JP2012224551A - Composition for sustentation of bifidobacterium or lactobacillus - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a plant-originated safe composition for sustaining life of Bifidobacterium or Lactobacillus which are useful enteric bacteria.SOLUTION: This composition for sustaining life of Bifidobacterium or Lactobacillus includes polyphenols, preferably contains punicalin. Also the composition includes a part or its extract of at least one plant selected from the group consisting of genus Punica plant, genus Quisqualis plant, genus Cistus plant, genus Terminalia plant and genus Combretum plant. The composition further includes an extract with hydrous organic solvents of Punica granatum, Osmanthus fragrans, Murraya koenigii or Linum usitalissimum.

Description

  The present invention relates to a composition for maintaining the survival of bifidobacteria or lactic acid bacteria.

  It has become recognized that it is important to keep the intestinal flora optimal in order to prevent the onset of the disease and maintain health. Probiotics are defined as living bacteria (useful bacteria) themselves that work beneficially to the host, such as the genus Lactobacillus and the genus Bifidobacterium. These bacteria are resident in human intestines. Probiotics have beneficial effects on human health, including degradation of indigestible components, promotion of the immune system, suppression of food allergies and tumors, vitamin production, metabolism of cholesterol and fat, and promotion of in vivo use of minerals.

  Prebiotics are defined as indigestible food ingredients that serve as nutrient sources for specific useful bacteria in the intestine, promote their growth, and have beneficial effects on the host, such as oligosaccharides and dietary fiber. This is the case. Currently, oligosaccharides (galactooligosaccharide, fructooligosaccharide, lactulose, etc.) and some dietary fibers (inulin, etc.) are recognized as prebiotics. In addition, most prebiotics are saccharides, but recently, 1,4-dihydroxy-2-naphthoic acid (DHNA), which is a non-saccharide propionic acid fermentation-derived component (Patent Document 1). Etc. are also permitted. Furthermore, it has been reported that Tellimagrandin II, a tannin contained in the flowers of Hermanus, has a slight growth effect on bifidobacteria (Non-patent Document 1), and a prebiotic effect has also been found in things other than carbohydrates. ing.

  The intestinal flora is stable when healthy, but is affected by host physiology, disease, drugs, stress, food, and so on. In particular, ingredients in foods affect the intestinal flora by increasing or decreasing the number of useful bacteria. Polyphenols such as plant-derived hydrolyzed tannins, condensed tannins, and flavonoids are said to be ingested 1 g per day by meal. They are contained in fruits, vegetables, teas, etc. that are usually eaten, and are known to selectively inhibit growth against pathogenic bacteria in the intestines and exhibit antibacterial properties.

  Pomegranate peel contains a large amount of tannins such as punicalin and punicalagin, and is used in China for antistasis, hemostasis, and anthelmintics. Moreover, punicalagin has a strong antibacterial activity against Clostridium bacteria represented by Staphylococcus aureus and Clostridium cerevisiae, while having a growth promoting action against Bifidobacterium breve (Bridobacterium breve), Punicalin has been reported to have a slight growth promoting effect on Bifidobacterium longum (Non-patent Document 2). However, the survival maintenance action of polyphenols against useful intestinal bacteria is not known.

JP 2002-193903 A

Manjiro Kamijo et al., Biosci. Biotechnol. Biochem., 2008, Vol. 72, No. 3, p. 773-777 Dobroslawa Bialonska et al., J. Agric. Food Chem., 2009, vol. 57, p. 8344-8349

  An object of the present invention is to provide a plant-derived safe composition for maintaining the survival of bifidobacteria or lactic acid bacteria, which are useful enteric bacteria.

  As a result of repeated studies to solve the above-mentioned problems, the present inventors have found that polyphenols in plant extracts such as pomegranates maintain the survival of bifidobacteria or lactic acid bacteria, thereby completing the present invention.

  The present invention provides a composition for maintaining the survival of bifidobacteria or lactic acid bacteria containing polyphenols.

  In one embodiment, the polyphenol is punicalin.

