RU2360956C2 - Method for kvass preparation - Google Patents

Abstract

FIELD: food products.

SUBSTANCE: kvass preparation method provides for multiplication of pure cultures of yeasts, its adding to kvass wort, its attenuation and blending with recipe components. After attenuation kvass is disengage from crust, pausterised, cooled down and bifidus bacteria or lactic acid bacillus previously grown up during 24-28 hours at 37°C in hydrolizate-soya medium till dilution not less than 10⁹ CFC are also added to it Bifidus bacteria or lactic acid bacillus are added together with culture medium in quantity 10-15% from final drink volume. Composition's peptic hydrolizate-soya medium is used as culture medium for bifidus bacteria or lactic acid bacillus. Soya-protein substrate - 75 g, water - 1000 ml, pepsin- 10 g, which is fermented during 4 hours at 37-38°C, filtered, sterilised and final pH 7.2-7.4 is set. Strain Bifidobacterium bifidum No 1 is used as bifidus bacteria (acquisition source - Research and Development Establishment of G. N. Gabrichevsky), Strain Lactobacillus acidophilus No NK-1 is used as lactic acid bacillus (acquisition source - Research and Development Establishment of G. N. Gabrichevsky).

EFFECT: simplified technology of kvass production and exception of need to add chemical preservatives into ready-made drink.

2 dwg, 1 tbl, 2 ex

Description

FIELD OF THE INVENTION

The invention relates to the food industry and biotechnology and can be used in the manufacture of beverages for medical purposes.

In particular, the invention is intended for the production of beverages combining the gustatory properties of the Russian national drink - kvass and the therapeutic and prophylactic properties of probiotics - preparations based on live microorganisms of the genera Bifidobacterium and Lactobacillus. At the same time, the kvass base acts as a “vector” ensuring the comfort of taking such a drink, and the probiotic component provides the formation of positive effects in relation to the microbiocenosis of the gastrointestinal tract of the consumer.

Thus, the inventive method is intended for use in enterprises of the food and biotechnological industries, solving the issues of production of functional food products.

State of the art

Historically, a large number of folk methods and recipes for the preparation of kvass are known, mainly based on the yeast fermentation of various food bases with the addition of sugars [1]. As a result of this, a specific taste, acidity and other advantages of Russian kvass are formed. However, the disadvantage of using such poorly controlled technologies is the non-standard nature of the resulting finished product, as well as the possibility of accumulation of extraneous microflora in it, potentially capable of adversely affecting the consumer’s gastrointestinal microbiocenosis.

In order to eliminate such shortcomings in the industrial production of kvass, the use of starter cultures from pure yeast cultures has been proposed, which makes it possible to obtain a standard high-quality drink with a small amount of extraneous microflora [2]. In the case of the production of bottled drinks, an additional introduction of chemical preservatives (lactic acid, sodium benzoate) allowed for use in kvass may be provided, which excludes the possibility of microbiological damage to the product during its long-term storage [3]. At the same time, such kvass does not have specific prophylactic or therapeutic properties, which does not allow its use as a functional food product.

To solve this problem, the introduction of pure cultures of lactic acid bacteria into the starter cultures has been proposed [4, 5], which makes it possible to give the produced beverage additional therapeutic and prophylactic properties. Moreover, the latter are mainly provided due to the presence of exoproducts of lactic acid bacteria in kvass that have a positive effect on the human body through an antagonistic effect against pathogens of intestinal and purulent-inflammatory infections, lowering serum cholesterol, antitumor and antiallergic effects, etc. [ 6].

In particular, a method for the production of kvass is known [4], which involves introducing a mixture of pure cultures of yeast and bacteria Streptococcus faecium 770 and Lactobacillus plantanum AN 11/16 into kvass wort, after which it is fermented to produce a finished drink. Another known method for preparing kvass [5] along with a pure yeast culture involves the use of bacteria Bifidobacterium longum CMPM B-2000 and / or Bifidobacterium bifidum CMPM B-3300, and / or Bifidobacterium bifidum CMPM B 3190 (in a mixture or monoculture). At the same time, not simultaneous but sequential fermentation of kvass wort is stipulated by two groups of microorganisms (yeast and bifidobacteria), the last of which are added to pre-prepared kvass in the form of starter culture in the amount of 3-5% by volume with the number of viable cells at least 10 ⁸ in 1 ml then carry out additional cultivation.

