Inoculate used for direct inoculation of lactic substratum bifidobacterium animalis lactis and containing l-cystein, and its use

FIELD: medicine.

SUBSTANCE: inoculate contains in a mixture or in a combination, main L-cystein of formula HSCH2CH(NH2)CO2H and at least one B.animalis lactis strain. Said cysteine and at least one B.animalis lactis strain are contained or have the form of at least one frozen granule and/or at least one lyophilizate. The pH value of the solution produced by thawing of at least one granule and/or dissolution, of at least one lyophilizate in the relation making 1 to 2 g of the lyophilizate per 8-10 ml of H2O, makes at least 4. L-cysteine contains in the inoculate in an amount within 1 g per 1·1014 CFU to 1 g per 3.5·1010 CFU, of at least one B.animalis lactis strain or all used B.animalis lactis strains whereas the same are numerous. Using the offered inoculate provides stimulation of B.animalis lactis growth and/or metabolism on a lactic substratum to produce a fermented diary product.

EFFECT: high probiotic value of the product.

21 cl, 9 dwg, 3 tbl, 3 ex

 

The technical field

The present invention relates to granules of cysteine and their use as activators growth Bifidobactehum animalis ssp. lactis.

The level of technology

The market of probiotics in the field of dairy products is in a constant state of growth. When receiving products containing increased amounts of probiotic strains, while giving them desirable organoleptic properties, manufacturers of dairy products face problems associated with mixed fermentation, which is used as a so-called technical or technological and probiotic microorganisms.

In practice, strains, called probiotics, are species that grow slowly in milk substrate (the duration of the cell cycle from 1 to 2 h for Bifidobacterium), technological microorganisms obtained from yogurt cultures, characterized by relatively rapid growth (the duration of the cell cycle from 15 to 30 min for S.thermophilus and L.bulgaricus).

A satisfactory compromise between the growth of technological and probiotic microorganisms is difficult to achieve, and most manufacturers prefer improved organoleptic properties to increase the number of probiotics: usually observed number of probiotics is 106-107CFU/g fermentive the aqueous product. Currently, the majority of hygiene studies using probiotics perform on products containing not less than 108CFU/g of probiotic strains.

The search for compounds which are activators of growth of probiotics based on in-depth studies of strain on nutrient needs and abilities of the dairy substrate to the satisfaction of such needs. On the other hand, you must make sure that these connections do not have negative effects and do not stimulate the growth of technological strains have quality satisfactory for food, and do not adversely affect the organoleptic properties of the product.

However, the number of connections-activators, which must be used to provide a stimulating action and which satisfies these requirements is usually very small compared to the amount of milk substrate. Similar doses (usually less than 100 mg/l) and tolerances incompatible with dosing systems and input used in the manufacture of dairy substrates, which makes industrial application of such compounds is problematic.

The problem becomes even more complicated when applied technological strains should operate in symbiosis several strains of lactic acid bacteria is, as in the case of fermented milk, for which in addition to the fermentation of milk under the action of probiotics applied symbiotic fermentation microorganisms S.thermophilus and L.bulgaricus.

In the prior art there are known various methods of determining the conditions conducive to the growth of probiotics. However, most of these tests were performed on pure cultures of probiotic bacteria without taking into account the possible presence and activities of other strains of lactic acid bacteria during the production of the product on an industrial scale.

One solution to this problem is described in patent application FR France 0507529 filed by the company Compagnie Gervais Danone July 15, 2005 In this application as an activator of bifidobacteria is proposed to apply the sulfur-containing amino acid.

In the present application proposed a further improvement of this invention, which is specially adapted for Bifidobacterium animalis ssp.lactis. Accordingly, in the present patent application proposed the inoculum, which allows to avoid the above described problems existing in the prior art, and provides a simple, reliable and efficient activation and/or stimulation Century animalis lactis during the industrial production of fermented milk product with probiotic value, intended for consumption by the person who kω, in particular, the fermented milk product in which, in addition to fermentation under the action of the probiotic Century animalis lactis, applied symbiotic fermentation under the action of S.thermophilus and L.bulgaricus.

Brief description of the invention

The present invention relates to improved inoculate Bifidobacterium animalis ssp. lactis, to the application of the above inoculum and fermented dairy products, obtained using the above-mentioned supernatant. The inoculum according to the present invention is specially adapted for direct inoculation of the milk substrate with at least one strain of C. animalis lactis to turn on the specified substrate in fermented milk product suitable for human consumption, in particular in the milk product with probiotic value, containing at least one strain Century animalis lactis.

The inoculum contains in the form of a mixture or combination of frozen granules and/or lyophilisate(s), which include a special form of L-cysteine and at least one strain Century animalis lactis. The authors of the present invention have chosen a particular form of L-cysteine and L-cysteine, granules and/or lyophilisate(s)obtained on the basis of which after freezing and/or dissolution have a pH value of not less than 4. An example of a suitable L-cysteine L-cysteine, usually called the basic form of the cyst is on (the formula can be presented as follows: HSCH 2CH(NH2)CO2H). This special form of cysteine is more effective than other commonly used forms of cysteine (usually cysteine-HCl in the form of the monohydrate hydrochloride cysteine). Despite the poor solubility of this special form of L-cysteine, selected by the authors of the present invention, the latter managed to get the inoculum in the form of frozen granules and/or lyophilisate (liofilizatow), in which cells .animalis lactis and specified a special form of L-cysteine combined or merged without prejudice to the physiological state and/or metabolism of .animalis lactis.

In the so-called "merged (combined)" implementation inoculum according to the present invention at least one pellet or freeze-dried inoculum at the same time contains cells .animalis lactis and found a special form of L-cysteine.

In the so-called "combined" version of the implementation of some of the granules or one of liofilizatow contains a special form of L-cysteine according to the present invention, and the other part of the granules or other freeze-dried cells .animalis lactis.

Thus, in the present invention proposed a "combination" or "combined" form inoculum .animalis lactis containing cysteine as an activator. It should be noted that the inoculum according to the present invention allows to obtain a specified population is about at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the number of which does not fall below 5·107SOME, preferably not lower than 108CFU per gram of fermented product within at least 20 days, preferably for at least 30 days of storage of the specified product at a temperature of from 4 to 11°C (see Fig and 9).

The inoculum according to the present invention is specially adapted for the production of fermented milk with a high probiotic value, because it allows you to maintain a very high content .animalis lactis during storage of the product specified in the conditions of cooling and also due to the fact that it contains as activator specific form of L-cysteine in such doses that have no adverse effects either on the symbiosis of S.Thermophilus and L.bulgaricus, nor on the sensory quality of fermented milk.

Brief description of Figures

This patent application contains references to the following figures:

Figure 1 - schematic diagram:

- "combined" version of the implementation, according to which it is proposed to get a combination of granules of at least one probiotic microorganism, and granules of cysteine, which then may mix;

- "combined" version of the implementation, according to which it is proposed to obtain pellets, which together contain at least about the in probiotic microorganism and cysteine.

Figure 2 - schematic diagram of receipt of the granules according to the "combined" implementation variant (cysteine-HCl + at least one probiotic microorganism): microorganism Bifidobacterium animalis lactis strain I-2494 or W® in the form of granules, together containing probiotic microorganism and cysteine, the latter in this case is a cysteine-HCl.

Figure 3 - change in time of the pH value of the milk mixture, inoculated:

granules Bifidobacterium animalis lactis strain 1-2494 (control experiment, lower curve), or

- granules, together containing Bifidobacterium animalis lactis strain 1-2494 and cysteine-HCl (upper curve).

Shows the deterioration in the physiological condition of the probiotic Bifidobacterium animalis lactis during its introduction in the composition of the granules together with cysteine-HCl (composition obtained by the "combined" version, containing the cysteine-HCl).

4 is a schematic diagram of producing granules by "combined" option (basically cysteine + at least one probiotic microorganism): microorganism Bifidobacterium animalis lactis strain 1-2494 or W® in the form of granules, together containing probiotic microorganism and cysteine, the latter in this case is a major cysteine.

Figure 5 - change in time of the pH value of the milk mixture, inoculated:

granules Bifidobacterium animalis lactis strain I-2494 (control the experiment, the lower curve), or

- granules, together containing Bifidobacterium animalis lactis strain 1-2494 and basically cysteine (top curve).

It is shown that the selection of the preferred cysteine for the introduction of the granules obtained by the "combined" option (Bifidobacterium animalis lactis + cysteine), provides no negative impact on the physiological state of the specified probiotic.

Fig.6-9: comparison of infant formula,

- inoculated frozen granules technological microorganisms (mixture of granules S.thermophilus 1-1630, L.bulgaricus 1-1632 and L.bulgaricus I-1519) and frozen granules probiotic microorganism Bifidobacterium animalis lactis (strain W® or 1-2494) in an amount of 0.5 g/l, and

which basically added cysteine in the form of pellets than granules probiotic microorganism (combined solution G4), or in the form included in the granules probiotic microorganism (combined variant G4I) compared with the control mixture, in which instead of cysteine added water.

6 and 7, a pH change depending on the time (in hours).

Fig and 9 - changes the amount of biomass (CFU/ml) depending on time (in days) during storage of the fermented milk product at 10°C.

6 and 8, data from probiotic microorganism Bifidobacterium animalis lactis strain VV®:

- milk formula W® T - milk mixture on odorou produced by seeding a mixture of granules of technological microorganisms and control granules microorganism Bifidobacterium W® (instead of cysteine granules contain water) in an amount of 0.5 g/l;

- milk formula W® G4 - milk formula, which produced seeding is similar to planting on milk mixture W® T, i.e. the mixture of granules of technological microorganisms and control granules microorganism Bifidobacterium W® in the amount of 0.5 g/l with additional add primary cysteine in the form of frozen pellets than granules probiotic, to obtain the concentration of cysteine in the mixture, equal to 7.5 mg/l;

- milk formula W® G4I - milk formula, which is produced by seeding a mixture of granules of technological microorganisms and granules obtained by the combined variant containing the microorganism Bifidobacterium BB12® in the amount of 0.5 g/l, and these granules contain probiotic basically cysteine, to obtain the concentration of cysteine in the mixture, equal to 7.5 mg/L.

