KR20170120264A - Fermented chestnut puree of probiotic lactic acid bacteria and foods composition, medicinal composition including the same - Google Patents

Abstract

More particularly, the present invention relates to a food composition and a pharmaceutical composition characterized by a night fermented puree utilizing probiotic lactic acid bacteria. More specifically, the present invention relates to a food composition and a pharmaceutical composition which are prepared by mixing 1 to 3% (v / w) of Lactobacillus plantarum (KCTC 21004) Wherein the Lactobacillus rhamnosus, Lactobacillus plantarum, and Lactobacillus strain of the present invention are inoculated. acidophilus, Lactobacillus. sakei and Lactococcus lactics, the overnight fermented puree fermented by inoculation with at least one selected lactic acid bacterium has a high productivity of γ-aminobutyric acid (GABA) by antioxidant, acid resistance, bile bain and free amino acid, So that it can be usefully used for the development of functional reinforcing products.

Description

[0001] The present invention relates to a fermented chestnut puree of probiotic lactic acid bacteria and foods composition,

The present invention relates to a night fermented puree utilizing probiotic lactic acid bacteria, and a food composition containing the same. More particularly, the present invention relates to a method of producing a night fermented puree by mixing 1 to 3% (v / w) of lactobacillus plantarum (KCTC 21004) The present invention relates to a night fermented puree utilizing a probiotic lactic acid bacterium, which can be used to develop a functional enhancing product using chestnut.

Chestnut is the fruit of chestnut tree and has been cultivated for a long time. Thirteen species are distributed in temperate regions such as Asia, Europe, North America and northern Africa. Korea is the second shortest-term income in the world, and it is suitable for the climate of Korea. Since 1970, the Korean Forest Service has been actively promoting it as a high income earner. According to the Korea Forest Service Report, the consumption rate of domestic consumers decreased from 82,450 tons in 2006 to 68,630 tons in 2010. Except for 25 ~ 30% of exports, domestic consumers account for 18 ~ 20% %, 12 ~ 15% for raw materials and 3 ~ 5% for night. On the other hand, the amount of corruption or deterioration is about 12 ~ 15%, and joint researches on night-related industries such as the establishment of nighttime storage and distribution system and the development of nighttime demand are underway.

 Chestnut has higher starch content and lower water content than ordinary fruit, and contains 5 nutrients such as protein, lipid, vitamins and minerals. There are differences according to regional varieties. In general, the nutrient content of chestnut is 26 ~ 34% of starch, 2.5 ~ 3.8% of protein, 0.9 ~ ~ 1.3% of ash, 0.8 ~ 1.1% of fiber, 0.4 ~ ~ 0.8% of crude fat and 61 ~ . Night is good not only for the symbol but also nutritionally weak, good for the poor nutrition, good for the muscles and the bones, so the young child or lower body is beneficial for the weak person, strengthen the stomach and spleen, protect the kidney, Beauty, fatigue, prevention of cold is effective in preventing the food is also widely used as a herbal medicine. There is antioxidant activity of superoxide dismutase (SOD) -like activity and 1.1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity in gall bladder and catechine.

  As the eating habits of modern people have become more convenient and the convenience foods have increased, there has been an increasing interest in functional ingredients capable of controlling the human body rhythm, and thus, functional reinforced foods are actively consumed.

On the other hand, attention is paid to various physiological activities of γγ-aminobutyric acid (GABA), which is known as inhibitory neurotransmitter. γγ-aminobutyric acid (GABA) is a nonprotein amino acid distributed in the natural environment. It exists in the brain or spinal cord of vertebrate animals. It is used for improving cerebral blood flow, increasing oxygen supply, promoting brain metabolic function, reducing blood pressure, relieving anxiety, , Insomnia, relieving depression, relieving stress, preventing stroke and dementia, and improving memory, and is called brain food that improves brain function. Among various studies to improve the content of γγ-aminobutyric acid (GABA) in natural materials, fermentation methods using microorganisms have been studied. Among them, studies on the enhancement of γγ-aminobutyric acid (GABA) content by lactic acid bacteria have been actively studied.

  Some strains of Lactobacilli decarboxylate glutamate to produce γγ-aminobutyric acid (GABA) and CO₂₂ as final products, and Lactobacillus brevis, Lactobacillus plantarum, Γ-aminobutyric acid (GABA) is produced at high concentration, and γγ-aminobutyric acid (GABA) is produced in cheese and fermented milk produced by lactic acid bacteria starter.