  In one embodiment, the composition comprises a part of at least one plant selected from the group consisting of pomegranate plant, cicanthus plant, gojiaia plant, momotamana plant and yotsubankazura plant, including punicalin. Contains extract.

  In one embodiment, the mass ratio of punicalin to punicalagin in the composition is 2 or more.

  In one embodiment, the composition comprises an extract from a pomegranate, antrum, plantain or flax solvent comprising polyphenols.

  In one embodiment, the composition is for prevention and / or treatment of gastrointestinal diseases.

  In one embodiment, the composition is used by being added to food and drink, pharmaceuticals, feed or pet food.

  In one embodiment, the composition is used by being added to a strain storage medium.

  The present invention also provides a food or drink containing punicalin, or an extract of pomegranate, guinea pig, plantain or flax, and bifidobacteria or lactic acid bacteria.

  The present invention further provides a method for maintaining the survival of bifidobacteria or lactic acid bacteria in the intestine of a pet or livestock, the method comprising punicalin, or an extract with a pomegranate, buffalo, corn borer or flax solvent Administering a edible composition to a pet or livestock.

  The present invention further provides a method for producing a fermented food or drink, the method comprising the step of adding punicalin or an extract of a pomegranate, cinnamon moth, moth, or flax.

  According to the present invention, a plant-derived safe composition for maintaining the survival of bifidobacteria or lactic acid bacteria, which are useful enteric bacteria, can be provided. The composition of the present invention can maintain the survival of bifidobacteria or lactic acid bacteria and keep the intestinal flora in a suitable state. And by maintaining the gut microbiota appropriately, the onset of disease can be prevented and health can be maintained. The survival maintaining action of the bifidobacteria or lactic acid bacteria of the present invention is different from the growth promoting action. Since the bifidobacteria and lactic acid bacteria can live long in the intestine due to the survival maintaining action, the intestinal regulating effect and immune enhancing effect of these bacteria are exhibited.

  The composition of the present invention can also prolong the survival period of bifidobacteria or lactic acid bacteria in food and drink. By adding to the strain storage medium, the number of surviving bacteria during storage can be increased. By adding to the strain culture medium, the culture efficiency of bifidobacteria or lactic acid bacteria can be increased. Furthermore, killing of bacteria during the production process can be suppressed by adding at the time of producing the fermented food or drink.

It is process drawing which shows an example of the extraction process of the extract of this invention. It is a graph which shows the survival maintenance effect with respect to the bifidobacteria of a pomegranate extract. It is a graph which shows the survival maintenance effect with respect to bifidobacteria of punicalin and punicalagin.

  The polyphenol contained in the composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention is a component contained in plants, and has a plurality of phenolic hydroxy groups in the molecule. Preferable examples of the polyphenols of the present invention include polyphenols contained in pomegranate plants, stalk genus plants, gojiaia genus plants, peach genus plants, euphorbia genus plants, cinnamon moths, giant grasshoppers or flax.

  The polyphenol contained in the composition of the present invention preferably includes punicalin. Punicalin has the following structure:

  Punicalin is contained in the pomegranate plant, the plant of the genus Pleurotus, the plant of the genus Godiaeus, the plant of the genus Momotamana, the plant of the genus Yotsubankazura, and can be obtained by extraction and purification from these plants. It can also be obtained by synthesis.

  The pomegranate plant belongs to the family Pomegranate. Pomegranate (Punica granatum) is preferably used. Examples of the site used for extraction include fruit, stem bark, branch bark or root bark, and a suitable site includes fruit skin. For example, it is extracted from dried pomegranate peel with water-containing acetone, water-containing ethanol, water or the like.

  The above-mentioned plant of the genus Skunkun belongs to the family Skunkunushi. Preferably, Quisqualis indica is used. The mature fruit of this plant is called “Akiko” and has been used for medicinal purposes since ancient times. It is especially famous as an anthelmintic. Examples of the site used for extraction include fruits and seeds.

  The plant of the genus Godiai belongs to the family Hannichiaceae. The genus Goliai includes 20 species. Preferably, Cistus ladaniferus and Cistus populifolius are used.

