Gastric acid-stable probiotic formulation and method for producing the same

Abstract

To provide a gastric acid-stable probiotic formulation and a method for producing the formulation.SOLUTION: A gastric acid-stable probiotic formulation contains (a) lactic acid bacterium, (b) edible oil, (c) emulsifier and (d) nuts as essential components, and furthermore, can contain (e) prebiotic and (f) adhesive force-imparting agent. In a method for producing the formulation, steps including the followings are performed: a first step of, after heating 1 to 90 pts.wt. of the edible oil at 70 to 95°C, mixing and stirring the emulsifier at 0.1 to 5 pts.wt. to create a mixture of the edible oil and the emulsifier; a second step of cooling and stirring the mixture of the first step to a temperature 2 to 20°C to create a gel-shaped aggregation mixture; and a third step of slowly inputting 1 pt.wt. of lactic acid bacteria into the gel-shaped aggregation mixture and stirring the mixture at low speed to create a lactic acid bacterium mixture.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to a gastric acid stable probiotic formulation containing edible fats and oils, an emulsifier and nuts, and a method for producing the same. [Background technology]

[0002] Generally, lactic acid bacteria settle in the intestines and exert various physiologically active effects, such as activating intestinal motility, suppressing harmful bacteria, and promoting vitamins and immune-enhancing substances. However, due to the structure of the human body, after a human ingests lactic acid bacteria, they pass through the stomach before reaching the intestines, and are often killed by gastric acid secreted from the intestines, and therefore do not exert their original physiologically active functions.

[0003] For this reason, there have been attempts to prevent the lactic acid bacteria from dying in the body by coating the acid bacteria powder with various proteins or carbohydrates, etc., and as a result, lactic acid bacteria powders coated with various substances and methods for producing them have been developed.

[0004] There are about 400 kinds of bacteria present in the human large intestine and feces, of which more than 90% are anaerobic strains. These maintain a certain balance to form a flora in the intestine, and are affected by the health condition of the human body, stress, aging, physiological changes, as well as food and drugs from the outside. To maintain physical health and extend lifespan, the intestinal flora must have more beneficial bacteria and less harmful bacteria. In 2001, a joint expert committee of the World Health Organization (WHO) and the Food and Agriculture Organization of the United Nations (FAO) defined probiotics as "bacteria that are beneficial to health when ingested in appropriate amounts as living microorganisms" (Non-Patent Document 1), and refers to live bacterial preparations that can help improve intestinal microorganisms. In addition, prebiotics refers to substances that can be used by beneficial intestinal microorganisms and help the growth and function of beneficial intestinal strains. The prebiotics known to date are non-starch polysaccharides and oligosaccharides, which are not easily decomposed by human enzymes but can be utilized by microorganisms (Non-Patent Document 2). Recently, the concept of such prebiotics has been broadened due to the knowledge of the effects of polyphenols, which can have a beneficial effect on intestinal microorganisms (Non-Patent Document 3).

[0005] Lactic acid bacteria are a representative bacterium widely used as a probiotic. They are widely present in natural systems and are easily found in the intestines of humans and animals, as well as in fermented foods. They are microorganisms recognized as safe by the U.S. Food and Drug Administration. Lactic acid bacteria attach to intestinal epithelial cells and parasitize them, improving the properties of the intestinal flora and providing many benefits to the host animal, such as stabilizing the intestinal flora, reducing the production of putrefactive products by inhibiting the establishment of harmful bacteria, preventing diseases, activating the immune system, anticancer effects, and lowering cholesterol. The fact that lactic acid bacteria have a growth inhibitory effect on various putrefactive and pathogenic microorganisms is due to several metabolic characteristics, and lactic acid bacteria are used to produce organic acids, hydrogen peroxide, reuterin, diacetyl, acetaldehyde, bacteriocin, and other antibacterial active factors as metabolic products (Non-Patent Document 4; Patent Document 1).

[0006] Currently, probiotics are mainly used as health functional foods, but are also used in some cases for therapeutic purposes. Specifically, probiotics increase the production of immunoglobulin A antibodies, reduce rotavirus shedding, and shorten the duration of diarrhea. Therefore, there have been active attempts to apply probiotics as preventive and therapeutic agents for antibiotic-associated diarrhea, therapeutic agents for inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, therapeutic agents for abdominal bloating and constipation, therapeutic agents for irritable bowel disease, therapeutic agents for radiation-induced enteritis, and adjunctive therapeutic agents for Helicobacter infections. Probiotics have been reported to be particularly effective as a therapeutic agent for chronic inflammatory bowel disease (Patent Document 2).

[0007] The prior patent documents are as follows: Patent Document 3 (applicant: Velvet Care Co., Ltd.) discloses a composition for inducing spontaneous intake of solid preparations in companion animals, characterized in that the composition comprises an oil containing unsaturated fatty acids, a tackifier, a sweetener, a mixture of lactic acid bacteria, a flavoring agent, a preservative and an emulsifier.

[0008] Patent Document 4 discloses a lactic acid bacteria preparation in which lactic acid bacteria are not killed by gastric acid and are active in the small and large intestines, and which is produced by heating and mixing fat and an emulsifier to approximately 60°C, heating and emulsifying two mixed solutions in which a protective agent and lactic acid bacteria are mixed at approximately 50°C, and spraying the emulsified solution under high pressure into a dispersion liquid cooled to 4°C. The microcapsule preparation of lactic acid bacteria is disclosed as being effective in promoting intestinal health by preventing the lactic acid bacteria from being killed by gastric acid, thereby increasing the colonization rate of intestinal lactic acid bacteria.

[0009] Patent Document 5 discloses a complex functionalized enteric soft capsule that contains high amounts of lactic acid bacteria and edible fats and oils, while still guaranteeing the number of lactic acid bacteria even during the expiration date. The complex contains 5.0 to 25.0% by weight of lactic acid bacteria powder, 5.0 to 20.0% by weight of a suspending agent, 0.5 to 2.0% by weight of an emulsifier, and the remaining amount is edible fats and oils.

[0010] Patent Document 6 discloses an oral paste containing about 80 to 90% oil, about 0.5 to 20% emulsifier, and about 1 to 2% probiotic lactic acid bacteria that are effective against tartar and plaque.

[0011] However, none of the above documents discloses or describes the substitution of gastric acid stable probiotic formulations containing edible fats and oils, emulsifiers and nuts, and the method of making same.

[0012] Therefore, the present inventors conducted research to develop a lactic acid bacteria preparation superior to the lactic acid bacteria complex containing lactic acid bacteria, edible oil and fat, and an emulsifier disclosed in the conventional prior art Patent 5 (Patent Document 5).