7 and 9: data for probiotic microorganism Bifidobacterium animalis lactis strain I-2494:

- milk formula I-2494 T - milk formula, which is produced by seeding a mixture of granules of technological microorganisms and control granules microorganism Bifidobacterium I-2494 (instead of cysteine granules contain water) in an amount of 0.5 g/l;

- milk formula I-2494 G4 - milk formula, which produced seeding is similar to planting in formula I-2494 T, i.e. the mixture of granules of technological microorganisms and control granules microorganism Bifidobacterium I-2494 in the amount of 0.5 g/l to complete inim introduction the main cysteine in the form of frozen granules, other than granules probiotic, to obtain the concentration of cysteine in the mixture, equal to 7.5 mg/l;

- milk formula I-2494 G4I - milk formula, which is produced by seeding a mixture of granules of technological microorganisms and granules obtained according to the "combined" version, containing the microorganism Bifidobacterium I-2494 in the amount of 0.5 g/l, and these granules contain probiotic basically cysteine, to obtain the concentration of cysteine in the mixture, equal to 7.5 mg/L.

Detailed description of the invention

Some definitions

In this application, all terms used in the meaning and value that they normally have in the field of breast and/or food industry. So, when mentioned "lactic fermentation" refers to the lactic acid fermentation, which results in the acidification due to the formation of lactic acid, which may be accompanied by formation of other acids, carbon dioxide and various substances, including exopolysaccharides (EPS) and substances that have a smell, including diacetyl and acetaldehyde. Under "lactic acid bacterium" (lactobacteria) understand living or viable microorganism or a microorganism strain capable of the specified lactic acid fermentation in the dairy substrate. The term "fermented milk" has the meaning commonly used in Molo the Indus, i.e. products intended for human consumption and animals, including man, which are the result of lactic acid fermentation of milk substrate. Such products may contain other ingredients, including fruits, plants, sugar, flavorings, etc.

The name "fermented milk" meets the strict official regulations. So, you can refer to the Codex Alimentarius Commission (code of Rules for the production and distribution of food prepared by the Codex Alimentarius Commission at FAO and who, published by the Information division FAO and available on the Internet at the address http://codexalimentarius.net; see, in particular, volume 12 Codex Alimentarius "Terminology for milk and dairy products" standard "CODEX STAN A-11(a)-1975", the contents of which are incorporated into this description by reference). In particular, you can refer to the Decree of the government of France No. 88-1203 of December 30, 1988 relating to fermented milk and yoghurt (aurton), published in Journal Officiel de la République Française December 31, 1988, the content of the Resolution is incorporated into this description by reference.

Thus, in this application, the term "fermented milk" refers to the product made using the milk substrate is subjected to processing at least equivalent to pasteurization, which is produced by sowing is mikroorganizmov, i.e. which was inoculable microorganisms belonging to the species characteristic of each product. If this is unacceptable collapse "fermented milk" paths other than the paths resulting from the activity of the microorganisms used.

"Fermented milk" is not subjected to any processing, allowing to extract constituent element used dairy substrate, in particular, the separation of the clot. To "fermented milk" can be added one or more extracts of odoriferous substances, one or more of the natural flavors, and one or more sugars, and other special food, which gives a special taste, or grains in an amount not exceeding 30% by weight of the finished product. The introduction of the similar products substitutes fats and/or proteins of non-dairy origin is unacceptable. The amount of free lactic acid contained in fermented milk when selling to the consumer, should not be less than 0.6 g per 100 g; the content of additional proteins in relation to the proportion of milk should not be lower than conventional milk.

The term "milk substrate" in this application means "milk" in the sense commonly used in the dairy industry, i.e. the substrate, containing only milk and/or components of milk and having calisota, that lactic fermentation is similar to the milk substrate strains of at least one lactic bacteria, including S.thermophilus and/or L.bulgaricus, yields a product suitable for human consumption, in particular, of the substrate corresponding to the term "fermented milk". Thus, the term "dairy substrate" encompasses the milk of animal origin in all forms and in all variants of the composition, including fat free or do not skim, condensed or normal, subjected or not subjected to ultrafiltration, fresh or not, powder or plain, restored or unrestored, modified or unmodified, enriched or not enriched any components manufactured with additives process agent or agents, to improve its quality, including food additives, flavorings, sugars, etc., or manufactured without adding any milk. For example, a dairy substrate, intended for the manufacture of fermented milk, may contain milk, cream and skim milk powder (see example 1 below).

However, the meaning of the term "milky substance" does not include "nutrient medium". In practice, the term "culture medium" means a medium that is designed to accelerate or stimulate the growth of lactic acid bacteria is s and/or received using inoculum of lactic acid bacteria, while the term "milk substrate" means an environment designed for the conversion by fermentation product intended for human consumption. Thus, many compounds which can be added to the culture medium to stimulate or accelerate the growth of lactic acid bacteria and/or cannot be added to a dairy substrate when receiving the fermented milk or yogurt.

Specified applies to the following compounds:

numerous surfactants and/or emulsifiers, and/or solubilizing agents and/or detergents, including polyoxyethylenesorbitan-20-monooleate (also known as Polysorbate-80 or tween-80);

- acid like lemon and vinegar;

extracts of meat;

vegetable gelatin;

- glycerophosphate.

The most commonly used dairy microorganisms include the following kislomolochniy bacteria:

- Streptococcus thermophilus (for example, strain 1-1630 available in CNCM);

- Lactobacillus delbrueckii ssp.bulgahcus, or Lactobacillus bulgaricus (for example, strains 1-1519 and 1-1632 available in CNCM);

- Pediococcus acidilacti;

- Leuconostoc, including Leuconostoc cremoris, Leiconostoc dextranicum and Leuconostoc lactis;

- Lactobacillus strains, including Lactobacillus acidophilus (for example, strain I-0967 available in CNCM), Lactobacillus casei (for example, strain I-518, available at CNCM), Lactobacillus helveticus and Lactobacillus delbrueckii ssp.lactis (n is an example, strain I-2843, available at CNCM);

- Lactococcus, including Lactococcus cremoris, Lactococcus lactis ssp. lactis (for example, strain I-1631, available at CNCM) and Lactococcus lactis ssp. lactis species diacetylactis (for example, strain I-2806 available in CNCM);

- Bifidobactehum, including Bifidobactehum animalis lactis (for example, strain I-52494 available in the CNCM, or strain, supplied to the market by the company Chr. Hansen under the name W®), Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium longum and BifidobacteriLim infantis.

Probiotic lactic acid bacteria are defined as live lactic acid bacteria, which when used in sufficient quantities to have a positive effect on the human body, in addition to their original nutritional value (the official definition of the world health organization). Probiotic lactic acid bacteria include, in particular, the following lactic acid bacteria:

- Lactobacillus strains, including Lactobacillus acidophilus (for example, strain I-0967 available in CNCM), Lactobacillus casei (for example, strain I-518, available at CNCM), Lactobacillus helveticus and Lactobacillus delbrueckii ssp. lactis (for example, strain I-2843, available at CNCM);

- Bifidobacterium, including Bifidobacterium animalis lactis (for example, strain I-202494 available in the CNCM, or strain, supplied to the market by the company Chr. Hansen under the name W®), Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium longum and Bifidobacterium infantis.

Probiotic lactic acid bacteria is not just nutritional supplements, which are in the larger number is the number to add to the milk product. They are live bacteria that carry out lactic acid fermentation, and their metabolism is affected by the conditions in which they are located during the manufacture of fermented dairy product and then during storage of the fermented milk product prior to the expiration date. The retention period depends on the period of conservation (date), established by the current legislation. For fresh fermented dairy products, including fermented milk, the shelf life is 30 days from the date of manufacture.

Under the technology microorganism, a bacterium or strain in this application understand the microorganism, bacterium or strain, affecting the structure and/or texture of fermented dairy product in contrast to microorganisms, bacteria or strains having first of all probiotic function. For example, fermented milk can be produced by symbiotic fermentation L.bulgaricus - S.thermophilus while using at least one strain .animalis lactis as a probiotic strain. In this case, technological strains are strains S.thermophilus and L.bulgaricus.

Tasks and objects of the invention

The present invention relates to inoculate, specially adapted for direct inoculation of milk substr is that at least one strain of Bifidobacterium animalis ssp. lactis with the conversion of the specified dairy substrate fermented milk product suitable for human consumption and having a probiotic value, as well as to the application of the said inoculum in the production of such dairy products. In particular, the inoculum according to the present invention is suitable for the production of fermented milk product obtained by fermentation of a dairy substrate probiotic .animalis lactis and symbiotic system S.thermophilus - L.bulgaricus. This product contains at least one strain S.thermophilus, at least one strain L.bulgaricus and at least one strain .animalis lactis in live form.

The inoculum according to the present invention contains at least one strain .animalis lactis.

The inoculum according to the present invention contains a special form of cysteine and L-cysteine which has such a shape that the granules and/or lyophilisate(s), obtained using a specified form of L-cysteine, have such a composition that the pH of a solution obtained by:

defrost (defrost) indicated at least one of pellets and/or dissolution (re-solubilization) of the specified at least one lyophilisate in respect of 1 to 2 g of freeze-dried to 8-10 ml of H2O is not less than 4.

The specified pH value preferably does not exceed 8. To do this, suppose that the equipment is the choice of cysteine in such form, which ensures that the pH of the containing solution was more than 4. For example, a solution containing 1.25 g of the specified form of cysteine in 50 ml of water, has a pH value of from 4.5 to 5.5. An example of this form of cysteine L-cysteine, usually called the main cysteine (or L-cysteine in the main form). The formula L-cysteine can be represented as follows: HSCH2CH(NH2)CO2H. Major cysteine different from cysteine-HCl and is neither a cysteine hydrochloride or hydrochloride monohydrate cysteine. Selection of a specified special form of cysteine is the result of a choice among different available forms of cysteine. Specified special form of cysteine absorbed .animalis lactis and is an activator of growth B.animalis lactis in milk substrate. This cysteine is suitable for human consumption. It complies with legislation relating to fermented milk products intended for human consumption. In addition, this special form of cysteine helps to stimulate the growth or at least the metabolism .animalis lactis and does not affect the metabolism of other lactic acid bacteria, which are not probiotics, in particular, on the fermentation of milk carried out by symbiotic system S.thermophilus - L.bulgaricus. In addition to all these properties specified special form of cysteine allows peluchatte called the United inoculate, containing together the specified at least one strain .animalis lactis and cysteine in the same physical structure in the absence of negative effects on growth or at least on the metabolism of the specified strain .animalis lactis. In the most preferred case, the inoculum according to the present invention does not contain a cysteine in the acidic form, such as the cysteine-HCl (hydrochloride of L-cysteine formula HSCH2CH(NH2)CO2H·HCl).