The reasons hindering the achievement of the desired result in this case can be reduced to two main points: 1) with the joint fermentation of kvass wort with yeast and any of the representatives of lactic acid microorganisms between them, it is possible to establish competitive or antagonistic interactions that adversely affect organoleptic and other properties of the finished drink; 2) the same reasons can prevent the multiplication and accumulation of lactic acid bacteria, including the sequential fermentation of kvass wort with yeast and lactic acid microorganisms.

A partial solution to this problem was proposed in the already mentioned method for preparing kvass [5], in accordance with one embodiment of which it is proposed to introduce a concentrate of bifidobacteria into ready-made and pasteurized kvass obtained after conventional yeast fermentation (prototype method). It is assumed that the concentrate of bifidobacteria will be obtained when they are cultured not on kvass wort of plant origin, but on a medium based on animal protein (e.g. milk and its derivatives), which is most suitable for propagation of this group of microorganisms. However, the intermediate product obtained with this in its organoleptic properties is very far from kvass, which before blending requires its transformation into a “concentrate” of bifidobacteria with a cell content of at least 10 ¹⁰ in 1 ml, for example, by centrifugation with separation of bacterial biomass and exoproducts, the last of which are removed and do not fall into the finished product. At the same time, it is the exoproducts of lactic acid bacteria that are attributed a significant proportion of the therapeutic effect of drinks made with their use (see above).

Thus, the main reason hindering the achievement of the desired result is a fundamental difference in the initial substrates for the production of kvass (plant-based with a predominance of sugars) and the cultivation of lactic acid bacteria (based on milk with a predominance of amino acids), which makes the production of “hybrid” drinks , combining the food and taste properties of kvass and the therapeutic and prophylactic properties of probiotics - preparations based on live microorganisms of the genera Bifidobacterium and Lactobacillus, is a difficult task.

Alternative approaches leading to the elimination of this drawback are not described in available sources of scientific and scientific-technical information. From the foregoing, it follows that the claimed method is not known from the prior art, i.e. is new.

SUMMARY OF THE INVENTION

The main task to be solved by the claimed method is aimed at simplifying the technology for the production of drinks that combine the food and taste properties of kvass and the therapeutic and prophylactic properties of probiotics - preparations based on live microorganisms of the genera Bifidobacterium and Lactobacillus. An additional problem solved by the implementation of the proposed method is the elimination of the need to add chemical preservatives to the finished drink.

The essence of the proposed method, formulated at the level of functional generalization and underlying it, is the following:

- as a trophic basis for the production of a concentrate of bifidobacteria or lactobacilli, subsequently introduced into kvass prepared using traditional technology by yeast fermentation, a hydrolyzate of vegetable, namely, soybean raw materials is used, which makes it possible to obtain an intermediate product for subsequent blending with organoleptic properties similar to kvass .

Thus, the goal is achieved in that when replacing an animal protein with a vegetable (soy) protein, it becomes possible to introduce live bacterial cells grown on it (for example, Bifidobacterium bifidum or Lactobacillus acidophilus) together with their exoproducts in an amount of 10-15% of the final volume drink without a significant change in its organoleptic, but with imparting therapeutic properties. An additional result achieved by the implementation of the proposed method is the effective conservation of the resulting beverage, which is also a consequence of the introduction of exoproducts Bifidobacterium and Lactobacillus with antibacterial properties.

Accordingly, when implementing the proposed method for the preparation of kvass, the characteristics of the actions, the order of their implementation and the conditions for implementation in comparison with the prototype method are presented as follows (table).

At the first stage, a food-taste kvass base is prepared.