 Lactobacillus such as Lactobacillus producing γγ-aminobutyric acid (GABA) has been reported to produce γγ-aminobutyric acid (GABA) at a high concentration in lactic acid bacteria isolated from rice, soybean fermented food, kimchi and cheese.

  Lactic acid bacteria, which are typical probiotics, are anaerobic or anaerobically proliferating gram-positive bacteria that decompose sugars to produce lactic acid. They are used in traditional fermented foods such as fermented milk, cheese, kimchi, and soybean paste. The World Health Organization (WHO) defined probiotics as "live microorganisms" that are used against antibiotics and that have beneficial effects on the host's health by improving the capacity of the host's intestinal flora. Currently, Lactobacillus, Bifidobacterium and Lactococcus are widely used in probiotics, and lactic acid bacteria in fermented foods cause fermentation of lactic acid, And bacteriocin, which is an antimicrobial substance, is secreted to inhibit food poisoning bacteria and lower the intestinal pH of humans, thereby suppressing the proliferation of spoilage bacteria.

 Studies on the nighttime composition of the chestnut have been performed in many researches and studies on the chestnuts. Studies on the product development using chestnuts include studies on beverage production, preparation and characterization of chestnut yogurt, And a study on the quality characteristics, which is a study on palatability. Recently, studies on functional foods have been actively carried out with increasing interest in biological control functions in addition to natural foods, nutritious foods and preferential foods using chestnuts. However, antioxidative activity and free amino acids of night purée fermented with probiotic lactic acid bacteria (Γ-aminobutyric acid) (GABA) production activity has not been reported.

Therefore, it is expected that it will contribute to the enhancement of the availability of the night through the night purée fermented from the lactic acid bacteria proposed by the present invention, and to the development of functional enhancing products using chestnut.

 Accordingly, the present inventors have found that antioxidant, acid-fast bacilli and gamma-gamma -butyrolactone for the study of functional foods of night fermented puree through fermentation of probiotic lactic acid bacteria, Aminobutyric acid (GABA) production activity was confirmed through experiments, and a functional strengthening product using night was developed. At the same time, the present invention was completed to enhance the added value by using night.

Korean Patent Laid-Open Publication No. 2015-0128373, Anti-Wrinkle and Antibacterial Composition Containing Fermented Balsam Extract Purified Product as Active Ingredient, Published Nov. 18, 2015

It is an object of the present invention to provide a process for preparing Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus. acidophilus, Lactobacillus. sakei and Lactococcus lactis. The fermented overnight fermented puree is inoculated with at least one lactic acid bacterium selected from the group consisting of sakei and lactococcus lactis.

Another object of the present invention is to provide a food composition comprising a night fermented puree, characterized in that the lactic acid bacterium is inoculated with 1 to 3% (v / w) of the overnight puree.

It is still another object of the present invention to provide a method for the treatment and prophylaxis of inflammatory diseases and functional intestinal diseases of the large intestine and small intestine including the night fermented puree, which comprises inoculating the lactic acid bacteria with 1 to 3% (v / w) And to provide a pharmaceutical composition for preventing or treating one or more selected diseases.

In order to achieve the above object, the present invention provides a method for producing Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus. acidophilus, Lactobacillus. sakei and Lactococcus lactis to provide a night fermented puree fermented by inoculation with at least one selected lactic acid bacterium.

The Lactobacillus plantarum according to one embodiment of the present invention provides Lactobacillus plantarum (KCTC 21004), a night fermented puree.

Also, the present invention provides a night fermented puree characterized in that the lactic acid bacterium of the present invention is inoculated with 1 to 3% (v / w) of chestnut puree.

In order to accomplish the above object, the present invention provides a food composition comprising a night fermented puree, wherein the lactic acid bacteria are inoculated with 1 to 3% (v / w) of a night purée. The composition according to another embodiment of the present invention provides a food composition having at least one activity selected from the group consisting of inflammatory diseases of the large intestine and small intestine and ability to improve functional intestinal diseases. In addition, the dairy product, the confectionery, the seasoning, the beverage and the drink, the snack, the candy, the ice cream and the frozen dessert, the morning grains, the nutrition bar, the snack bar chocolate product, the processed food, Dairy and beverage dairy products, soy dairy-like products, frozen foods, cooked foods and alternative foods, meat products, pasta, soups, sauces and dressings, confectionery products, oil and fat products, dairy drink and milk drinks, Cheese, yogurt, bread and buns, yeast products, cakes, cookies and crackers.

