KR20160117029A - Method for culturing lactic acid bacteria having improved immunoactivity and stability - Google Patents
Method for culturing lactic acid bacteria having improved immunoactivity and stability Download PDFInfo
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- KR20160117029A KR20160117029A KR1020150045599A KR20150045599A KR20160117029A KR 20160117029 A KR20160117029 A KR 20160117029A KR 1020150045599 A KR1020150045599 A KR 1020150045599A KR 20150045599 A KR20150045599 A KR 20150045599A KR 20160117029 A KR20160117029 A KR 20160117029A
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- lactic acid
- acid bacteria
- cultured
- seaweed
- culturing
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Images
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
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- A—HUMAN NECESSITIES
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- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
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- C12N2500/00—Specific components of cell culture medium
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- C12N2500/76—Undefined extracts from plants
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Abstract
Description
The present invention relates to a method for culturing a lactic acid bacterium, and more particularly, to a method for culturing a lactic acid bacterium having improved immunological activity and stability.
Lactic acid bacteria is one of the microorganisms that have been used in ancient human beings since ancient times. It has been widely used in fermented foods such as kimchi and soybean in Korea and fermented foods in dairy industry such as cheese in the west. In recent years, And it is used as health functional foods and medicines as well as general foods. Specifically, the lactic acid bacteria are widely distributed in the natural world from human and animal digestive tracts to dozens of agricultural products, and are widely distributed in foods such as yogurt, lactobacillus, cheese, fermented butter, soybean paste, soy sauce, kimchi, sausage, They are actively used for manufacturing and feed additives.
Foods fermented using these lactic acid bacteria are known to have various physiological effects as well as nutritional value. In addition, the produced abortion enhances the shelf life of the product, gives a unique refreshing sour taste, exhibits a growth inhibitory function against harmful microorganisms, enhances the digestibility of milk proteins, and promotes absorption of calcium, iron and phosphorus It has advantages. So far, the health effects of lactic acid bacteria have been found to be diarrhea, diarrhea, prevention of constipation, inhibition of harmful bacteria, prevention of cancers and aging, promotion of vitamins and development of cholesterol, prevention of adult diseases and improvement of immunity. In addition, lactic acid bacteria inhibit the growth of pathogens in the intestines, induce intestinal epithelial cells or immune cells to produce substances that regulate immune capacity, compete with pathogens to inhibit the growth of pathogens, and, secondarily, It has been reported that it maintains homeostasis of intestinal flora against environmental changes.
Recently, the function of the intestinal beneficial bacteria, that is, the intestinal acidity, is promoted to promote the intestinal motility, thereby helping digestion of food and absorption of nutrients, inhibiting the production of harmful substances and decomposing, synthesizing vitamins and amino acids, Various efforts have been actively made to isolate lactic acid bacteria and commercialize them as health functional foods or medicines by paying attention to an important role of lactic acid bacteria in protecting intestinal infections and increasing immunity.
Thus, the lactic acid bacteria exert various physiological activity effects in the intestines. In the structure of the human body, after ingesting the lactic acid bacteria, the lactic acid bacteria are killed due to the stomach acid secreted from the stomach until they reach the intestines. I can not. There have been attempts to prevent the death of lactic acid bacteria in the body by coating the lactic acid bacteria powder with various substances such as various proteins or carbohydrates. However, since such coated lactic acid bacteria powder is a powder coated with lactic acid bacteria, not.
In addition, lactic acid bacteria may be produced in the form of capsules or sugar beads, but lactic acid bacteria are unlike general organic substances and drugs, so they are not viable methods for preserving lactic acid bacteria in a living form.
In the case of the MRS medium which is generally used, the pH of the lactic acid bacterium is lowered to about 3 after about 24 hours of culture, A problem occurs. To solve this problem, an artificial pH control is attempted by adding basic chemicals such as ammonia and sodium hydroxide to the MRS medium to control the acidity. Since the lactic acid bacteria are used by freeze-drying the cells immediately, And the high price of the product causes the increase of the price of the lactic acid bacteria itself. In particular, the addition of the basic chemical is an undesirable problem for the functional use of the lactic acid bacteria as the fermented food.