To do this, in pre-purified water dilute the kvass wort concentrate to a solids content of 1.4 g in 100 g of wort. Wort prepared in this way is pasteurized at 78-80 ° C for 30-35 min, cooled to 28 ± 2 ° C, after which pre-prepared sugar syrup with a concentration of 60-65% is added to it to the amount provided by the recipe. Propagated pure yeast culture with a content of 5 * 10 ⁶ cells in 1 ml of yeast wiring set in kvass must in the amount of 3% of its volume. Fermentation is carried out at 30-35 ° C for 10-12 hours until the true solids content of the wort is reduced by 1 g in 100 ml of fermented wort. After fermentation, the kvass is separated from the yeast by settling in a fermentation tank at 5-7 ° C and pumped to the blending tank.

In parallel, one of the production strains of Bifidobacterium bifidum and / or Lactobacillus acidophilus is cultivated on a peptic hydrolyzate of soya raw materials.

To do this, soy protein hydrolyzate is initially prepared by diluting the dry soy protein substrate in distilled water at the rate of 75 g of powder per 1 liter of water. The resulting liquid is boiled for 5-10 minutes, cooled to 38-40 ° C, adjusted to pH 3.0 by adding 10.0% HCl (hydrochloric acid) and pepsin is added at a rate of 10 g / l, after which fermentation is carried out (hydrolysis) at 37-38 ° C for 4 hours. After fermentation is complete, the pH is reversed to 4.5 by adding 20% NaOH (sodium hydroxide). The resulting hydrolyzate is boiled for 15 minutes, filtered and diluted with distilled water in a ratio of 1: 1. After heating to 80 ° C, the substrate is combined with molten food agar to a final concentration of the latter of 0.075% and boiled for 15 minutes. After settling for 20 min, the precipitate is drained, the volume of the substrate is brought to the initial distilled water and filtered mechanically to release from undissolved particles. The prepared hydrolyzate-soybean medium is sterilized at a pressure of 0.5 MPa (t = + 112-115 ° C) for 30 minutes with preliminary heating with steam for 30 minutes. The pH of the finished medium is set in the range of 7.2-7.4.

In the hydrolyzate-soybean medium prepared by the described method, one of the strains of bifidobacteria or lactobacilli with proven therapeutic and prophylactic properties (for example, Bifidobacterium bifidum and / or Lactobacillus acidophilus) allowed for use in the food industry is added at the rate of 5-10% or 1-3% liquid culture to the volume of inoculated medium, respectively. Cultivation is carried out at 37-38 ° C for 24-48 hours (in the case of using bifidobacteria) or 24 hours (in the case of using lactobacilli).

This stage determines the main difference between the proposed technical solution and the prototype method, since the resulting intermediate product containing living cells of bifidobacteria or lactobacilli in a titer of 10 ⁹ -10 ¹⁰ CFU / ml, as well as their metabolic products, by their organoleptic properties It turns out to be extremely close to kvass, which allows it to be added to the kvass base without any additional processing or concentration and, thus, directly proceed to the stage of making the finished drink.

Accordingly, at the final stage, the finished drink is made by blending the kvass base and probiotic biomass grown on hydrolyzate-soy medium, the last of which is added at a rate of 10-15% of the final volume, so that the final content of living cells of bifidobacteria or lactobacilli is at least 10 ⁸ CFU / ml. Kvass is cooled to 4 ° C and consumed without subsequent fermentation.

Obtained by this option, a fundamentally new product not only does not lose the inherent flavoring characteristics of kvass, but also has all the therapeutic and prophylactic properties of probiotic preparations, as it contains not only live bifidobacteria or lactobacilli, but also a full range of metabolic products, which account for a significant proportion of their therapeutic effect (in contrast to the prototype method).

In addition, an additional result achieved by the method is the elimination of the need for chemical preservatives, the function of which is performed by antibacterial substances produced in the culture medium by bifidobacteria or lactobacilli.