  The plant of the genus Momotamana is a plant belonging to the family Sequimidae as well as Circulatum. Preferably, Terminalia arborea, Terminalia calamansanai, Terminalia catappa, Terminalia chebula, Terminaria horrida, Terminalia triflora are used.

  The above-mentioned plant of the genus Vulgaris also belongs to the family Sequimidae. Preferably, Combretum glutinosum is used.

  As a raw material of punicalin, the above-mentioned plant or a part of the plant, a processed product thereof (dried product, ruptured product, shredded product, or a dry powder obtained by pulverizing these, a product obtained by pulverizing a dried product, etc.), Alternatively, these crude extracts can be used. In the present specification, a plant simply includes a processed product of a plant (a dried product, a ruptured product, a shredded product, or a dry powder obtained by pulverizing these, a product obtained by pulverizing a dried product, etc.). A plant part is a part of a plant such as a fruit, pericarp, flower, stem, leaf, stem, or seed. In the present specification, the extract means an extract obtained by extracting the plant or a processed product thereof with a solvent, a diluted solution or a concentrated solution thereof, or a dried product thereof.

  The plant extract is obtained by extracting a plant or a processed product thereof with a solvent. Examples of the solvent used for extraction include water, lower alcohols such as ethanol and methanol, lower esters such as ethyl acetate and methyl acetate, acetone, and a mixture thereof (water-containing organic solvent). Among these, it is preferable to use water alone, ethanol alone or a mixture of ethanol and water (so-called water-containing ethanol) because it is ingested by humans. In particular, it is more preferable to use an ethanol / water mixture containing ethanol at a concentration of 30% (v / v) or more.

  For example, in the examples described later, in the extraction from pomegranate, an acetone / water mixture (70:30 v / v) is used, but instead of this, ethanol of 30% (v / v) or more is used. It is also possible to use a / water mixture.

  The extraction method is not particularly limited, but it is preferable to perform the extraction under as mild a condition as possible from the viewpoint of safety and convenience in use. For example, a raw material plant part or a dried product thereof is pulverized, crushed or shredded, a 5 to 20 times mass solvent is added thereto, and the temperature ranges from 0 ° C. to the reflux temperature of the solvent for 30 minutes to several weeks, for example under heating. Extraction is performed under any conditions such as standing, shaking, stirring or refluxing for 30 to 48 hours. It may be extracted by homogenization after adding a solvent. After extraction, separation operations such as filtration and centrifugation are performed to remove insoluble matters, and dilution and concentration operations are performed as necessary to obtain an extract. Furthermore, if necessary, the same operation may be repeated for the insoluble matter to further extract, and the extract may be used together.

  For the purification procedure and conditions of punicalin preferred in the present invention, for example, silica gel column chromatography, reverse phase ODS column chromatography, preparative HPLC, etc. are combined, and under suitable conditions, eluent (for example, water or hexane, It is carried out using various organic solvents such as ethyl acetate, chloroform, methanol and n-butanol, or a mixture thereof, but is not particularly limited thereto.

  One embodiment of the composition for maintaining the survival of the bifidobacteria or lactic acid bacteria of the present invention comprises a pomegranate-containing plant, a prunus genus plant, a gossiod plant, a momotana plant, a part of the genus Yotsubankazura or an extract thereof. contains. Considering the point of using the composition of the present invention as a food or medicine, the composition of the present invention preferably contains an extract.

  About a plant, a part of plant, an extract, and an extraction method, it is the same as that of the above, and has the same concept and meaning.

  The extract contained in the composition of the present invention is used in the form of a liquid product, a concentrated product, a paste, or a dried product obtained by drying or concentrating the extract. Drying is performed by a method commonly used by those skilled in the art, such as spray drying, freeze drying, reduced pressure drying, and fluidized drying. The extract may be further purified by a purification method commonly used by those skilled in the art.

