Gastric acid-stable probiotic preparation and method for manufacturing 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 preparation containing edible oil, an emulsifier, and nuts, and a method for producing the same.

Background Art

[0002] Generally, lactic acid bacteria colonize in the intestine and exhibit various physiological activity effects such as activation of intestinal motility, suppression of harmful bacteria, promotion of vitamins and immune-enhancing substances. However, due to the structure of the human body, after humans ingest lactic acid bacteria, they pass through the stomach before reaching the intestine, and lactic acid bacteria are killed by gastric acid secreted from the intestine, and in many cases, the original physiological activity function of lactic acid bacteria cannot be exerted.

[0003] Therefore, there have been attempts to coat acid bacteria powder with various proteins or carbohydrates to prevent the death of lactic acid bacteria in the body, and thus, lactic acid bacteria powder coated with various substances and a method for producing the same have been developed.

[0004] The human large intestine and feces contain over 400 species of bacteria, more than 90% of which are anaerobic strains. These bacteria form a microbiome in the gut, maintaining a certain balance, and are affected not only by the body's health, stress, aging, and physiological changes, but also by external factors such as food and drugs. For physical health and extended lifespan, it is necessary to maintain a state in the gut microbiome where there are more beneficial bacteria and fewer 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 Literature 1), meaning live bacterial preparations that can help improve gut microbiota. Prebiotics, on the other hand, refer to substances that can be used by beneficial gut microorganisms and that help the growth and function of beneficial gut bacterial strains. Prebiotics known to date are non-starch polysaccharides and oligosaccharides, which are not easily broken down by human enzymes but are usable by microorganisms (Non-Patent Literature 2). Recently, the concept of such prebiotics has broadened with the recognition of the effects of polyphenols, which can have beneficial effects on intestinal microorganisms (Non-Patent Literature 3).

[0005] Lactic acid bacteria are representative bacteria widely used as probiotics. They are abundant in nature, easily found in the intestines of humans and animals, and in fermented foods, and are microorganisms recognized as safe by the U.S. Food and Drug Administration. Lactic acid bacteria attach to and parasitize intestinal epithelial cells, 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 suppressing the colonization of harmful bacteria, preventing disease, activating the immune system, anticancer effects, and lowering cholesterol. The fact that lactic acid bacteria have growth-inhibiting effects on various putrefactive and pathogenic microorganisms is due to several metabolic characteristics, but lactic acid bacteria produce antibacterial active factors such as organic acids, hydrogen peroxide, reuterin, diacetyl, acetaldehyde, and bacteriocins as metabolites (Non-patent Literature 4; Patent Literature 1).

[0006] Probiotics are currently mainly used as functional foods, but they are also used to some extent for therapeutic purposes. Specifically, they increase the production of immunoglobulin A antibodies, reduce rotavirus shedding, and shorten the duration of diarrhea. Therefore, their application is being actively explored as a preventative and therapeutic agent for antibiotic-associated diarrhea, a treatment for inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, a treatment for abdominal bloating and constipation, a treatment for irritable bowel syndrome, a treatment for radiation-induced colitis, and an adjunct to the treatment of Helicobacter infections. They have been reported to be particularly effective as a treatment 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 the spontaneous intake of solid formulations by companion animals, characterized by comprising an oil containing unsaturated fatty acids, a tackifier, a sweetener, a lactic acid bacteria mixture, 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 stomach acid and exhibit activity in the small and large intestines. This preparation is manufactured by heating and emulsifying two mixed solutions of fat and emulsifier (heated to approximately 60°C) and a protective agent and lactic acid bacteria (heated to approximately 50°C), and then spraying the emulsified solution under high pressure into a dispersion cooled to 4°C. This microcapsule preparation of lactic acid bacteria prevents the death of lactic acid bacteria by stomach acid, thereby increasing the colonization rate of lactic acid bacteria in the intestines and being effective in promoting intestinal health.

[0009] Patent Document 5 discloses a complex functional enteric-coated soft capsule that contains a high content of lactic acid bacteria and edible oils and fats, while also guaranteeing the number of lactic acid bacteria throughout the shelf life. The complex contains 5.0-25.0% by weight of lactic acid bacteria powder, 5.0-20.0% by weight of a suspending agent, 0.5-2.0% by weight of an emulsifier, and the remainder of edible oils and fats.

[0010] Patent document 6 discloses an oral paste containing approximately 80-90% oil, approximately 0.5-20% emulsifier, and approximately 1-2% probiotic lactic acid bacteria effective against tartar and plaque.

[0011] However, none of the aforementioned documents disclose or describe alternatives to gastric acid-stabilized probiotic preparations containing edible oils and fats, emulsifiers, and nuts, or methods for producing the same.

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

[0013] Experiments were conducted on the lactic acid bacteria preparation of the present invention, including: emulsification liquid production experiments at different heating temperatures (Experimental Example 1); effects on the growth of beneficial intestinal bacteria (Experimental Example 2); lactic acid bacteria survival rate experiments in artificial gastric juice (pH 2) (Experimental Example 3); constipation relief efficacy experiments of the composition (Simplified Clinical Example 1); and palatability evaluation experiments based on the type of nuts contained in the composition (Simplified Clinical Example 2). As a result, it was confirmed that the lactic acid bacteria preparation of the present invention strongly promotes the growth of beneficial intestinal bacteria, exhibits excellent survival rate of lactic acid bacteria in gastric acid, and shows remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of constipation relief and palatability. This confirmed that an excellent probiotic preparation can be provided, thus completing the present invention. [Prior art documents] [Patent Documents]

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

[0015] [Non-Patent Document 1] Jung 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

Non-Patent Document 4

Summary of the Invention

Problems to be Solved by the Invention

[0016] The present invention has been made to overcome the problems as described above. The present invention provides a gastric acid-stable probiotic preparation containing edible oil, an emulsifier, and nuts, which strongly proliferates beneficial bacteria in the intestine, shows excellent viability against gastric acid, and exhibits remarkable efficacy superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability, as well as a method for producing the same.