The ability to obtain a technical result when performing the indicated actions in the indicated ranges of values is determined by the following set of cause-and-effect relationships:

1. For the first time it was established that the product obtained by culturing bifidobacteria or lactobacilli on hydrolyzate-soybean medium (GSS) has organoleptic properties similar to kvass, which allows it to be added to the gustatory base without additional purification and concentration.

Evidence of this was obtained during the tasting with the involvement of 60 volunteers from among potential consumers of the drink being developed, who were offered its options with the content of bifidobacteria or lactobacilli on GSS 1%, 2.5%, 5%, 10% and 25% of the final volume. Moreover, as the presence of the bacterial additive increases, the assessment of the drink as “sweet” progressively decreased, so that the transition from the zone of “average” to the zone of “weak” values occurred at the calculated level of 17.0 ± 2.0% of the relative content of bifidobacteria or lactobacilli by GSS. In contrast, the assessment of the drink as “acidic” increased with exceeding the “strongly acidic” threshold at a calculated level of 19 ± 2.2% of the relative content of bifidobacteria or lactobacilli. Against this background, although the assessment according to the criterion of “specific kvass taste” had a slight tendency to decrease, in comparison with the organoleptic indicators given above it turned out to be more stable and did not go beyond the permissible limits in any of the proposed blending recipes.

Based on the data obtained, the level of 15% of the final volume was established as the upper limit of the relative content of bifidobacteria or lactobacilli on GSS in the composition of the proposed drink, which allows us to maintain its consumer rating as a “sweet and sour drink with a specific kvass taste,” corresponding to the established view about Russian kvass.

2. It has been shown for the first time that the addition of bifidobacteria or lactobacilli to the KVS base on GSS containing the metabolic products of these microorganisms allows the product to be preserved equivalent to the level achieved using chemical preservatives.

Evidence of this was obtained by studying the bactericidal level of the drinks on offer with respect to Escherichia coli K12, used as an example of a sanitary-indicative microorganism of the intestinal group bacteria. Moreover, as a necessary and sufficient level of preservatives in the drink, it was considered as such in accordance with the TU and TI of Zhivaya Voda OJSC for Old Russian kvass, achieved by the presence of 0.0177% sodium benzoate in it. In particular, kvass preserved in a similar manner at a 30-minute exposure ensures the development of a bactericidal effect with respect to at least 99% of Escherichia coli K 12 cells and at least 95% at 16-fold dilution. Against this background, the analysis of bifido- and lacto-containing kvass drinks of various blend formulations made it possible to demonstrate a direct proportionality of their bactericidal potential to the relative content of the added additive. So, drinks with a relative content of bacterial additives of 1% and 2.5% of the final volume provided the formation of 95%, and with a content of 5%, 10% and 25% - more than 99% of the bactericidal effect against Escherichia coli K 12. When they however, a consistent 2-, 4-, 8- and 16-fold dilution of the bactericidal potential progressively lost, remaining at the level of 95% and higher only in drinks with bifidobacteria or lactobacilli on GSS from 10% or more.

From the above considerations, the level of at least 10% of the final volume is set as the lower limit of the relative content of the bacterial additive in the composition of the proposed drink, which allows for the complete replacement of chemical preservatives with antagonistic substances that accumulate in the hydrolyzate-soybean medium when bifidobacteria or lactobacilli are grown on it , provide the necessary and sufficient preservation of the finished drink.

3. It was also shown that when cultivated on a hydrolyzate-soybean medium, the achieved concentration of bifidobacteria or lactobacilli is not less than 10 ⁹ CFU / ml, which, when this intermediate product is added to the kvass drink in an amount of 10% or more of the final volume, ensures the presence of these microorganisms at a level of at least 10 ⁸ CFU / ml, regulated as a sufficient presence of probiotic microorganisms in the composition of functional nutrition products [7].