In order to achieve the above-mentioned further object, the present invention provides a method for preventing inflammation of the small intestine and small intestine and functional intestinal diseases including a night fermented puree by mixing lactic acid bacteria with chestnut puree at 1 to 3% (v / w) The present invention provides a pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of:

The inflammatory disease of the large intestine and the small intestine according to another embodiment of the present invention is selected from the group consisting of acute enteritis, Crohn's disease, ulcerative colitis, and ischemic colitis. Also, the functional bowel disease is selected from the group consisting of diarrhea, constipation, enteric diverticulum, and irritable bowel syndrome.

The present invention relates to a night fermented puree utilizing probiotic lactic acid bacteria and a food composition containing the same, wherein Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus. acidophilus, Lactobacillus. sakei and Lactococcus lactis were inoculated with 1 ~ 3% (v / w) mixture of night fermented overnight fermented puree and Lactobacillus plantarum (KCTC 21004) The present invention relates to purée, which is characterized by high antioxidant, acid resistance, biliary properties and high productivity of gamma-gamma -aminobutyric acid (GABA) by free amino acid, and thus can be usefully used in the development of functional enhancing products using chestnut.

1 is a GABA (gamma -aminobutricacid) content according to a kind of a lactic acid bacterium at a GABA (gamma -aminobutricacid) concentration of a night fermentation product obtained by culturing a lactic acid bacterium according to an embodiment of the present invention,
FIG. 2 shows the effect of pH and titratable acidity on the fermentation of night fermented puree according to the inoculation amount of Lactobacillus plantarum KCTC 21004,
FIG. 3 shows the viscosity effect of fermented overnight fermented puree according to the inoculation amount of Lactobacillus plantarum KCTC 21004,
FIG. 4 shows pH and acidity changes of night fermented puree by Lactobacillus plantarum KCTC 21004 according to the fermentation temperature.

The present invention may be embodied in many different forms and should not be construed as limited to the embodiments set forth herein.

Specific embodiments are illustrated in the drawings and described in detail in the detailed description. It should be understood, however, that the invention is not intended to be limited to the particular embodiments, but includes all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Terms including ordinals such as first, second, etc. may be used to describe various elements, but the elements are not limited by such terms. These terms are used only to distinguish one component from another.

When an element is referred to as being "connected" or "connected" to another element, it may be directly connected or connected to the other element, but other elements may be present in between . On the other hand, when an element is referred to as being "directly connected" or "directly connected" to another element, it should be understood that there are no other elements in between.

The terminology used in this application is used only to describe a specific embodiment and is not intended to limit the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. In this application, the terms " comprises ", or " having ", and the like are used interchangeably with a feature, a number, a step, an operation,

One or more other features, integers, steps, operations, elements, components, or combinations thereof, as a matter of principle, without departing from the spirit and scope of the invention.

Hereinafter, a preferred embodiment of the present invention will be described in detail with reference to the accompanying drawings.

The present invention relates to the use of Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus sakei and Lactococcus lactis, ), Which is a fermented chestnut fermented overnight, which is inoculated with at least one selected lactic acid bacterium.

The Lactobacillus plantarum of the present invention may be characterized as a night fermented puree which is Lactobacillus plantarum (KCTC 21004). The following experiment was conducted to confirm the fermentation characteristics of lactic acid bacteria according to the kind of lactic acid bacteria in selecting the lactobacillus plantarum (KCTC 21004) of the present invention. It is known that the proper pH value of the fermented product can improve the flavor. The lactic acid bacteria are known to inhibit the growth of harmful bacteria by increasing the acidity and decreasing the pH in the intestines. To produce lactic acid fermented puree products using chestnut paste, lactic acid bacteria ferment the chestnut to produce a large amount of lactic acid, and it is most important to determine the quality of the fermented product.

Further, it can be characterized as a night fermented puree characterized in that the lactic acid bacterium of the present invention is inoculated with 1 to 3% (v / w) of the overnight puree by mixing. In order to confirm the growth characteristics of lactic acid bacteria by night, the same amount of 2% (v / w) of each lactic acid bacterium was inoculated after sterilization of the night paste prepared by mixing the same amount of water. The fermentation was carried out at 150 rpm for 48 hours at < RTI ID = 0.0 > 150 C. < / RTI > and the pH and acidity of the night pasted fermented product sampled every 12 hours were measured (see FIG. The changes of pH and titratable acidity of fermentation period according to kinds of lactic acid bacteria using chestnut were confirmed. The pH of the 12 kinds of lactic acid bacteria used in the experiment decreased with fermentation time and the acidity was increased. The initial pH of the chestnut paste was 5.75 ~ ~ 5.83, and the initial pH was 0.15 ~ ~ 0.18%. When the chestnut paste was fermented for 48 hours, the acidity was increased in all bacteria, but the bacteria increased to 1.0% in pH were Lactobacillus plantarum subsp. Plantarum KCTC 3103, plantarum KCTC 3104, Lactobacillus subsp. Sakei KCTC 3598 and L. plantarum KCTC 21004 were found to have higher acid production than other fungi. The Lactobacillus plantarum KCTC 21004 was reduced to pH 2.87 after 48 hours of fermentation and the acidity increased up to 1.46%, indicating higher acid production than other fungi [see Table 3]. As a result, L. plantarum KCTC 21004 was found to produce more organic acid with faster fermentation speed and acidity, and Lactobacillus plantarum (KCTC 21004) .