On the other hand, seaweed is a very rich and quantitatively rich food. Generally, it contains about 10% of protein and it contains about 30 ~ 40% of saccharides, but it is not expressed by calorie because it is vegetable fiber. Seaweeds contain many inorganic minerals that are essential for health, and at the same time contain sperm-forming nutrients such as proteins. Seaweeds such as seaweed, seaweed, and kelp are rich in potassium ions and are used as highly alkaline foods chemically. In addition, seaweeds are known to have various pharmacological actions such as facilitating intestinal motility, so that modern people who are interested in health are attracting attention not only as edible foods but also as health functional foods, and their demand is steadily increasing.
The amount of seaweed produced in Korea is about 500,000 tons per year based on the edible part of the seaweed, but about 40 ~ 50% of the amount of seaweed is roots, stems, young leaves, It is drowned in the sea as it is and causes environmental pollution. That is, the dumped waste is piled under the farm to kill offspring such as wastes in the seabed, and it is pushed by the birds in the coastal rock gaps to eliminate fish migration and spawning ground. In addition, , And the phenomenon that the seaweed of the seaweed is lowered in the seaweed in the cultivation and it becomes to be drowned out even before it grows is intensified. However, there have been no reports on the use of such discarded seaweeds in microorganisms, particularly in the culture medium for lactic acid bacteria, and furthermore, regarding the improvement of the immunological activity and stability of the lactic acid bacteria cultured in the medium supplemented with seaweeds Not known.
[Prior Patent Literature]
- Registration Patent No. 0453376 (Oct. 15, 2004)
- Registration Patent Publication No. 0623626 (September 19, 2006)
Accordingly, the present invention provides a method for replacing the wastes to be discarded with a synthetic chemical added to a culture medium for microbial culture, particularly a culture medium for lactic acid bacteria culture, as a buffer for conventional pH control.
It was also found that the lactic acid bacteria cultured using the culture medium for culturing the Lactobacillus with added seaweed improves the immunological activity and the resistance to gastric acid, bile acid and heat is improved. In the case of the composition containing the cultured Lactobacillus, Or a pharmaceutical composition.
According to an aspect of the present invention, there is provided a method for culturing a lactic acid bacterium with a culture medium for culturing a lactic acid bacterium comprising a carbon source, a nitrogen source and a buffer, wherein the buffer contains seaweeds, Wherein the microorganism is augmented compared to the lactic acid bacteria cultured in a culture composition containing no seaweed.
And the resistance of the cultured lactic acid bacteria to gastric acid, bile acid, and heat is enhanced compared to the cultured lactic acid bacteria with the culture composition not containing the seaweed.
Characterized in that the buffer does not contain synthetic chemicals as a pH controlling component.
And the seaweed is seaweed or kelp.
And the seaweed or kelp is contained in at least one form selected from the group consisting of dry powder, juice extract and extract.
The lactic acid bacteria may be selected from the group consisting of Lactobacillus sp., Lactococcus sp., Streptococcus sp., Leuconostoc sp. And Pediococcus sp. ). ≪ / RTI >
In order to solve the above-mentioned problems, the present invention provides a pharmaceutical composition for enhancing immunity, which comprises lactic acid bacterium cultured by the above method as an active ingredient.
In order to solve the above-mentioned problems, the present invention provides a food composition for enhancing immunity, which comprises lactic acid bacteria cultured by the above method as an active ingredient.
According to the present invention, there is provided a method for culturing a lactic acid bacterium with a medium composition for culturing a lactic acid bacterium comprising a carbon source, a nitrogen source and a buffer, wherein a buffer containing seaweeds is used to culture a lactic acid bacterium There is an effect that a satisfactory culture can be performed without lowering the pH at the time of culturing, and culturing a lactic acid bacterium with enhanced immunological activity compared with the conventional method.
In addition, the tolerance to gastric acid, bile acid and heat is enhanced, and when the lactic acid bacteria are consumed, the lactic acid bacteria having improved stability can be cultured by increasing the content of the lactic acid bacteria which reach to the intestine.
In addition, the main purpose of the functional lactic acid bacterium is to regulate the function. For this purpose, the dietary fiber of seaweeds can exhibit a synergistic effect on the suicidal action when cultured by using seaweeds such as wakame.
It also has the effect of preventing environmental pollution as well as creating new added value by utilizing abandoned natural plant resources.