[0013] As a result of carrying out an emulsion production experiment at different heating temperatures using the lactic acid bacteria preparation of the present invention (Experimental Example 1); the effect on the proliferation of beneficial intestinal bacteria (Experimental Example 2); an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the composition's efficacy in improving constipation (Simplified Clinical Example 1); and an experiment to evaluate the palatability of different nuts contained in the composition (Simplified Clinical Example 2), it was confirmed that the lactic acid bacteria preparation of the present invention strongly proliferates beneficial intestinal bacteria, exhibits excellent survival ability of lactic acid bacteria in gastric acid, and exerts remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability. It was thus possible to provide an excellent probiotic preparation, which led to the completion of the present invention. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] Korean Patent Publication No. 10-2015-0075447 [Patent Document 2] Korean Patent Publication No. 10-2008-0075971 [Patent Document 3] Korean Patent Publication No. 10-2022-0068518 [Patent Document 4] Korean Patent Publication No. 10-1997-0025405 [Patent Document 5] Korean Patent Publication No. 10-2023-0001175 [Patent Document 6] Korean Patent Publication No. 10-2021-0083423 [Non-patent literature]

[0015] [Non-Patent Document 1] Jeon Hye Mi et al., 2011, J Korean Soc. Food. Sci. Nutr. 40(4), pp. 500-508 [Non-Patent Document 2] Manning TS.Gibson GR.2004.Prebiotics.Best Practice & Research Clinical Gastroenterology.18(2):287-298 [Non-Patent Document 3] Cardona et al.,2013,Journal of Nutritional biochemistry,24,1415-1422 [Non-Patent Document 4] Fuller, K., 1989, Probiotics in man and animals, J. Appl. Bacteriol., 66, 365-378 Summary of the Invention [Problem to be solved by the invention]

[0016] The present invention has been made to overcome the above-mentioned problems, and aims to provide an excellent probiotic preparation that is stable to gastric acid and contains edible oils and fats, an emulsifier, and nuts, and that strongly proliferates beneficial intestinal bacteria, exhibits excellent survival ability in gastric acid, and exhibits remarkable efficacy in improving constipation and in terms of palatability, superior to conventionally known lactic acid bacteria preparations, and a method for producing the same. [Means for solving the problem]

[0017] The characteristic configuration of the present invention for achieving the object of the present invention as described above and realizing the characteristic effects of the present invention described below is as follows.

[0018] In one preferred embodiment of the present invention, there is provided a gastric acid-stable probiotic preparation obtained by carrying out steps including: a first step of heating 1 to 90 parts by weight of edible fats and oils at 70 to 95°C, and then mixing and stirring the same with 0.1 to 5 parts by weight of an emulsifier to produce a mixture of edible fats and oils and an emulsifier; a second step of cooling and stirring the mixture from the first step until the temperature reaches 2 to 20°C to produce an aggregated mixture in the form of a gel; and a third step of slowly adding 1 part by weight of lactic acid bacteria to the aggregated mixture in the form of a gel and stirring at low speed to produce a lactic acid bacteria mixture.

[0019] In one preferred embodiment of the present invention, a gastric acid stable probiotic preparation is provided by carrying out the steps further comprising: a fourth step of adding ground nuts to the mixture obtained in the third step and stirring to produce a nut lactic acid bacteria mixture; and a fifth step of optionally adding one or more additional ingredients selected from prebiotics and adhesive agents to the mixture obtained in the fourth step and stirring the mixture.

[0020] In a preferred embodiment of the present invention, the ground nuts have a particle size of 0.01 to 0.5 mm, preferably 0.01 to 0.1 mm, and are stirred at a stirring speed of 30 to 50 rpm for 30 minutes to 3 hours until all materials are uniformly mixed to produce a nut lactic acid bacteria mixture.

[0021] In one preferred embodiment of the present invention, the gastric acid-stable probiotic preparation is characterized by containing, as essential ingredients, (a) lactic acid bacteria, (b) edible oils and fats, (c) an emulsifier, and (d) nuts.

[0022] In a preferred embodiment of the present invention, the gastric acid-stable probiotic preparation is characterized in that the weight ratios of the individual components are (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 90 parts by weight of edible fats and oils, (c) 0.1 to 5.0 parts by weight of an emulsifier, and (d) 5.0 to 90.0 parts by weight of nuts.

[0023] In a preferred embodiment of the present invention, the (a) lactic acid bacteria is one or more strains selected from microorganisms of the genera Lactobacillus, Bifidobacterium, Bacillus, Streptococcus, and Enterococcus.

[0024] In a preferred embodiment of the present invention, the edible oil (b) is one or more components selected from the group consisting of avocado oil, canola oil, evening primrose oil, palm oil, Brazil nut oil, corn oil, cottonseed oil, linseed oil, grape seed oil, hemp oil, olive oil, palm oil, safflower oil, soybean oil, peanut oil, sunflower oil, krill oil, anchovy oil, salmon oil, rice bran oil, brown rice oil, coconut oil, rosehip oil, and tuna oil.

[0025] In a preferred embodiment of the present invention, the (c) emulsifier is one or more components selected from the group consisting of propylene glycol fatty acid esters, glycerin fatty acid esters, sucrose fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, organic acid monoglycerides, polyoxyethylene fatty acid esters, propylene glycol fatty acid esters, polyoxyethylene fatty acid esters, polysorbates, egg yolk lecithin, soybean lecithin, carboxymethylcellulose, glycerin, soybean phospholipids, and calcium stearyl lactylate.

[0026] In one preferred embodiment of the present invention, the nuts (d) are one or more components selected from the group consisting of peanuts, almonds, walnuts, pine nuts, pistachios, pecans, macadamia nuts, hazelnuts, cashew nuts, Brazil nuts, chestnuts, soybeans, cacao nibs, coconuts, sunflower seeds, and pumpkin seeds.

[0027] In one preferred embodiment of the present invention, the gastric acid-stable probiotic formulation contains, in addition to the essential components of (a) lactic acid bacteria, (b) edible fats and oils, (c) an emulsifier, and (d) nuts, one or more additional components selected from (e) prebiotics and (f) adhesives.

[0028] In a preferred embodiment of the present invention, the gastric acid-stable probiotic formulation has a weight ratio among the individual components thereof as follows: (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 60 parts by weight of edible fats and oils, (c) 0.1 to 5.0 parts by weight of emulsifier, (d) 5.0 to 90.0 parts by weight of nuts, (e) 0.01 to 5.0 parts by weight of prebiotics, and (f) 0.1 to 5.0 parts by weight of adhesive strength imparting agent.

[0029] In a preferred embodiment of the present invention, the prebiotic (e) is one or more components selected from the group consisting of pentoses such as xylose and arabinose, glucose, mannose, fructose, galactose; lactulose, lactitol, sucrose, lactose, maltose, trehalose; fructo-oligosaccharides, raffinose, stachyose, maltodextrin; amylose, amylopectin, starch, cellulose, and pectin.

[0030] In one preferred embodiment of the present invention, the (f) tackifier is one or more components selected from the group consisting of sodium aluminosilicate, xanthan gum, carrageenan, guar gum, diutan gum, cellulose gum, gellan gum, pectin, and carboxymethylcellulose.

[0031] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings in order to enable those skilled in the art to easily carry out the present invention.

[0032] The gastric acid-stable probiotic preparation according to the present invention is produced by carrying out steps including: a first step of heating 1 to 90 parts by weight of edible oil or fat to a temperature in the range of 70 to 95°C, preferably 75 to 85°C, and then mixing and stirring with 0.1 to 5 parts by weight of the emulsifier to produce a mixture of edible oil or fat and emulsifier; a second step of producing a gel-shaped aggregate mixture while cooling and stirring the mixture from the first step until the temperature reaches 2 to 20°C, preferably 4 to 10°C; and a third step of slowly adding 1 part by weight of lactic acid bacteria to the gel-shaped aggregate mixture and stirring at low speed to produce a lactic acid bacteria mixture.

[0033] Therefore, the present invention provides a method for producing a gastric acid-stable probiotic preparation according to the present invention, which comprises the steps of: a first step of heating 1 to 90 parts by weight of edible fats and oils to a temperature in the range of 70 to 95°C, preferably 75 to 85°C, and then mixing and stirring with 0.1 to 5 parts by weight of the emulsifier to produce a mixture of edible fats and oils and an emulsifier; a second step of cooling and stirring the mixture from the first step to a temperature of 2 to 20°C, preferably 4 to 10°C, while producing an aggregated mixture in the form of a gel; and a third step of slowly adding 1 part by weight of lactic acid bacteria to the aggregated mixture in the form of a gel and stirring at low speed to produce a lactic acid bacteria mixture.