The present invention may be a food composition comprising the night fermented puree. In addition, it may be a food composition having at least one activity selected from the group consisting of inflammatory diseases of small intestine and small intestine and ability of improving functional intestinal diseases. In addition, the food composition of the present invention can be used in dairy products, confectionery products, seasonings, beverages and drinks, snacks, candies, ice cream and frozen desserts, breakfast cereals, nutritional bars, snack bars, chocolate products, Soy sauce and dressing, confectionery products, oil and fat products, dairy drink and milk drinks, soy dairy-like product, frozen foods, cooked foods and alternative foods, meat products, cheese, yogurt , A bread and a bun, a yeast product, a cake, a cookie, and a cracker, but the present invention is not limited thereto. The food composition in the form of tablets can be prepared by granulating the food composition as it is or by mixing it with an excipient, binder, disintegrant or other additives in an appropriate manner, and then extruding the composition into a tablet, compressing the composition, A binder, a disintegrant, or other suitable additives, or by directly compressing the mixture, or by mixing the food composition directly or with a suitable additive into a pre-made granule, mixing the mixture uniformly by compression molding, or adding the excipient, Add the additives, wet the powder evenly mixed with the solvent, wet the powder into a mold at low pressure, and dry it by a suitable method. The hard capsules in the capsule form of the food composition are usually prepared by mixing the capsules with the food composition or an appropriate excipient suitable for the food composition or by granulating the mixture in a suitable manner, The soft capsule is usually prepared by filling capsules with an appropriate excipient for a food composition or a food composition into a suitable capsule base such as gelatin or the like with glycerin or sorbitol, If necessary, a coloring agent preservative or the like may be added to the capsule base.

The food composition of the ring form is usually prepared by mixing an excipient, a binder, a disintegrant, etc. into an ordinary food composition, shaping it into a spherical form by an appropriate method, and then adding a starch, starch, or a suitable substance It is possible that the The granular food composition is usually prepared by mixing the food composition as it is or the food composition with an excipient, a binder, a disintegrant, etc., and then granulating the mixture into granules by an appropriate method. And the like.

The food composition of the beverage formulation may contain various flavors or natural carbohydrates as an additional ingredient such as a conventional beverage. Examples of the above-mentioned natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. A functional ingredient such as antler extract, fructooligosaccharide which helps defecation activity and absorption of calcium, acacia honey, complex gold extract as a natural preservative, and gellan gum which is a precipitation improving thickener may be added, but not limited thereto, The components can be suitably used.