The inoculum according to the present invention are presented in a specific physical form. It can be in the form of at least one frozen granules and/or at least one lyophilisate. Such a specific form of inoculum is particularly suitable for direct seeding (direct inoculation). The choice of these specific forms of cysteine allows to obtain the inoculum specified in the particular physical form, which is particularly active. The choice concretos form of L-cysteine according to the present invention allows a mix between a specified at least one strain .animalis lactis and specified cysteine with getting frozen granules and/or at least one lyophilisate, together with the specified strain .animalis lactis and specified cysteine within the same granules and/or the same freeze-dried in the absence of negative effects on growth or at least metabol the ZM specified strain .animalis lactis. Similar results cannot be obtained with the use of cysteine in the acidic form, such as cysteine-HCl (hydrochloride of L-cysteine formula HSCH2CH(NH2)CO2H·HCl) instead of the basic L-cysteine.

Thus, the authors of the present invention proposed a frozen pellet or freeze-dried, together containing cells of the specified at least one strain .animalis lactis and found a special form of L-cysteine. Similar to the frozen pellet or lyophilisate can be called inoculate .animalis lactis, combined with an activator. Similar to the frozen pellet or lyophilisate are preferred implementation of the present invention.

Another way that is different from the combination with an activator, is not the introduction of the activator and one pellet or in one lyophilisate together with strain Century animalis lactis, and in obtaining separate frozen pellets or liofilizatow.

It is possible, for example, obtaining and combining among themselves in the form of a mixture or combination ("kit parts"):

- frozen granules containing cells of the specified at least one strain .animalis lactis, and other frozen pellets containing the specified special form of L-cysteine (obtaining "combined" granules in contrast to the "United");

the lyophilisate containing cells specified at the ore one strain S.animalis lactis, and another lyophilisate containing the specified special form of L-cysteine (obtaining a "combined" freeze-dried in contrast);

- frozen granules containing cells of the specified at least one strain B.animalis lactis, and lyophilisate containing the specified special form of L-cysteine or Vice versa (a combination of granules or lyophilisate (s));

the combination of these implementation options.

This inoculum are a direct alternative to the frozen pellets "combined" option or lyophilizates "combined" option, because they combine the means according to the present invention in various physical forms between them. In the present description such inoculum called "combined" and are the subject of this application.

Figure 1 shows a method of manufacturing granules "combined" option and a method of manufacturing composite pellets according to the present invention.

The technical result of the present invention is achieved through the use of a particular form of cysteine (preferably, main L-cysteine) in combination with the creation of a special physical form (frozen granules and/or lyophilisate(s)), specially adapted for direct inoculation.

In addition, the inoculum according to the present invention is especially adapted for growth stimulation is tommow .animalis lactis. It may not be suitable for stimulating the growth of other species or genera of probiotic bacteria.

To stimulate growth of strains .animalis lactis essential activator of growth that have a positive impact on growth or at least on the metabolism .animalis lactis in the absence of a negative or detrimental effect on the texture, structure, as well as sanitary and sensory quality of fermented dairy product. This problem is especially difficult in the case of obtaining fermented dairy products, which, in addition to .animalis lactis, require the use of other lactic acid bacteria. In fact, all types of lactic acid bacteria is not necessarily characterized by the same metabolism, and conditions favorable for one species or genus, are not necessarily favorable for other species or genus. Therefore, finding the conditions for the fermentation of milk, suitable for aggregate used strains can be difficult. To this is added the fact the fact that to successfully produce high quality and meets sanitary standards fermented dairy products fermentation of milk must meet specific limitations on the kinetics of the formation of acids. To limit the risk of various kinds of infection is desirable as it is possible more than the rapid achievement of pH, 5.5. However, the high rate of formation of acids is an obstacle to rapid growth and/or metabolism normal .animalis lactis.

The case of fermented milk produced by fermentation under the action of S.thermophilus and L.bulgaricus while fermenting probiotic .animalis lactis, is particularly difficult because, in addition to fermentation .animalis lactis, requires symbiotic fermentation of milk, which use at least one strain of Streptococcus thermophilus and at least one strain of Lactobacillus delbrueckii ssp. bulgaricus. B.animalis lactis is a form of slow-growing at a dairy substrate (the duration of the cell cycle is of the order of 1-2 h), whereas technological microorganisms S.thermophilus and L.bulgaricus a high growth rate (duration of the cell cycle is of the order of 15-30 min). In such circumstances, maintaining the set of parameters at the proper level is particularly difficult.

The inoculum according to the present invention is specially adapted to ensure that it does not adversely affect the fermentation is carried out technological strains of lactic acid bacteria, in particular, on symbiotic fermentation under the action of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. In particular, the inoculum according to the present invention is specially adapted for the application of the Oia in the production of fermented milk, containing at least one strain .animalis lactis as a probiotic strain and produced by fermenting a milk substrate with at least one strain of Streptococcus thermophilus and at least one strain of Lactobacillus delbrueckii ssp. bulgaricus, in addition to fermentation, carried out .animalis lactis.

The inoculum according to the present invention corresponds to the different constraints and allows to obtain a fermented dairy product with a high probiotic value with exceptional quality.

Despite the fact that the inoculum according to the present invention is optimized for particularly complex cases symbiotic fermentation under the action of S.thermophilus and L.bulgaricus simultaneous fermentation .animalis lactis, it is obvious that it can be used in less complex cases, for example, in fermenting the milk substrate with the use of only one genus or species of lactic acid bacteria other than .animalis lactis, in addition to the specified at least one strain .animalis lactis, or when using only .animalis lactis.

One of the advantages of inoculum according to the present invention is that it allows you to get the population of the specified at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the size of which substantially exceeds the size of the population is odorou can be obtained in the same conditions using conventional inoculum of the same strain .animalis lactis, not containing activator, growth, and population size that can be obtained in the same conditions using the inoculum of the same strain .animalis lactis containing as an activator of growth cysteine-HCl. Thus, the inoculum according to the present invention allows to obtain the population of the specified at least one strain .animalis lactis (or all applied strains Century animalis lactis, if there are several), which upon completion of the fermentation is at least 5·107CFU per 1 g of fermented product, preferably at least 108CFU per 1 g of fermented product. The particular advantage of inoculum according to the present invention is the ability to generate populations of the specified at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the amount of which does not fall below 5·107CFU per gram of fermented product after at least 20 days, preferably after at least 30 days of storage of the specified fermented product at a temperature of from 4 to 11°C, preferably at temperatures from 4 to 10°C, more preferably at a temperature of from 4 to 9°C. the Greatest advantage of inoculum according to the present invention is the possibility of obtaining a specified population at least on the aqueous strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the amount of which does not fall below 108CFU per gram of fermented product after at least 20 days, preferably after at least 30 days of storage of the specified fermented product at a temperature of from 4 to 11°C, preferably at temperatures from 4 to 10°C, more preferably at a temperature of from 4 to 9°C (see Fig and 9, showing the preservation of the population size .animalis lactis in fermented milk (S.thermophilus - L.bulgaricus + .animalis lactis))made using the inoculum according to the present invention, after 30 days of storage compared with the control samples fermented milk, population size .animalis lactis which quickly falls starting from the 21st day.

Another advantage of inoculum according to the present invention is that it has a shape suitable for application in industry. In fact, the inoculum according to the present invention has an improved form allowing us to use his special, reliable and simple way within the industrial production of fermented dairy product. Doses of growth promoters and tolerances often difficult to determine with the help of dosing systems and input used in the manufacture of fermented dairy products in a production environment. N the example, excess activator favorable for probiotic strain, usually has a very negative impact on the quality of the obtained fermented milk product; it can give him Neplatny taste (for example, the taste of sulfur in excess addition of cysteine)to have a negative or detrimental impact on the fermentation under the influence of technological strains (that is, for example, when an excess of cysteine in the case of S.thermophilus). Control of the number of added activator is also of interest from the point of view of sanitation, since this control allows you to limit human intervention, as well as the modification and improvement of technological units, sanitary quality which must be absolute. Control of the number of added activator, of course, is also of economic interest. Thus, it appears possible situation in which a specialist in this field of technology has a suitable activator of growth, but in its application in the production environment it is facing difficulties, because in light of these facts the number of added activator is difficult. The inoculum according to the present invention is very easy to apply on an industrial scale, and it allows reliable and easy to control applied to the number of activator. Thus, the inoculum according to the present invention is easier to apply; it can effectively and reliably produce in production of fermented dairy products with exceptional performance consistency, structure and taste and contains probiotic .animalis lactis in especially large numbers.

The inoculum according to the present invention is the result of a difficult compromise, in particular, between the need to preserve the exceptional characteristics for the consumer (structure, texture, health and taste) fermented dairy product; a desire to improve the content .animalis lactis order to enhance the probiotic properties of the product; the desire to obtain the inoculum in an improved form, specially adapted for direct seeding and allows you to control the added amount of the activator in a simple way while complying with security requirements.

The inoculum according to the present invention can solve these tasks.

The inoculum according to the present invention contains in the form of a mixture or combination (set of parts):

at least one strain B.animalis lactis and

- L-cysteine free form (i.e. L-cysteine, is not associated with other amino acids of the peptide bond), which, in addition, there is t such form, what it allows you to get these granules and/or lyophilisate(s)that the pH of the solution resulting from:

- defrost the specified at least one of the granules; and/or

- dissolution of the specified at least one lyophilisate in respect of 1 to 2 g of freeze-dried to 8-10 ml of H2O is not less than 4.

The specified pH value, preferably not greater than pH 8. To solve these problems, it is preferable selection of free L-cysteine in such form that upon its introduction into the solution pH latter is more than 4. For example, a solution containing 1.25 g of the specified form of cysteine in 50 ml of water, has a pH value of from 4.5 to 5.5. A preferred example of such form of cysteine L-cysteine, usually called the main cysteine (or L-cysteine in the main form). The basic formula of L-cysteine can be represented in the form HSCH2CH(NH2)CO2H.

The number of the special form of L-cysteine according to the present invention and cells .animalis lactis contained in the inoculum according to the present invention, distributed within a solid physical structures, namely frozen granules and/or at least one lyophilisate.

These cysteine and at least one strain .animalis lactis can work together to be part of the same solid physical structure or part of various the solid structures. The latter can be manufactured separately and be part of the set or to be in the mix. Respectively can be obtained a mixture of any one of the physical structure, containing together a special form of L-cysteine according to the present invention and cells .animalis lactis with various other or the same solid structures.