Means for Solving the Problems

[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 later is as follows.

[0018] As a preferred embodiment of the present invention, in the first step, 1 to 90 parts by weight of edible oil is heated at 70 to 95 °C, and then 0.1 to 5 parts by weight of the emulsifier is mixed and stirred to produce a mixture of edible oil and emulsifier; in the second step, while cooling and stirring the mixture of the first step until the temperature reaches 2 to 20 °C, a gel-shaped aggregated mixture is produced; in the third step, 1 part by weight of lactic acid bacteria is slowly added to the gel-shaped aggregated mixture and stirred at a low speed to produce a lactic acid bacteria mixture. A gastric acid-stable probiotic preparation obtained by performing the steps including these is provided.

[0019] As a preferred embodiment of the present invention, in the fourth step, a nut pulverized product is added to and stirred in the mixture obtained by performing the third step to produce a nut lactic acid bacteria mixture; in the fifth step, one or more additional components arbitrarily selected from prebiotics and adhesives are added to and stirred in the mixture of the fourth step. A gastric acid-stable probiotic preparation obtained by performing the steps further including these is provided.

[0020] As a preferred embodiment of the present invention, the nut pulverized product has a particle size of 0.01 to 0.5 mm, preferably 0.01 to 0.1 mm, and is 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] As a preferred embodiment of the present invention, the gastric acid-stable probiotic preparation contains (a) lactic acid bacteria, (b) edible oil, (c) emulsifier, and (d) nuts as essential components.

[0022] As a preferred embodiment of the present invention, the blending weight ratio between the individual composition components of the gastric acid-stable probiotic preparation is composed of (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 90 parts by weight of edible oil, (c) 0.1 to 5.0 parts by weight of emulsifier, and (d) 5.0 to 90.0 parts by weight of nuts.

[0023] In a preferred embodiment of the present invention, the lactic acid bacteria (a) are characterized by being 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 characterized by being one or more components selected from the group consisting of avocado oil, canola oil, evening primrose oil, coconut 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 emulsifier (c) is characterized by being 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, polysorbate, egg yolk lecithin, soy lecithin, carboxymethylcellulose, glycerin, soy phospholipids, and calcium stearyl lactate.

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

[0027] In a preferred embodiment of the present invention, the gastric acid-stable probiotic preparation contains essential components consisting of (a) lactic acid bacteria, (b) edible oils and fats, (c) emulsifiers and (d) nuts, and further contains one or more additional components selected from (e) prebiotics and (f) tackifiers.

[0028] In a preferred embodiment of the present invention, the weight ratio of the individual components of the gastric acid-stable probiotic preparation is as follows: (a) 1.0 part by weight of lactic acid bacteria, (b) 1 to 60 parts by weight of edible oils and fats, (c) 0.1 to 5.0 parts by weight of emulsifiers, (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 tackifiers.

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

[0030] In a preferred embodiment of the present invention, the (f) tackifier is characterized by being 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] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings, so that persons with ordinary skill in the art to which the present invention pertains can easily implement the present invention.

[0032] The gastric acid-stable probiotic preparation according to the present invention is manufactured by performing the following steps: a first step of heating 1 to 90 parts by weight of edible oil and fat to 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 and fat and 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, to produce a gel-like aggregated mixture; and a third step of slowly adding 1 part by weight of lactic acid bacteria to the gel-like aggregated mixture and stirring at a low speed to produce a lactic acid bacteria mixture.

[0033] Accordingly, the present invention provides a method for producing a gastric acid-stable probiotic preparation according to the present invention, comprising the steps of: a first step of heating 1 to 90 parts by weight of edible oil and fat to 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 and fat and emulsifier; a second step of cooling and stirring the mixture from the first step until the temperature reaches 2 to 20°C, preferably 4 to 10°C, to produce a gel-like aggregated mixture; and a third step of slowly adding 1 part by weight of lactic acid bacteria to the gel-like aggregated mixture and stirring at low speed to produce a lactic acid bacteria mixture.

[0034] Through this process, the emulsifier is dissolved and completely mixed with heated edible oils and fats. Subsequently, cooling is performed to produce a stable, aggregated gel-like mixture. By mixing lactic acid bacteria into this mixture, the ability of the lactic acid bacteria to maintain their presence in the intestines can be significantly improved.

[0035] The gastric acid-stable probiotic preparation according to the present invention may be manufactured further comprising: a fourth step of adding crushed 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 components selected from prebiotics and tackifiers to the mixture obtained in the fourth step and stirring.

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

[0037] In this invention, "(a) lactic acid bacteria" refers to a strain of bacteria that aids digestion and absorption, inhibits the growth of harmful bacteria in the intestines, and maintains a favorable intestinal environment. The form of the lactic acid bacteria is not limited, but various forms such as dried powder and dried coarse powder are preferred, and the strain size may be in the range of 0.1 to 8.0 μm, more preferably 0.2 to 4.0 μm.

[0038] In this invention, the content of individual components is expressed in parts by weight, based on 1 part by weight of lactic acid bacteria.