[0034] In this process, the emulsifier is dissolved and thoroughly mixed in the heated edible oil and fat, and then cooled to produce a stably aggregated gel-like mixture. Lactic acid bacteria are then mixed into this, significantly improving the intestinal retention of the lactic acid bacteria.

[0035] The gastric acid-stable probiotic preparation according to the present invention may be prepared by further comprising a fourth step of adding ground nuts to the mixture obtained in the third step and stirring to produce a nut lactic acid bacteria mixture; and a fifth step of optionally adding one or more additional ingredients selected from prebiotics and adhesives to the mixture obtained in the fourth step and stirring the mixture.

[0036] In a preferred embodiment of the present invention, the probiotic preparation prepared in step 5 can be sealed under nitrogen gas and stored at 25° C. or less. The temperature of the probiotic preparation can be controlled to 30° C. or less, and the decrease in lactic acid bacteria content due to temperature and physical friction can be minimized by slow stirring.

[0037] The term "(a) lactic acid bacteria" as defined in the present invention means a bacterial strain that aids digestion and absorption, inhibits the growth of harmful intestinal bacteria, and maintains a favorable intestinal environment. There are no limitations on the form of the lactic acid bacteria, but various forms such as a dry powder or a dry crude powder are preferred, and the size of the strain may be in the range of 0.1 to 8.0 μm, more preferably 0.2 to 4.0 μm.

[0038] In the present invention, the content of each component is expressed in parts by weight based on 1 part by weight of lactic acid bacteria.

[0039] Representative strains are preferably any lactic acid bacteria widely used in probiotics, and can be readily available in the art, for example, American Type Culture Collection (ATCC), Korean Collection for Type Cultures (KCTC), Korean Culture Center of Microorganisms (KCCM), Korean Agricultural Culture Center (KACC), Rural Development Administration, Korea Institute of Bioscience and Biotechnology, Korea Institute of ... The strains are those available from the Streptococcus Collection and the like, specifically, microorganisms of the genus Lactobacillus, Bifidobacterium, Bacillus, Streptococcus, and Enterococcus; preferably, they are strains of the genus Bifidobacterium, Lactobacillus, or Streptococcus, more preferably, they are strains of the genus Bifidobacterium or Streptococcus; more preferably, they are strains of the genus Bifidobacterium, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium animalis subsp. lactis, and the like. ssp. lactis), Bifidobacterium adolescentis, Bifidobacterium pseudocatenulatum, Bifidobacterium catenulatum or Bifidobacterium infantis, Bifidobacterium thermophilum (B.Bifidobacteria such as Lactobacillus thermophilum species; Bifidobacterium strains such as Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus casei, Lactobacillus casei ssp. paracasei, Lactobacillus rhamnosus, Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus delbrueckii ssp. bulgaricus ssp.bulgaricus, Lactobacillus delbrueckii ssp.delbrueckii, Lactobacillus fermentum, Lactobacillus gasseri, Lactobacillus reuteri, Lactobacillus brevis, Lactobacillus cellobiosus, Lactobacillus crispatus, Lactobacillus GG, Lactobacillus johnsonii, Lactobacillus lactis, Lactobacillus reuteri, Lactobacillus salivariusLactobacillus species such as Bacillus salivarius species; Bacillus species such as Bacillus cereus toyoi or Bacillus cereus species; Leuconostoc species such as Leuconostoc citreum and Leuconostoc mesenteroides; Pediococcus species such as Pediococcus acidilactici and Pediococcus pentosaceus; Propionibacterium acidifaciens Propionibacterium species such as Propionibacterium acidifaciens and Propionibacterium acidipropionici; Streptococcus species such as Streptococcus thermophilus, S. cremoris, S. infantarius, S. intermedius, S. lactis, S. salivarius subsp. thermophilus; Enterococcus faecalis and Enterococcus faecium.Enterococcus species such as Enterococcus faecium; Saccharomyces species such as Saccharomyces cerevisiae and Saccharomyces boulardii, preferably Lactobacillus or Bifidobacterium strains, most preferably Lactobacillus acidophilus, Lactobacillus rhamnosus GG, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium animalis Lactobacillus ssp.Lactis, and more preferably a mixture of two or more strains selected from Lactobacillus acidophilus, Lactobacillus rhamnosus GG, Lactobacillus plantarum, Bifidobacterium longum, and Bifidobacterium animalis ssp.Lactis.

[0040] The term "(b) edible oil" as defined in the present invention includes one or more components selected from the group consisting of avocado oil, canola oil, evening primrose oil, palm oil, Brazil nut oil, corn oil, cottonseed oil, linseed oil, grape seed oil, hemp oil, olive oil, palm oil, safflower oil, soybean oil, peanut oil, sunflower oil, krill oil, anchovy oil, salmon oil, rice bran oil, brown rice oil, coconut oil, rosehip oil, and tuna oil.

[0041] The edible oil and fat wraps the lactic acid bacteria complex and protects it from gastric acid. The edible oil and fat contains 1 to 90 parts by weight, and preferably 15 to 30 parts by weight. The edible oil and fat is heated to 75 to 95°C, preferably 75 to 85°C, and then the emulsifier is mixed and stirred to produce a mixture of the edible oil and fat and the emulsifier. The mixture is stirred in a circulating cooling water tank maintained at 0 to 10°C, preferably 0 to 5°C, until the temperature of the mixture reaches 2 to 20°C, preferably 4 to 10°C, to form a stable gel-shaped aggregate.

[0042] The "emulsifier (c)" defined in the present invention includes one or more components selected from the group consisting of propylene glycol fatty acid esters, glycerin fatty acid esters, sucrose fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, organic acid monoglycerides, polyoxyethylene fatty acid esters, propylene glycol fatty acid esters, polyoxyethylene fatty acid esters, polysorbates, egg yolk lecithin, soybean lecithin, carboxymethylcellulose, glycerin, soybean phospholipids, and calcium stearyl lactylate.

[0043] The emulsifier serves to increase the miscibility between the components in the lactic acid bacteria complex. When the edible oil or fat is heated to 75 to 95°C, preferably 75 to 85°C, and then the emulsifier is mixed, the emulsifier melts and the miscibility with the edible oil or fat is greatly increased.

[0044] A process has been known in the past in which edible oils and fats are heated to 60°C and then mixed with an emulsifier. However, even when the edible oils and fats are actually heated to 60°C, there is a problem in that the emulsifier does not dissolve and cannot be mixed.

[0045] Experimental results confirmed that emulsifiers dissolve in edible oils and fats when the oils and fats are heated to at least 70°C, and that rancidity may occur when the oils and fats are heated to more than 100°C. Therefore, it is more preferable to heat the oils and fats to a temperature of 75-85°C. (See Figure 1)

[0046] The emulsifier content may be 0.1 to 5.0 parts by weight, preferably 0.20 to 1.60 parts by weight, and more preferably 0.30 to 1 part by weight, based on the total weight of the lactic acid bacteria complex. If the emulsifier content is less than 0.1 parts by weight, there may be a problem of reduced miscibility between the lactic acid bacteria mixed powder and edible oils and fats, and even if it is used in excess of 5.0 parts by weight, there is no further increase in miscibility between the compositions, so it is appropriate to use it within the above range.

[0047] According to a preferred embodiment of the present invention, there is provided a gastric acid-stable probiotic preparation comprising, as essential ingredients, (a) lactic acid bacteria, (b) edible oils and fats, (c) an emulsifier, and (d) nuts.

[0048] The probiotic preparation is characterized in that the weight ratios of the individual components are (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 90 parts by weight of edible oils and fats, (c) 0.1 to 5.0 parts by weight of an emulsifier, and (d) 5.0 to 90.0 parts by weight of nuts.