Thus, these cysteine and at least one strain B.animalis lactis can be represented as:

- a set of frozen granules combined composition and/or lyophilisate of the joint staff and/or

- separate system in the form of a "set" of certain frozen pellets of different composition and/or lyophilisate(s).

Thus, according to the preferred implementation variant of the present invention indicated a special form of L-cysteine fully or part thereof and cells .animalis lactis fully or part of them can work together to be part of the same frozen pellets or the same freeze-dried. The inoculum according to the present invention in this case contains at least one of the granules and/or lyophilisate, together with a special form of L-cysteine according to the present invention and cells .animalis lactis. Preferably a special form of L-cysteine fully and cells .animalis lactis entirely contained in the granules of the combined composition or in one of lyophilize the E.

A special form of L-cysteine according to the present invention completely or partially and cells .animalis lactis fully or partially can be physically separated and included in the set, including a variety of frozen granules and/or different lyophilizate. In this case, the inoculum according to the present invention contains either at least two frozen granules, or at least two different freeze-dried, or at least one frozen granule and at least one lyophilisate, each of which is either a special form of cysteine according to the present invention, or cells .animalis lactis, and for each of these pairs of physical structures (pellet + pellet, freeze-dried + another lyophilisate or pellet + lyophilized) one of the two physical structures contains only the specified special form of L-cysteine or its part and does not contain cells .animalis lactis, and the other physical structure contains cells .animalis lactis and does not contain the specified special form of L-cysteine.

Accordingly, the total number of the special form of L-cysteine according to the present invention and the total number of cells Century. animalis lactis contained in the inoculum may be contained in different granules and/or different lyophilisate or lyophilizate or to form any.

The inoculum according to the present from which retenu, thus, may include or consist of the following elements:

a) frozen pellets containing a special form of L-cysteine according to the present invention and cells Century. animalis lactis;

b) freeze-dried, contains a special form of L-cysteine according to the present invention and cells .animalis lactis;

C) frozen pellets containing a special form of L-cysteine according to the present invention, but not containing cells .animalis lactis, and frozen pellets containing cells .animalis lactis, but not containing a special form of L-cysteine according to the present invention;

d) freeze-dried, contains a special form of L-cysteine according to the present invention, but not containing cells .animalis lactis, and lyophilisate containing cells .animalis lactis, but not containing a special form of L-cysteine according to the present invention;

d) frozen pellets containing a special form of L-cysteine according to the present invention, but not containing cells .animalis lactis, and lyophilisate containing cells .animalis lactis, but not containing a special form of L-cysteine according to the present invention;

e) freeze-dried, contains a special form of L-cysteine according to the present invention, but not containing cells .animalis lactis, and frozen pellets containing cells .animalis lactis, but not containing a special form of L-cysteine according to the present invention;

<> g) a mixture of at least two of the described compositions.

The inoculum according to the present invention contains a cysteine in the so-called free form (i.e., not associated with other amino acids of the peptide bond), which is the result of a special selection made by the authors of the present invention. In fact, this cysteine free form is a free L-cysteine, which, in addition, has a form that allows you to get these granules and/or at least one lyophilisate that the pH of the solution obtained after thawing these granules (for example, by placing them in a water bath at 37°C for 3 min with manual agitation), and/or dissolution of these freeze-dried or liofilizatow in the ratio of 1 to 2 g of freeze-dried to 8-10 ml of water (for example, dissolution in water for 30 min at 25°C under stirring with a magnetic stirrer), is not less than 4. This is the preferred choice of cysteine in such form, in which the pH of a solution consisting of 1-1,5 g of such cysteine and 50-100 ml of water is at least 4. An example of a suitable cysteine is a cysteine, a is usually called the main cysteine, the formula can be presented as follows: HSCH2CH(NH2)CO2H. Example of unsuitable cysteine is a cysteine-HCl (hydrochloride cysteine or monohydrate guide is ochloride cysteine).

Preferably none of the compounds used in the preparation of the inoculum according to the present invention, does not have the form of the hydrochloride.

Contained in the inoculum special form of L-cysteine according to the present invention is not acidic form of cysteine, including the hydrochloride of L-cysteine (HSCH2CH(NH2)CO2H·HCl) or monohydrate hydrochloride L-cysteine (HSCH2CH(NH2)CO2H·HCl·H2O). Monohydrate hydrochloride L-cysteine is not a special form of cysteine according to the present invention, since a solution containing 1.0 g of the monohydrate of cysteine hydrochloride in 100 ml of H2O, has a pH of 1.5-2.0. Thus, the defrosting of frozen granules or dissolving the lyophilisate, which could be derived from cysteine, which is not a special form of cysteine according to the present invention, would give a solution with a low pH value, in particular, with a pH value less than 4, usually less than 3.5. The most strongly acidic forms of cysteine represent a cysteine in the form of salts, including the hydrochloride, cysteine or monohydrate hydrochloride cysteine. Ion or ions included in the composition of this salt are included in the frozen pellets or freeze-dried. The presence of such ion or ions can sometimes be an indicator of the use of cysteine in combination with mo is ecoloy acid (which is usually used to increase the solubility). For example, such is the case with chlorine ions contained in the hydrochloride of L-cysteine or monohydrate hydrochloride cysteine. In fact, in the case of acidic cysteine in the form of a hydrochloride of L-cysteine or monohydrate hydrochloride L-cysteine can be stated that the pellet or lyophilisate contains a large number of chloride ions (Cl) taking into account the fact that the monohydrate hydrochloride L-cysteine contains more than 19% of the chloride ions, while basically L-cysteine typically contains less than 0.05%, in particular less than 0.04% of the chloride ions. High chloride content in the pellet or the lyophilisate can be an indirect measure of the utilization of cysteine, which is not the main cysteine. So, the form of cysteine used in the inoculum according to the present invention typically contains less than 60 micrograms of chloride ions per gram of granules with a solids content equal to 16-17%.

For these reasons, the main criterion is pH. In fact, the measurement of pH is a direct criterion for the presence of a special form of cysteine according to the present invention: pH thawed pellets or dissolved lyophilized in this case is at least 4. Usually it is not less than 5, for example between 5 and 6.5. The specified pH value preferably does not exceed 8.

The value of pH of at least 4 and not more than 8, not class aetsa stressful for B.animalis lactis. Accordingly, the latter can be mixed with a solution of cysteine, not subjecting them to stress, and to enter into the frozen pellets or freeze-dried, in which cysteine and cell .animalis lactis United in one solid physical structure (mixed). Thus obtained inoculate .animalis lactis contains cells .animalis lactis, are in good physiological and/or metabolic state and gives quick response to the activation of the cysteine. Frozen pellets or freeze-dried "United" composition according to the present invention, together containing cells .animalis lactis and a special form of L-cysteine according to the present invention, therefore, represent a special way improved inoculate .animalis lactis, because in this way cannot enter the activator .animalis lactis in the containing structure in the absence of a negative impact on the physiological state and/or metabolism .animalis lactis.

The choice of a special form of L-cysteine according to the present invention allows to obtain granules and/or lyophilisate, which is more efficient than granules and/or lyophilisate obtained with the use of strong acid L-cysteine, including the hydrochloride of L-cysteine. Specified selection also allows you to effectively introduce cysteine into pellets or freeze-dried, contains cells .animalis lactis. In fact, p is and the use of cysteine in strongly acidic form, including the hydrochloride of L-cysteine to obtain frozen pellets or freeze-dried, together containing cysteine-HCl and cells .animalis lactis obtained pellet or a lyophilisate of the combined composition is ineffective as an inoculum .animalis lactis no comparison with similar granule or lyophilisate obtained on the basis of a special form of L-cysteine according to the present invention.

Thus, in a preferred implementation of the present invention, the inoculum according to the present invention does not contain strongly acidic forms of cysteine, in particular the hydrochloride of L-cysteine, and/or received on the basis of such cysteine.

The inoculum according to the present invention may not contain cystine. According to the present invention preferably do not add cystine in the inoculum. However, it can be found traces of cystine, which can be formed, for example, in the oxidation of the cysteine contained in the inoculum.

Specified special form of L-cysteine according to the present invention preferably is present in the inoculum in a quantity amounting to:

from 1 g of 1·1014SOME of the specified at least one strain .animalis lactis (or all applied strains Century animalis lactis, if there are several), up to 1 g 3.5·1010SOME of the specified at least one strain B.animalis lactis (or su is x used strains .animalis lactis, if there are several), and/or in the quantity constituting:

from 0.01 mg to one billion CFU of the indicated at least one strain B.animalis lactis (or all applied strains .animalis lactis, if there are several);

up to 30 mg per one billion CFU of the indicated at least one strain B.animalis lactis (or all applied strains .animalis lactis, if there are several).

In this application all mass values for cysteine calculated on the basis of cysteine formula HSCH2CH(NH2)CO2H.

More preferably, the specified special form of L-cysteine according to the present invention is present in the specified inoculum in a quantity amounting to:

from 1 g 0.2·1014SOME of the specified at least one strain B.animalis lactis (or all applied strains B.animalis lactis, if there are several), up to 1 g 10·1010SOME of the specified at least one strain B.animalis lactis (or all applied strains .animalis lactis, if there are several), and/or in the quantity constituting:

from 0.05 mg to one billion CFU of the indicated at least one strain B.animalis lactis (or all applied strains .animalis lactis, if there are several);

up to 10 mg per one billion CFU of the indicated at least one strain B.animalis lactis (or all applied strains .animalis lactis, if there are several).

According to a special variant of " the present invention indicated a special form of L-cysteine according to the present invention preferably is present in the specified inoculum in a quantity of 4,90 up to 144 mg / 5·109-5·1011SOME of the specified at least one strain of C. animalis lactis.

Preferably specified special form of L-cysteine according to the present invention is present in the specified inoculum in a quantity of 25 to 50 mg, more preferably from 30 to 40 mg, more preferably from 31 to 39 mg, for example about 31, 32, 33, 34, 35, 36, 37, 38 or 39 mg, even more preferably 35 to 37 mg, for example, about 35, 36 or 37 mg, most preferably about 36,9 mg per 5·109-5·1011SOME of the specified at least one strain .animalis lactis.