The present invention relates to a method for the treatment and prophylaxis of inflammatory diseases and functional intestinal diseases of the large intestine and small intestine including the night fermented puree, which comprises inoculating Lactobacillus plantarum (KCTC 21004) lactic acid bacteria with 1 to 3% (v / w) And a pharmaceutical composition for preventing or treating one or more diseases selected from the group consisting of The microbial characteristics of the fermented overnight fermented puree according to the inoculation amount of Lactobacillus plantarum were confirmed to confirm its function as a pharmaceutical composition for the prevention and treatment of intestinal diseases. The most common cause of bacterial inoculation of lactic acid bacteria fermented food is the activity of bacteria, which is generally inoculated in the range of 2.0% when milk is fermented. Therefore, the range of 0.5 ~ 3.0% (v / w) , And the pH and titratable acidity were measured during lactic acid fermentation progress of the 50% water content paste [see FIG. 2]. As a result, the initial pH was measured at 5.5 to 6.0 and the initial pH was 0.20 to 0.35%. When the inoculum was 3% (v / w), the initial acid production was 1.10% at 12 hours of culture, and the highest acid production was 1.71% at 48 hours in the final culture, , The pH was 1.65 ~ ~ 1.70%, and the pH and titratable acidity were not significantly different. In addition, the number of live bacteria per hour of the night fermented puree according to the inoculation amount was measured (see Table 7). L. plantarum KCTC 21004 was cultivated in a solid medium (MRS) for 24 hours, and 0.5 to 3.0% (v / w) was inoculated in a 50% . The initial number of bacteria after inoculation was 8.34 ~ 9.28 log CFU / mL, and the number of viable cells increased steadily until 36 hours of fermentation and decreased at 48 hours after fermentation. And 48 hours after fermentation, it showed 10.85 ~ ~ 12.93 log CFU / mL in all experimental groups, indicating probiotic function. Further, in the present invention, to confirm the function of the night fermented puree according to the inoculation amount of Lactobacillus plantarum as a pharmaceutical composition for the prevention and treatment of intestinal diseases, changes in the basic properties of the night fermented puree before fermentation and after fermentation were examined Respectively. The changes in color before fermentation and after fermentation of 50% of the cooked paste fermented for 48 hours at 0.5-3.0% of L. plantarum KCTC 21004 inoculated 24 hours before in the solid medium (MRS) Observed [Table 8]. In all experimental groups, the lightness (L) value increased and the lightness (a) value decreased and the yellowness (b) value increased after the fermentation, indicating that the fermented night fermented puree showed a bright yellow color. In addition, the results of viscosity change before and after fermentation of the chestnut paste according to the inoculum amount were confirmed (see FIG. 3). When L. plantarum KCTC 21004 was inoculated, the amount of puree in 50% of cooked overnight was decreased as the inoculation amount was increased. In the inoculation rate of 0.5% ~ 2.0% (v / w), it decreased after fermentation. However, in the inoculation amount of 3.0% (v / w), the increase from 124,000cp to 131,000cp before fermentation showed that the starch (V / w) of the inoculated amount was found to be appropriate because it is considered that the probability of inoculation exceeding 3.0% (v / w) is lower than that reported by hydrolysis. The present invention also provides a method for preventing or treating intestinal diseases, including a night fermented puree, which comprises admixing 1 to 3% (v / w) of lactobacillus plantarum (KCTC 21004) Antioxidative activity of chestnut fermented puree according to the inoculation amount of Lactobacillus plantarum KCTC 21004 was confirmed in order to confirm its function as a composition. In the present invention, changes in total phenol content and electron donating ability after Lactobacillus plantarum KCTC 21004 cultured for 24 hours before the fermentation and 48 hours after fermentation inoculated with 50% (See Table 9). Gallic acid was used as a reference substance in total phenol content, and the amount of inoculation was 1.0 ~ 3.0% (v / w) in the 0.5% of the experimental group, from 21.78 mg% before fermentation to 19.91 mg% In the experimental group, total phenol increased after fermentation, and the antioxidant activity was increased due to hydrolysis of chestnut puree by lactic acid bacteria according to inoculation amount. The total phenol content of 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity was similar to that of total phenol contents. In the 0.5% (v / w) In the case of 1.0% (v / w) and 2.0% (v / w) before and after fermentation, the amount of radical scavenging activity was decreased from 17.78% before fermentation to 20.0% . As a result, in order to maintain antioxidative activity upon lactic acid fermentation at night, it was judged that the inoculation amount of 1.0% (v / w) or more was appropriate in the above inoculation method. In the present invention, the inoculum amount was set to 2.0% (v / w) And fermented.

The inflammatory disease of the large intestine and small intestine of the present invention may be a pharmaceutical composition selected from the group consisting of acute enteritis, Crohn's disease, ulcerative colitis, and ischemic colitis. In addition, the present invention may be a pharmaceutical composition wherein the functional bowel disease is selected from the group consisting of diarrhea, constipation, enteric diverticulum, and irritable bowel syndrome. In the case of a pharmaceutical composition comprising the night fermented puree of the present invention, it may be prepared by using a pharmaceutically suitable and physiologically acceptable adjuvant. Examples of the adjuvant include an excipient, a disintegrant, a sweetener, a binder, a coating agent, A lubricant, a lubricant or a flavoring agent can be used. In addition, the pharmaceutical composition may be suitably formulated into a pharmaceutical composition containing one or more pharmaceutically acceptable carriers for administration. The pharmaceutical preparation form can be a granule, a powder, a tablet, a coating, a capsule, a suppository, a liquid, a syrup, a juice, a suspension, an emulsion or the like. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methylcellulose, agarose, bentonite, xanthan gum and the like. Acceptable pharmaceutical carriers for pharmaceutical compositions that are formulated into liquid solutions include those suitable for sterilization and in vivo such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol And one or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders and lubricants can be additionally added and formulated into pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.

