SK172897A3 - Composition based on probiotics, method for the preparation of probiotic bacteria being contained in the composition, and usage of said composition - Google Patents

Abstract

Disclosed is a composition based on probiotics, which composition can be characterised in that it contains lyophilised probiotic bacteria utilising lactic acid fermentation of Enterococcus or Bifidobacterium strain with organically bound selenium in an amount ranging from 410 to 420 ug Se/1g of lyophilised bacteria, with the proviso that the amount of living germs in 1g of lyophilised bacteria ranges from 2.3 to 9.0 . 10exp(11). Said probiotic bacteria contained in the composition is prepared by adding 0.00033 - 0.00046 wt.% of selenium inorganic salt into cultivation medium that contains lactic acid fermentation bacteria of Enterococcus or Bifidobacterium strain, at a temperature between 37 and 42 degrees Celsius, during 15 and 18 hours. The product thus obtained is isolated, washed by sterile physiological solution and, at last, it is subjected to lyophilisation. The composition is usable for the preparation of foodstuffs with a probiotic activity.

Description

A composition based on probiotics, a process for preparing the probiotic bacteria contained in the composition and the use of the composition.

Technical field

The present invention relates to a composition based on probiotics, to a process for preparing the probiotic bacteria contained in the composition and to the use of the composition.

BACKGROUND OF THE INVENTION

Probiotics are living organisms mainly found in certain foods, for example in dairy products prepared by fermentation, which have a beneficial effect on the animal and human organism by ensuring optimal composition of the intestinal microflora. There is ample evidence that lactic acid bacteria are characterized by antimutagenic, anticarcinogenic, immunostimulatory and anticholesterolemic properties, as a result of which they have been implicated in the prevention of infectious, tumor and cardiovascular diseases as well as in delaying the onset of senescence (Ebr. , et al .: Anti-mutagenic and immuno-stimulatory properties of lactic acid bacteria World J. Microbiol Biotechnol., 11, 294-298, 1995; Bakalinsky, AT, et al. 350, 199-200, 1996, Pool-Zobel, BL, et al .: Antigenotoxic Properties of Lactic Acid Bacteria in the Salmonella Typhimurium Mutagenicity Assay Nutr.Cancer 20, 261-270, 1993, Singh, J., et al. : Bifidobacterium longum, a lactic acid producing intestinal bacterium inhibiting colon cancer and modulates the intermediate biomarkers of colon carcinogenesis, Carcinogenesis 18, 4, 833-841, 1997; Zacconi, c., Et al .: Serum cholesterol level with inocular mice colonized with Enterococcus faecium and Lactobacillus acidophilus. Microbiologica 15: 413-418 (1992); Challa, A., et al .: Bifidobacterium longum and lactulose supress azoxymethane-induced colonic aberant crypt foci in rats. Carcinogenesis 18: 517-521, 1997; Orhage, K., et al .: Binding of mutagenic heterocyclic amines by intestinal and lactic bacteria. J. Dairy Sci. 78: 491-497 (1995); Hadault, S., et al. Antagonistic activity exerted in vitro and in vivo by Lactobacillus casei (strain GG) against Salmonella typhimurium C5 infection. Appl. Environ. Microbiol. 62: 513-518 (1997).

Selenium belongs to the group of essential trace elements required to maintain optimal glutathione peroxidase activity. This enzyme plays an important role in protecting biological membranes against the damaging effects of reactive oxygen species. The lack of this microelement in food is one of the important risk factors for increased cancer incidence (Combs, GF, and Combs, SB: The role of selenium in nutrition, Academic Press, Orlando, 1986; Tex, G., et al.,: Low Plasma Selenium as a Risk Factor for Cancer Death in Middleaged Men, Nutr.Cancer 10: 221-229, 1987; Lee, BJ, et al., Molecular Biology of Selenium and Its Role in Human Health Molecules and Cells 6: 509 -520, 1996; Fleet, JC and Mayer, J.,: Dietary selenium repletion may reduce cancer incidence in people at high risk who live in areas with low soil selenium (Nutr. Rev. 55: 277-279, 1997).

In general, food intake of selenium is relatively low. It is likely that part of the population does not receive even the lower recommended dose (50 pg / day) in the diet. Selenium deficiency in the diet is addressed by fertilization with inorganic selenium soles, which enters this element into the food chain. Another method consists in preparing tablets containing inorganic salts of selenium, but selenium in inorganic form is more difficult to utilize than protein-bound selenium.

SUMMARY OF THE INVENTION

The above-mentioned disadvantages are largely eliminated by the probiotics-based composition consisting of lyophilized probiotic lactic acid bacteria with organically bound selenium in an amount of 410 to 420 pg Se / lg of lyophilized bacteria, with live germ content in 1 g of lyophilized bacteria is 2.3 to 9.0. 10 ¹¹

The probiotic lactic acid bacteria are preferably Enterococcus spp.

Lactobacillus or Bifidobacterium.