[0049] Of these, (a) lactic acid bacteria, (b) edible fats and oils, and (c) emulsifiers have been explained above and will not be explained here.

[0050] The "(d) nuts" defined in the present invention includes one or more ingredients selected from the group consisting of peanuts, almonds, walnuts, pine nuts, pistachios, pecans, macadamia nuts, hazelnuts, cashew nuts, Brazil nuts, chestnuts, soybeans, cacao nibs, coconuts, sunflower seeds, and pumpkin seeds, as additives for forming an appropriate viscosity of the composition, enhancing the flavor / texture, and further enhancing the activity of lactic acid bacteria.

[0051] The nuts contain a large amount of unsaturated fatty acids, which are essential components that play a beneficial role in various aspects of health, including healthy blood circulation. The nuts are preferably in a pulverized form, and are characterized by being pulverized to a particle size of 0.01 to 0.5 mm, preferably 0.01 to 0.1 mm, and having a paste-like shape.

[0052] The content of the nuts is 5.0 to 90.0 parts by weight, and preferably 10.0 to 30.0 parts by weight.

[0053] It has been confirmed through experiments that the nuts not only improve the palatability and organoleptic properties of the probiotic preparation, but also have an excellent effect of maintaining the viability of lactic acid bacteria in gastric acid.

[0054] The present invention provides a gastric acid-stable probiotic formulation which contains, in addition to the essential ingredients, one or more additional ingredients selected from (e) prebiotics and (f) adhesives.

[0055] In this case, the weight ratio of the individual components of the probiotic preparation is (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 90 parts by weight of edible oils and fats, (c) 0.1 to 5.0 parts by weight of emulsifier, (d) 5.0 to 90.0 parts by weight of nuts, (e) 0.01 to 5.0 parts by weight of prebiotics, and (f) 0.1 to 5.0 parts by weight of adhesive strength imparting agent.

[0056] More preferably, the composition is characterized by comprising (a) 1.0 part by weight of lactic acid bacteria, (b) 10 to 30 parts by weight of edible fats and oils, (c) 0.2 to 1.6 parts by weight of an emulsifier, (d) 15.0 to 30.0 parts by weight of nuts, (e) 0.5 to 2.0 parts by weight of prebiotics, and (f) 2.0 to 5.0 parts by weight of an adhesive strength imparting agent.

[0057] The "(e) prebiotics" as defined in the present application are ingredients that are used as food for lactic acid bacteria, are not digested by the digestive enzymes of animals, and increase the numbers of lactic acid bacteria such as bifidobacteria, which are intestinal bacteria, in the lower part of the small intestine, thereby exhibiting various physiological effects including an intestinal regulating effect. The monosaccharides, disaccharides, oligosaccharides, and polysaccharides that can be used as prebiotics well known in the art are possible, and preferably, monosaccharides containing pentoses such as xylose and arabinose, or hexoses such as glucose, mannose, fructose, and galactose; disaccharides such as lactulose, lactitol, sucrose, lactose, maltose, and trehalose; fructo-oligosaccharides, raffinose, and sucrose; and oligosaccharides such as amylose, amylopectin, starch, cellulose, pectin, etc.; and more preferably, sugar compounds such as glucose, fructose, galactose, sucrose, lactose, fructo-oligosaccharide, etc., and more preferably, glucose, fructose, lactulose, lactitol, or fructo-oligosaccharide.

[0058] The content of the (e) prebiotic component can be adjusted in a dosage well known in the art depending on the type and content of the (a) lactic acid bacteria, and is usually added in an amount of 0.01 to 5 parts by weight, preferably 0.5 to 2.0 parts by weight, per 1 weight of the lactic acid bacteria.

[0059] The "(f) tackifier" defined in the present application is contained in an amount of 0.1 to 5 parts by weight based on the total weight of the composition and serves to impart tackiness to the composition. It is preferable that the tackifier is one or more selected from the group consisting of sodium aluminosilicate, xanthan gum, carrageenan, guar gum, diutan gum, cellulose gum, gellan gum, pectin, and carboxymethylcellulose.

[0060] If the content of the tackifier is less than 0.1 parts by weight, the above effect is minimal, and if the content of the tackifier is more than 5 parts by weight, the viscosity may increase excessively, resulting in reduced workability.

[0061] In addition, the present invention may further add one or more additives selected from a suitable amount of coloring matter, flavoring, etc., to the composition in the range of about 0 to 10 parts by weight per 1 weight of the total lactic acid bacteria.

[0062] The present invention also provides a pharmaceutical composition, a functional health food, and a dietary supplement for promoting the proliferation of beneficial intestinal bacteria, which contain as an active ingredient the gastric acid-stable probiotic preparation produced by the above-mentioned production method.

[0063] The probiotic preparation of the present invention has been subjected to an emulsion production experiment at different heating temperatures (Experimental Example 1); an effect on the proliferation of beneficial intestinal bacteria (Experimental Example 2); an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the composition's efficacy in improving constipation (Simplified Clinical Example 1); and an experiment to evaluate the palatability of different nuts contained in the composition (Simplified Clinical Example 2). As a result, it has been confirmed that the probiotic preparation of the present invention strongly proliferates beneficial intestinal bacteria, exhibits excellent viability of lactic acid bacteria in gastric acid, and exerts remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability, and is useful as an excellent probiotic preparation for pharmaceutical compositions, health functional foods, and dietary supplements.

[0064] In order to achieve the above object, the present invention provides a pharmaceutical composition for promoting the proliferation of beneficial intestinal bacteria or for preventing and treating diseases caused by the proliferation of harmful intestinal bacteria, which comprises the probiotic preparation as an active ingredient.

[0065] "Diseases caused by the proliferation of harmful intestinal bacteria" as defined in the present application include, but are not limited to, antibiotic-associated diarrhea, ulcerative colitis, inflammatory bowel diseases such as Crohn's disease, abdominal bloating, constipation, acute or chronic irritable bowel disease, radiation-induced enteritis, Helicobacter infection, and the like.

[0066] The composition of the present invention contains the probiotic preparation in an amount of 0.01 to 99% by weight based on the total weight of the composition.

[0067] However, the composition is not necessarily limited to the above, and may vary depending on the condition of the patient and the type and progression of the disease.

[0068] The compositions containing the probiotic preparations of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

[0069] The compositions containing the probiotic preparations according to the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions by a conventional method, and examples of carriers, excipients, and diluents that can be included therein include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, and mineral oil. When formulated, the compositions are prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing the extract with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to water and liquid paraffin, which are frequently used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, etc. can be used.

[0070] The preferred dosage of the probiotic preparation of the present invention varies depending on the condition and weight of the patient, the degree of the disease, the drug form, the route and duration of administration, and can be appropriately selected by those skilled in the art. However, for a favorable effect, the probiotic preparation is administered at 0.01 mg / kg to 10 g / kg per day, preferably at 1 mg / kg to 1 g / kg per day. The administration can be once a day or in several divided doses. Moreover, the above dosage does not limit the scope of the present invention in any aspect.

[0071] The composition of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc., by various routes. All modes of administration can be envisaged, for example, oral and rectal, or intravenous administration.

[0072] The present invention also provides a method for treating a patient suffering from a disease caused by the promotion of proliferation of beneficial intestinal bacteria or the proliferation of harmful intestinal bacteria, comprising administering the probiotic preparation to the patient suffering from the disease caused by the promotion of proliferation of beneficial intestinal bacteria or the proliferation of harmful intestinal bacteria.

[0073] The present invention also provides use of the probiotic preparation for producing a medicament for promoting the proliferation of beneficial intestinal bacteria or for treating a disease caused by the proliferation of harmful intestinal bacteria.