In the present invention, the lactic acid bacteria used as a probiotics should have stability against gastric acid and bile acid since the lactic acid bacteria consumed to show useful functions in the intestine must stick to the intestines through the digestive tract. When a person eats food, the pH of gastric acid secreted from the stomach is about 2.0 to 3.0, indicating a strong acid, and the time it takes for the food to pass through the stomach is about 2 to 4 hours. Since gastric acid-induced microbial killing affects the survival of lactic acid bacteria, the gastrointestinal transit time of fermented milk is 50% after 90 minutes, 80% after 90 minutes, 3 hours after ingestion All go through the stomach and the gastric pH remains below 2.7. In order for lactobacillus to reach the human field and to be effective as a probiotics, it is necessary to overcome the harsh environment of the human digestive organs in which gastric juice is secreted from the stomach and secreted from the pancreas. These probiotic lactic acid bacteria have been reported to inhibit harmful bacteria caused by acid tolerance and biliary properties resistant to survival inhibition, and to improve constipation and diarrhea, as well as to treat diarrhea and irritable bowel syndrome. Therefore, in the present invention, the fermentation of L. plantarum KCTC 21004 by fermentation used as a probiotics in the present invention is a pharmaceutical composition for preventing or treating intestinal diseases, and it should contain acid-fast bacilli do. In the present invention, the acidity of four kinds of lactic acid bacteria having an acidity of 1.0% or more after 48 hours of fermentation of the chestnut paste inoculated with 2% (v / w) of each lactic acid bacterium was examined (see Table 4). The solid medium (MRS) was adjusted to pH 2.0 with 0.1 N HCl to inoculate the bacteria and the number of bacteria was measured over time. The survival rate of KCTC 3598 was 32.4% lower than that of other lactic acid bacteria. KCTC 3103, 3104 and KCTC 21004 showed high survival rate of over 50%. On the other hand, the measurement of biliary ductility is intended to investigate whether the bile released from the upper part of the small intestine has resistance against acidic properties because it acts to sterilize microorganisms having a membrane composed of fat, because it promotes the digestion and absorption of the lipid food. It has been discovered that bile salt hydrolase (BSH), an enzyme in many species of lactic acid bacteria, has been found to hydrolyze bile salts to reduce solubility, thereby reducing the action of bile acids. However, the mechanism of bile bile acid action has not yet been established. In the present invention, bile acid tolerance was measured while each bacterium was cultured in a (0-1.0%) MRS medium with different concentrations of bile (see Table 5). In the four tested strains, the survival rate was over 50% in the solid medium (MRS) containing 1.0% bile, and KCTC 3104 and KCTC 21004 were not significantly affected by bile Respectively. Although the concentration of bile in the intestines varies, the concentration of bile used in the selection of probiotics strains is known to be about 0.3%. Therefore, it was confirmed that many fungi can reach the large intestine through the stomach and small intestine when the fermented product using KCTC 3104 and KCTC 21004 were ingested. Aminobutyric acid (GABA) content of lactic acid bacteria was analyzed by night fermented puree using probiotic lactic acid bacteria, and the content of γ-aminobutyric acid (GABA) The content of free amino acid in white flesh was 593.4mg% and glutamic acid was found to be 250.8mg%. Therefore, in the present invention, the content of γ-aminobutyric acid (GABA) in the fermented milk obtained by culturing the lactic acid bacteria using glutamic acid-containing chestnut was confirmed (see FIG. 1). The amount of γ-aminobutyric acid (GABA) produced in the fermented broth fermented for 72 hours after inoculation with 2% of lactic acid bacteria cultured in a solid medium (MRS) at 37 ° C and 150 rpm for 24 hours in a 20% (w / w) A small amount of γ-aminobutyric acid (GABA) was detected in 9 kinds of lactic acid bacteria, ranging from 9.32 to 14.31 mg / 100 g. To confirm the conversion of γ-aminobutyric acid (GABA) according to the kind of bacteria, a 20% (w / w) paste of boiled nut added with 1% (v / w) sodium L-glutamate (MSG) Aminobutyric acid (GABA) production was increased when 1% MSG was added to four kinds of lactic acid bacteria. Lactobacillus plantarum KCTC 21004 had a GABA content of 136.4 mg / 100 g, Γ-aminobutyric acid (GABA) production was higher than that of γ-aminobutyric acid (GABA), and the production of γ-aminobutyric acid (GABA) was increased with glutamate content. In the present invention, antioxidative activity as a pharmaceutical composition for the treatment of night fermented puree using the probiotic lactic acid bacterium was also measured. Total phenolic compounds are widely distributed in plants and have various structures and molecular weights. Because they contain many hydroxyl (OH) groups, they easily bind to proteins and other macromolecules, And such an effect has an efficacy as a therapeutic drug composition. Total phenol contents of chestnut paste after lactic acid fermentation were measured before and after fermentation. Gallic acid was used as reference material. Among the four strains used in the present invention, the antioxidant activity and the total phenol content were measured using a night fermented puree at a night content of 20% fermented for 48 hours (see Table 6). The total phenol contents of the four strains were not significantly different from those of the control before fermentation. The increase of total phenol content after lactic acid fermentation showed that the antioxidant activity was increased by hydrolysis of chestnut puree by lactic acid bacteria. In the case of 1.1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, 1.1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability was higher than that of the fermented L. plantarum KCTC 21004 (DPPH) radical scavenging ability, but the difference of radical scavenging ability according to strain was not significant. When compared to the overnight purée before fermentation, the radical scavenging activity was slightly increased in the fermented puree after fermentation. Therefore, lactic acid bacteria used in the night-fermented puree which has a high probiotic function and can enhance the physiological activity of the night fermented puree as a medicinal composition for treatment using the probiotic lactic acid bacterium as described above, (L. plantarum) KCTC 21004 was found to be suitable.