In general, summarizing the above materials about the nature of the proposed method, the characteristics of the actions, the order of their implementation and the conditions for implementation, we can state that the claimed method does not follow from the prior art and should be evaluated as meeting the criterion of "inventive step". At the same time, the application of the technology for culturing bifidobacteria and lactobacilli on hydrolyzate-soybean medium with their subsequent blending with the food-taste kvass base obtained using traditional technology allows one to obtain a fundamentally new product with a therapeutic and prophylactic effect and preserving all the advantages of Russian kvass.

Example 1. Preparation of kvass with bifidobacteria

In accordance with the Technological Instructions of OJSC Living Water (Orenburg, Russia) for the production of bread kvass according to GOST 28188-89 by fermentation (TI 10-11925600-0007-99), kvass wort concentrate is diluted in pre-treated water to a solids content of 1, 4 g per 100 g of wort. Wort prepared in this way is pasteurized at 78-80 ° C for 30-35 min, cooled to 28 ± 2 ° C, after which pre-prepared sugar syrup is added to the amount specified in the kvass recipe.

A pure yeast culture is propagated until yeast accumulates 50 million cells in 1 ml of yeast wiring, which is given in the kvass wort in an amount of 3% of its volume. Fermentation is carried out at 25-30 ° C for 12-16 hours until the true solids content of the wort is reduced by 1 g in 100 ml of fermented wort.

After fermentation, the kvass is separated from the yeast by settling in a fermentation tank at 5-7 ° C and carefully, without touching the yeast sediment, is pumped into the blending tank.

In parallel, a soy protein hydrolyzate is prepared by diluting the dry soy protein substrate in distilled water at the rate of 75 g of powder per 1 liter of water. The resulting liquid is boiled for 5-10 minutes, cooled to 38-40 ° C, adjusted to pH 3.0 by adding 10.0% HCl (hydrochloric acid) and pepsin is added at a rate of 10 g / l, after which fermentation is carried out (hydrolysis) at 37-38 ° C for 4 hours. After fermentation is complete, the pH is reversed to 4.5 by adding 20% NaOH (sodium hydroxide). The resulting hydrolyzate is boiled for 15 minutes, filtered and diluted with distilled water in a ratio of 1: 1. After heating to 80 ° C, the substrate is combined with molten food agar to a final concentration of the latter of 0.075% and boiled for 15 minutes. After settling for 20 min, the precipitate is drained, the volume of the substrate is brought to the initial distilled water and filtered mechanically to release from undissolved particles. The prepared hydrolyzate-soybean medium is sterilized at a pressure of 0.5 MPa (t = + 112-115 ° C) for 30 minutes with preliminary heating with steam for 30 minutes. The pH of the finished medium is set in the range of 7.2-7.4.

Bifidobacterium bifidum No. 1 (source of acquisition - NIIEM named after G.N.Gabrichevsky) is added to the hydrolyzate-soybean medium prepared by the described method at the rate of 5-10% of the liquid culture to the volume of inoculated medium and cultivation is carried out at 37-38 ° С for 24 -48 hours before the accumulation of bacterial cells to a titer of 10 ⁹ -10 ¹⁰ CFU / ml

At the final stage, the finished drink is made by mixing the kvass base that has been fed into the blending tank and the biomass of bifidobacteria grown on soybean medium, the last of which is added in the calculation of 10-15% of the final volume, so that the final content of living bifidobacteria cells is at least 10 ⁸ CFU / ml

The finished drink is thoroughly mixed and cooled to 4 ° C, then poured into containers, packaged and labeled in accordance with GOST 28188-89 “Non-alcoholic drinks. General specifications. " Transportation and storage of the finished drink is carried out at 4 ± 2 ° C for up to 30 days.

Example 2. Preparation of kvass with lactobacilli

In the hydrolyzate-soybean medium prepared according to Example 1, Lactobacillus acidophilus No. NK-1 (source of acquisition - NIIEM named after G.N.Gabrichevsky) is added at the rate of 1-3% of liquid culture to the volume of inoculated medium and cultivation is carried out at 37-38 ° С within 24 hours before the accumulation of bacterial cells to a titer of 10 ⁹ -10 ¹⁰ CFU / ml

The resulting biomass of lactobacilli is introduced into the blending tank with the kvass base prepared according to Example 1 in an amount of 10-15% of the final volume, so that the content of living lactobacillus cells in the finished drink is at least 10 ⁸ CFU / ml.