[0074] The present invention also provides a health functional food containing a probiotic preparation as an active ingredient for promoting the proliferation of beneficial intestinal bacteria or for preventing and improving diseases caused by the proliferation of harmful intestinal bacteria.

[0075] The composition of the present invention contains the probiotic preparation in an amount of 0.01 to 99% by weight based on the total weight of the composition.

[0076] However, the composition is not necessarily limited to the above, and may vary depending on the condition of the patient and the type and progression of the disease.

[0077] The compositions containing the probiotic preparations of the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.

[0078] The compositions containing the probiotic preparations according to the present invention can be formulated and used in the form of oral preparations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, and sterile injection solutions by a conventional method, and examples of carriers, excipients, and diluents that can be included therein include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, and mineral oil. When formulated, the compositions are prepared using diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and such solid preparations are prepared by mixing the probiotic preparation with at least one or more excipients, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, oral liquids, emulsions, syrups, etc., and may contain various excipients such as wetting agents, sweeteners, flavoring agents, preservatives, etc. in addition to water and liquid paraffin, which are frequently used simple diluents. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a suppository base, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin, etc. can be used.

[0079] The preferred dosage of the probiotic preparation of the present invention varies depending on the condition and weight of the patient, the degree of the disease, the drug form, the route of administration and the period of administration, but can be appropriately selected by those skilled in the art. However, for a favorable effect, the probiotic preparation is administered at 0.01 mg / kg to 10 g / kg per day, preferably at 1 mg / kg to 1 g / kg per day. The administration can be once a day or in several divided doses. Moreover, the above dosage does not limit the scope of the present invention in any aspect.

[0080] The compositions of the present invention can be administered to mammals such as rats, mice, livestock, humans, etc., by various routes, for example, orally, rectally, or intravenously, although any mode of administration can be envisaged.

[0081] The present invention also provides a health food containing the probiotic preparation as an active ingredient for promoting the proliferation of beneficial intestinal bacteria or for preventing and ameliorating diseases caused by the proliferation of harmful intestinal bacteria.

[0082] "Food with health claims" as defined in this application means food that is manufactured and processed using raw materials or ingredients that have functional properties that are beneficial to the human body as defined by Law No. 6727 on Food with Health Claims, and "functional" means that the food is consumed for the purpose of obtaining beneficial effects for health purposes, such as regulating nutrients or achieving physiological effects on the structure and functions of the human body.

[0083] The health functional food of the present invention for promoting the proliferation of beneficial intestinal bacteria or preventing and improving diseases caused by the proliferation of harmful intestinal bacteria contains the above-mentioned components in a weight percentage of 0.01 to 95%, preferably 1 to 80%, based on the total weight of the composition.

[0084] Furthermore, the composition of the present invention may be a health food intended to promote the proliferation of beneficial intestinal bacteria or to treat or ameliorate diseases caused by the proliferation of harmful intestinal bacteria.

[0085] In addition, for the purpose of promoting the proliferation of beneficial intestinal bacteria or preventing and improving diseases caused by the proliferation of harmful intestinal bacteria, the composition can be manufactured and processed into health functional foods in the form of pharmaceutical dosage forms such as powders, granules, tablets, capsules, pills, suspensions, emulsions, syrups, etc., or in the form of tea bags, infused tea, health drinks, etc.

[0086] The present invention also provides a dietary supplement containing the probiotic preparation as an active ingredient for promoting the proliferation of beneficial intestinal bacteria or for preventing and ameliorating diseases caused by the proliferation of harmful intestinal bacteria.

[0087] In the dietary supplement defined in this application, the main ingredient means that the content of the active ingredient is in the range of about 30% to 99% by weight of the total composition, preferably in the range of 50% to 99%, and more preferably in the range of 70% to 99%.

[0088] The health functional beverage composition of the present invention contains the probiotic preparation as an essential component in the indicated ratio, and other components are not particularly limited, and may contain various flavorings or natural carbohydrates, etc., as additional components, like ordinary beverages. Examples of natural carbohydrates as mentioned above are monosaccharides, e.g., glucose, fructose, etc.; disaccharides, e.g., maltose, sucrose, etc.; and polysaccharides, e.g., ordinary sugars such as dextrin, cyclodextrin, etc., and sugar alcohols such as xylitol, sorbitol, erythritol, etc. As flavorings other than those mentioned above, natural flavorings (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.) can be advantageously used. The ratio of the natural carbohydrates is generally about 1 to 20 g, preferably about 5 to 12 g, per 100 ml of the composition of the present invention.

[0089] When the probiotic preparation according to the present invention is used as a food additive, the probiotic preparation can be added as it is or together with other foods or food ingredients, and can be used appropriately in a conventional manner. Examples of foods to which the substance can be added include meat, sausages, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, energy drinks, alcoholic beverages, and vitamin complexes, including all health foods in the conventional sense.

[0090] In addition to the above, the composition of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavorings such as synthetic flavorings and natural flavorings, coloring agents and enhancers (cheese, chocolate, etc.), pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonation agents used in carbonated drinks, etc. In addition, the composition of the present invention may contain natural fruit juices and fruit pulp for the production of fruit juice drinks and vegetable drinks. Such ingredients may be used independently or in combination. The proportion of such additives is not particularly important, but is generally selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

[0091] The probiotic preparation of the present invention can be added to food or beverages for the purpose of promoting the proliferation of beneficial intestinal bacteria or preventing diseases caused by the proliferation of harmful intestinal bacteria. In this case, the amount of the probiotic preparation in the food or beverage can be 0.01 to 15% by weight of the total food weight, and the health drink composition can be added in a proportion of 0.02 to 5 g, preferably 0.3 to 1 g, based on 100 ml.

[0092] The present invention also provides a food or food additive for promoting the proliferation of beneficial intestinal bacteria or for preventing and ameliorating diseases caused by the proliferation of harmful intestinal bacteria, which contains the probiotic preparation as an active ingredient.

[0093] Examples of items listed in the "Japanese Standards for Food Additives" include chemically synthesized products such as ketones, glycine, potassium citrate, nicotinic acid, and cinnamic acid; natural additives such as licorice coloring, licorice extract, crystalline cellulose, and guar gum; and mixed preparations such as sodium L-glutamate preparations, alkaline agents added to noodles, preservative preparations, and tar color preparations.

[0094] Examples of functional foods containing the probiotic preparation of the present invention include confectioneries such as bread, rice cakes, dried fruits, candies, chocolates, chewing gums, and jams; ice cream products such as ice cream, frozen desserts, and ice cream powders; dairy products such as milk, low-fat milk, lactose-decomposed milk, processed milk, goat's milk, fermented milk, buttermilk, concentrated milk, dairy cream, buttermilk, natural cheese, processed cheese, milk powder, and whey; processed meat products, processed eggs, and meat products such as hamburgers; kamaboko, hayashi Examples of suitable foods include, but are not limited to, fish products such as processed fish products such as sausages, bacon, and the like; noodles such as ramen, dried noodles, fresh noodles, milky noodles, gelatinized dried noodles, improved aged noodles, frozen noodles, and pastas; beverages such as fruit drinks, vegetable drinks, carbonated drinks, soy milk, and lactic acid bacteria drinks such as yogurt, and mixed drinks; soy sauce, miso, gochujang, chunjang, cheonggukjang, mixed jang, vinegar, sauces, tomato ketchup, curry, and seasonings such as dressings; margarine, shortening, and pizza.