The present invention may be better understood by the following examples, which are for the purpose of illustrating the invention and are not intended to limit the scope of protection defined by the appended claims.

< Example  1> Night Puree  Materials for manufacturing

In the experiment, white flesh was used for the experiment. For the preparation of chestnut paste, it was steamed at 90 ° C for 90 minutes using autoclave, mixed with water, and then sterilized at 121 ° C for 15 minutes.

< Example  2> Used strain  And culture

The lactic acid bacteria used in the study were Lactobacillus rhamnosus KCTC 3237, KCTC 5033, L. kimchicus KCTC 12976, L. plantarum KCTC 21004 and L. plantarum subsp., Which were purchased from the Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology. plantarum KCTC 3103, KCTC 3104, L. brevis KCTC 3102, KCTC 13094, L. acidophilus KCTC 3164, L. sakei subsp. sakei KCTC 3598, Lactococcus lacitics subsp. lactics KCTC 2013 and KCTC 3899 were used.

  Cultures of lactic acid bacteria were cultured at 37 ° C. using a solid plate culture medium (MRS broth, Difico Co., MD, USA).

< Example  3> Night Paste  Fermentation of lactic acid bacteria

Lactobacillus cultured overnight in a solid plate culture medium (MRS) was inoculated with 2% (v / w) and then fermented at 37 ° C for a certain period of time.

end. Measurement of pH and titratable acidity

  The pH was measured at room temperature using a pH meter (DocuMeter, Sartorius, USA). The titratable acidity was determined by neutralizing 10 mL of 10-fold diluted sample with 0.1 N NaOH standard solution until the pH reached 8.2, and converting it into lactic acid.

Titratable acidity (%) =

V: Proper consumption of 0.1 N NaOH (mL)

F: factor of 0.1N NaOH

A: Amount of organic acid equivalent to 1 mL of 0.1 N NaOH (latic acid: 0.009)

D: Dilution factor

S: Sample weight

I. Measure the number of lactic acid bacteria

  The number of lactic acid bacteria was measured by standard agar culture method. To measure the number of viable bacteria in lactic acid bacteria, samples were diluted up to 10 with 0.85% NaCl solution, and 0.1 mL was added to MRS agar medium. And cultured for 36 hours. After culturing, the number of colony counts (number of fungi visible on the solid medium) was selected and the number of viable cells per unit volume (CFU / mL) was calculated by multiplying the number of colonies by the number of dilutions.

All. Determination of soluble solids content

  The fermented product of lactic acid fermented overnight was diluted 2 times and 1 mL of the filtrate was collected and measured using a portable refractometer (PAL-1, ATAGO, Japan).

la. Viscosity measurement

  Viscosity was measured using a viscometer (Brookfield DV-II II + pro, Brookfield Engineering Laboratories, Inc., MA, USA). The spindle was selected by No. 64, and the results were repeated three times.

hemp. Chromaticity measurement

  Chromaticity measurements were carried out in a petri dish (glass jar with a pair of plates and a single lid fitting to it) and measured with a color (Colori-meter, Model JC 801S, Color Techno System, Japan). Hunter? S values L, a and b were measured.