  The extract contained in the composition of the present invention is a water or solvent extract obtained by the above procedure and a supercritical carbon dioxide extract, and these obtained extracts are vacuum-dried or spray-dried known in the art. The solid content obtained by removing the solvent by a method such as (spray drying) and the fraction obtained by further purifying these are also included.

  For preferred purification procedures and conditions in the present invention, for example, silica gel column chromatography, reverse phase ODS column chromatography and preparative HPLC are combined, and eluent (for example, water or hexane, ethyl acetate) under suitable conditions. , Various organic solvents such as chloroform, methanol and n-butanol, or a mixture thereof), but is not particularly limited thereto.

  The punicalin in the composition of the present invention is preferably highly pure. It is particularly preferred that the content is higher than that of punicalagin. Preferably, the mass ratio of punicalin to punicalazine is 2 or more, more preferably 5 or more, and still more preferably 10 or more. Most preferred is a composition that is substantially free of punicalagin. When the mass ratio of punicalin to punicalazine is 2 or more, the survival maintaining effect of bifidobacteria or lactic acid bacteria is particularly high.

  The composition for maintaining the survival of Bifidobacteria or lactic acid bacteria of the present invention also contains an extract of a pomegranate, cinnamon moss, large white kite or flax containing polyphenol. The composition also includes a composition containing two or more of an extract from a pomegranate solvent, an extract from a beetle solvent, an extract from a papaver solvent, or an extract from a flaxseed solvent.

  The pomegranate used in the present invention is a pomegranate plant, Punica granatum. Examples of the site used for extraction include fruit, stem bark, branch bark or root bark. Suitable skin includes the pericarp. For example, it is extracted from dried pomegranate peel with water-containing acetone, water-containing ethanol, water or the like.

  As for this invention, the Oxmanaceae plant is Osmanthus fragrans. Also called Keika. Suitable sites used for extraction include flowers and leaves.

  The giant grasshopper (Ohatsuki Tachibana) used in the present invention is a citrus plant, Murraya koenigii. Another name is curry tree, and its leaves are called curry leaf. A suitable part used for extraction includes leaves (curry leaf).

  The flax used in the present invention is the flaxaceae plant, Linum usitatissimum. Flaxseed oil is a yellowish dry oil obtained from mature flax seeds, which is obtained by pressing flaxseed (flaxseed) or crushing it and extracting it with a solvent. A suitable site used for extraction includes seeds.

  Examples of the solvent used in the present invention include water and a water-containing organic solvent. Preferably, a water-containing organic solvent is used. Examples of the water-containing organic solvent include lower alcohols such as ethanol and methanol, lower esters such as ethyl acetate and methyl acetate, or a mixture of acetone and water.

  The meaning and concept of the extract of the pomegranate, cinnamon moth, sorghum or flax's solvent are the same as described above, and the extraction method and the purification procedure are the same as those described above.

  Bifidobacteria and lactic acid bacteria that can maintain the survival of the composition of the present invention are not particularly limited as long as they are useful bacteria that form intestinal flora. As Bifidobacterium, for example, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium infantis ( Bifidobacterium adolesentis), Bifidobacterium breve, Bifidobacterium bifidum, Bifidobacterium animalis, Bifidobacterium catenulatum, Bifido Examples include Bifidobacterium pseudocatenulatum.

  Examples of the lactic acid bacteria include Lactococcus lactis subsp. Lactis, Lactococcus lactis subsp. Cremoris, Streptococcus salvarius subsp. Thermosp. thermophilus, Leuconostoc mesenterides subsp. cremoris, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei ii subsp. bulgaricus), Lactobacillus delbrueckii subsp. lactis, Lactobacillus gasseri (Lactobacillus) gasseri), Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus brevis, Tetragenococcus halophilus, Pedococcus pentosus cc Can be mentioned.