The number Century. animalis lactis is preferably 5·109-5·1011CFU, more preferably 2·1010-5·1011CFU, even more preferably 2·1010-1·1011CFU, even more preferably 2·1010-7·1010SOME.

One gram of the granules of the present invention and containing 16-17% solids (dried at 105°C for 40 min), preferably contains 5·109-5·1011SOME of the specified at least one strain .animalis lactis. One gram of the granules of the present invention and containing 16-17% solids (dried at 105°C for 40 min), preferably contains 4,90 144 mg of the specified special form of L-cysteine according to the present invention, preferably the main L-C is Stein.

The number of lyophilisate according to the present invention, equal to from 160 to 170 mg, preferably contains 5·109-5·1011SOME of the specified at least one strain .animalis lactis. The number of lyophilisate according to the present invention, equal to from 160 to 170 mg, preferably contains 4,90 144 mg of the specified special form of L-cysteine according to the present invention.

For counting colony forming units (CFU) .animalis lactis may use any means it deems appropriate specialist in the art, for example, the method of counting bacteria described in Grand et al. 2003 (Eur. Food Res. Technol. 217: 90-92). It is preferable to apply the method described in S.N.Thitaram, G.R.Siragusa and A.Hinton, 2005 (Jr Letters In Applied Microbiology, Volume 41, Page 355 - October 2005; "Bifidobacterium-selective isolation and enumeration from chicken caeca by a modified oligosaccharide antibiotic-selective agar medium").

To determine the number of cysteine contained in the liquid, you can use any method that is deemed appropriate specialist in the art, for example, gas chromatography coupled with mass spectrometry or high performance liquid chromatography with fluorimetric detection. It is also possible to determine the quantity of cysteine using analyzer amino acids like L-8800 High Speed Amino Acid Analyzer (Hitachi High Technologies).

It is advisable to inoculate the agreement is but the present invention contain not more than 0.5 wt.% yeast the yeast extract or yeast autolysate. The inoculum according to the present invention preferably contains not more than 0.5 wt.% yeast, yeast extract or yeast autolysate. More preferably the specified inoculum contains no yeast, yeast extract or yeast autolysate.

An advantage of the present invention is the fact that it allows you to get very active inoculum .animalis lactis, forming a stable in time the population .animalis lactis (during storage of the fermented milk product at a temperature of refrigerator) without the use of preservative in the inoculum and add it to the milk substrate or in fermented dairy product. Thanks to the application of the present invention population .animalis lactis in fermented dairy product is especially beneficial physiological and/or metabolic condition, which ensures the maintenance of its high abundance.

The inoculum according to the present invention contains at least one strain .animalis lactis, for example, strain I-2494 or strain VV® Chr.Hansen. Of course, the inoculum according to the present invention may contain several strains .animalis lactis, for example, strain 1-2494 and strain VV® Chr.Hansen.

In addition, the inoculum according to the present invention may contain at IU is e one another strain of lactic acid bacteria, other than .animalis lactis. The specified at least one other strain of dairy bacteria may be part of a physical structure different from the structure containing cells .animalis lactis. Specified different physical structure, for example, may be a solid structure like a frozen pellets or freeze-dried, mixed with the granules and/or lyophilisate .animalis lactis and a special form of L-cysteine according to the present invention or located separately from the latter and forming part of the set. The specified at least one other strain of lactic acid bacteria may, for example be in the form of frozen pellets or freeze-dried, non-granular or lyophilisate (liofilizatow)containing .animalis lactis and a special form of cysteine according to the present invention. The specified at least one other strain of lactic acid bacteria may represent a strain of Bifidobacterium or strain of a different kind of bacteria. The strain of another kind of bacteria can be a strain with probiotic activity, for example, at least one strain selected from Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei. It can also be a technological strain, including S.thermophilus and/or L.bulgaricus. The inoculum according to the present invention mainly contains, in addition to pellets(granules) and/or lio is ilizat (liofilizatow) .animalis lactis and a special form of L-cysteine, the inoculate S.thermophilus and L.bulgaricus, for example, in the form of frozen pellets or granules and/or lyophilisate or liofilizatow adapted for direct seeding. The inoculum according to the present invention preferably contains several strains other than .animalis lactis, for example, at least two, at least three or at least four strains other than .animalis lactis. These other strains preferably represent at least one strain S.thermophilus (for example, strain I-1630 available in CNCM) and at least two strains L.bulgaricus (for example, strains of I-1632 and I-1519 available in CNCM). This inoculum may be a mixture or combination of frozen granules and/or at least one lyophilisate containing .animalis lactis and a special form of L-cysteine according to the present invention, and frozen granules and/or at least one lyophilisate containing S.thermophilus and/or L.bulgaricus. This system is specially adapted to direct seeding in the dairy substrate with the aim of turning it into fermented dairy product with a high probiotic value of preserving the past during storage in the refrigerator, by symbiotic lactic acid fermentation under the action of S.thermophilus - L.bulgaricus and fermentation under the action of .animalis lactis.

This application also applies separately to one with echannel granule according to the present invention as such and separately to mixed lyophilisate according to the present invention. As shown and described above and/or below in the Examples section, one mixed granule or mixed lyophilisate according to the present invention contains at least one strain .animalis lactis and a special form of L-cysteine according to the present invention in the dose and/or relationships. Preferably such a granule or a lyophilisate not contain cysteine in strongly acidic form like the hydrochloride of L-cysteine. Thus, the present invention relates to frozen granule containing at least one strain .animalis lactis and L-cysteine that is not associated with other amino acids, peptide bond, with the specified L-cysteine has such a shape that the pH of the solution resulting from defrosting of said pellets or granules is not less than 4, usually more than 5. The present application also relates to the lyophilisate containing at least one strain .animalis lactis and L-cysteine that is not associated with other amino acids, peptide bond, with the specified L-cysteine has such a shape that the pH of a solution obtained by dissolving the above freeze-dried or liofilizatow in the amount of 1 to 2 g of freeze-dried to 8-10 ml of H2O is at least 4, usually more than 5. The pH of the pellets or freeze-dried (the pH of a solution obtained by thawing the pellets or dissolving the lyophilisate), is less than 8.

This is the invention also relates to methods of applying the inoculum according to the present invention. In particular, the present invention relates to its use in lactic acid fermentation, and, more specifically, for use in the manufacture of fermented milk products intended for human consumption.

Thus, the present invention refers to any way in which apply at least one inoculate, and/or at least one granule or at least one lyophilisate according to the present invention.

In particular, an object of the present invention is a method of promoting the growth and/or metabolism of .animalis lactis in milk substrate, which is applied at least one inoculate, and/or at least one granule, and/or at least one lyophilisate according to the present invention. The advantage of this method of stimulation is high efficiency stimulation .animalis lactis. As shown above and illustrated below, the method of stimulation according to the present invention allows to obtain a population .animalis lactis, metabolism and/or growth which ensures the preservation of living cells .animalis lactis over time. To implement this method, the specified inoculate contribute in doses and/or the relationships mentioned above and/or described in the following examples and drawings. These inoculate, and/or granules and/or lyophilisate according to the present izopet is to mainly make in quantities when the milk substrate contains from 5 to 20 mg of cysteine per 1 liter of milk substrate, preferably 6-15 mg/l, more preferably 7 to 11 mg/l; 109up to 1011SOME .animalis lactis on 1 l of the substrate, preferably of 0.4·1010-1·1011SOME B.animalis lactis, more preferably of 0.4·1010to 1.2·1010SOME .animalis lactis, most preferably of 0.4·1010-1·1010SOME .animalis lactis. Inoculated substrate is preferably supported in the temperature and atmospheric conditions conducive to metabolism .animalis lactis, for example, at a temperature of from 37°C to 42°C in a closed vessel.

The object of the present invention is also a method of obtaining a fermented milk product, preferably a milk product obtained by lactic acid fermentation, and in the latter case, in addition to the fermentation process carried out .animalis lactis, is at least one process symbiotic fermentation under the action of S.thermophilus-L.bulgaricus. The method according to the present invention provides the use of at least one of inoculum, and/or at least one of granules, and/or at least one lyophilisate according to the present invention Obtained fermented milk product is a fermented dairy product with a high probiotic value, p is because it contains .animalis lactis in large quantities and retains a high content of the latter during storage in the refrigerator. This method has the advantage of very efficient activation and/or stimulation .animalis lactis. In addition, its advantage is the absence of negative effects on the symbiosis of S.thermophilus and L.bulgahcus. As shown above and illustrated below, the method of stimulation according to the present invention allows to obtain a population .animalis lactis, metabolism and/or growth which ensures the preservation of living cells .animalis lactis over time. To implement this method, the specified inoculate contribute in doses and/or the relationships mentioned above and/or described in the following examples and drawings. These inoculate, and/or granules and/or lyophilisate according to the present invention mainly contribute in quantities at which the milk substrate contains from 5 to 20 mg of cysteine per 1 liter of milk substrate, preferably 6-15 mg/l, more preferably 7 to 11 mg/l; from 10° to 1011SOME .animalis lactis on 1 l of the substrate, preferably of 0.4·1010-1·1011SOME .animalis lactis, more preferably of 0.4·1010to 1.2·1010SOME .animalis lactis, most preferably of 0.4·1010-1,1·1010SOME .animalis lactis.

Examples of doses that are suitable for depositing in 1 l of milk substrate, are dose mixed granule according to the present invention, 0.05 to 1 g, for example, approximately 0.2 g

Inoculated substrate preferably the support is more in the atmospheric and temperature conditions, favorable for the metabolism .animalis lactis, for example, at a temperature of from 37°C to 42°C in a closed vessel. The specified at least one strain .animalis lactis plays the role of a probiotic strain. In view of this requirement, you must provide the lactic acid fermentation of milk substrate indicated at least one strain .animalis lactis. Therefore, the specified at least one strain B.animalis lactis are not a simple additive to the substrate or the product is not as simple additives, without which the conditions to carry out lactic acid fermentation, is not supported.