FIG. 1 is a graph showing the results of evaluation of tolerance to artificial gastric juice of cultured lactic acid bacteria cultured according to an embodiment of the present invention,
FIG. 2 is a graph showing the results of evaluating resistance to artificial bile acids of cultured lactic acid bacteria according to an embodiment of the present invention,
FIG. 3 is a graph showing the results of evaluating the thermal stability of the cultured lactic acid bacteria according to the embodiment of the present invention,
4 is a graph showing experimental results on immune cell viability of cultured lactic acid bacteria according to an embodiment of the present invention,
5 and 6 are graphs showing experimental results on the enhancement of immune activity of the cultured lactic acid bacteria according to an embodiment of the present invention,
7 is a graph showing the results of measurement of viable cell counts of lactic acid bacteria using seaweed extract and medium containing juice.
Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.
In view of the fact that synthetic chemicals such as ammonia and sodium hydroxide are added as a buffer solution in order to prevent the pH from decreasing in the conventional method of culturing lactic acid bacteria with a medium composition for culturing a lactic acid bacterium comprising a carbon source, a nitrogen source and a buffer, As a result of intensive researches on the additive materials that can realize high concentration culture of lactic acid bacteria while replacing chemical substances with natural substances, it is possible to prevent inhibition of lactic acid bacteria growth due to decrease of acidity without addition of synthetic chemicals, In addition, when the lactic acid bacteria are consumed, the stability of the lactic acid bacteria is improved by increasing the survival rate of the lactic acid bacteria, and the effect of culturing the lactic acid bacteria with enhanced immunity activity And arrived at the present invention.
Accordingly, the present invention provides a method for culturing a lactic acid bacterium with a medium composition for culturing a lactic acid bacterium comprising a carbon source, a nitrogen source and a buffer, wherein the buffer contains seaweeds and the immune activity of the cultured lactic acid bacterium is Which is enhanced compared to the cultured lactic acid bacteria.
In the present invention, the culture medium contains basically the components required for culturing the lactic acid bacteria, such as carbon source and nitrogen source, and may be based on known MRS medium, for example.
Wherein the carbon source is selected from the group consisting of glucose, maltose, lactose, cellobiose, mannose, trehalose, fructose, melibiose, But are not limited to, galactose, raffinose, sucrose, stachyose, mannitol, arabinose, sorbitol, xylose, esculin, esculin, rhamnose and salicin, and preferably glucose, maltose, lactose, galactose, sucrose, sorbitol, fructose or xylose may be used, For example, glucose, maltose, lactose, galactose, sucrose, fructose or sorbitol may be used, and glucose, maltose or lactose may be most preferably used.
As the nitrogen source, various sources such as casein, animal nitrogen source such as beef extract, vegetable nitrogen source such as peptone, and nitrogen source such as yeast extract may be used in various ways, and various other trace elements may be contained. Herein, the term " trace element " means the minimum amount of ingredient required for proper growth, proliferation and physiology of a lactic acid bacterium. Examples thereof include sodium, magnesium, phosphorus, potassium, calcium, manganese, iron, cobalt, nickel, , Cerium, molybdenum, and the like.
In the present invention, the buffer solution is added to prevent the inhibition of the growth of lactic acid bacteria caused by lowering the pH of the culture medium to about 3 when the buffer solution is not contained in the culture of the lactic acid bacteria. In the past, addition of basic chemicals such as ammonia and sodium hydroxide And a buffer solution containing seaweed is used.
The above seaweeds may be used as seaweeds, kelp, mackerel, mackerel, mackerel, mackerel, pulses, parasites, hearing, mackerel, mackerel, mackerel, rhubarb and carrageenin. Can be cultured, and most preferably, seaweed can be used. Seaweed or sea tangle may be added to the medium composition in various ways. For example, it may be added in the form of a dry powder or a juice, or may be added in the form of an extract which can be prepared using various extraction methods.
The lactic acid bacteria that can be cultured using a culture medium composition according to the invention is Lactobacillus genus which is generally used (Lactobacillus sp.), Lactobacillus nose kusu in (Lactococcus sp.), Streptococcus genus (Streptococcus sp.), Flow Leuconostoc sp. And Pediococcus sp. Lactobacillus can be applied. Lactobacillus sp. Lactic acid bacteria can be preferably used.
Hereinafter, the present invention will be described in more detail by way of examples.
Culture of lactic acid bacteria using existing medium
Basic lactic acid bacteria culture medium composition (
Cultivation of lactic acid bacteria using a seaweed powder-containing medium
Culturing was carried out in the same manner as in the cultivation of lactic acid bacteria using the conventional culture medium, except that the culture medium prepared by adding 1 part by weight of the dried seaweed powder to 100 parts by weight of the basic culture medium for lactic acid bacteria culture was used.