[0095] The content of the probiotic preparation according to the present invention added to foods including beverages during the process of producing the health food can be appropriately adjusted as required. Effect of the Invention

[0096] The gastric acid-stable probiotic preparation of the present invention has been subjected to various experiments, including an emulsion production experiment at different heating temperatures (Experimental Example 1); an experiment on the effect on the proliferation of beneficial intestinal bacteria (Experimental Example 2); an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the composition's efficacy in improving constipation (Simplified Clinical Example 1); and an experiment to evaluate the palatability of different nuts contained in the composition (Simplified Clinical Example 2). As a result, it has been confirmed that the lactic acid bacteria preparation of the present invention strongly proliferates beneficial intestinal bacteria, exhibits excellent survival ability of the lactic acid bacteria in gastric acid, and exerts remarkable effects in terms of both constipation improvement and palatability that are superior to conventionally known lactic acid bacteria preparations.

[0097] Therefore, the present invention can provide an excellent gastric acid-stable probiotic preparation and a method for producing the same. [Brief description of the drawings]

[0098] [Figure 1] FIG. 1 is a diagram showing the results of an experiment observing the properties of emulsions of the emulsifier of the present invention at various temperatures. [Diagram 2] FIG. 2 shows the results of an experiment measuring the effect of the lactic acid bacteria preparation of the present invention on the growth of bacterial strains in a selective medium (MRS agar plate). [Diagram 3] FIG. 2 is a graph showing the results of an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) according to the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0099] It will be apparent to those skilled in the art that various modifications and variations can be made to the compositions, uses and manufacture of the present invention without departing from the spirit or scope of the invention.

[0100] However, the following examples and experimental examples are merely illustrative and do not limit the scope of the present invention, and the contents of the present invention are not limited to the following examples and experimental examples.

[0101] Example 1. Production of emulsifier-added lactic acid bacteria preparation (composition: Table 1) 20 g of sunflower oil (vegetable oil, SAJO DAERIM Co., Ltd.) was placed in a stirring heating device (Hotplate Stirrer. MSH-20D) and heated to 75-85°C. 0.4 g of glycerin fatty acid ester (emulsifier, ES food ingredient) was then added and mixed with stirring.

[0102] The mixture of vegetable oil and emulsifier from the above step was emulsified while stirring in a circulating cooling water bath (low-temperature circulating thermostatic water bath, SH-WB-7CDR) maintained at 0-4°C until the temperature of the mixture reached 4-10°C. (Test-1: See Experimental Example 1 below for the experimental process and results of emulsion production at each temperature of 40°C, 60°C, 85°C, and 100°C)

[0103] The gel-shaped emulsified oil was mixed with the mixed lactic acid bacteria, i.e., Lactobacillus acidophilus DDS-1 (All Lab Co., Ltd.), Lactobacillus rhamnosus GG (All Lab Co., Ltd.), Lactobacillus plantarum VI-07 (Bitec Co., Ltd.), Bifidobacterium longum BL5 (Bitec Co., Ltd.), and Bifidobacterium animalis ssp. lactis BLC1 (All Lab Co., Ltd.), at the same bacterial count (2 x 10 9 cfu / g) was used. (Test-2: For the activity test process and results in artificial gastric juice (pH 2) to observe the viability of lactic acid bacteria, please refer to Experimental Example 2 below.)

[0104] Fructooligosaccharides (food for lactobacilli; prebiotics), a natural sweetener, and peanuts (nuts, Daisho Foods), which contain large amounts of unsaturated fatty acids and play various beneficial roles in health, including healthy blood circulation, were added as additives to form the appropriate viscosity of the composition, enhance flavor / texture, and further strengthen the activity of lactobacilli. The mixture was then stirred at 30-50 rpm in a mixer (OFM-1507B, Ofel Co., Ltd.) for at least 1 hour until all ingredients were uniformly mixed.

[0105] In this case, in the case of nuts, the best method is to crush them with a crushing device (ultra-high speed vacuum blender, KCB-001W, KUCHAM Co., Ltd.) to a particle size of 0.01 to 0.1 mm (enough to form a paste) (Test-3: Peanuts, almonds, and walnuts, which were selected and evaluated as the most palatable peanuts, were mixed into the composition, and the ease of drinking and palatability were evaluated. See Experimental Example 3 for the experimental results).

[0106] [Table 1]

[0107] Example 2. Production of a lactic acid bacteria preparation without the addition of an emulsifier (composition: Table 1) The emulsification step using vegetable oil and an emulsifier as described in the preparation method of composition A in Example 1 was omitted, and sunflower oil (vegetable oil), five kinds of mixed lactic acid bacteria, and the remaining additives were simply mixed under the same conditions to prepare a lactic acid bacteria preparation (composition C) without adding ground nuts, which was used as a sample in the following experimental examples.

[0108] Example 3. Production of comparative lactic acid bacteria preparation A comparative enteric-soluble lactic acid bacteria complex containing 10.0 wt% lactic acid bacteria powder, 11.5 wt% yellow lead (suspending agent), 1.0 wt% soybean lecithin (emulsifier) ​​and the remaining rTG-type refined fish oil (edible oil) as disclosed in a conventional document (Korean Patent Publication No. 10-2023-0001175) was prepared by the manufacturing method of Example 1 of the conventional document, and this was used as a sample in the following experimental examples.

[0109] Experimental example 1. Experiment to produce emulsion at different temperatures 2.0 g of emulsifier (glycerin fatty acid ester, ES food ingredient) was added to 100 g of sunflower oil (vegetable oil, SAJO DAERIM Co., Ltd.), heated at four different temperatures, i.e., 40°C, 60°C, 85°C, and 100°C, and emulsified with stirring in a circulating cooling water bath (low-temperature circulating thermostatic water bath, SH-WB-7CDR) maintained at 0-4°C until the temperature of the mixture reached 4-10°C, and the optimal temperature for producing an emulsion was measured.

[0110] The experimental results are shown in Figure 1, which shows the shape of the emulsion formed at each temperature. At 40°C and 60°C, the emulsifier was not fully dissolved and the emulsion was not well formed, but at 85°C and 100°C, the emulsifier was fully dissolved and the emulsion was uniformly formed. (See Figure 1: Experiment on emulsion properties at various temperatures and emulsifier)

[0111] Experimental Example 2. Effect on the proliferation of beneficial intestinal bacteria In order to confirm the effect of the samples of the present invention obtained from the above examples on the proliferation of beneficial intestinal bacteria, the following experiment was carried out with reference to the lactobacillus count test method announced in the Food Regulations Microbiology Testing Method Manual (published by the Food and Drug Safety Evaluation Service of the Ministry of Food and Drug Safety).

[0112] 2-1. Experimental strains The strains were provided by the Korean Culture Center of Microorganisms (KCCM, Korea), and Lactobacillus acidophilus ATCC4356 (ATCC: American Type Culture Collection) was used as the beneficial intestinal bacteria. Lactobacilli MRS broth (288130, BD difco, USA) was used as the growth medium for culturing the beneficial intestinal bacteria, and the strains were activated by subculture at least three times before use in the experiment.

[0113] 2-2. Experimental process The number of lactobacilli produced was measured by counting the number of lactobacillus colonies using MRS plate measurement medium, and the average colony number was multiplied by the dilution factor to calculate the number of live bacteria per mL of culture liquid. A 9-fold dilution solution (sterile saline) was added to 10 mL of the test sample and homogenized (10 -1 Solution). Test solution (10 -1 Add diluent to 1 mL of solution to make 10 mL. -2 After the test solutions are made, they are diluted in the same manner.

[0114] Each diluted test solution was inoculated at 1 (v / v)% into Lactobacillus selective medium (MRS agar plaste), and then anaerobically cultured at 37°C for 72±3 hours. After that, plates that produced 15 to 300 colonies per plate were selected and the number of colonies was calculated. The measured number of colonies was multiplied by the dilution factor to calculate the number of bacteria.