A method of obtaining a fermented dairy product mainly can include applying at least one strain different from the at least one strain .animalis lactis. Specified strain is preferably a lactic acid bacteria strain. The last function, in particular, is the implementation of lactic acid fermentation. So it inoculant on the milk substrate in such a way as to provide lactic acid fermentation, i.e. at the moment of time and in quantities that ensure the effective implementation of lactic acid fermentation. The method of fermentation according to the present invention, therefore, conforms to see what shanna fermentation, carry out the specified at least one strain of lactic acid bacteria and the specified at least one strain .animalis lactis, and is not the usual process of lactic fermentation, which .animalis lactis add as usual additive in the absence of detectable enzymatic activity. Specified at least one strain of lactic acid bacteria and at least one strain B.animalis lactis, thus, are brought together in the dairy substrate. According to a preferred implementation variant of the present invention is a method of obtaining may also not be a two-step: adding the specified at least one strain .animalis lactis and possibly other strains of lactic acid bacteria can be performed simultaneously or at least one stage of the technological process, namely at the beginning of the process, so there is no need to wait and monitoring the achievement of a certain stage of fermentation, carried out by one of the strains, with the purpose of making decisions about adding at least one strain.

Fermentation of the milk substrate is carried out at a temperature favorable to the applied strains, which typically ranges from 36°C to 42°C. This process is usually carried out to achieve a pH value equal to 4,2-4,8.

A method of obtaining a fermented dairy product according to the present invention may also include at least one stage, for example, at least one phase selected from the known stages of the receipt of the fermented dairy product. For example, it is possible to implement at least one stage selected from the following stages:

- preparation of a dairy substrate;

- heat treatment of the milk substrate (equivalent to at least pasteurization, for example, at 95°C for 5 min;

at least one inoculation of the milk substrate;

- implement at least one process of lactic fermentation inoculated substrate under conditions favorable for the inoculated strains (usually to achieve a pH value equal to 4,2-4,8);

possible flavorings, fruits, sweeteners, dyes or preservatives;

possible mixing, including after cooling;

packaging fermented product, for example, in sealed containers;

- storage of the fermented milk product in a refrigerated environment.

The inoculum according to the present invention allows to obtain the population of the specified at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the amount of which at the end of fermentation is at least 5·107CFU per gram of fermented product, preferably at least 108 CFU per gram of fermented product. It should be noted that the advantage of inoculum according to the present invention is that it allows you to get the population of the specified at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the amount of which does not fall below 5·107CFU per gram of fermented product after at least 20 days, preferably after at least 30 days of storage of the specified fermented product at a temperature of from 4 to 11°C, preferably from 4 to 10°C, more preferably from 4 to 9°C. it Should also be noted that a particular advantage of inoculum according to the present invention is that it allows you to get the population of the specified at least one strain .animalis lactis (or all applied strains .animalis lactis, if there are several), the amount of which does not fall below 108CFU per gram of fermented product after at least 20 days, preferably after at least 30 days of storage of the specified fermented product at a temperature of from 4 to 11°C, preferably from 4 to 10°C, more preferably from 4 to 9°C (see Fig and 9, showing the conservation of populations B.animalis lactis in fermented milk obtained by the use of inoculum agreement is but the present invention (S.thermophilus - L.bulgaricus + .animalis lactis) no expiration at least 30 days of storage of the specified milk at 10°C compared with the control samples of fermented milk, population size which falls sharply beginning with the 21st day).

In this application under the weight of the fermented dairy product understand the mass fraction of milk for the specified product.

The present application also relates to a fermented dairy product as such. This product is different population sizes .animalis lactis, which is higher than the population sizes .animalis lactis in the products known in the prior art, even after 20 days, preferably after 30 days of storage of the specified fermented product at a temperature of from 4 to 11°C, preferably from 4 to 10°C, more preferably from 4 to 9°C. Therefore, the composition of this product is significantly different from the products known in the prior art. Thus, the present invention relates to a fermented milk product obtained by lactic acid fermentation of milk substrate using at least one of inoculum according to the present invention to achieve pH values, usually lying between a 4.2 and 4.8. The specified inoculate preferably has the form of mixed frozen granules and/or mixed lyophilisate according to this the mu invention (inoculum, in which a special form of L-cysteine and microorganism .animalis lactis is part of the same physical structure). This inoculum mainly may also contain at least one strain S.thermophilus and/or at least one strain L.bulgaricus.

In this application the reduction CNCM means the National collection of microorganism cultures (CNCM, Institut Pasteur, 25, rue du Docteur Roux, F-75724 PARIS CEDEX 15). The strains stored in the CNCM, have a number starting with the letter I, for example, "I-2494" (strain .animalis lactis), " I-1630 (S.thermophilus strains), I-1632" or "I-1519 (strains L.bulgaricus).

The term "containing", which is synonymous with the term "comprising"is an open term and does not exclude the presence of one or more elements, one or more parts or additional stages in ways that are not marked explicitly, while the term "consisting of" is a closed term that excludes the presence of any other additional elements, stages or components that are not marked explicitly. The term "consisting essentially of" is partially open term that does not exclude the presence of one or more elements, one or more parts or more stages, provided that these elements, components, or additional stages do not affect the basic characteristic of the invention. Therefore, the term "containing" or "including", "include" or "includes" encompasses the terms "consisting of" or "consist of"and " consisting essentially of".

The contents of all documents, publications or publications cited in this application, is hereby incorporated into this description by reference.

The following examples are given solely for illustrative purposes and in no way limit the present invention.

Examples

Example 1. Cysteine-HCl is unsuitable for use in manufacturing composite pellets, together containing cysteine and B. animalis lactis.

Materials and methods:

In this example followed the scheme of manufacture of the granules, as shown in figure 2.

In this example, we used the following probiotic strains:

- Bifidobacterium animalis lactis strain W®, commercially available from Chr. Hansen A/S, Hoersholm (Hoersholm, Denmark;

- Bifidobacterium animalis lactis strain I-2494, available at CNCM (date of receipt at the CNCM on June 20, 2000). Cm. publication WO 02/02800 from 4 July 2001 (Compagnie Gervais Danone) and national and regional counterparts specified applications, in particular in respect of analogues of the specified application to the United States should make reference to U.S. patent No. 7008785 B2. If necessary, the strains before using thawed.

Used cysteine represented cysteine-HCl, which is th is usually used as an additive cysteine due to its very good solubility (approximately 110 g per 100 g of H 2O at 20°C). Cysteine-HCl was a monohydrate hydrochloride cysteine (or monohydrate monohydrochloride (R)-2-amino-3-mercaptopropionic acid). The formula of this cysteine C3H7NO2S·HCl·H2O (or L-cysteine HCl·H2O). CAS-like cysteine 7048-04-6; its molar mass is 175,63. Cysteine-HCl can be purchased in the company Ajinomoto Aminoscience LLC, Raleigh NC 27610, USA.

A method of producing granules (see figure 2):

50 g of each strain Century animalis lactis was mixed with:

- 15 g cysteine-HCl at a concentration of recent 325 g/l (mixed granules G4I Cys-HCl), either:

to 15 g of water (control pellets I-2494 T and W T).

The mixture was obtained by stirring for better mixing with a solution of cysteine-HCl (or water in the case of the control samples).

The formation of the frozen pellet was performed by infusion of a mixture dropwise in liquid nitrogen. The device, which gave a mixture, which consisted of several needles syringe connected by tubing with a peristaltic pump, was provided by the falling of drops formed at the exit of the needle, insulated container filled with liquid nitrogen. In this way received frozen granules probiotic microorganism G4I Cys-HCl (granules .animalis lactis 1-2494 or W®containing cysteine-HCl) and control granules containing water instead of cysteine-HCl.

Preparation of milk the mixtures (dairy substrates).

The formula consisted of the following ingredients: skim milk with a fat content of 0%, cream with 40% fat and skim milk powder with a protein content of 33%. At first, all the ingredients were combined together to obtain standardized values of protein concentration, equal to 4.4%, fat content, equal to 3.5%, and the content of dry substances, 15.8%, by stirring the mixture for 60 minutes at about 750 rpm using a stirrer HEIDOLPH® with the aim of hydration of proteins. The control of homogeneity was performed using an IR detector MILKOSAN FT 120® FOSS®. The following is an example of the amounts of each ingredient needed to obtain the target characteristics of milk.

Table 1
Ingredientsin %
Skim milk, 0% IRN.87,5
Cream, 40% of IRN.8,7
Protein skim milk, 33%the 3.8
Only100

The milk is then heated at a temperature from 50°C to 60°C for maximally complete melting of fat globules. Upon reaching a predetermined temperature 10 litres of milk is homogenized in the installation MICROFLUIDIZIER®. Similar processing was provided by the splitting of fat globules in the transmittance of milk through the capillaries of the lattice under pressure of 350 bar. Prepare a water bath company MEMMERT® and tuned it to 103°C. the Milk was placed in 8 bottles of 1 liter; the amount of milk that is poured into each bottle, accurately weighed. The bottle was immersed to the bottom of the bottle in a water bath at a temperature of 103°C for 35 min, then in the same bath at a temperature of 95°C for 10 minutes, the Bottle was cooled in a bath with running cold water, then kept in a refrigerator at 4°C for 12-24 h before use.

Inoculation infant formula:

Milk bottles were removed from the refrigerator for 45 min prior to inoculation with microorganisms and placed in a water bath at a temperature of fermentation (37°C). Used four different versions of inoculation of infant formula:

the pellets I-2494 G41 Cys-HCl;

control pellets I-2494 T;

granules W G41 Cys-HCl;

control pellets BB 12 So

For each inoculation, the number of seed inoculum was 0.2 g per 1 liter of formula. The bottle was again immersed in a water bath (37°C) and monitored the change in pH over time (by setting CINAC® YSEBAERT®).

Results

As for .animalis lactis I-2494, and .animalis lactis BB12® watched the deterioration of the physiological state probity is due to the presence of cysteine-HCl granules (physiological time = time, necessary to ensure that the bacteria reduced the pH by 0.08 pH units). Describes the results shown in figure 3 (I-2494 G4I compared to I-2494 T). Adding cysteine-HCl had a strong effect on the activity of the probiotic .animalis lactis (activity was increased). Cysteine-HCl was unsatisfactory source of cysteine for the manufacture of granules of the combined composition containing .animalis lactis and cysteine in single grains (granules) and intended for the manufacture of fermented dairy products with a high probiotic value. "Combined" option implementation (granules, together containing .animalis lactis and cysteine) can be implemented using cysteine-HCl, since the latter makes probiotic microorganism incapable of normal metabolic activity.

From the above results show that the pH of the solution of cysteine-HCl is less than 3.5; in particular, the pH of the solution 1 g cysteine-HCl in 100 ml of H2O is 1.5-2.0, and that probiotic .animalis lactis is very sensitive to low pH even at lower contact times. Thus, the challenge of finding the source of cysteine, digestible .animalis lactis, fit for human consumption and having in solution a pH value that is not stressful for .animalis lactis.