Evaluation of growth activity of lactic acid bacteria
After culturing the lactic acid bacteria as described above, 9 ml of the sterilized water is mixed with 1 ml of the culture solution, and the mixture is continuously diluted 10-fold, and then 1 ml of each solution is dispensed. 9 ml of plate count agar is mixed and hardened, pour method for 48 hours. The number of colony formed was counted and the number of viable cells was measured with colony forming units per milli liter (CFU / ml). The results are shown in Table 1 below .
As shown in Table 1, it can be seen that the number of lactic acid bacteria was significantly increased by about 58 times as compared with the case of using the basic lactobacillus culture medium when the culture was carried out using the medium containing seaweed according to the present invention.
Assessment of artificial gastric tolerance
To evaluate the resistance of the cultured lactic acid bacteria to artificial gastric juice, artificial gastric juice was prepared by adjusting the pH to 2.5 with 1 mg / ml pepsin solution using hydrochloric acid (HCl) according to the method of Kobayasih et al. The lactic acid bacteria cultured in each culture medium was inoculated therein, followed by incubation at 37 ° C. for 6 hours. The number of live bacteria was measured after 1, 3 and 6 hours, and the results are shown in Table 2 and FIG.
The pH of the pure gastric juice is maintained at about 1.0 ~ 2.0, and most of the microorganisms are killed in the gastric juice. However, the pH of the gastric juice may be slightly increased due to the buffering effect of the ingested food. However, in order for the lactic acid bacteria to function physiologically, gastric acid should be viable in the stomach.
As shown in Table 2 and FIG. 1, the survival rate of the lactic acid bacteria cultured in the seaweed supplemented medium was high at 3 hours after the reaction, and the growth rate of the lactic acid bacteria cultured in the basic culture medium was 9.13 ± 1.61%, while the survival rate of lactic acid bacteria cultured in seaweed supplemented medium was 26.6 ± 3.27%. Since the lactic acid bacteria cultured in the seaweed supplemented medium showed a higher survival rate than the basic culture medium, many of the lactic acid bacteria cultured in the seaweed supplemented medium could survive the gastric juice.
Evaluation of tolerance to artificial bile acids
To evaluate the resistance of cultured Lactobacilli to artificial bile acids, 0.02 M cholic acid, 0.02 M deoxycholic acid, 0.05 mg / mL lipase and 0.02 mg / mL pacreatin The prepared solution was adjusted to pH 8.0 with 1N hydrochloric acid (HCl) and sodium hydroxide (NaOH) to prepare artificial bile acid. The lactic acid bacterium cultured in each culture medium was inoculated therein, followed by incubation at 37 ° C. for 6 hours. The number of viable cells was measured after 1, 3, and 6 hours, and the results are shown in Table 3 and FIG.
As shown in Table 3 and Fig. 2, the strains did not show any significant difference until 1 hour after the initial reaction. However, after 3 hours of the reaction, the lactic acid bacteria cultured in seaweed supplemented medium In the case of artificial bile acids.
Thermal stability evaluation
In order to evaluate the thermal stability of the cultured Lactobacillus, the Lactobacillus cultured in each culture medium was inoculated into the MRS broth and allowed to stand at 55 ° C for 6 hours. The viable counts of lactic acid bacteria survived after 1, 3 and 6 hours were examined. Are shown in Table 4 and FIG.
When Porbiotics is used as a product, it may be exposed to a temperature change such as a high temperature which is far from the optimal growth condition of the microorganism. In addition, during spray-drying, which is a formulation method, the viable cell power may be drastically reduced when exposed to a high-temperature environment, so it is important to understand the tolerance of the cells to heat.
As shown in Table 4 and FIG. 3, it was confirmed that the lactic acid bacteria cultured in the seaweed supplemented medium were more excellent in thermal stability than the lactic acid bacteria cultured in the basic culture medium, as the lactic acid bacteria growth rate was increased.
Effect on Immune Cell Viability
In order to confirm the effect on the cell viability of the cultured samples, the MTT assay described in Carmichael et al. (1987) was applied as follows. RAW 264.7 cells (ATCC, USA) were cultured in DMEM medium (Gibco, USA) supplemented with 10% heat-inactivated fetal bovine serum and penicillin-streptomycin at 37 ° C and 5% CO 2 Lt; / RTI > and 95% air. Lactic acid bacteria cultured in each culture medium were treated at 10, 10 2 , 10 3 and 10 4 CFU / ml in the state of live cells and dead cells (100 ° C, 15 minutes) and cultured for 24 hours. And FIG. 4, respectively.
