JP2008120789A - Inhibitor or bacteriostatic agent of helicobacter pylori - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inhibitor or a bacteriostatic agent of Helicobacter Pylori inhibiting the increase of Helicobacter Pylori and being safe to human bodies. <P>SOLUTION: The inhibitor or the bacteriostatic agent of Helicobacter pylori contains water-soluble saccharide which is a polysaccharide, in which a plurality of saccharides are chemically bonded, having at least one β bond in the structure, and the polysaccharide contains at least one glucose and/or xylose as a constituent saccharide and β1-4 bond is preferably β1-4 glucoside bond and/or β1-4 xyloside bond. <P>COPYRIGHT: (C)2008,JPO&INPIT

Description

The present invention provides a H. pylori inhibitor or bacteriostatic agent in the food and pharmaceutical fields, and more specifically, a composition containing cellooligosaccharide and / or xylooligosaccharide having a specific sugar composition as an active ingredient. It is related with the inhibitor or bacteriostatic agent of Helicobacter pylori (Helicobacter pylori) which suppresses the increase in Helicobacter pylori which is harmful bacteria by ingesting this.

Cellooligosaccharide is a generic name for cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose, and is a polysaccharide in which 1 to 6 glucopyranose units are β1-4 linked. In the human digestive tract, Not decomposed.

  Xylooligosaccharide is a generic name for xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose, and xyloheptaose, and is a polysaccharide in which 1 to 7 xylose units are β1-4 linked. Like the cellooligosaccharide, it is hardly degraded in the human digestive tract.

Cellooligosaccharides and xylooligosaccharides are useful as low-calorie sweeteners, as well as general foods, functional foods, cosmetics, pharmaceuticals and their additives, other chemical conversion raw materials, and fermentation raw materials.
In recent years, cellooligosaccharides, like other oligosaccharides, are becoming clearer in physiological function and are expected as new materials for functional foods (Non-patent Document 1).

  H. pylori (Helicobacter pylori, or Helicobacter pylori) is a Gram-negative microaerobic bacilli. Helicobacter pylori is considered to live in a strongly acidic environment in the stomach by neutralizing gastric acid by producing ammonia from urea by its urease activity. It has been clarified that the ammonia produced by H. pylori damages the gastric mucosa and is associated with gastric diseases such as gastritis, ulcers, gastric cancer, and lymphoma. It is recognized as an effective preventive or ameliorating measure against the disease.

  As a conventional method for inhibiting Helicobacter pylori, administration of antibiotics has been attempted. This method certainly has the effect of suppressing Helicobacter pylori. However, there is a problem that the inhibitory action of antibiotics affects not only H. pylori but also enteric bacteria useful for the human body. In addition, diarrhea occurs as a side effect, and there are problems that affect the human body. Accordingly, there has been a demand for an inhibitor that is safe for the human body and that selectively acts on H. pylori.

As a prior art, the following preventive agents for Helicobacter pylori containing saccharides as an active ingredient are listed below.
Patent Document 1 discloses an antitumor agent containing fucoidan as an active ingredient. The anti-tumor agent of this document uses fucoidan's inhibition of colonization of Helicobacter pylori on the stomach wall and is effective in preventing infection of Helicobacter pylori, but does not directly inhibit Helicobacter pylori.
In addition, a human safer H. pylori inhibitor or bacteriostatic agent that suppresses an increase in Helicobacter pylori by ingestion has not been known.

Cellulose Communications, 5, No 2,91-97 (1998) JP 7-138166 A

  An object of the present invention is to provide a human safe H. pylori inhibitor or bacteriostatic agent that suppresses an increase in Helicobacter pylori.

The present inventors have found that a composition containing a specific cellooligosaccharide and / or xylooligosaccharide as an active ingredient suppresses an increase in Helicobacter pylori and makes the present invention. It came to.
That is, the present invention is as follows.