[0039] As representative strains, preferably any lactic acid bacteria widely used in probiotics, readily available in the industry, such as those from the American Type Culture Collection (ATCC), the Korea Biotechnology Research Institute's Center for Type Cultures (KCTC), the Korean Culture Center of Microorganisms (KCCM), and the Korean Agricultural Genetic Resources Center (KACC) of the Korea Rural Development Administration. The microorganisms are available from collections such as the Collection, and specifically include microorganisms of the genera Lactobacillus, Bifidobacterium, Bacillus, Streptococcus, and Enterococcus; preferably Bifidobacterium, Lactobacillus, or Streptococcus strains, and more preferably Bifidobacterium or Streptococcus strains; more preferably Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, and Bifidobacterium animalis subspecies lactis. ssp. lactis), Bifidobacterium adolescentis, Bifidobacterium pseudocatenulatum, Bifidobacterium catenulatum, or Bifidobacterium infantis, Bifidobacterium thermophyllum (B.Bifidobacteria such as *Lactobacillus thermophilum*; strains of the genus *Bifidobacterium*, such as *Lactobacillus plantarum*, *Lactobacillus pentosus*, *Lactobacillus casei*, *Lactobacillus casei ssp. paracasei*, *Lactobacillus rhamnosus*, *Lactobacillus acidophilus*, *Lactobacillus delbrueckii*, and *Lactobacillus delbrueckii* subspecies *Lactobacillus delbrueckii*. 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 salivarius (Lb.Lactobacillus strains such as *Lactobacillus salivarius*; Bacillus strains such as *Bacillus cereus toyoi* or *Bacillus cereus*; Leuconostoc strains such as *Leuconostoc citreum* and *Leuconostoc mesenteroides*; Pediococcus strains such as *Pediococcus acidilactici* and *Pediococcus pentosaceus*; Propionibacterium assidiaceens Propionibacterium species such as acidifaciens and Propionibacterium acidipropionici; Streptococcus species such as Streptococcus thermophilus, S. cremoris, S. infantarius, S. intermedius, S. lactis, and S. salivarius subsp. thermophilus; Enterococcus faecalis and Enterococcus faecium (E.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*, *Lactobacillus plantarum*, *Bifidobacterium longum*, and *Bifidobacterium animalis*. It comprises one or more single or mixed strains, such as Lactobacillus ssp. Lactis, more preferably two or more mixed strains selected from Lactobacillus acidophilus, Lactobacillus rhamnosus GG, Lactobacillus plantarum, Bifidobacterium longum, and Bifidobacterium animalis ssp. Lactis.

[0040] In the present invention, "(b) edible oils and fats" means one or more components selected from the group consisting of avocado oil, canola oil, evening primrose oil, coconut 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 plays a role in enveloping the lactic acid bacteria complex and protecting it from stomach acid. The edible oil is mixed in an amount of 1 to 90 parts by weight, preferably 15 to 30 parts by weight. First, the edible oil is heated to 75 to 95°C, preferably 75 to 85°C, and then the emulsifier is mixed in and stirred to form a mixture of edible oil and emulsifier. This mixture is then 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, thereby forming a stable gel-like aggregate.

[0042] In the present invention, "(c) emulsifier" as defined herein 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, polysorbate, egg yolk lecithin, soy lecithin, carboxymethylcellulose, glycerin, soy phospholipids, and calcium stearyl lactate.

[0043] The emulsifier plays a role in increasing the miscibility between components within the lactic acid bacteria complex. When the emulsifier is mixed after heating the edible oil to 75-95°C, preferably 75-85°C, the emulsifier dissolves, greatly increasing its miscibility with the edible oil.

[0044] Traditionally, a process involving heating edible oils and fats to 60°C and mixing in an emulsifier has been known. However, even when edible oils and fats are actually heated to 60°C, there is a problem in that the emulsifier does not dissolve and therefore does not mix.

[0045] Experimental results confirmed that emulsifiers dissolve in edible oils when heated to a minimum of 70°C or higher, and that rancidity may occur when heated above 100°C. Therefore, it is preferable to heat edible oils at 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, of the total weight of the lactic acid bacteria complex. In this case, 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 amounts exceeding 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 one preferred embodiment of the present invention, a gastric acid-stable probiotic preparation is provided which contains (a) lactic acid bacteria, (b) edible oils and fats, (c) emulsifiers and (d) nuts as essential components.

[0048] The aforementioned probiotic preparation is characterized in that the weight ratio of the individual components 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 emulsifiers, and (d) 5.0 to 90.0 parts by weight of nuts.

[0049] Of these, explanations regarding (a) lactic acid bacteria, (b) edible oils and fats, and (c) emulsifiers have been given above and will be omitted here.

[0050] In the present invention, "(d) nuts" means one or more components selected from the group consisting of peanuts, almonds, walnuts, pine nuts, pistachios, pecans, macadamia nuts, hazelnuts, cashews, Brazil nuts, chestnuts, soybeans, cocoa nibs, coconuts, sunflower seeds, and pumpkin seeds, as additives to further enhance the viscosity formation of the composition, increase the flavor / texture, and enhance the activity of lactic acid bacteria.

[0051] The aforementioned nuts contain a large amount of unsaturated fatty acids, which are essential components that play a beneficial role in various aspects of health, including blood circulation. The pulverized form is preferred, and the pulverized material is characterized by being pulverized to the extent that it has a particle size of 0.01 to 0.5 mm, preferably 0.01 to 0.1 mm, and exhibits a paste-like consistency.

[0052] The amount of nuts is mixed in an amount of 5.0 to 90.0 parts by weight, preferably 10.0 to 30.0 parts by weight.

[0053] Experiments have confirmed that the aforementioned nuts not only improve the palatability and sensory appeal of probiotic preparations, but also have an excellent effect in maintaining the survival rate of lactic acid bacteria in stomach acid.