The finished drink is mixed, cooled to 4 ± 2 ° C and consumed without subsequent fermentation.

Information sources

1. Kvass for every taste. N-Novgorod, 1991, 13 pp.

2. Koscheev A.K. Russian kvass and other drinks. Perm, 1983, 190 p.

3. Technological instructions for the production of bread kvass according to GOST 28188-89 by fermentation (TI 10-11925600-0007-99).

4. Patent SU No. 1738838. Method for the production of kvass (authors: Pribylsky V.L. et al.), Publ. 06/07/92. Bull. 21.

5. Patent RU No. 2061392. The method of preparation of kvass (authors: Shenderov B.A., Manvelova M.A., Stepanchuk Yu.B.), publ. 07/07/1993.

6. Shenderov B.A. Medical microbial ecology and functional nutrition, Moscow, 1998. - 285 p.

7. SanPiN 2.3.2.1078-01 "Hygienic requirements for safety and nutritional value of food products."

Comparative characteristics of the actions, the order of their implementation and the conditions for the implementation of the preparation of kvass using the proposed method and the prototype method Compare Features Method - prototype The inventive method Stage I - the creation of a food taste kvass base Cultivation medium Plant-based kvass wort, sugar syrup Also Leaven Pure yeast culture Also Fermentation conditions 30-35 ° C Also Fermentation time 10-12 hours Also Stage II - receiving supplements based on bifidobacteria or lactobacilli Cultivation medium Animal (Milk) Protein Based Medium Peptic Soy Protein Hydrolyzate Medium Used strains Bifidobacterium longum CMPM B-2000, and / or Bifidobacterium bifidum CMPM B-3300. and / or Bifidobacterium bifidum TsMPMV-3190 (in monoculture or in a mixture) Monocultures of production strains of bifidobacteria and lactobacilli, approved for use in the food industry Cultivation conditions 37-38 ° C Also Cultivation time 24-48 hours Also Separation of biomass from the culture medium with the creation of a "concentrate" of bifidobacteria or lactobacilli + Not required Stage III - the creation of a finished drink by blending a food taste kvass base and an additive based on bifidobacteria or lactobacilli The relative volume of the additive based on bifidobacteria or lactobacilli About 1% 10-15% Achieved final concentration of bifidobacteria or lactobacilli in 1 ml of drink At least 10 ⁸ CFU Also The introduction of chemical preservatives There is no data Not required

Claims (3)

1. A method of preparing kvass, involving the propagation of pure yeast cultures, introducing them into kvass wort, fermenting and blending with the recipe components, separating fermentation of kvass from the sediment, pasteurizing, cooling and introducing fermentation of pure cultures of lactic acid bacteria, characterized in that they use bifidobacteria or lactobacilli, previously grown for 24-48 hours at 37 ° C on a hydrolyzate-soybean medium to a titer of at least 10 ⁹ CFU / ml, while bifidobacteria or lactobacilli are added to naturally with the culture medium in the amount of 10-15% of the final volume of the drink. 2. The method according to claim 1, characterized in that as a medium for the cultivation of bifidobacteria or lactobacilli, a peptic hydrolyzate-soybean medium of the composition is used: soy protein substrate 75 g, water 1000 ml, pepsin 10 g, which is fermented for 4 hours at 37 -38 ° C, filtered, sterilized and set the final pH of 7.2-7.4. 3. The method according to claim 1, characterized in that the strain Bifidobacterium bifidum No. 1 is used as the bifidobacterium (source of acquisition is NIIEM named after G.N. Gabrichevsky), and the strain Lactobacillus acidophilus No. NK-1 is used as the lactobacillus (source of acquisition - NIIEM named after G.N. Gabrichevsky).