[0115] 2-3.Experimental results Based on the viability results of the above-mentioned example samples, it was confirmed that the best method for preparing the emulsion was at 85°C (see Table 2 and Figure 2).

[0116] [Table 2]

[0117] Experimental Example 3. Lactic acid bacteria viability experiment in artificial gastric juice (pH 2) In order to confirm the effect of the samples of the present invention obtained from the above examples on the viability of lactic acid bacteria in artificial gastric juice (pH 2), the following experiment was performed by applying the method disclosed in the literature (Jeon et al., 2007, Identification and Characterization of Lactic Acid Bacteria Starters Isolated from the Commercial Drink-Yogurt Products, Korean J. Food Sci. Ani. Resour. 27(4), 509-516).

[0118] 3-1. Experimental strains and experimental procedures In order to confirm the viability of the lactic acid bacteria in the composition, composition A was stirred in artificial gastric juice (pH 2) for a certain time (1 hour), and the collected composition was immediately plated and cultured on a lactobacillus selection medium (MRS agar plate, MB-M1024-P50, KisanBio) and cultured anaerobic at 37°C for 72±3 hours. Plates that produced 15 to 300 colonies per plate were selected, and the number of colonies was calculated by multiplying the measured number of colonies by the dilution factor. For comparison, a composition control group in which a lactic acid bacteria mixture was mixed with non-emulsified oil was used.

[0119] 3-2.Experimental results From the experimental results, it can be seen that the emulsified oil and lactic acid bacteria mixture has a higher lactic acid bacteria survival rate than the control group. In addition, when ground peanuts were further added as an additive to the emulsified oil and cultured under the same conditions, it was observed that the lactobacillus survival rate in the lactic acid bacteria mixture further containing ground peanuts was significantly higher (see Table 3 and Figure 3).

[0120] [Table 3]

[0121] Simple clinical example 1. Efficacy test of the composition for improving constipation In order to confirm the efficacy of the samples of the present invention obtained from the above examples in improving constipation, the following experiment was carried out.

[0122] Composition A, which was used in the test for improving constipation, was evaluated for its effect of improving constipation on 50 Seoul-area supporters in their 20s to 50s who experience inconvenience in their daily lives due to constipation.

[0123] The subjects were instructed to take 5 g of composition A twice a day (morning and evening) for 10 days and to answer the following questions. As a control group, 50 subjects were instructed to take composition B in the same manner as the subjects.

[0124] As a result of the above experiments, it can be confirmed that the composition A of the present invention has an excellent effect of improving constipation (see Tables 4 and 5).

[0125] [Table 4]

[0126] [Table 5]

[0127] Simple clinical example 2. Preference evaluation experiment based on the type of nuts contained in the composition In order to confirm the preference depending on the type of nuts contained in the sample compositions of the present invention obtained from the above examples, the following experiment was carried out.

[0128] Compositions were prepared using peanuts, almonds, and walnuts as ground nuts, and the taste, aroma, texture, and overall preference of each type of nut were evaluated by having 20 adult men and women ingest 3g of each.

[0129] The criteria for the preference evaluation were a 5-point rating method followed by the average value (very good 5 points, good 4 points, normal 3 points, poor 2 points, very poor 1 point) (see Table 6).

[0130] [Table 6]

[0131] From the above experimental results, it can be seen that the composition using peanut powder has the highest preference.

[0132] As a result of the above experiments, which were conducted on the lactic acid bacteria preparation of the present invention, including the effect on the proliferation of beneficial intestinal bacteria (Experimental Example 2); an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the composition's efficacy in improving constipation (Simplified Clinical Example 1); and an experiment to evaluate the palatability of the type of nuts contained in the composition (Simplified Clinical Example 2), it was confirmed that the lactic acid bacteria preparation of the present invention strongly proliferates beneficial intestinal bacteria, exhibits excellent survival ability of lactic acid bacteria in gastric acid, and exerts remarkable effects in terms of improving constipation and palatability that are superior to conventionally known lactic acid bacteria preparations. EXAMPLES

[0133] Hereinafter, examples of formulations of compositions containing the probiotic preparation of the present invention will be described, but the present invention is not intended to be limited thereto, and is merely intended to specifically illustrate the same.

[0134] Formulation example 1. Manufacturing of powder Composition A---------------------- 20mg Lactose----------------------100mg Talc------------------------10mg

[0135] The above ingredients are mixed and filled into an airtight bag to produce a powder.

[0136] Formulation Example 2. Tablet Production Composition A---------------------- 10mg Cornstarch--------------------100mg Lactose----------------------100mg Magnesium stearate---------------2mg The above ingredients are mixed and then compressed into tablets by a conventional tablet manufacturing method.

[0137] Formulation Example 3. Production of capsules Composition A---------------------- 10mg Crystalline cellulose-------------------3mg Lactose----------------------14.8mg Magnesium stearate---------------0.2mg

[0138] The above ingredients are mixed according to a conventional capsule manufacturing method, and the mixture is filled into a gelatin capsule to produce a capsule.

[0139] Formulation Example 4. Manufacture of injections Composition A---------------------- 10mg Mannitol--------------------180mg Sterile distilled water for injection ----------------- 2974mg Na2HPO4·12H2O --------------- 26mg

[0140] It is prepared according to the usual manufacturing method for injections, with the above-mentioned ingredient contents per ampoule (2 ml).

[0141] Formulation Example 5. Preparation of liquid formulation Composition A---------------------- 20mg Isomerized sugar---------------------- 10g Mannitol---------------------5g Purified water ------------------------ Appropriate amount

[0142] According to a typical liquid preparation method, each ingredient is dissolved in purified water, and an appropriate amount of lemon scent is added. The ingredients are mixed, and then purified water is added to make a total of 100 ml. The liquid is then filled into a container and sterilized to produce the liquid.

[0143] Formulation example 6. Manufacturing of health foods Composition A---------------------- 1000mg Vitamin Mixture-------------------- Dosage Vitamin A acetate----------------- 70μg Vitamin E----------------------1.0 mg Vitamin B1--------------------0.13mg Vitamin B2---------------------0.15mg Vitamin B6--------------------0.5mg Vitamin B12--------------------0.2μg Vitamin C----------------------10mg Biotin-----------------------10μg Nicotinamide-------------------1.7mg Folic acid------------------------- 50μg Calcium pantothenate----------------0.5mg Inorganic mixture---------------------- Appropriate amount Ferrous sulfate----------------------1.75mg Zinc oxide----------------------- 0.82mg Magnesium carbonate-------------------25.3mg Potassium phosphate monobasic ------------------ 15mg Dicalcium phosphate----------------- 55mg Potassium citrate-------------------90mg Calcium carbonate--------------------100mg Magnesium chloride-------------------24.8mg

[0144] The composition ratio of the above vitamin and mineral mixture is a mixture of ingredients relatively suitable for health foods in a preferred embodiment, but the mixing ratio may be modified as desired. After mixing the above ingredients using a normal health food manufacturing method, granules can be produced and used to manufacture a health food composition using a normal method.

[0145] Formulation example 7. Manufacturing of health drinks Composition A---------------------- 1000mg Citric acid-------------------1000mg Oligosaccharides-----------------------100g Plum extract-------------------2g Taurine-----------------------1g Add purified water ----------------- Total 900ml

[0146] The above ingredients are mixed according to a conventional method for producing health drinks, then stirred and heated at 85°C for about 1 hour. The resulting solution is then filtered and placed in a sterilized 2L container, which is then sealed and sterilized, refrigerated, and then used to produce the health drink composition of the present invention.