[0054] The present invention provides a gastric acid-stabilizing probiotic formulation that further contains the essential components plus one or more additional components selected from (e) prebiotics and (f) adhesives.

[0055] In this case, the weight ratio of the individual components of the probiotic preparation is as follows: (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 emulsifiers, (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 tackifiers.

[0056] More preferably, the mixture is characterized by comprising (a) 1.0 part by weight of lactic acid bacteria, (b) 10 to 30 parts by weight of edible oils and fats, (c) 0.2 to 1.6 parts by weight of 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 tackifier.

[0057] The term "(e) prebiotics" as defined in this application refers to components that are used as food for lactic acid bacteria, are not digested by enzymes in the digestive tract of animals, and promote the growth of intestinal bacteria such as Bifidobacterium in the lower part of the small intestine, thereby exhibiting various physiological effects, including intestinal regulation. Monosaccharides, disaccharides, oligosaccharides, and polysaccharides that are well-known in the industry as prebiotics are possible, 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 and raffinose. Oligosaccharides such as raffinose, stachyose, and maltodextrin; polysaccharides such as amylose, amylopectin, starch, cellulose, and pectin; more preferably sugar compounds such as glucose, fructose, galactose, sucrose, lactose, and fructo-oligosaccharides, and more preferably glucose, fructose, lactulose, lactitol, or fructo-oligosaccharides.

[0058] The amount of the (e) prebiotics component can be adjusted in amounts well known in the industry depending on the type and amount of the (a) lactic acid bacteria, and it is generally preferable to add 0.01 to 5 parts by weight, preferably 0.5 to 2.0 parts by weight, per 1 weight of lactic acid bacteria.

[0059] The "(f) tackifier" as defined in this application is contained in an amount of 0.1 to 5 parts by weight relative to the total weight of the composition and plays a role in imparting tackiness to the composition, and preferably consists of 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 amount of the tackifier is less than 0.1 parts by weight, the effect is minimal, and if the amount of the tackifier exceeds 5 parts by weight, the viscosity may increase excessively, reducing workability.

[0061] Furthermore, the present invention allows for the addition of one or more additives selected from pigments, fragrances, etc., in appropriate amounts, within a range of about 0 to 10 parts by weight per 1 weight of total lactic acid bacteria, to the composition.

[0062] Furthermore, the present invention provides a pharmaceutical composition, a health functional food, and a health supplement for promoting the growth of beneficial intestinal bacteria, which contain a gastric acid-stabilized probiotic preparation produced by the above manufacturing method as an active ingredient.

[0063] The probiotic preparation of the present invention was subjected to experiments such as emulsification liquid production experiments at different heating temperatures (Experimental Example 1); effects on the growth of beneficial intestinal bacteria (Experimental Example 2); lactic acid bacteria survival experiments in artificial gastric juice (pH 2) (Experimental Example 3); constipation relief efficacy experiments of the composition (Simplified Clinical Example 1); and palatability evaluation experiments based on the type of nuts contained in the composition (Simplified Clinical Example 2). The results confirmed that the probiotic preparation of the present invention strongly promotes the growth of beneficial intestinal bacteria, exhibits excellent survival ability of lactic acid bacteria in gastric acid, and shows remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of constipation relief and palatability. Therefore, it is useful as an excellent probiotic preparation for pharmaceutical compositions, health functional foods, and health supplements.

[0064] To solve the above-mentioned objectives, the present invention provides a pharmaceutical composition for promoting the growth of beneficial intestinal bacteria or for preventing and treating diseases caused by the growth of harmful intestinal bacteria, the probiotic preparation being used as an active ingredient.

[0065] The term "diseases caused by the proliferation of harmful intestinal bacteria" as defined in this application is not limited to these, but includes, for example, antibiotic-associated diarrhea, inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, abdominal distension, constipation, acute or chronic irritable bowel syndrome, radiation-induced colitis, and Helicobacter infections.

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

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

[0068] A composition comprising the probiotic formulation of the present invention may further contain suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions.

[0069] The compositions containing the probiotic formulations according to the present invention can be prepared by conventional methods in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injection solutions. Examples of carriers, excipients, and diluents that may be included 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 formulation, these are prepared using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules. Such solid formulations are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose, or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as humectants, sweeteners, fragrances, and preservatives may be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suitable suppository bases include witepsol, macrogol, tween 61, cocoa butter, lauric acid butter, and glycerogenous gelatin.

[0070] The preferred dosage of the probiotic preparation of the present invention varies depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the duration, but can be appropriately selected by those skilled in the art. However, for desirable effects, it is preferable to administer the probiotic preparation at a dose of 0.01 mg / kg to 10 g / kg per day, preferably 1 mg / kg to 1 g / kg. The administration can be done once a day or in several divided doses. Furthermore, the above dosage does not limit the scope of the present invention in any way.

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

[0072] Furthermore, the present invention provides a therapeutic method for treating patients with diseases caused by the promotion of the growth of beneficial intestinal bacteria or the growth of harmful intestinal bacteria, comprising administering the probiotic preparation to patients with diseases caused by the promotion of the growth of beneficial intestinal bacteria or the growth of harmful intestinal bacteria.

[0073] Furthermore, the present invention provides applications for the probiotic preparation for producing agents for treating diseases caused by the proliferation of beneficial intestinal bacteria or harmful intestinal bacteria.

[0074] Furthermore, the present invention provides a health functional food containing a probiotic preparation as an active ingredient for promoting the growth of beneficial intestinal bacteria or for preventing and improving diseases caused by the growth 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 relative to the total weight of the composition.