Example 2. The selection of the preferred L-cysteine allows to achieve a very effective "is umestnyh" granules, despite the problems associated with low solubility such form cysteine.

Performed the sequence of operations similar to that described in example 1, except that instead of cysteine-HCl was chosen basically L-cysteine (L-cysteine, C3H7NO2S, CAS 52-90-4, molar mass 121,16).

Basically L-cysteine can be obtained, for example, the company Ajinomoto Aminoscience LLC, Raleigh NC 27610, USA. The solubility of the basic cysteine at 20°C is 16 g per 100 g of N2O. the pH of a solution containing 1,15 g main cysteine in 50 ml of H2O is from 4.5 to 5.5.

Preparing the mother liquor primary cysteine concentration of 160 g/l and mixed 3 g of the specified stock solution with 10 g of the microorganism Century animalis lactis (B. animalis lactis strain I-2494 or W®). Of the mixture has been frozen pellets (1-2494 G4I Cys-base and W G4I Cys-base), as described above in example 1. Control pellets I-2494 T and W T was obtained using 3 g of water instead of the cysteine. Scheme for these pellets is shown in figure 4.

Performed four surgeries inoculation, at each of which the number of seed inoculum was 0.2 g/l, as described above in example 1.

Thus, the number of primary cysteine in the granules was G4I(160×3)/(3+10)=36,92 mg primary cysteine per gram of frozen granules G4I. In this experiment, the content of the Century. animalis lactis was approximately 2·1010CFU per gram of frozen granules G4I or G4 (I-2494 or W). The number of cysteine C3H7NO2S per unit of size of the colonies of the microorganism .animalis lactis was 1,85 mg per one billion CFU. The mass of the main cysteine added to milk mixture to granules G4I inoculated in an amount of 0.2 g/l, 0.2×36,92=7,384 mg of cysteine per litre milk mixture.

Results incubation of the mixtures at 37°C show that the selection of the preferred cysteine greatly to avoid deterioration in the physiological condition used probiotic microorganism. These results are shown in figure 5 (strain I-2494).

Combined variant of implementation of granules combined with the cysteine" ((granules containing .animalis lactis and cysteine) can be implemented using the main L-cysteine, as in this case .animalis lactis maintains the normal metabolic activity.

Example 3. Comparison of the characteristics obtained for the combined system G4I Cys-base, the combined system G4 (including separate granules cysteine and granules probiotic microorganism), and a control system (granules of T that do not contain cysteine).

In this example, the used microorganism granules .animalis lactis obtained according to example 2, namely:

- granules is 12 T (control granules containing .animalis lactis BB12® and water);

- granules BB12 G4I (granules combined composition containing .animalis lactis BB12® and is basically L-cysteine in an amount of 1.85 mg L-cysteine C3H7NO2's one billion CFU probiotic microorganism);

- granules-I-2494 T (control granules containing .animalis lactis I-2494 and water);

- granules-I-2494 G4I (granules combined composition containing .animalis lactis I-2494 and basically L-cysteine in an amount of 1.85 mg L-cysteine C3H7NO2's one billion CFU probiotic microorganism).

In the present example also used technological microorganisms, which represented the microorganisms S.thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. Used technological microorganisms was a mixture of three technological strains, namely strain I-1630 S.thermophilus and strains I-1632 and I-1519 L.bulgaricus (strains available at CNCM). Technological microorganisms was made in the form of frozen granules and inoculable in milk mixture in an amount of about 1·106SOME S.thermophilus I-1630 and about 2.7-105SOME L.bulgaricus (approximately 5% strain I-1632 and 95% of the strain I-1519) per gram of the milk substrate.

Prepared six infant formula, as described in example 1, which was inoculable as follows:

the mixture W T and a mixture of I-2494 T (control mix) - inoculable mixture g is zero technological microorganisms and granules W T or 1-2494 m, respectively (granules, containing probiotic .animalis lactis and not containing cysteine);

the mixture W G4 and a mixture of I-2494 G4 (mixture "combined method") was inoculable similar to the control, except that they also added frozen pellets cysteine;

the mixture W G4I and a mixture of I-2494 G4I: (combined mixture composition) was inoculable mixture of granules of technological microorganisms and granules W G4I or I-2494 G4I respectively (granules .animalis lactis containing cysteine in the form of a joint staff).

For each inoculation, the number of seed inoculum was 0.5 g of a mixture of granules per 1 liter (0.3 g/l of technological microorganisms and 0.2 g/l of frozen granules probiotic microorganism .animalis lactis).

The mixture G4 granules cysteine was introduced so that the concentration of cysteine in the mixture was 7.5 mg/l, i.e. in the mix were a number of cysteine, comparable to that made with granules G4I (the calculated value is 7,384 mg/l cysteine; see example 2 above). Six inoculated mixtures were placed in incubation at 37°C as described in example 1. The bottle was cooled and kept at 20°C. Then the clot was removed from the bottles by hand. The clot was Packed up into the cups with a volume of 125 ml, which was sealed by thermo-sealing machine DNV-100-25 PPV-A® company FESTO®. The products were stored in a cold chamber at 10°C during the whole time the experimental is the same.

When storing milk product at 10°C during fermentation was measured pH and watched the development of biomass (CFU/ml depending on the elapsed time in days). The results are shown in Fig.6-9.

The results for strain W .animalis lactis was sudeste close to the results for strain I-2494. From Fig.6 and 7 (the pH change depending on the time in hours) shows that the presence of cysteine causes a change in the Vmax values compared to the control mixture. Thus, it is possible to notice the effect of cysteine on the kinetics of the formation of acids under the action of .animalis lactis. It is also seen that the mixture G4I (basically L-cysteine, "entered" into granules .animalis lactis) is characterized by the kinetics, is essentially identical to the kinetics of the mixture G4 (primary granules of L-cysteine separately from granules .animalis lactis). This indicates the advantage of the introduction of the activator growth. In the following tables 2 and 3 presents the values of some kinetic parameters of formation of the acids shown in Fig.6 and 7.

On Fig and 9 illustrate the dynamics of the population of cells of probiotic microorganism .animalis lactis strain W (Fig) and strain I-2494 (Fig.9) during storage of the fermented milk product (symbiosis S.thermophilus - L.bulgaricus, in addition to .animalis lactis) at 10°C. Data curves of the cell population is podtverjdayut results who observed when studying the kinetics of the formation of acids, presented in Fig.6 and 7. The presence of L-cysteine significantly stimulates cell population of probiotic; integration of basic L-cysteine for each of the two strains .animalis lactis was performed successfully.

So, when compared with the "combined" system G4 (granules probiotic microorganism .animalis lactis + individual granules of the main L-cysteine) granules G4I "joint structure"containing basic L-cysteine, are equally effective: they allow symbiotic lactic fermentation (S.thermophilus + L.bulgaricus)providing at least the same high quality and grow the population of cells of probiotic microorganism .animalis lactis almost comparable size, as long-lasting over time during storage of fermented milk. Thus, the selection of the preferred L-cysteine has enabled us to achieve successful integration of growth activator specific for .animalis lactis.

Granules G4I United composition containing probiotic microorganism .animalis lactis and is basically L-cysteine, and "combined" granules G4 (primary granules of L-cysteine and separately granules probiotic), are much more effective than a control system that does not contain cysteine (granules T): they allow auth to implement symbiotic fermented milk fermentation (S.thermophilus + L.bulgaricus), providing at least equally high quality and allows to obtain a population of cells of probiotic microorganism .animalis lactis significantly larger, much longer persisting over time during storage of fermented milk.

1. The inoculate, specially adapted for direct inoculation of the milk substrate with at least one strain of Bifidobacterium animalis ssp.lactis for the conversion of the specified dairy substrate fermented milk product suitable for human consumption, and the specified inoculum contains a mixture or in combination:
cysteine that is not associated with other amino acids, peptide bond, and
at least one strain .animalis lactis, thus:
these cysteine and at least one strain .animalis lactis contain or have the form of at least one frozen granules and/or at least one lyophilisate;
specified cysteine is a basically L-cysteine formula HSCH2CH(NH2)CO2H, whereupon the pH of the solution obtained after
thawing of the specified at least one of pellets and/or
dissolution of the specified at least one lyophilisate in respect of 1 to 2 g of freeze-dried to 8-10 ml of H2O is not less than 4;
specified cysteine contained in the said inoculum in a quantity are what you eat from 1 g to 1·10 14KOE specified at least one strain .animalis lactis or all used strains B.animalis lactis, if there are few, up to 1 g 3.5·1010SOME of the specified at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are few.

2. The inoculum according to claim 1, characterized in that the cysteine is an L-cysteine, the presence of which in the solution provides a pH greater than 4.

3. The inoculum according to any one of claims 1 to 2, characterized in that it contains at least one frozen grains and/or at least one lyophilisate, simultaneously containing:
all of this cysteine or part of cells and the specified at least one strain B.animalis lactis.

4. The inoculum according to any one of claims 1 to 2, characterized in that it contains either at least two frozen granules, or at least two different freeze-dried, or at least one frozen granule and at least one lyophilisate, and in each of these pairs of physical forms one of the forms contains all the specified cysteine or its part and does not contain cells B.animalis lactis, and the other form contains cells B.animalis lactis and does not contain cysteine.

5. The inoculum according to any one of claims 1 to 2, characterized in that the cysteine is present in the specified inoculum in a quantity amounting to:
from 0.01 mg per one billion TO the specified at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are several
up to 30 mg per one billion CFU of the indicated at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are few.

6. The inoculum according to any one of claims 1 to 2, characterized in that the cysteine is present in the specified inoculum in a quantity of 1 g 0.2·1014KOE specified at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are few, up to 1 g 10·1010KOE specified at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are few.

7. The inoculum according to any one of claims 1 to 2, characterized in that the cysteine contained in the said inoculum in a quantity amounting to:
from 0.05 mg to one billion CFU of the indicated at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are several
up to 10 mg per one billion CFU of the indicated at least one strain B.animalis lactis or all used strains B.animalis lactis, if there are few.

8. The inoculum according to any one of claims 1 to 2, characterized in that the cysteine contained in the said inoculum in a quantity of 4,90 up to 144 mg / 5·109-5·1011KOE specified at least one strain B.animalis lactis.

9. The inoculum according to any one of claims 1 to 2, characterized in that 1 g of these is renal with solids content, equal 16-17%, contains 5·109-5·1011KOE specified at least one strain B.animalis lactis.

