immobilised. Orthophosphoric acid or its salts are preliminarily dissolved in sewage water as biogenic additive in order to obtain pH 7-8, optimal for phenol destruction by bacteria strain.

EFFECT: invention ensures increased quality of purification of sewage water, which contain sulfide-ions and sulfate-ions, from phenol compounds to 0-50 mg/l.

1 dwg, 8 tbl, 7 ex

FIELD: biotechnologies.

SUBSTANCE: invention may be used to extract carotenoids, in particular, deinoxanthine, which is used to develop new antioxidant and radioprotector preparations to increase adaptation capabilities of humans and aminals, prevention and treatment of diseases. The method provides for extraction of carotenoids from the bacterial mass Deinococcus radiodurans by the mixture acetone : ethanol (at the 1:1 ratio). Separation of carotenoids with extraction of deinoxanthine on preparative columns of a liquid low-pressure chromatograph, where the sorbent is hydroxyapatite, and the eluent - ethanol.

EFFECT: invention makes it possible to increase quantity and quality of extracted deinoxanthine.

2 dwg, 1 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: invention relates to the field of microbiology and genetic engineering. A new yeast strain Yarrowia lipolytica RNCIM Y-3600 has been produced - a producer of cell-bound lipase.

EFFECT: higher activity.

3 ex

FIELD: biotechnologies.

SUBSTANCE: strain Trichoderma harzianum Rifai, having L-lysine-alpha-oxidase activity is deposited in the Russian National Collection of Industrial Microorganisms (RNCIM) under the registration number RNCIM F-180 and may be used in agricultural biotechnology and plant growing.

EFFECT: invention makes it possible to reduce losses of decorative and vegetable crops.

2 ex

FIELD: food industry.

SUBSTANCE: method envisages bacteria suspension production in a liquid nutrient medium, the bacteria separation from the suspension by way of centrifugation to produce a biomass, the produced biomass mixing with a protective medium containing sucrose and glycerine and the produced mixture freezing at a temperature of (-18)-(-20)°C. The liquid nutrient medium contains components at the following ratio, wt %: (10 - 12) - sucrose, (3 - 5) - malt sprouts and (4 - 6) - chalk. Before freezing the biomass mixture with the protective medium is cooled at a temperature of 4-6°C, maintained at the said temperature during 1 hour.

EFFECT: enhancement of biosynthetic activity of Lactobacillus delbrueckii lactic acid bacteria during preservation by freezing and subsequent storage.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA10247-A0137 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-GCTGCTTCGCCCACTTCG-3'-BmVAT8-Ch2s 5'-AGCATCGGATTTCATCGTCGTC-3-BmVAT8-Ch2as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA10247-A0137 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA2089 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CACCCTGTTTAGGACGGAG-3' - BmVAT3-Ch1s 5'-GATTGCGTGAACACGAAG-3' - BmVAT3-Ch1as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA2089 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA10247-A1008 of an equinia agent genome possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CGAACCCACACGAGCCTA-3' - BmVAT9-Ch2s 5'-GCGAAGATTCCGAAGATGC-3' - BmVAT9-Ch2as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA 10247_A1008 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA2262 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CGCCGTCACCCGCCAACTCAG-3'-BmVAT4-Chls 5'-AACAACCGCCGCCCGACGAC-3'-BmVAT4-Chlas. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA2262 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: presented primers are complementary to a differentiation fragment BMA0958 of an equinia agent genome possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-GCAGGACG AGTTCGGCGAGC-3'-BmVAT2-Ch1s 5'-GACGAAACCCACGCCCATTCC-3'-BmVAT2-Ch1as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA0958 of the equinia agent.

EFFECT: higher primer activity.

1 dwg, 2 ex

FIELD: biotechnologies.

SUBSTANCE: invention may be used to extract carotenoids, in particular, deinoxanthine, which is used to develop new antioxidant and radioprotector preparations to increase adaptation capabilities of humans and aminals, prevention and treatment of diseases. The method provides for extraction of carotenoids from the bacterial mass Deinococcus radiodurans by the mixture acetone : ethanol (at the 1:1 ratio). Separation of carotenoids with extraction of deinoxanthine on preparative columns of a liquid low-pressure chromatograph, where the sorbent is hydroxyapatite, and the eluent - ethanol.

EFFECT: invention makes it possible to increase quantity and quality of extracted deinoxanthine.

2 dwg, 1 tbl, 4 ex

FIELD: biotechnologies.

SUBSTANCE: probiotic lacto-amylovorin is produced by means of depth growth of the strain Lactobacillus amylovorus RNCIM B-6253 on the liquid nutrient medium, which contains the following: corn extract - 10 ± 0.01 ml, lactose - 20 ± 0.005 g, chemically deposited chalk - 0.5 ± 0.01 g, triple-substituted citric-acid sodium - 7.5 ± 0.05 g, 3-water acetous sodium - 2 ± 0.01 g, iron sulfate - 0.1± 0.001 g, 5-water manganese sulfate - 0.16 ± 0.002 g, hydrolysate of dry nonfat milk - up to l. Subsequent extraction of the target product in liquid condition or in the form of a dry powder is carried out with preliminary addition of a stabiliser to a cultural liquid or to a protective medium used when producing the probiotic in the form of the dry powder. The stabiliser is selected from: sodium metabisulfite, sodium hydrosulfite and ascorbic acid.