(1) A Helicobacter pylori inhibitor or bacteriostatic agent comprising a water-soluble saccharide having a structure in which a plurality of sugars are chemically bonded and having at least one β bond in the structure.
(2) The inhibitor or bacteriostatic agent of H. pylori according to claim 1, wherein the β bond is a β1-4 bond.
(3) The polysaccharide comprises at least one glucose and / or xylose as a constituent sugar, and the β1-4 bond is a β1-4 glucoside bond and / or a β1-4 xyloside bond. 2. The suppressor or bacteriostatic agent of Helicobacter pylori according to 2.
(4) The cellobiose content is 50% by mass or more, the content of one or more selected from the group consisting of cellotriose, cellotetraose, cellopentaose and cellohexaose is 0 to 50% by mass, and the glucose content is 30 The suppressor or bacteriostatic agent of Helicobacter pylori according to any one of claims 1 to 3, comprising a cellooligosaccharide that is not more than mass% as an active ingredient.
(5) 30% by mass of at least one selected from the group consisting of xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose, xyloheptaose and water-soluble xylan having a degree of polymerization exceeding 7. The suppressor or bacteriostatic agent of Helicobacter pylori according to any one of claims 1 to 3, which contains, as an active ingredient, a xylooligosaccharide having a xylose content of 70% by mass or less.
(6) The inhibitor or bacteriostatic agent according to any one of claims 1 to 5 and a food material or a pharmaceutical active ingredient are dispersed or dissolved in water or a mixed liquid of water / organic solvent. An aqueous composition, wherein the content of the bacteriostatic agent is 0.1% by mass or more.
(7) The aqueous composition according to claim 6, further comprising at least one selected from the group consisting of a surfactant, a thickener and a gelling agent.
(8) The aqueous composition according to claim 6 or 7, which is in any state of an aqueous solution, an aqueous dispersion, a paste form, and a gel form.
(9) The inhibitor or bacteriostatic agent according to any one of claims 1 to 5 and at least one selected from a food material, a pharmaceutical medicinal ingredient, a food additive and a pharmaceutical additive. A solid composition, wherein the content of the fungicide is 0.1% by mass or more.
(10) The solid composition according to claim 9, wherein the solid composition is in the shape of any one of a powder, a granule, and a molded body.
(11) The solid composition according to claim 10, wherein the surface is coated.

The cellooligosaccharide composition and the xylooligosaccharide composition of the present invention have the effect of suppressing or bacteriostatically increasing the increase in Helicobacter pylori.
The cellooligosaccharide composition and xylooligosaccharide composition of the present invention directly inhibits Helicobacter pylori regardless of colonization on the stomach wall, and thus has an improvement effect in addition to prevention, and is completely different from conventional preventive agents.

The present invention will be specifically described below, particularly focusing on preferred embodiments thereof.
The H. pylori inhibitor or bacteriostatic agent of the present invention must contain a water-soluble saccharide in which a plurality of sugars are β-bonded.
Examples of the sugar include known hexoses such as glucose, fructose, galactose and mannose or isomers thereof, derivatives, pentoses such as xylose and arabinose or isomers thereof, derivatives, tetroses such as erythrose and threose, or isomers and derivatives thereof. Is mentioned.

  Water-soluble saccharides in which a plurality of sugars of the present invention are β-bonded, and known ones include, for example, cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose, etc. Gentiooligosaccharides, galactooligosaccharides in which a plurality of glucoses and / or galactoses are β-bonded, xylooligosaccharides in which a plurality of xyloses are β-bonded, chitin oligosaccharides in which N-acetylglucosamine, which is a glucose derivative, is β-linked, N- And chitosan oligosaccharides in which multiple glucosamines are β-bonded. Moreover, the sugar which comprises a polysaccharide does not need to be single like lactose which beta-bonded galactose and glucose, and if it has one or more beta bonds in a polysaccharide structure, others The bond may be other than β bond.

However, for those in which a plurality of sugars are β-bonded and the effects of the present invention can be obtained, in addition to the above known water-soluble saccharides, newly discovered or synthesized saccharides are also included in the water-soluble saccharides of the present invention. included. The β bond of the present invention includes β1-4 bond, β1-6 bond, and β1-3 bond.