10. The inoculum according to any one of claims 1 to 2, characterized in that 1 g of these granules with a solids content equal to 16-17%, contains from 4,90 up to 144 mg of the indicated cysteine.

11. The inoculum according to any one of claims 1 to 2, characterized in that the strain B.animalis lactis is a strain 1-2494.

12. The inoculum according to any one of claims 1 to 2, characterized in that it also contains at least one other strain of lactic acid bacteria other than B.animalis lactis.

13. The inoculum according to item 12, wherein the specified at least one other strain of lactic acid bacteria contained in the physical structure different from the structure containing cells B.animalis lactis.

14. The inoculum according to item 12, wherein the specified at least one other strain of lactic acid bacteria comprises at least one strain S.thermophilus and at least one strain L.bulgaricus.

15. The frozen pellet, obtained from the supernatant according to any one of claims 1 to 14, which contains at least one strain B.animalis lactis and L-cysteine that is not associated with other amino acids, peptide bond, with the specified L-cysteine has such a shape that the pH of the solution obtained by thawing the specified at least one of the granules is not less than 4.

16. The lyophilisate, suitable for use in ka is este inoculum according to any one of claims 1 to 14 or obtained from this inoculum, which contains at least one strain B.animalis lactis and L-cysteine that is not associated with other amino acids, peptide bond, with the specified L-cysteine has such a shape that the pH of a solution obtained by dissolving the specified at least one lyophilisate in a quantity of 1 to 2 g of freeze-dried to 8-10 ml of H2O is not less than 4.

17. The method of promoting the growth and/or metabolism of B.animalis lactis in milk substrate, including:
- inoculation of the milk substrate at least one inoculate according to any one of claims 1 to 14, and/or at least one pellet in 15, and/or at least one lyophilisate for P16;
the content of the inoculated substrate in the temperature and atmospheric conditions conducive to metabolism B.animalis lactis.

18. The method according to 17, characterized in that the inoculum added to the milk substrate in an amount in which the content of L-cysteine is from 7 to 11 g per 1 l of milk substrate, and in which the number of cells B.animalis lactis is from 109up to 1011CFU per 1 liter of milk substrate.

19. A method of manufacturing a fermented dairy product with a high probiotic value, including:
inoculation of the milk substrate at least one inoculate according to any one of claims 1 to 14, and/or at least one pellet in 15, and/or less than the least one lyophilisate according to article 16;
fermentation specified substrate at least one specified inoculum.

20. The method according to claim 19, further including:
inoculation specified dairy substrate at least one strain of S.thermophilus and at least one strain of L.bulgaricus; and specified fermentation carry out the specified at least one inoculate and these strains S.thermophilus and L.bulgaricus.

21. The method according to PP or 20, characterized in that the inoculum added to the milk substrate in an amount in which the content of L-cysteine is from 7 to 11 g per 1 l of milk substrate, and in which the number of cells B.animalis lactis is from 109up to 1011CFU per 1 liter of milk substrate.



 

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4 tbl, 5 ex

FIELD: food industry.

SUBSTANCE: invention relates to food industry, in particular, to methods of production of high-protein dairy-and-vegetal food products of functional-purpose with soya application. The method envisages mixing defatted cow milk and a soya component with subsequent introduction of a starter, ripening of the produced mixture and introduction of additional recipe ingredients. One introduces the soya component represented by flour produced of insoluble soya remains containing: 9.4% of water, 28.1% of protein, 13.5% of lipids, 3.9% of mineral substances and 45.1% of carbohydrates in an amount of 10% of defatted milk weight. The soya component is produced by way of two-stage dehydration till granules moisture content is 9.4-10.0% and the granules flouring. One introduces the starter based on symbiosis of thermophilic streptococcus and acidophilous bacterium (Str. thermophilus +L. acidophilus) at a ratio of (1:1) with addition of a coagulant i.e. calcium chloride solution (CaCl2). One introduces additional ingredients represented by berries puree.

EFFECT: invention allows to produce high quality food products of functional purpose, with synbiotic properties, produced only from natural ingredients, with enhanced organoleptic indices and nutritive value.

4 cl, 2 tbl, 4 ex

FIELD: food industry.

SUBSTANCE: method involves standardisation of milk raw material represented by whole milk, dry skimmed milk, pasteurisation, cooling to the temperature of souring, YO-MTX 511 LYO ferment introduction, ripening, cooling, introduction of a filler represented by pumpkin puree, a sweetener represented by tagatose containing a sweetener with 20% of weight fraction of dry substances, introduction of Palsgaard 5913 stabiliser, homogenisation, freezing and hardening.

EFFECT: invention allows to improve quality of the product, its organoleptic characteristics, enhance nutritive value and impart synbiotic properties to it.

2 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: invention is related to a food product. The food emulsion contains: (a) whole fresh fruits and/or pieces thereof, (b) water composition that wholly or partly fills the spaces between whole fresh fruits and/or pieces thereof and containing: (i) solid substances selected from the group consisting of: sugars, sugar alcohols, sugar substitutes, fructo-oligosaccharides, polydextroses, ballast substances and mixtures thereof in a total amount of over 40 wt % per the water composition total amount, (ii) one or more galactomannans for the composition thickening under storage conditions due to bonding water from fruits and/or pieces thereof.

EFFECT: invention enables production of a food product containing fresh fruits that preserve their appearance and consistency for a long time.

16 cl, 1 dwg

FIELD: food industry.

SUBSTANCE: probiotic composition for infant food includes Bifidobacterium breve, undigestible saccharide A and undigestible saccharide B with a degree of polymerisation between 2 and 200, additionally contains Lactobacillus paracasei. The composition has a viscosity 1-6 MPa·s, a caloric content 10 and 250 kcal per 100 ml. The said composition is applied for food composition production for usage in a method for provision of infant food, for production of a composition for prophylactic and/or treatment of gastrointestinal diseases, immune diseases and/or endocrinopathy, for prophylactic and/or treatment of allergy, atopic dermatitis, eczema and/or infection.

EFFECT: invention ensures possibility to imitate infant indestinal flora at breastfeeding, optimal composition caloric content promotes to reduce diarrhea, immune diseases, endocrinopathy and allergy.

17 cl, 1 dwg, 4 tbl, 3 ex

FIELD: food industry.

SUBSTANCE: invention is related to dairy industry. The method envisages milk normalisation, homogenisation, warming the normalised mixture, introduction of an additive represented by "Lactobel ED" demineralised bifidogenic food concentrate containing (12.9±2)% of lactulose in an amount of 8%, stirring, homogenisation, filtration, pasteurisation of the produced mixture, cooling to the temperature of starting, addition of starter, fermentation at a temperature of 40-45°C during 4-5 hours till production of 75-90°T milk coagulum, cooling and packing. The starter is represented by a starter of thermophilic lactic streptococcus and acidophilous bacterium at a ratio of 4:2.

EFFECT: invention allows to enrich the drink with valuable proteins, lactulose and probiotic microorganisms and to produce a product with sinbiotic properties and an anticancer activity promoting suppression of pathogenic flora.

3 ex

FIELD: food industry.

SUBSTANCE: invention is related to dairy industry. The method envisages addition of wheat malt extract in an amount of 8% into normalised milk with fat content 1.5%, pasteurised at a temperature of 90 - 94°C, with maintenance during 15-20 sec. The produced milk-vegetable mixture is cooled to 20-25°C, a kefir fungi starter in an amount of 5% is added to it, the mixture is ripened at a temperature of 20-25°C during 8-12 hours, upon ripening completion the milk coagulum is stirred, cooled till temperature is 12-16°C; at this temperature the product is left to maturing during 9-13 hours. Then the product is additionally cooled to a temperature of no more than 8°C and packed.

EFFECT: invention allows to produce a beverage characterised by improved food and biological value.

1 tbl, 4 ex

FIELD: food industry.

SUBSTANCE: invention is related to dairy industry. The composition contains cow milk, vegetable oil, JTL Stabisol stabiliser, a starter containing Streptococcus thermophilus and Lactobacillus delbrueckii, a bulgaricus subspecies and B. bifidum and/or B. longum and/or B. adolescentis, phytocomponents, "Okeanol" or "Omeganol" biologically active supplement, Fibregum food fibres.

EFFECT: invention allows to increase nutritional and biological value of the product and to impart synbiotic properties to it.

3 cl, 3 tbl

FIELD: food industry.

SUBSTANCE: method involves whole camel milk normalisation by way of addition of common licorice water extract, homogenisation, chilling to starting temperature, introduction of a starter containing lactic acid streptococci. Then one proceeds with chilling to starting temperature, introduction of a starter containing lactic acid streptococci, Lactobacillus bulgaricus and yeast, ripening till acidity is 65-72°T, two-stage addition of whole camel milk after a 3 h interval, bottling and additional ripening during 12 h, chilling in refrigeration chamber to a temperature of 4°C and maturing and self aeration during 24 h.

EFFECT: invention enables enhancement of prophylactic properties of the drink, in particular - its immunomodulating, antibacterial and general tonic properties as well as extension of marketability period.

FIELD: food industry.

SUBSTANCE: method involves addition of cedar nut cake in an amount of 0.5% (preliminarily pulverised and added to cold milk pasteurised at a 92-95°C temperature with maintenance during 30 minutes and chilled to 20°C) to the ripened product prior to bottling combined with chilling and stirring, as well as addition of lactulose syrup in a quantity of 1.0%, preliminary pasteurised at a 75±2° temperature with maintenance during 15 min and chilled to 20°C.

EFFECT: high organoleptic indicators, stability, bifidogenic properties, increased biological and food value and storage ability.

2 tbl, 5 ex

FIELD: food industry.

SUBSTANCE: food composition contains a food component with a moisture activity in excess of 0.7 and a bakery product such as a biscuit or a cake. Content of fat in the bakery product is less than 10% of the baked bakery product weight, the protein to fat weight ratio being at least 0.8:1.

EFFECT: invention allows to improve visual and organoleptic recognition of bakery product pieces in liquid or moist food product.

9 cl, 4 dwg, 1 tbl, 4 ex

FIELD: food industry.

SUBSTANCE: invention is related to dairy industry. Amaranth seed flour in an amount of 1.5% of the used raw material weight is applied as a stabiliser for yoghurt.

EFFECT: invention allows to prevent yoghurt syneresis process and extend the range of natural stabilisers.

2 dwg, 3 tbl

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