As shown in Table 5 and Fig. 4, the effect on cell viability was evaluated in the condition of live cells treated with 10 4 CFU / ml It was confirmed that cytotoxicity was not observed in the lactobacillus treatment group in the seaweed supplemented medium, although it showed cytotoxicity in the basic culture. In addition, it was confirmed that all the groups did not show cytotoxicity in the dead cells state.
Enhancement of immunological activity (confirmation of secretion effect of NO and TNF-α)
(Nakai M, Sudo K, Yamada Y, Kojima Y, Kato T, Saito K, Moriwaki H, Seishima M, Dig Dis Sci. pp1669-1676, 2005), the following experiment was conducted. RAW 264.7 cells (ATCC The Global Bioresource Center ™ ) were dispensed into 96-well microplates at a rate of 1 × 10 5 CFU / ml per well. Lactic acid bacteria cultured in each culture medium were inoculated into a 96- 10, 10 2 , 10 3, and 10 4 CFU / ml were treated at 37 ° C and 5% CO 2 for 24 hours. The supernatant was taken and 50 μl of the cell culture supernatant was mixed with an equal amount of a grease reagent (Griess reagent, 0.1% (w / v) N- (1-naphthyl) ethylenediamine dihydrochloride + 1% (w / v) sulfanilamide in 5% (v / v) phosphoric acid) And the absorbance was measured at 540 nm using a spectrophotometer. The results are shown in Table 6 and FIG. 5, respectively.
As shown in Table 6 and FIG. 5, in the RAW 264.7 cells, the concentration of nitrite ion (NO 2 - ) was significantly increased in the seaweed supplemented medium compared to the control, It was confirmed that NO secretion was increased.
In addition, 50 μl of the cell culture supernatant was taken under the same conditions and ELISA test for TNF-alpha (inflammatory mediator) was performed (R & D Systems Quantikine Mouse TNF-α / TNFSF1A kit. Vilcek J. and TH Lee, J. Biol. Chem., 266, p7313, 1991). The results are shown in Table 7 and FIG.
As shown in Table 7 and FIG. 6, in the RAW 264.7 cells, the tumor necrosis factor TNF-α was significantly increased in the seaweed supplemented medium compared with the control, and thus the immunopotentiating effect of the lactic acid bacteria cultured through the seaweed supplemented medium .
Culture of lactic acid bacteria using seaweed extract
In order to confirm that cultivation of lactic acid bacteria can be smoothly performed even when seaweeds are used as an extract form other than the powder form as described above, the seaweed extract extracted in the following manner is added to L. plantarum isolated from kimchi, And 1% by weight. The culture was carried out in an incubator in the same manner as in the cultivation of lactic acid bacteria using the conventional medium, and the number of live cells was measured. The results are shown in Table 8 and FIG.
[Method for preparing seaweed extract]
The seaweeds in the dry state were crushed and extracted with a mixed solvent of water and ethanol as an extraction solvent at 30 ° C for 1 hour, followed by filtration, concentration under reduced pressure and drying to obtain a seaweed extract.
Referring to Table 8 and FIG. 7, in the cultivation of lactic acid bacteria using the culture medium containing the seaweed extract, the number of viable cells (11.0 ± 0.8) × 10 8 CFU / ml after 24 hours of culture was high and cultivation was possible without addition of chemicals such as ammonia Can be confirmed.
Seaweed The juice Cultivation of lactic acid bacteria using
In order to confirm that the cultivation of lactic acid bacteria can be performed smoothly even when the seaweeds are used in the form of powders or extracts as well as in the form of juices, the Lactobacillus plantarum isolated from kimchi is subjected to The lactic acid bacteria were cultured by the following experimental method, and the number of lactic acid bacteria was measured. The results are shown in Table 8 and FIG.
[Preparation method of seaweed juice]
The seaweed was heated at 80 ° C for 30 minutes and then poured. The juice was sterilized and filtered.
Referring to Table 8 and FIG. 7, in the cultivation of lactic acid bacteria using the culture medium containing the seaweed juice, the number of viable cells (36.3 ± 1.8) × 10 8 CFU / ml after 24 hours of culture was high and cultivation was possible without addition of chemicals such as ammonia .