Among the water-soluble saccharides in which a plurality of sugars are β-bonded, the most effective effects of the present invention are as follows. Water-soluble saccharides in which a plurality of glucoses are β1-4 glucoside-bonded and a plurality of xyloses are β1-4. Xyloside-linked water-soluble polysaccharides are suitable.
Water soluble saccharides with β1-4 glucoside linkages of multiple glucoses and β1-4 xyloside linkages of multiple xyloses can be easily dissolved as a composition in the food / pharmaceutical field and have a molecular weight that can be easily incorporated into harmful fungi. From the viewpoint of having it, oligosaccharides are preferable, and cellobiose and xylobiose which are disaccharides are more preferable.

Below, cellooligosaccharide which is one of the preferable polysaccharides used by this invention is explained in full detail.
The cellooligosaccharide of the present invention preferably contains 50% by mass or more of cellobiose. Cellobiose has the effect of suppressing the growth of H. pylori in an aqueous medium. Therefore, a higher cellobiose content is preferable because growth of H. pylori is suppressed. As a preferable content, it is 70 mass% or more, More preferably, it is 90 mass% or more, and 95 mass% or more is preferable.

  The cellooligosaccharide of the present invention may contain 50% by mass or less of one or more selected from cellotriose, cellotetraose, cellopentaose and cellohexaose. These saccharides also have the effect of inhibiting the growth of Helicobacter pylori, similar to cellobiose. However, as the number of glucose residues increases, the solubility in an aqueous medium decreases. Therefore, in order to obtain the effect of the present invention as a cellooligosaccharide, these contents must satisfy the above-mentioned range. The content range is preferably 30% by mass or less, more preferably 10% by mass or less, still more preferably 5% by mass or less, and particularly preferably 0.5 to 5% by mass.

  The cellooligosaccharide of the present invention has a glucose content of 30% by mass or less. Glucose acts as a carbon source for H. pylori and contributes to its growth and increase. Therefore, when it is contained in a large amount, the inhibitory effect of cellooligosaccharide on H. pylori is reduced. Therefore, the glucose content needs to satisfy the above range. The smaller the glucose content is, the more effective the suppression effect of Helicobacter pylori is promoted. The preferred range is 20% by mass or less, the more preferred range is 10% by mass or less, and the most preferred range is 5% by mass or less. The lower limit is not particularly set, but the range obtained by a simple operation is 0.1% by mass or more.

Next, xylo-oligosaccharide, which is one of the preferred polysaccharides used in the present invention, will be described in detail.
The xylo-oligosaccharide of the present invention is 30% by mass or more of at least one selected from xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose, xyloheptaose and water-soluble xylan having a degree of polymerization exceeding 7. Can be included. These saccharides also have the effect of inhibiting the growth of Helicobacter pylori, similar to cellobiose. The content range is preferably 40% by mass or more, more preferably 50% by mass or more, still more preferably 60% by mass or more, and particularly preferably 70% by mass or more.

  The xylooligosaccharide of the present invention preferably has a xylose content of 70% by mass or less. Xylose acts as a carbon source for H. pylori and contributes to its growth and increase. Therefore, when it is contained in a large amount, the effect of Xylooligosaccharide to suppress H. pylori is reduced. Therefore, the xylose content needs to satisfy the above range. The smaller the xylose content, the more effective the suppression effect of Helicobacter pylori is promoted. The preferred range is 50% by mass or less, the more preferred range is 40% by mass or less, and particularly preferred is 30% by mass or less. The lower limit is not particularly set, but the range obtained by a simple operation is 10% by mass or more.

  The Helicobacter pylori is classified as Helicobacter pylori, and the Helicobacter pylori, for example, conserved strains such as Helicobacter pylori ATCC 43504 and 43526, and clinical Isolates are included.