  The composition for maintaining the survival of bifidobacteria or lactic acid bacteria in the present invention maintains the survival of bifidobacteria or lactic acid bacteria present in the intestinal flora, food and drink, culture medium, storage medium, and the like. The survival maintenance action is an action that suppresses the death of bifidobacteria or lactic acid bacteria and prolongs the survival period, and is different from the proliferation action that activates the division of bacteria. In order to grow bifidobacteria or lactic acid bacteria, it is necessary to become a nutrient source selective to those bacteria. However, the survival maintaining action of the present invention is not a nutrient source, and it is necessary for these bacteria to survive. It is presumed to protect bacteria from oxygen and enzymes that become obstacles, and is completely different from the proliferation effect.

  The shape of the composition for maintaining the survival of Bifidobacterium or lactic acid bacteria of the present invention is not limited. Examples are solid, semi-solid and liquid. The composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention includes those directly eaten, consumed, or administered to humans or animals. Moreover, what is added to food-drinks etc. is included. Examples of the target to be added include foods and drinks, pharmaceuticals, feed or pet food bacteria culture medium, bacteria preservation medium, bifidobacteria or lactic acid bacteria-containing food and drink. Moreover, it is added during the manufacturing process of fermented food.

  Compositions that are eaten and consumed directly by humans or animals include health foods and supplements that are specially processed to maintain the survival of bifidobacteria or lactic acid bacteria. For example, beverages, noodles, confectionery, jelly-like foods and the like are included. It may be formulated for food and drink. Examples thereof include powders, granules, tablets, capsules, liquids (including suspensions and emulsions). It can be formulated by mixing with additives usually used by those skilled in the art (for example, excipients such as dextrin, starch, saccharides, calcium phosphate, solvents, fragrances, perfume oils, etc.). For pharmaceutical compositions to be administered to humans or animals, punicalin or the above extract is mixed with additives usually used by those skilled in the art (for example, excipients such as dextrin, starch, saccharides, calcium phosphate, solvents, fragrances, perfume oils, etc.) And can be formulated. Examples thereof include powders, granules, tablets, capsules, liquids and the like.

  As a method for producing a capsule, a conventionally known method for producing a capsule may be used except that punicalin or the extract is used as the content. For example, as a method for producing a soft capsule, a capsule film sheet is used to enclose the contents with a rotary filling machine and a capsule preparation is molded, or a seamless capsule is produced by a dropping method.

  As for the granules, various known wet and dry granulation methods can be applied, and the granules are molded together with appropriate binders and excipients. Tablets are prepared by known tableting methods by adding appropriate binders, excipients, disintegrants and, if necessary, lubricants to the granules containing the plant or the extract or the extract itself. be able to. Furthermore, about a drink, an appropriate saccharide | sugar, an acid, a fragrance | flavor, etc. can be added, a flavor can be adjusted, and it can prepare by a well-known manufacturing method.

  The composition added to food and drink is not limited as long as it is a shape that can be easily added, such as powder or liquid. Examples include food additives, nutritional supplements, food supplements, and the like. Composition depending on the addition method and application, such as a composition containing a shape and amount that can be added to and mixed with food and drink by a person who eats and drinks as a sprinkle, and a composition containing a shape and amount for addition during processing of food and drink However, the composition of the present invention includes all of them.

  One embodiment of the composition of the present invention is a composition for the prevention and / or treatment of gastrointestinal diseases. For example, it is formulated into powders, granules, tablets, capsules, liquids and the like by methods commonly used by those skilled in the art. The administration method is preferably oral administration, and is administered once or multiple times a day. Gastrointestinal diseases can be prevented and / or treated by the survival maintaining action of bifidobacteria or lactic acid bacteria.

  The composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention can be used by being added to foods and drinks, pharmaceuticals, feeds or pet foods. The food or drink to be added is not particularly limited. Examples include processed foods, beverages, seasonings, powdered milk, health foods, and supplements. Feed and pet food are food given to livestock and pets, including livestock and pet drinks.

  The composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention can be used by adding to a strain storage medium. The composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention is used as an auxiliary for strain preservation, and suppresses the death of bifidobacteria or lactic acid bacteria during storage.