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

[0077] A composition comprising the probiotic formulation of the present invention may further contain suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions.

[0078] The compositions containing the probiotic formulations according to the present invention can be prepared by conventional methods in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, and sterile injection solutions. Examples of carriers, excipients, and diluents that may be included 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 formulation, these are prepared using commonly used fillers, bulking agents, binders, wetting agents, disintegrants, surfactants, and other diluents or excipients. Solid formulations for oral administration include tablets, pills, powders, granules, and capsules. Such solid formulations are prepared by mixing the probiotic formulation with at least one excipient, such as starch, calcium carbonate, sucrose, or lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, oral solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as humectants, sweeteners, fragrances, and preservatives may be used. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized formulations, and suppositories. Non-aqueous solvents and suspensions that can be used include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suitable suppository bases include witepsol, macrogol, tween 61, cocoa butter, lauric acid butter, and glycerogenous gelatin.

[0079] The preferred dosage of the probiotic preparation of the present invention varies depending on the patient's condition and weight, the severity of the disease, the drug form, the route of administration, and the duration of treatment, but can be appropriately selected by those skilled in the art. However, for desirable effects, it is preferable to administer the probiotic preparation at a dose of 0.01 mg / kg to 10 g / kg per day, preferably 1 mg / kg to 1 g / kg. The administration can be done once a day or in several divided doses. Furthermore, the above dosage does not limit the scope of the present invention in any way.

[0080] The compositions of the present invention can be administered to mammals such as rats, mice, livestock, and humans via various routes. All methods of administration are conceivable, but for example, they can be administered orally, rectally, or intravenously.

[0081] Furthermore, the present invention provides a health functional food containing the above-mentioned probiotic preparation as an active ingredient for promoting the growth of beneficial intestinal bacteria or for preventing and improving diseases caused by the growth of harmful intestinal bacteria.

[0082] In this application, "health functional foods" means foods manufactured and processed using raw materials or components that have beneficial functional properties for the human body, as defined in Act No. 6727 on Health Functional Foods. "Functionality" means being ingested for the purpose of obtaining beneficial effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body.

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

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

[0085] Furthermore, it can be manufactured and processed as a health functional food in the form of a drug administration such as a powder, granules, tablet, capsule, pill, suspension, emulsion, or syrup, or in the form of a tea bag, infused tea, or health beverage, for the purpose of promoting the growth of beneficial intestinal bacteria or preventing and improving diseases caused by the growth of harmful intestinal bacteria.

[0086] Furthermore, the present invention provides a health supplement containing the above-mentioned probiotic preparation as an active ingredient for promoting the growth of beneficial intestinal bacteria or for preventing and improving diseases caused by the growth of harmful intestinal bacteria.

[0087] In the health supplements as defined in this application, the main component means that the content of the active ingredient relative to the weight of the whole composition is in the range of about 30% to 99%, 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 proportion, but there are no special restrictions on other components, and various flavorings or natural carbohydrates, as in ordinary beverages, can be included as additional components. Examples of the aforementioned natural carbohydrates include monosaccharides, e.g., glucose, fructose; disaccharides, e.g., maltose, sucrose; and ordinary sugars such as polysaccharides, e.g., dextrin, cyclodextrin; and sugar alcohols such as xylitol, sorbitol, and erythritol. Natural flavorings (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavorings (saccharin, aspartame, etc.) can be advantageously used as flavorings other than those mentioned above. The proportion of the aforementioned 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 is or used together with other foods or food ingredients, and can be used as appropriate in the usual way. Examples of foods to which the substance can be added include meats, sausages, bread, chocolate, candies, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and include all health foods in the usual sense.

[0090] In addition to the above, the compositions of the present invention may contain various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic and natural flavoring agents, colorants and enhancers (for cheese, chocolate, etc.), pectinic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. Furthermore, the compositions of the present invention may contain fruit pulp for the production of natural fruit juices, fruit juice beverages, and vegetable beverages. Such components can be used independently or in combination. The proportion of such additives is not particularly important, but it is common to select them in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.

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

[0092] Furthermore, the present invention provides a food or food additive containing the above-mentioned probiotic preparation as an active ingredient for promoting the growth of beneficial intestinal bacteria or for preventing and improving diseases caused by the growth of harmful intestinal bacteria.

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

[0094] Functional foods containing the probiotic preparation of the present invention include confectionery such as bread, mochi, dried fruits, candies, chocolates, chewing gum, and jams; ice cream products such as ice cream, frozen desserts, and ice cream powders; milk products such as milk, low-fat milk, lactose-free milk, processed milk products, goat's milk, fermented milk products, buttermilk, concentrated milk products, milk creams, buttermilk, natural cheese, processed cheese, milk powders, and whey products; processed meat products, processed egg products, meat products such as hamburgers; kamaboko, ha Examples of prohibited items include, but are not limited to, fish products such as fish products like sausages and bacon, noodles such as ramen, dried noodles, fresh noodles, milk noodle soup, gelatinized dried noodles, improved aged noodles, frozen noodles, and pasta, beverages such as fruit drinks, vegetable drinks, carbonated drinks, soy milk, yogurt and other lactic acid bacteria drinks, and mixed drinks, condiments such as soy sauce, miso, gochujang, chunjang, cheonggukjang, mixed sauces, vinegar, sauces, tomato ketchup, curry, and dressings, margarine, shortening, and pizza.