EFFECT: invention provides for stable production of a probiotic with high values of a titre of antimicrobial bodies not only in a cultural liquid, but also after its treatment required for production of different pharmaceutical forms of a preparation.

3 cl, 2 tbl, 3 ex

FIELD: biotechnologies.

SUBSTANCE: bacteria strain Planomicrobium koreense 78k, is extracted from soils and provides for production of site-specific endonuclease PkrI, which recognises and splits both chains of nucleotide sequence of DNA, which contains three or four C5-methylcytosine bases in recognition site 5'-GCNAGC-3' with formation of a single-nucleotide 3'-protruding end. The strain is deposited in the Russian National Collection of Industrial Microorganisms FGUP GosNIIgenetika under the number B-10627. The provided strain may be used for extraction of the new site-specific endonuclease PkrI, which may be applied in detection and splitting of methylated DNA sections.

EFFECT: production of site-specific endonuclease PkrI.

3 dwg, 3 ex

FIELD: biotechnologies.

SUBSTANCE: invention represents a bacteria strain Microbacterium testaceum 17B, deposited under the number RNCIM B-10628 in the Russian National Collection of Industrial Microorganisms FGUP GosNIIgenetika, being the producent of the site-specific endonuclease MteI. The invention makes it possible to produce the site-specific endonuclease, which recognises and splits both chains of the nucleotide DNA sequence 5'-G(m5C)G(m5C)NG(m5C)G(m5C)-3'/3'-(m5C)G(m5C)GN(m5C)G(m5C)G-5', where m5C-5-methylcytosine, with production of the single-nucleotide 5'-protruding end, and does not split the methylated DNA sequence 5'-G(m5C)NG(m5C)-3'/3'-(m5C)GN(m5C)G-5'.

EFFECT: method improvement.

2 dwg, 1 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: method envisages bacteria suspension production in a liquid nutrient medium, the bacteria separation from the suspension by way of centrifugation to produce a biomass, the produced biomass mixing with a protective medium containing sucrose and glycerine and the produced mixture freezing at a temperature of (-18)-(-20)°C. The liquid nutrient medium contains components at the following ratio, wt %: (10 - 12) - sucrose, (3 - 5) - malt sprouts and (4 - 6) - chalk. Before freezing the biomass mixture with the protective medium is cooled at a temperature of 4-6°C, maintained at the said temperature during 1 hour.

EFFECT: enhancement of biosynthetic activity of Lactobacillus delbrueckii lactic acid bacteria during preservation by freezing and subsequent storage.

1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA10247-A0137 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-GCTGCTTCGCCCACTTCG-3'-BmVAT8-Ch2s 5'-AGCATCGGATTTCATCGTCGTC-3-BmVAT8-Ch2as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA10247-A0137 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA2089 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CACCCTGTTTAGGACGGAG-3' - BmVAT3-Ch1s 5'-GATTGCGTGAACACGAAG-3' - BmVAT3-Ch1as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA2089 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA10247-A1008 of an equinia agent genome possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CGAACCCACACGAGCCTA-3' - BmVAT9-Ch2s 5'-GCGAAGATTCCGAAGATGC-3' - BmVAT9-Ch2as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA 10247_A1008 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: primers are complementary to a differentiation fragment BMA2262 of an equinia agent genome, possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-CGCCGTCACCCGCCAACTCAG-3'-BmVAT4-Chls 5'-AACAACCGCCGCCCGACGAC-3'-BmVAT4-Chlas. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA2262 of the equinia agent.

EFFECT: higher primer activity.

3 dwg, 3 ex

FIELD: medicine.

SUBSTANCE: presented primers are complementary to a differentiation fragment BMA0958 of an equinia agent genome possess activity of upstream and downstream primers in an amplification reaction and has the following structure: 5'-GCAGGACG AGTTCGGCGAGC-3'-BmVAT2-Ch1s 5'-GACGAAACCCACGCCCATTCC-3'-BmVAT2-Ch1as. The presented invention may be used in practical healthcare for detection of the differentiation DNA fragment BMA0958 of the equinia agent.

EFFECT: higher primer activity.

1 dwg, 2 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to medicine and pharmaceutical industry and represents a composition with anti-infectious activity presented in the solid dosage form for oral administration in the form of a powder, representing a balanced complex containing a probiotic agent, an adsorbent and an excipient; the probiotic agent is sterilised cultural fluid containing metabolites of the strain Bacillus subtilis VKPM (Russian National Collection of Industrial Microorganisms) No. B-2335; the adsorbent is zeolite; the excipient is calcium stearate or aerosil; the composition is characterised by the fact that it additionally contains a high-active immunomodulatory agent representing a complex of polysaccharides - products of polymer enzymatic hydrolysis of internal and external layers of brewers' yeast cell coating in the form of β-glucane and mannan with the ingredients of the composition taken in specific proportions, wt %.

EFFECT: invention provides high effectiveness and versatility with respect to dangerous bacterial and viral infections; it is storage safe and stable.

11 ex, 10 tbl, 10 dwg

Up!