[0147] The above composition ratio is a mixture of ingredients that are relatively suitable for luxury drinks in a preferred embodiment, but the blending ratio may be arbitrarily modified depending on regional and ethnic preferences such as demand class, demand country, and intended use. [Industrial Applicability]

[0148] As described above, the probiotic preparation of the present invention has been subjected to an emulsion production experiment at different heating temperatures (Experimental Example 1); an effect on the proliferation of beneficial intestinal bacteria (Experimental Example 2); an experiment on the viability of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the composition's efficacy in improving constipation (Simplified Clinical Example 1); and an experiment to evaluate the palatability of different nuts contained in the composition (Simplified Clinical Example 2). As a result, it has been confirmed that the lactic acid bacteria preparation of the present invention strongly proliferates beneficial intestinal bacteria, exhibits excellent viability of lactic acid bacteria in gastric acid, and exerts remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability. Therefore, the lactic acid bacteria preparation of the present invention can be usefully used as an excellent probiotic preparation in various pharmaceutical compositions, health functional foods, and dietary supplements.

Claims

1. A first step of heating 1 to 90 parts by weight of edible fats and oils at 70 to 95°C, and then mixing and stirring with 0.1 to 5 parts by weight of an emulsifier to produce a mixture of edible fats and oils and an emulsifier; A second step of cooling and stirring the mixture of the first step until the temperature reaches 2-20°C to produce a gel-shaped coagulated mixture; A method for producing a gastric acid-stable probiotic preparation, comprising the steps of: (a) slowly adding 1 part by weight of lactic acid bacteria to the gel-form aggregate mixture and stirring at low speed to produce a lactic acid bacteria mixture; and (b) carrying out a step of:

2. A fourth step of adding ground nuts to the mixture obtained in the third step and stirring to produce a nut-based lactic acid bacteria mixture; 2. The method for producing the gastric acid stable probiotic formulation of claim 1, further comprising a fifth step of adding one or more additional ingredients selected from prebiotics and adhesives to the mixture of the fourth step and stirring the mixture.

3. 3. The method for preparing a gastric acid-stable probiotic formulation according to claim 2, wherein the ground nuts in the fourth step have a particle size of 0.01-0.5 mm, and are stirred at a stirring speed of 30-50 rpm for 30 minutes to 3 hours until all ingredients are uniformly mixed.

4. A gastric acid-stable probiotic preparation comprising, as essential ingredients, (a) lactic acid bacteria, (b) edible oils and fats, (c) an emulsifier, and (d) nuts.

5. 5. The gastric acid-stable probiotic preparation according to claim 4, characterized in that the weight ratios of the individual components in the probiotic preparation are: (a) 1.0 part by weight of lactic acid bacteria, (b) 1.0 to 90.0 parts by weight of edible oils and fats, (c) 0.1 to 5.0 parts by weight of an emulsifier, and (d) 5.0 to 90.0 parts by weight of nuts.

6. 5. The gastric acid-stable probiotic preparation according to claim 4, wherein the (a) lactic acid bacteria is one or more strains selected from microorganisms of the genus Lactobacillus, the genus Bifidobacterium, the genus Bacillus, the genus Streptococcus, and the genus Enterococcus.

7. 5. The gastric acid-stable probiotic formulation of claim 4, wherein the (b) edible oil is one or more components selected from the group consisting of avocado oil, canola oil, evening primrose oil, palm oil, Brazil nut oil, corn oil, cottonseed oil, linseed oil, grape seed oil, hemp oil, olive oil, palm oil, safflower oil, soybean oil, peanut oil, sunflower oil, krill oil, anchovy oil, salmon oil, rice bran oil, brown rice oil, coconut oil, rosehip oil, and tuna oil.

8. The gastric acid-stable probiotic formulation of claim 4, characterized in that the (c) emulsifier is one or more components selected from the group consisting of propylene glycol fatty acid esters, glycerin fatty acid esters, sucrose fatty acid esters, polyglycerin fatty acid esters, sorbitan fatty acid esters, organic acid monoglycerides, polyoxyethylene fatty acid esters, propylene glycol fatty acid esters, polyoxyethylene fatty acid esters, polysorbates, egg yolk lecithin, soybean lecithin, carboxymethylcellulose, glycerin, soybean phospholipids, and calcium stearyl lactate.

9. 5. The gastric acid-stable probiotic formulation of claim 4, wherein the nuts (d) are one or more components selected from the group consisting of peanuts, almonds, walnuts, pine nuts, pistachios, pecans, macadamia nuts, hazelnuts, cashew nuts, Brazil nuts, chestnuts, soybeans, cocoa nibs, coconuts, sunflower seeds, and pumpkin seeds.

10. 5. The gastric acid-stable probiotic formulation according to claim 4, further comprising, in addition to the essential ingredients, one or more additional ingredients selected from (e) prebiotics and (f) adhesives.

11. 11. The gastric acid-stable probiotic preparation according to claim 10, wherein the weight ratios of the individual components are: (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 60 parts by weight of edible fats and oils, (c) 0.1 to 5.0 parts by weight of emulsifier, (d) 5.0 to 90.0 parts by weight of nuts, (e) 0.01 to 5.0 parts by weight of prebiotics, and (f) 0.1 to 5.0 parts by weight of adhesive strength imparting agent.

12. The (e) prebiotics may be a pentose such as xylose, arabinose, or glucose, mannose, fructose, galactose, lactulose, lactitol, sucrose, lactose, maltose, trehalose, etc.

11. The gastric acid stable probiotic formulation according to claim 10, characterized in that the probiotic is one or more components selected from the group consisting of saccharides, saccharides (saccharides), ...

13. 11. The gastric acid-stable probiotic formulation of claim 10, wherein the (f) adhesive agent is one or more components selected from the group consisting of sodium aluminosilicate, xanthan gum, carrageenan, guar gum, diutan gum, cellulose gum, gellan gum, pectin and carboxymethylcellulose.

14. A pharmaceutical composition for promoting the proliferation of beneficial intestinal bacteria or for preventing and treating diseases caused by the proliferation of harmful intestinal bacteria, comprising the gastric acid-stable probiotic preparation according to claim 4 as an active ingredient.

15. The pharmaceutical composition according to claim 14, characterized in that the disease caused by the proliferation of harmful intestinal bacteria is antibiotic-associated diarrhea, watery diarrhea, vomiting, ulcerative colitis accompanied by nausea, inflammatory bowel disease such as Crohn's disease, abdominal distension, constipation, acute or chronic irritable bowel disease, radiation-induced enteritis, or Helicobacter infection.

16. A health food with functional properties for promoting the proliferation of beneficial intestinal bacteria or for preventing or improving diseases caused by the proliferation of harmful intestinal bacteria, comprising the gastric acid-stable probiotic preparation according to claim 4 as an active ingredient.

17. The health functional food according to claim 16, characterized in that the health functional food is in the form of a pharmaceutical dosage form such as powder, granules, tablets, capsules, pills, suspensions, emulsions, or syrups, or in the form of tea bags, infused tea, or health drinks.

18. A dietary supplement for promoting the proliferation of beneficial intestinal bacteria or for preventing or improving diseases caused by the proliferation of harmful intestinal bacteria, comprising the gastric acid-stable probiotic preparation according to claim 4 as an active ingredient.

19. A food additive for promoting the proliferation of beneficial intestinal bacteria or for preventing or ameliorating diseases caused by the proliferation of harmful intestinal bacteria, comprising the gastric acid-stable probiotic preparation according to claim 4 as an active ingredient.

20. 5. Use of the gastric acid-stable probiotic preparation according to claim 4 for the production of a medicament for promoting the proliferation of beneficial intestinal bacteria or for treating a disease caused by the proliferation of harmful intestinal bacteria.

Images