RU2430157C1 - Growth stimulant for lactic-acid bacteria in milk - Google Patents

Abstract

FIELD: food industry. ^ SUBSTANCE: growth stimulant for lactic-acid bacteria in milk contains MnSO4, FeSO4, KJ, ZnSO4 and CuSO4 at preset components ratios. ^ EFFECT: invention allows to decrease the quantity of introduced starter microorganisms and reduce the period of clot formation. ^ 2 tbl, 2 ex

Description

The invention relates to biotechnology and can be used in the dairy industry in the production of bacterial starter cultures and fermented milk products.

In the industrial use of microorganisms, it is necessary to constantly develop methods for intensifying their growth and reproduction. Methods of growth regulation include various stimulants.

Known application of yeast, corn extracts, other organic compounds [Bannikova L.A., Koroleva N.S., Semenikhina V.F. Microbiological basis of dairy production. - M .: Agropromizdat, 1987 .-- 400 p .; RF patent No. 2233875, 2000]. An aqueous extract from amaranth leaves is introduced into milk as a stimulant for growing a culture strain of Lactobacillus acidophilus NK-1 [RF Patent No. 2233322, 2001].

The disadvantage in the stimulating effect of compounds and substances is the lack of a stable composition, which determines the results in industrial production.

The closest in technical essence to the invention is a method of cultivation of lactic acid microorganisms, involving the use of a solution of individual salts: ZnSO ₄ at a concentration of 5 mg / l; KJ 1.25 mg / L; FeSO ₄ - 1 mg / l; CuSO ₄ - 5 mg / l; MnSO ₄ - 0.25 mg / l (RF Patent No. 2316585, 2008].

The disadvantage of using mineral salts separately is that they have a different stimulating effect, which, when using multi-strain and multi-species starter cultures, can have a different effect in industrial production.

The purpose of the invention is the development of a method for the development of lactic acid microorganisms in milk using a mixture of mineral salts as a growth promoter, which have a stable positive result in industrial production and reduce the amount of fermented milk products produced in the production.

The technical result of the invention is to increase the reproductive properties of lactic acid bacteria when grown in milk.

The essence of the invention lies in the fact that as the proposed stimulants for lactic acid bacteria used a mixture of mineral salts containing manganese, iron, zinc, potassium, copper.

One of the most important reasons for the stimulating effect of manganese ions is associated with nucleic acids, which are part of many compounds that increase enzymatic activity. Iron ions are necessary for the synthesis of iron porphyrin enzymes. Zinc ions stimulate the synthesis of cellular protein due to the intense incorporation of nitrogen into synthetic reactions, enhance fission processes. Potassium and copper ions contribute to the utilization of glucose for the growth and reproduction of microorganisms.

Strains of mesophilic lactic acid cultures of Lactococcus lactis subsp. lactis 378, Lactobacillus acidophilus 5 e, Lactobacillus bulgaricus 245, Streptococcus salivarius subsp. thermophilus 124, obtained from the collection of the Museum of Pure Microorganism Cultures of the Vologda State Dairy Academy named after N.V. Vereshchagin.

Microorganisms are cultured in pasteurized milk, to which a mixture of mineral salts is added, and the dose of each salt in the total combination is different and ranges from 0.2 to 4.5 mg / dm ³ .

The invention is illustrated by the following examples.

Example 1

To implement the proposed method, skim milk is used, pasteurized at a temperature of (87 ± 2) ° C with an exposure of (10-15) min, cooled to (28-30) ° C and used as a nutrient medium for the cultivation of mesophilic lactic acid bacteria.

Strain Lactococcus lactis subsp. lactis is seeded in pasteurized milk in amounts of 1 and 3% of the volume of skim milk.

Mineral salts are introduced into milk in the form of a mixture containing 3.1% MnSO ₄ , 12.5% FeSO ₄ , 15.6% KJ, 62.5% ZnSO ₄ and 6.3% CuSO ₄ in an amount of 7.2 mg / DM ³ , previously dissolved in distilled water and subjected to boiling. The fermented mixture is stirred and fermented at a temperature of (28-30) ° C until a clot forms at a pH value (5.3-5.4). Control options do not contain additives of a mixture of mineral salts, while the amount of yeast is 1 and 3%.