[0095] The amount of the probiotic preparation according to the present invention, which is added to food products including beverages during the manufacturing process of the aforementioned health functional foods, can be appropriately adjusted as needed. [Effects of the Invention]

[0096] The gastric acid-stable probiotic preparation of the present invention was subjected to experiments such as the production of emulsified liquid at different heating temperatures (Experimental Example 1); the effect on the growth of beneficial intestinal bacteria (Experimental Example 2); the survival rate of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); the efficacy of the composition in improving constipation (Simplified Clinical Example 1); and the evaluation of palatability based on the type of nuts contained in the composition (Simplified Clinical Example 2). The results confirmed that the lactic acid bacteria preparation of the present invention strongly promotes the growth of beneficial intestinal bacteria, exhibits excellent survival rate of lactic acid bacteria in gastric acid, and shows remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability.

[0097] Therefore, the present invention can provide a gastric acid-stable probiotic formulation and a method for producing the same. [Brief explanation of the drawing]

[0098] [Figure 1] This figure shows the experimental results of observing the properties of the emulsified solution of the emulsifier of the present invention at various temperatures. [Figure 2] This figure shows the experimental results of measuring the effect of strain growth on the selective medium (MRS agar plate) of the lactic acid bacteria preparation of the present invention. [Figure 3] This figure shows the results of an experiment on the survival rate of lactic acid bacteria in the artificial gastric juice (pH 2) of the present invention. [Modes for carrying out the invention]

[0099] Those skilled in the art will see that various modifications and variations can be used in the compositions, uses, and manufactures of the present invention, as long as they do not deviate 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 content of the present invention is not limited by the following examples and experimental examples.

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

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

[0103] To the emulsified oil in gel form formed as described above, a mixture of lactic acid bacteria, namely Lactobacillus acidophilus DDS-1 (Alllove Co., Ltd.), Lactobacillus rhamnosus GG (Alllove Co., Ltd.), Lactobacillus plantarum VI-07 (Bitec Co., Ltd.), Bifidobacterium longum BL5 (Bitec Co., Ltd.), and Bifidobacterium animalis ssp. lactis BLC1 (Alllove Co., Ltd.), were added in equal numbers (2 × 10⁻¹⁰) of each type. 9 1 g of mixed lactic acid bacteria, uniformly mixed at a ratio of cfu / g, was used. (Test-2: For the activity experiment process and results in artificial gastric juice (pH2) for observing the viability of lactic acid bacteria, please refer to Experimental Example 2 below.)

[0104] To achieve the appropriate viscosity of the composition, enhance the flavor / texture, and further strengthen the activity of lactic acid bacteria, fructooligosaccharides (a natural sweetener and prebiotic) and peanuts (a nut containing large amounts of unsaturated fatty acids, which play a beneficial role in various health aspects, including blood circulation) were added. The mixture was then stirred in a mixing and stirring machine (OFM-1507B, Ofer Co., Ltd.) at a speed of 30-50 rpm for more than one hour until all ingredients were uniformly mixed.

[0105] In this case, for nuts, the best method is to crush them using a crushing device (ultra-high-speed vacuum blender, KCB-001W, KUCHAM Co., Ltd.) until the particle size is 0.01 to 0.1 millimeters (mm) (sufficient to form a paste). (Test-3: Peanuts, almonds, and walnuts, which were selected as the most palatable, were mixed into the composition, and the results of the experiment evaluating drinkability and palatability were shown in Experiment Example 3.)

[0106] [Table 1]

[0107] Example 2. Production of a lactic acid bacteria preparation without emulsifiers (Composition: Table 1) Without using the emulsification step involving vegetable oil and emulsifier as described in the manufacturing method of Composition A in Example 1, a lactic acid bacteria preparation (Composition C) without crushed nuts was produced by simply mixing sunflower oil (vegetable oil), five types of mixed lactic acid bacteria, and the remaining additives under the same conditions, and this was used as a sample in the experimental example below.

[0108] Example 3. Production of a comparative lactic acid bacteria preparation A lactic acid bacteria complex containing 10.0% by weight of lactic acid bacteria powder, 11.5% by weight of yellow lead (suspending agent), 1.0% by weight of soy lecithin (emulsifier), and the remainder of rTG type refined fish oil (edible oil) was prepared using the manufacturing method described in Example 1 of the conventional literature. A comparative enteric-coated lactic acid bacteria complex was then prepared and used as a sample in the following experimental example.

[0109] Experiment Example 1: Experiment on producing emulsified liquid at different temperatures. 100g of sunflower oil (vegetable oil, SAJO DAERIM Co., Ltd.) was mixed with 2.0g of emulsifier (glycerin fatty acid ester, ES food ingredient), heated at four different temperatures including 40°C, 60°C, 85°C, and 100°C, and then emulsified while stirring in a circulating cooling water bath (low-temperature circulating constant-temperature water bath, SH-WB-7CDR) maintained at 0-4°C until the mixture reached a temperature of 4-10°C. The optimal temperature for producing the emulsion was then determined.

[0110] As shown in Figure 1, the experimental results indicate that each photograph shows the shape of the emulsion formed at different temperatures. At 40°C and 60°C, the emulsifier did not dissolve sufficiently, and the emulsion did not form well. However, at 85°C and 100°C, the emulsifier dissolved sufficiently, and the emulsion formed uniformly. (See Figure 1: Experiment on the properties of emulsions at various temperatures with an emulsifier)

[0111] Experimental Example 2: Effects on the Growth of Beneficial Intestinal Bacteria To confirm the effect of the samples of the present invention obtained from the above examples on the growth of beneficial intestinal bacteria, the following experiment was conducted, referring to the lactic acid bacteria count test method announced in the Commentary on Microbial Testing Methods for Food Regulations (published by the Food and Drug Safety Evaluation Institute of the Ministry of Food and Drug Safety).