The increase suppression rate of Helicobacter pylori of the cellooligosaccharide of the present invention is preferably 1% or more. The increase suppression rate here is defined as the number of viable bacteria when H. pylori is cultured in a state in which a test solution containing cellooligosaccharide and / or xylooligosaccharide is added (viable cell count 1). It is represented by the following formula when the viable cell count when cultivated with / or without adding xylooligosaccharide is (viable cell count 2).
Increase suppression rate (%) = [(viable cell count 2-viable cell count 1) / viable cell count 2] × 100
The higher the rate of increase, the greater the suppression of Helicobacter pylori or the bacteriostatic effect. Therefore, the preferred range is 10% or more, more preferably 40% or more, and particularly preferably 50% or more.

The viable count of Helicobacter pylori according to the present invention can be measured by the following method. A test strain (Helicobacter pylori ATCC 43504) was cultured in sheep blood agar medium K (BBL) at 35 ° C. for 3 days by microaerobic culture, and then subjected to McFarland No. 2 (about 10 ⁷ to 10 ⁸ CFU / mL). This is diluted 100 times with a sterilized physiological saline solution to obtain a bacterial solution for addition. A solution obtained by adding 5% horse serum to Brucella broth (Eiken Chemical) diluted 1/10 with water is used as a test solution without addition of cellooligosaccharide and / or xylooligosaccharide. Alternatively, a test solution containing cellooligosaccharide and / or xylooligosaccharide is added with xylooligosaccharide. 1 mL of the bacterial solution for addition is added to 9 mL of each test solution, and cultured under shaking at 35 ° C. under microaerobic conditions. Each test solution is collected 48 hours after culturing, and a 10-fold diluted series solution is prepared with sterile physiological saline. 50 μL of the stock solution and each dilution series solution are smeared on sheep blood agar medium K and cultured under microaerobic conditions at 35 ° C. for 4-5 days. After 0 hour, only the test solution is quantified. The number of bacteria that have grown after culturing is counted and the number of viable bacteria per mL is obtained.

Hereinafter, methods for analyzing cellooligosaccharide, glucose content, xylooligosaccharide, and xylose content in the present invention will be described. The cellooligosaccharide and / or xylooligosaccharide of the present invention is dissolved in pure water at a concentration of 1% by mass, and then subjected to high performance liquid chromatography (chromatography system: manufactured by Shimadzu Corporation, trade name: SCL-10A, column: Shimadzu Corporation) Product name Asahipak NH ₂ P-50, mobile phase: acetonitrile / water = 75/25 (volume ratio)). The sugar composition of the cellooligosaccharide is calculated by converting the peak areas of cellobiose, cellotriose, cellotetraose, cellopentaose, cellohexaose and glucose in the chromatogram obtained by the above-mentioned method into mass percentages of the total mass. It is represented by

The sugar composition of xylo-oligosaccharides is water-soluble having a degree of polymerization exceeding xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose, xyloheptaose in the chromatogram obtained by the above method. The peak area of xylan or xylose is converted into mass and expressed as a percentage by mass of the total mass.

Next, the manufacturing method of the cellooligosaccharide of this invention is demonstrated.
The origin of the cellooligosaccharide of the present invention is not particularly limited, and may be one produced by hydrolysis of a cellulosic material, one produced by condensation or sugar transfer of a monosaccharide such as glucose or a derivative thereof, What was obtained by the enzymatic decomposition method is preferable in terms of safety.

Cellulosic substances used for enzymatic degradation may be plant or animal, and may be derived from natural products such as wood, bamboo, cotton, ramie, squirts, bagasse, kenaf, wheat, rice, and bacterial cellulose. Fibrous substances, or regenerated cellulose that has been regenerated by dissolving them in a solvent, may be those obtained by subjecting them to chemical treatment to give cellulose derivatives, and one or more of the above may be used in combination. Among these, natural cellulosic substances that do not undergo dissolution or chemical treatment are preferable because the obtained cellooligosaccharides do not contain solvents or chemical substances harmful to the human body. Moreover, it is preferable to use a cellulosic substance in the state of a refined pulp, and there is no restriction | limiting in particular in the refinement | purification method of a pulp, You may use any pulp, such as a sulfite pulp, a kraft pulp, and NBKP pulp.