  The present invention also relates to an extract of punicalin or a pomegranate, guinea pig, lobster, or flax, and a food or drink containing bifidobacteria or lactic acid bacteria. By containing an extract of punicalin or a pomegranate, cinnamon moth, moth, or flax, the death of bifidobacteria or lactic acid bacteria in foods and drinks can be suppressed, and it can survive for a long period of time. Retained.

  Furthermore, the present invention provides a bifidobacteria or lactic acid bacterium in the intestine of a pet or livestock, comprising the step of administering to the pet or livestock a edible composition containing punicalin, or an extract of a pomegranate, cinnabar, lobster, or flax's solvent. It relates to a method for maintaining the survival of a child. The composition for eating and drinking is food and beverage, and can maintain the health by favoring the intestinal flora of pets and livestock.

  Furthermore, this invention relates to the manufacturing method of the fermented food / beverage products of bifidobacteria or lactic acid bacteria including the process of adding the extract by the solvent of punicalin or a pomegranate, a cinnamon moth, a prickly pear, or flax. By adding punicalin or the like, the survival of bifidobacteria or lactic acid bacteria in the production process and in fermented foods and drinks is maintained, and efficient production becomes possible. The amount of punicalin added to the fermented food or drink is preferably 0.00001 to 0.01 w / w%, more preferably 0.0001 to 0.001 w / w%, still more preferably 0.0002 to 0%. 0006w / w%. The amount of the extract added to the fermented food or drink is preferably 0.0001 to 1 w / w%, more preferably 0.001 to 0.1 w / w%, and still more preferably 0.005 to 0.00. 02 w / w%.

  EXAMPLES Next, although an Example is given and this invention is further demonstrated, this invention is not limited to these Examples.

Example 1
(Preparation of extracts from plants)
Asunaro (leaves), flax (seed), everlasting flower (flowers), young barley leaves (leaves), olives (leaves), oregano (leaves), coconuts (fruits), white fox (leaves), Butterflies (flowers), pomegranates (fruit skin), shaba (fruits), sunshuyu (fruits), German chamomile (flowers), Sukyo (stems), nigella (seed), noni (leaves), hibiscus (flowers), winter buds (flowers) Leaf), mokka (fruit), peach (flower), lemongrass (leaf), rose petal (petal), and roman chamomile (flower), respectively, 6.5 L of 70% (v / v) acetone aqueous solution was added. Mix and homogenize. After filtration, the filtrate was collected, concentrated under reduced pressure, and dried to obtain an extract.

(Example 2)
The pomegranate peel extract obtained in Example 1 was fractionated with ether, ethyl acetate, and butanol to obtain an ether extract, an ethyl acetate extract, a butanol extract, and a water extract (FIG. 1).

(Example 3)
500 mg of butanol extract obtained in Example 2 was added to water (150 mL), 10% methanol (550 mL), 20% methanol (250 mL) using an MCI® GEL (1.2 cm ID × 45 cm) column. ), And the active compound was isolated by sequential elution with 30% methanol. As a result, punicalin was obtained from fractions 2-8 of the 10% methanol elution part, and punicalazine was obtained from fractions 33-48. The punicalin solution did not substantially contain punicalagin. Based on the analysis results of spectral data of ¹ H-NMR, ¹³ C-NMR, and MS, respectively, it was identified as punicalin and punicalazine. In addition, assignment was performed by two-dimensional NMR analysis of each compound. NMR data of punicalin is shown in Table 1.

Example 4
(Evaluation test for survival of Bifidobacterium longum (B. longum))

(1) Survival maintaining action of the extract obtained in Example 1 The survival maintaining action of the extract obtained in Example 1 was evaluated by the following method 1 for measuring viable cell count.