< Example  4> Night fermentation Puree  Measurement of physiological activity

end. Determination of total phenolic compound content

 The total phenolic compound content was colorimetrically determined by the Folin-Denis method. 1 mL of a constantly diluted sample was mixed with 1 mL of a 50% phenol reagent (Folin-Ciocalteu? S reagent, Sigma-Aldrich, Co., St. Louis, Mo., USA) 1 mL of sodium carbonate (Na₂₂CO₃ ₃, SAMCHUN PURE CHEMICAL CO., LTD, Korea) was added, and the mixture was allowed to stand at room temperature for 1 hour. The absorbance was measured at 700 nm using an ultraviolet spectrophotometer. At this time, a standard curve was prepared using garlic acid (Sigma-Aldrich, Co., St. Louis, Mo., USA) as a reference material.

I. Measurement of DPPH radical scavenging activity

  DPPH radical scavenging activity was measured using α-diphenyl-1-picrylhydrazyl (DPPH, Sigma Aldrich, Co., St. Louis, Mo., USA). 5 mL of the DPPH solution adjusted to an absorbance of about 1.0 was mixed with 0.5 mL of the sample solution and allowed to stand at room temperature for 10 minutes. The absorbance at 517 nm was measured with an ultraviolet spectrophotometer. The absorbance difference between the sample and the non- (%).

DPPH radical scavenging contents (%) =

All. GABA (γ-aminobutricacid) content measurement

AccQ-Tag method was used for GABA (γ-aminobutricacid) analysis, and reversed phase column (Waters, ACCQ-TAG ULTRA C18 1.7um, 2.1 * 1000mm, USA) was used for amino acid derivatization using Waters AccQ-Tag Fluor Reagent Kit. (UltiMate 3000RS; UltiMate, USA) equipped with a magnetic stirrer. The amino acid analysis was carried out in the same manner. The analysis conditions are shown in Table 1, and the gradient conditions are shown in Table 2.

Table 1 shows amino acid analysis conditions using high performance liquid chromatography (HPLC).

Table 2 shows the free amino acid elution conditions of high performance liquid chromatography (HPLC).

Table 3 shows the change of pH and titratable acidity according to fermentation period according to kinds of lactic acid bacteria using chestnut.

Table 4 shows the acid resistance effect of the four kinds of lactic acid bacteria having high acid production.

Table 5 shows the results of measurement of bile acid tolerance in cultured bacterium with different concentrations of bile.

Table 6 shows the total polyphenol content and 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity of the night fermented puree against the lactic acid bacteria strain.

Table 7 shows the results of measuring the number of live bacteria per hour of night fermented puree according to inoculation amount.

Table 8 shows the change in color before fermentation and after fermentation of the fermented chestnut paste by varying the amount of lactic acid bacteria inoculated.

Table 9 shows the results of measurement of changes in total phenol content and electron donating ability before and after fermentation after inoculation with different doses.

Claims (9)

Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus. acidophilus, Lactobacillus. sakei and Lactococcus lactics. The fermented overnight fermented puree is inoculated with at least one selected lactic acid bacterium.
The method of claim 1, wherein the Lactobacillus plantarum is Lactobacillus plantarum (KCTC 21004).
[3] The night fermented puree according to claim 1, wherein the lactic acid bacterium is inoculated with 1 to 3% (v / w) of the overnight puree.
A food composition comprising the night fermented puree of any one of claims 1 to 3.
5. The food composition according to claim 4, wherein the composition has at least one activity selected from the group consisting of inflammatory diseases of the large intestine and small intestine and ability to improve functional intestinal diseases.
The food according to claim 4, wherein the food is selected from the group consisting of dairy products, confectionery products, seasonings, beverages and drinks, snacks, candies, ice cream and frozen desserts, breakfast cereals, nutritional bars, snack bars, chocolate products, Soy sauce and dressing, confectionery products, oil and fat products, dairy drink and milk drinks, soy dairy-like product, frozen foods, cooked foods and alternative foods, meat products, cheese, yogurt , Bread and buns, yeast products, cakes, cookies and crackers.
A pharmaceutical composition for preventing or treating at least one disease selected from the group consisting of inflammatory diseases of the large intestine and small intestine and functional intestinal diseases including the night fermented puree of any one of claims 1 to 3.
[Claim 7] The pharmaceutical composition according to claim 7, wherein the inflammatory disease of the large intestine and small intestine is selected from the group consisting of acute enteritis, Crohn's disease, ulcerative colitis, and ischemic colitis.
[Claim 7] The pharmaceutical composition according to claim 7, wherein the functional bowel disease is selected from the group consisting of diarrhea, constipation, enteric diverticulum and irritable bowel syndrome.

Images