As described above, it was confirmed that the growth rate of the lactic acid bacteria was increased in the cultivation of the lactic acid bacteria using the culture medium containing the seaweed extract or the seaweed extract, as compared with the result of culturing in the basic culture medium.
Hereinafter, a pharmaceutical composition for immuno-enhancement containing the lactic acid bacterium cultured by the method according to the present invention as an active ingredient or a preparation for application to food will be described, but the present invention is not limited thereto.
Formulation Example 1: Powder preparation
Table 9 below shows an acid composition containing lactic acid bacteria cultured by the method according to the present invention as an active ingredient. Powders can be prepared by mixing the following ingredients and filling the airtight container.
Formulation Example 2: Preparation of tablets
Table 10 below shows a tablet composition containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient. The tablet may be prepared by mixing the following ingredients and then tableting according to a conventional tablet preparation method.
Formulation Example 3: Capsule preparation
Table 11 below illustrates the composition of the capsule formulation containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient. The capsules may be prepared by mixing the following ingredients according to the conventional method for preparing a capsule, and filling the capsules with gelatin.
Formulation Example 4: Injection preparation
Table 12 below shows the composition of injections containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient. The injectable preparation can be prepared in the following ingredient contents per ampoule (2 ml) according to the usual injection preparation method.
Formulation Example 5: Liquid preparation
In Table 13 below, the composition of the liquid formulation containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient is illustrated. The liquid preparation is prepared by adding and dissolving each of the following ingredients in purified water according to a conventional liquid preparation method, mixing the following components after adding a proper amount of lemon flavor, adding purified water to adjust the total volume to 100 ml, filling and sterilizing in a brown bottle .
Formulation Example 6: Preparation of health food
Table 14 below shows the composition of a functional health food containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient. In the following composition, the composition ratio of the vitamin and mineral mixture shows that a composition suitable for health food is mixed and formulated as a preferable preparation. However, the blending ratio may be arbitrarily changed, and each ingredient may be mixed The granules can then be prepared and used in the manufacture of a health food composition according to conventional methods.
Formulation Example 7: Healthy Beverage Manufacturing
Table 15 below shows the composition of a functional health drink containing the lactic acid bacteria cultured by the method according to the present invention as an active ingredient. For example, the following ingredients are mixed in accordance with a conventional health drink manufacturing method, and the mixture is heated at 85 DEG C for about 1 hour under stirring, and the solution thus obtained is filtered to obtain a sterilized 2 liter container. After sealing sterilization, It can be stored and used. Although the following composition ratio is mixed with the ingredients suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preference such as the demand class, the demanding country, and the use purpose.
The preferred embodiments of the present invention have been described in detail with reference to the drawings. It will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the present invention as defined by the appended claims.
Accordingly, the scope of the present invention is defined by the appended claims rather than the foregoing detailed description, and all changes or modifications derived from the meaning, range, and equivalence of the claims are included in the scope of the present invention Should be interpreted.
Claims (8)
Wherein the buffer contains seaweeds so that the immunogenic activity of the cultured lactic acid bacteria is enhanced compared to the cultured lactic acid bacteria with the culture composition not containing the seaweeds.
Wherein the tolerance to gastric acid, bile acid and heat of the cultured lactic acid bacteria is enhanced compared to that cultured with a culture composition not containing the seaweeds.
Characterized in that the buffer does not contain a synthetic chemical as a pH controlling component
Characterized in that the seaweed is wakame or kelp.
Wherein the seaweed or kelp is contained in at least one form selected from the group consisting of dry powder, juice extract and extract.
The lactic acid bacteria may be selected from the group consisting of Lactobacillus sp., Lactococcus sp., Streptococcus sp., Leuconostoc sp. And Pediococcus sp. ). ≪ / RTI >
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KR20180084258A (en) * | 2017-01-16 | 2018-07-25 | (주)천호바이오 | Composition for enhancing immune activity comprising cinnamon extracts and lactic acid bacteria |
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KR20180084258A (en) * | 2017-01-16 | 2018-07-25 | (주)천호바이오 | Composition for enhancing immune activity comprising cinnamon extracts and lactic acid bacteria |
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CN110129217A (en) * | 2019-04-22 | 2019-08-16 | 华中农业大学 | A kind of method and application improving lactic acid bacteria heat resistance |
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