Viable count method 1
The test sample was dissolved in DMSO so that the final concentration of each sample was 0.01%. Bifidobacterium longum BB536 was precultured in MRS medium at 37 ° C. for 18 hours or more under anaerobic conditions. 1% volume of the pre-cultured bacteria was added to GAM medium (Nissui Pharmaceutical) and cultured at 37 ° C. under anaerobic conditions. The bacterial solution is diluted 12 hours after the start of the culture and then seeded on a BL agar medium (Nissui Pharmaceutical). The same operation is performed 24 hours after the start of the culture and the culture is performed under anaerobic conditions at 37 ° C. for 48 to 72 hours. The number of colonies on the BL agar medium was counted, and the number of viable bacteria was expressed in CFU / mL. The survival maintenance action was evaluated by comparing the number of viable cells 12 hours after the start of culture and the number of viable cells after 24 hours for the control and each extract in consideration of the growth of the number of viable cells. Relative residual bacteria values were calculated.

  The results are shown in Table 2. As can be seen from Table 2, the survival-sustaining effects against B. longum bacteria were flaxseed seed extract, cinnamon blossom flower extract, and grasshopper leaf extract.

  (2) The following viable count method 2 for the pomegranate peel extract obtained in Example 1 and the ether extract, ethyl acetate extract, butanol extract and water extract obtained in Example 2 Evaluated by.

Viable count method 2 (Method of adding test sample at the beginning of the decay period)
Bifidobacterium longum BB536 was seeded in MRS medium and pre-cultured at 37 ° C. under anaerobic conditions for 18 hours or more. Add 1% volume of pre-cultured bacteria to GAM medium (14.75 g / 250 mL water), culture under anaerobic conditions for 12 hours at 37 ° C., and dispense 10 mL each into a test tube sterilized at 121 ° C. for 20 minutes. Noted. A test sample (sample extract 1 mg / 100 μL DMSO) dissolved in DMSO was added to each test tube (the final concentration of the sample was 0.01%). Furthermore, it culture | cultivates on 37 degreeC and anaerobic conditions for 24 hours, Bacterial solution is diluted, and it seeds | inoculates on BL agar medium. The number of bacteria was expressed in CFU / mL and evaluated.

  In the ether extract, the number of viable bacteria was remarkably small compared to the control, and it was observed that B. longum was not maintained. In the ethyl acetate extract, butanol extract and water extract, the number of viable bacteria was larger than that of the control, and the survival maintenance action was stronger than that of the whole pomegranate extract, and the survival maintenance action of B. longum was observed. The degree of each survival maintaining action was 1.41 times for the ethyl acetate extract, 1.42 times for the butanol extract, and 1.43 times for the aqueous layer extract relative to the control. The results are shown in FIG.

(3) The acetone extract of the pomegranate peel obtained in Example 1 and the punicalin and punicalazine obtained in Example 3 were evaluated by the above viable count method 2. The results are shown in FIG.

  From FIG. 3, the number of viable bacteria decreased with respect to the control in punicaladine (0.77 times), and the number of viable bacteria with the punicalin was significantly increased (1.41 times) with respect to the control. A survival maintenance effect was observed against longum.

  The composition for maintaining the survival of bifidobacteria or lactic acid bacteria of the present invention has a strong bifidobacteria or lactic acid bacteria survival support action, optimizes the intestinal flora, and is useful for maintaining the health of humans and animals. It is also useful for addition to fermented foods, cell preservation media, and cell culture media.

Claims (8)

  A composition for maintaining the survival of bifidobacteria or lactic acid bacteria containing polyphenols.   The composition of claim 1, wherein the polyphenol is punicalin.   A part of at least one plant selected from the group consisting of the pomegranate plant, the genus Kunkushi, the plant of the genus Godiai, the plant of the genus Momota, and the plant of the genus Yotsuban, which contain the punicalin, or an extract thereof. The composition as described.   The composition of Claim 1 containing the extract by the solvent of a pomegranate, a hemlock, a green fox, or flax containing the polyphenol.   The composition according to any one of claims 1 to 4, which is used for prevention and / or treatment of gastrointestinal diseases.   The composition according to any one of claims 1 to 4, which is used by being added to food and drink, pharmaceuticals, feed or pet food.   The composition according to any one of claims 1 to 4, which is used by being added to a strain storage medium.   A food and drink containing punicarine, or an extract of pomegranate, cinnamon moth, plantain or flax, and bifidobacteria or lactic acid bacteria.

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