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

[0113] 2-2. Experimental Process The number of lactic acid bacteria produced was measured by counting the number of lactic acid bacteria colonies using MRS plate measurement medium, and the number of viable bacteria per mL of culture medium was calculated by multiplying the average number of colonies by the dilution factor. A 9-fold dilution (sterile physiological saline) was added to 10 mL of the test sample and homogenized (10 -1 Solution). Test solution (10 -1 Add the diluent to 1 mL of solution to make a total volume of 10 mL. -2 After preparing the test solution, dilute it using the same procedure.

[0114] Each diluted test solution was inoculated at 1(v / v)% on Lactobacillus Selective Medium (MRS agar plaste), followed by anaerobic incubation at 37°C for 72±3 hours. Plasts that produced 15 to 300 colonies per plate were selected, the number of colonies was calculated, and the bacterial count was calculated by multiplying the measured number of colonies by the dilution factor.

[0115] 2-3. Experimental Results Based on the viability results of the sample in the above example, it was confirmed that the best method for producing the emulsified solution is at 85°C. (See Table 2 and Figure 2)

[0116] [Table 2]

[0117] Experiment Example 3. Lactic acid bacteria survival rate experiment using artificial gastric juice (pH 2). 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 conducted using a 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 To confirm the viability of lactic acid bacteria in the composition, composition A was stirred with artificial gastric juice (pH 2) for a set period (1 hour). The collected composition was immediately cultured on Lactobacillus selective medium (MRS agar plate, MB-M1024-P50, KisanBio) and cultured anaerobically at 37°C for 72±3 hours. Plates that produced 15 to 300 colonies per plate were selected, the number of colonies was calculated, and the bacterial count was calculated by multiplying the measured number of colonies by the dilution factor. For comparison, a control group of compositions, in which the lactic acid bacteria mixture was mixed with unemulsified oil, was used.

[0119] 3-2. Experimental Results The experimental results show that the survival rate of lactic acid bacteria in the emulsified oil and lactic acid bacteria mixture is even better than that of the control group. Furthermore, when peanut powder was added as an additive to the emulsified oil and cultured under the same conditions, it was observed that the survival rate of Lactobacillus in the lactic acid bacteria mixture containing even more peanut powder was even more remarkably superior. (See Table 3 and Figure 3)

[0120] [Table 3]

[0121] Simple Clinical Case 1: Experiment on the Constipation-Relieving Efficacy of the Composition To confirm the constipation-relieving efficacy of the sample of the present invention obtained from the above-mentioned examples, the following experiment was conducted.

[0122] The constipation-improving effect of composition A, which was used in a test to improve constipation, was evaluated on 50 support workers in the Seoul area, ranging in age from their 20s to 50s, who experience inconvenience in their daily lives due to constipation.

[0123] The experimental subjects were given composition A twice a day (morning and evening) at a dose of 5g each time for 10 days and answered the following questionnaire. In addition, 50 subjects were given composition B as a control group in the same manner as the experimental subjects.

[0124] The experimental results confirm that composition A of the present invention has excellent constipation-relieving effects. (See Tables 4 and 5)

[0125] [Table 4]

[0126] [Table 5]

[0127] Simple Clinical Case 2: Experiment to Evaluate Palatability Based on the Type of Nuts Contained in the Composition To confirm the palatability of the composition of the sample of the present invention obtained from the above examples, depending on the type of nut contained in it, the following experiment was conducted.

[0128] Compositions were prepared using peanuts, almonds, and walnuts as crushed nuts, and then the taste, aroma, texture, and overall palatability of each nut type were evaluated after 20 adult men and women consumed 3g of each.

[0129] The criteria for evaluating preferences were based on a 5-point scale, with the average value being displayed after the initial survey. (Excellent: 5 points, Good: 4 points, Average: 3 points, Poor: 2 points, Very Poor: 1 point) (See Table 6)

[0130] [Table 6]

[0131] The experimental results confirm that the composition using peanut powder exhibits the best palatability.

[0132] Based on the aforementioned experimental results, we conducted experiments on the effects of the lactic acid bacteria preparation of the present invention on the growth of beneficial intestinal bacteria (Experimental Example 2); an experiment on the survival rate of lactic acid bacteria in artificial gastric juice (pH 2) (Experimental Example 3); an experiment on the efficacy of the composition in improving constipation (Simplified Clinical Example 1); and an experiment on the palatability of the composition based on the type of nuts contained in the composition (Simplified Clinical Example 2). As a result, we were able to confirm that the lactic acid bacteria preparation of the present invention strongly promotes the growth of beneficial intestinal bacteria, exhibits excellent survival rate of lactic acid bacteria in gastric acid, and demonstrates remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of improving constipation and palatability. [Examples]

[0133] The following describes examples of formulations of compositions containing the probiotic preparation of the present invention, but the present invention is not intended to limit itself, but merely to provide specific examples.

[0134] Formulation Example 1: Manufacturing of Powdered Formulations Composition A---------------------- 20mg Lactose---------------------- 100mg Talc ------------------------ 10mg

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

[0136] Formulation Example 2. Tablet Manufacturing Composition A---------------------- 10mg Corn starch -------------------- 100mg Lactose---------------------- 100mg Magnesium stearate --------------- 2mg After mixing the aforementioned components, tablets are manufactured by compressing them using a standard tablet manufacturing method.

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

[0138] The above-mentioned components are mixed using a standard capsule manufacturing method, and the mixture is filled into gelatin capsules to produce capsules.

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

[0140] The product is manufactured using a standard method for manufacturing injectable drugs, with the above-mentioned component content per ampoule (2 ml).