The determination of CFU is carried out by the method of seeding samples on a nutrient medium Lee.

The strain of Lactobacillus plantarum is cultured according to the above method. The number of CFU is determined on an MRS culture medium.

A technological assessment of the influence of mineral salts was carried out by the duration of fermentation and the number of lactic acid bacteria.

The data are shown in table 1.

The results obtained indicate that the mixture of mineral salts has a stimulating effect on the growth of mesophilic lactic acid microorganisms in milk, which reduces the amount of starter microorganisms introduced by 3 times, while the content of mineral salts in the mixture is mg / dm ³ : ZnSO ₄ - (4,5 ± 0.5); KJ - (1.2 ± 0.1); FeSO ₄ - (0.9 ± 0.1); CuSO ₄ - (0.45 ± 0.05); MnSO ₄ - (0.20 ± 0.05).

Example 2

To implement the proposed method using strains of thermophilic lactic acid bacteria.

To grow cells, the strain Lactobacillus acidophilus 5 e is introduced into pasteurized milk in the amount of 1 and 3% by weight of milk, the mineral mixture is introduced according to the method specified in example 1. Cultivation is carried out at a temperature of (38-40) ° C until a clot forms. Control options do not contain additives of a mixture of mineral salts, while the amount of yeast is 1 and 3%.

The culture of Lactobacillus bulgaricus 245 is cultivated in a mixture of pasteurized milk and a mixture of mineral salts at the parameters adopted for the cultivation of Lactobacillus acidophilus 5 e. The number of CFU is determined according to the data of culture on MRS medium.

Culture of Streptococcus salivarius subsp. thermophilus is grown in a mixture of pasteurized milk and a mixture of trace elements at 40 ° C until a clot forms. The number of CFU is determined on the medium of hydrolyzate-milk agar.

The results of the action of the addition of a mixture of mineral salts in milk for growing thermophilic lactic acid bacteria are shown in table 2.

The results obtained indicate that the mixture of mineral salts has a stimulating effect on the growth of thermophilic lactic acid microorganisms in milk, which reduces the amount of starter microorganisms introduced by 3 times, while the content of mineral salts in the mixture is mg / dm ³ : ZnSO ₄ - (4,5 ± 0.5); KJ - (1.2 ± 0.1); FeSO ₄ - (0.9 ± 0.1); CuSO ₄ - (0.45 ± 0.05); MnSO ₄ - (0.20 ± 0.05).

Table 1 The effect of a mixture of mineral salts on the growth of mesophilic lactic acid microorganisms The studied strains of cultures of lactic acid bacteria Reduction of ripening time in comparison with the control,% The change in the number of bacteria in comparison with the control, ln CFU / cm ³ Fermentation% one 3 one 3 Lactococcus lactis subsp. lactis 378 3.2 24.8 +0.41 +2.52 Lactobacillus plantarum 132-5 5,6 14.8 +0.23 +2.35

table 2 The effect of a mixture of mineral salts on the growth of thermophilic lactic acid microorganisms The studied strains of cultures of lactic acid bacteria Reduction of ripening time in comparison with the control,% The change in the number of bacteria in comparison with the control, ln CFU / cm ³ Fermentation% eleven 3 one 3 Lactobacillus acidophilus 5 e 3.2 10.5 +0.12 +1.98 Lactobacillus bulgaricus 245 3,5 10,4 +0.22 +2.1 Streptococcus salivarius subsp. thermophilus 124 3.4 24 +0.21 +1.95

Sources of information taken into account

1. Bannikova L.L., Koroleva N.S., Semenikhina V.F. Microbiological basis of dairy production. - M.: Agropromizdat, 1987 .-- 400 p.

2. RF patent No. 2233875, 2000.

3. RF patent No. 2233322, 2001.

4. RF patent No. 2316585, 2008.

Claims (1)

Lactic acid bacteria growth stimulator Lactococcus lactis subsp. lactis, Lactobacillus acidophilus, Lactobacillus bulgaricus, Streptococcus salivarius subsp.thermophilus in milk, consisting of mineral salts MnSO ₄ : FeSO ₄ : ZnSO ₄ : KJ: CuSO ₄ , characterized in that the introduced mixture of mineral salts has the following percentage ratio: 3.1 : 12.5: 62.5: 15.6: 6.3, respectively.