The essence of the process for preparing the probiotic bacteria contained in the composition is that the lactic acid bacteria are cultured in a culture medium with the addition of an inorganic salt of selenium in an amount of 0.00033 to 0.00046% by weight. at 37 ° to 42 ° C

After 15 to 18 hours, the resulting product was collected, washed with sterile saline and finally subjected to lyophilization.

In some species of bacteria, particularly Bifidobacterium, cultivation takes place under anaerobic conditions under a CO ₂ atmosphere.

The present invention also provides the use of the present composition for the preparation of food products having probiotic effects.

The composition of the invention has the properties of probiotics and has the ability to supply the organism with essential selenium bound to an organic form. The incorporation of selenium into the proteins of probiotic bacteria can be considered as an important means of reducing the risk of civilization diseases. Thus, in addition to the original beneficial effects on human health, lactic acid bacteria also become carriers of organic forms of selenium, which have more advantageous properties over inorganic selenium salts. The selenoamino acids found in lactic acid proteins become precursors to the biosynthesis of glutathione peroxidase and other protective enzymes with antioxidant effects in the human gastrointestinal tract.

DETAILED DESCRIPTION OF THE INVENTION

Example 1

Preparation of probiotic containing organically bound selenium by cultivation of Enterococcus faecium Bacteria of the genus Enterococcus faecium were stationary cultivated in a volume of 10 L in a glass container closed with a cotton wool plug on the soil having the following composition: peptone 10.0 g, meat extract 10.0 g, yeast extract 5.0 g, glucose 20.0 g, tween 80 1.0 g, ammonium citrate 2.0 g, sodium acetate 5.0 g, magnesium sulfate 0.1 g, manganese sulfate 0.05 g, potassium hydrogen phosphate 2.0 g, distilled water 1000 ml. The pH of the soil was adjusted to pH

6.5 ± 0.2 prior to sterilization. In particular, a sterile Na ₂ SeO · solution was prepared. . 5 H ₂ O at a concentration of 0.33 wt. in distilled water. The sterile culture broth was inoculated with an 11-12 hour culture of Enterococcus faecium and a sterile solution of Na ₂ SeCl 5 · 5 H ₂ O in an amount of 10 ml was added. The cultivation time at 37 ° C was 15 hours

The -4 cells were separated by centrifugation at 3000 rpm, washed three times with sterile saline, and finally lyophilized.

The lyophilized cells of the microorganism are stored in a sealed container at 4 ° C, and their viability does not change within 6 months and is 2.3. 10 * ¹ live bacteria / 1 g dry matter. The content of organically bound selenium in the bacteria was determined by atomic absorption spectrometry and had a value of 410 pg Se / 1g of lyophilized bacteria.

Example 2

Preparation of probiotic containing organically bound selenium by cultivation of Lactobacillus acidophilus.

Lactobacillus acidophilus was cultured according to the procedure of Example 1 except that 12 ml of sterile Na ₂ SeO ₃ solution was added. 5 H ₂ O and cultivated at 40 ° C for 17 hours.

The live germ content in 1g of lyophilized bacteria was 6.0. 10 ¹ 'and the organically bound selenium content was 415 µg Se / 1g of lyophilized bacteria.

Example 3

Preparation of probiotic containing organically bound selenium by cultivation of Bifidobacterium longum.

The bacteria of the species Bifidobacterium longum were cultured according to the procedure described in Example 1 except that 14 ml of sterile Na ₂ SeO ₂ solution was added. . 5 H ₂ O and cultivation under anaerobic conditions under CO ₂ atmosphere for 18 hours at 40 ° C.

The live germ content in 1g of lyophilized bacteria was 9.0. 10 ¹¹ and the organically bound selenium was 420 Pg / g lyophilised bacteria.

Industrial usability

The composition according to the invention has utility in the pharmaceutical industry for the production of preparations with preventive or therapeutic application in human and veterinary medicine, as well as in the food industry as an additive to various kinds of foods such as yoghurts, acidophilic milk, edible fats, confectionery products.

Claims (5)

PATENT CLAIMS A probiotics-based composition, characterized in that it consists of lyophilized probiotic lactic acid bacteria with organically bound selenium in an amount of 410 to 420 µg Se / 1g of lyophilized bacteria, wherein the content of live germs in 1 g of lyophilized bacteria is 2.3 to 9 , 0. 10 Composition according to claim 1, characterized in that the probiotic lactic acid bacteria are preferably Enterococcus, Lactobacillus or Bifidobacterium. A process for the preparation of the probiotic bacteria contained in the composition according to claims 1 and 2, characterized in that the lactic acid bacteria are cultured in the culture medium with the addition of an inorganic selenium salt in an amount of 0.00033 to 0.00046% by weight. at 37 to 42 ° C for 15 to 18 hours, the product formed is separated, washed with sterile saline and finally subjected to freeze-drying. 4. A method according to claim 3, wherein the cultivation takes place under anaerobic conditions under a CO2 atmosphere. Use of a composition according to the preceding claims for the preparation of food products with probiotic effects.