[0141] Formulation Example 5. Manufacturing of Liquid Formulations Composition A---------------------- 20mg Isomerized sugar---------------------- 10g Mannitol 5g Purified water ------------------------ Appropriate amount

[0142] The liquid preparation is manufactured by adding each component to purified water and dissolving them using a standard liquid preparation method, adding an appropriate amount of lemon fragrance, mixing the components, adding purified water to adjust the total volume to 100 ml, filling the container with brown disease-causing material, and sterilizing it.

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

[0144] The composition ratio of the aforementioned vitamin and mineral mixture was chosen in the preferred example to include components relatively suitable for health foods. However, the mixing ratio may be arbitrarily modified. After mixing the components using a conventional health food manufacturing method, granules can be produced and used in the manufacture of health food compositions using conventional methods.

[0145] Formulation Example 7. Manufacturing of Health Drinks Composition A---------------------- 1000mg Citric acid ----------------------- 1000mg Oligosaccharides ----------------------- 100g Plum extract ----------------------- 2g Taurine ----------------------- 1g Add purified water to make a total of 900ml

[0146] After mixing the above-mentioned ingredients using a conventional method for manufacturing health beverages, the mixture is stirred and heated at 85°C for approximately 1 hour. The resulting solution is then filtered, collected in a sterilized 2L container, sealed and sterilized, and then stored in a refrigerator before being used in the production of the health beverage composition of the present invention.

[0147] The aforementioned composition ratio is a mixture of ingredients relatively suitable for beverages, as in the preferred example. However, the blending ratio may be arbitrarily modified depending on regional and ethnic preferences such as the demand segment, the country of demand, and the intended use. [Industrial applicability]

[0148] As described above, the probiotic preparation of the present invention was subjected to experiments such as emulsification liquid production experiments at different heating temperatures (Experimental Example 1); effects on the growth of beneficial intestinal bacteria (Experimental Example 2); lactic acid bacteria survival experiments in artificial gastric juice (pH 2) (Experimental Example 3); constipation relief efficacy experiments of the composition (Simplified Clinical Example 1); and palatability evaluation experiments based on the type of nuts contained in the composition (Simplified Clinical Example 2). The results confirmed that the lactic acid bacteria preparation of the present invention strongly promotes the growth of beneficial intestinal bacteria, exhibits excellent survival ability of lactic acid bacteria in gastric acid, and shows remarkable effects superior to conventionally known lactic acid bacteria preparations in terms of constipation relief and palatability. As a result, it can be usefully used as an excellent probiotic preparation in various pharmaceutical compositions, health functional foods, and health supplements.

Claims

1. The first step involves heating 1 to 90 parts by weight of sunflower oil, an edible oil, at 70 to 95°C, and then mixing and stirring with 0.1 to 5 parts by weight of glycerin fatty acid ester, an emulsifier, to produce a mixture of edible oil and emulsifier; The second step involves cooling and stirring the mixture from the first step until its temperature reaches 2 to 20°C, thereby generating a gel-like aggregated mixture; The third step involves slowly adding 1 part by weight of Lactobacillus bacteria to the gel-shaped aggregated mixture and performing slow stirring to produce a lactic acid bacteria mixture; The fourth step involves adding peanut powder, which is a nut powder, to the mixture from the third step and stirring to produce a nut lactic acid bacteria mixture; A fifth step involves adding fructooligosaccharides, which are prebiotics, to the mixture from the fourth step and stirring; The process includes the following steps: The method for producing a gastric acid-stable probiotic preparation is characterized in that the fourth-stage pulverized nut material has a particle size of 0.01 to 0.5 mm, and is stirred at a stirring speed of 30 to 50 rpm for 30 minutes to 3 hours until all materials are uniformly mixed.

2. The individual compositional components are blended in the following weight ratios, containing (a) Lactobacillus bacteria 1.0 part by weight, (b) sunflower oil 1.0 to 90.0 parts by weight, (c) glycerin fatty acid ester 0.1 to 5.0 parts by weight, (d) peanut powder 5.0 to 90.0 parts by weight, and (e) fructooligosaccharides 0.01 to 5.0 parts by weight as essential components. The peanut powder is a gastric acid-stable probiotic preparation comprising nut powder with a particle size of 0.01 to 0.5 mm.

3. A pharmaceutical composition for promoting the growth of beneficial intestinal bacteria or preventing and treating diseases caused by the growth of harmful intestinal bacteria, comprising the gastric acid-stabilized probiotic preparation described in claim 2 as an active ingredient.

4. The pharmaceutical composition according to claim 3, 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 syndrome, radiation-induced colitis, or Helicobacter infection.

5. A health functional food for promoting the growth of beneficial intestinal bacteria or preventing or improving diseases caused by the growth of harmful intestinal bacteria, comprising the gastric acid-stabilized probiotic preparation described in claim 2 as an active ingredient.

6. The health functional food according to claim 5, characterized in that the health functional food is in the form of a drug administration form such as a powder, granules, tablets, capsules, pills, suspension, emulsion, or syrup, or in the form of a tea bag, infused tea, or health beverage.

7. A health supplement for promoting the growth of beneficial intestinal bacteria or preventing or improving diseases caused by the growth of harmful intestinal bacteria, comprising the gastric acid-stabilized probiotic preparation described in claim 2 as an active ingredient.

8. A food additive for promoting the growth of beneficial intestinal bacteria or preventing or improving diseases caused by the growth of harmful intestinal bacteria, comprising the gastric acid-stabilized probiotic preparation described in claim 2 as an active ingredient.

9. Use of the gastric acid-stable probiotic preparation according to claim 2 for manufacturing an agent for treating diseases caused by the proliferation of beneficial intestinal bacteria or harmful intestinal bacteria.