  In the case of enzymatically decomposing a cellulosic material, the cellulosic material used is a cellulosic material that is once hydrolyzed and partially hydrolyzed to an average polymerization degree of 700 or less. It is preferable in terms of improvement. Furthermore, it is possible to use a cellulosic material having a specific degree of polymerization with an average particle size of 100 μm or less and a colloidal cellulose component content of 10% by mass or more to improve the enzymatic degradation rate and select cellooligosaccharides. This is preferable because the rate is improved.

  In the present invention, an enzyme used for hydrolysis of a cellulosic substance is referred to as cellulase, and the cellulase used in the present invention is a general term for enzymes that decompose cellulose. It is included in the cellulase referred to in the invention. Cellulase enzyme sources include, for example, cellulase-producing living cells themselves, purified cellulase-producing bacteria, and formulated purified enzymes with additives such as excipients and stabilizers. . In the case of a cellulase preparation, the additive added thereto is not particularly limited, and the dosage form may be any of powder, granule, liquid and the like.

  The origin of cellulase is not particularly limited. For example, microorganisms that produce known cellulases include the genus Tricodederma, the genus Acremonium, the genus Aspergillus, the genus Bacillus, and the pseudomonas. (Pseudomonas genus), Penicillium genus, Aeromonus genus, Irpex genus, Sporotrichum genus, Humicola genus, Cellobibrio celioc Issued (1987)), and the fungus described in "Encyclopedia of Cellulose" (published by Asakura Shoten (2000)) is produced. It can be exemplified cellulase, if enzymes that degrade cellulose, not only the enzyme derived from the known bacteria, enzymes from novel microorganisms are also included in cellulase of the present invention.

The enzymatic decomposition method may be any known method, and is not particularly limited. For example, the cellulosic material is suspended in an aqueous medium, cellulase is added, and stirring or shaking is performed. And a method of performing a saccharification reaction by heating.
In the above method, the suspension method, the stirring method, the addition method / order of addition of cellulase / substrate, the concentration thereof, and other reaction conditions are appropriately adjusted so that the cellooligosaccharide can be obtained in a higher yield. In this case, the pH and temperature of the reaction solution may be within the range where the enzyme is not deactivated. Generally, when the reaction is performed at normal pressure, the temperature is 5 to 95 ° C., and the pH is 1 to 11. Range may be sufficient. Further, the pressure, temperature, and pH are appropriately adjusted so that the cellooligosaccharide can be obtained in a higher yield, as described above.

The cellooligosaccharide aqueous solution obtained by the above enzymatic decomposition can be subjected to purification treatment such as decolorization, desalting, enzyme removal, etc., if necessary. The purification method is not particularly limited as long as it is a known method. For example, activated carbon treatment, ion exchange resin treatment, chromatography treatment, microfiltration, ultrafiltration, reverse osmosis filtration and other filtration treatments, crystallization treatment, etc. are used. These may be used alone or in combination of two or more.
Among the cellooligosaccharide purification methods, the crystallization treatment is preferable because the composition of the cellooligosaccharide is easy to control.

  Below, the manufacturing method of the xylooligosaccharide of this invention is demonstrated. The xylo-oligosaccharide of the present invention can be obtained by enzymatic degradation of xylan, which is a component of plant cell walls. Moreover, as long as it has the saccharide | sugar composition of this invention, you may use the commercially available xylo-oligosaccharide for foods (Suntory xylo-oligo 35, xylo-oligo 70, xylo-oligo 90, xylo-oligo 95P, etc.).

Below, an example of the manufacturing method of the xylooligosaccharide by an enzymatic decomposition method is shown. The raw material of the xylooligosaccharide of the present invention needs to use a vegetable raw material. The plant raw material referred to here is derived from a plant and includes components constituting the plant.
The components constituting the plant contained in the raw material are, for example, polysaccharides such as cellulose and hemicellulose, oligosaccharides composed of xylose, glucose, arabinose, mannose, galactose, and the like, or monosaccharides. Among the above plant raw materials, cotton seed, bagasse or corn is preferably used in order to obtain a high yield of xylooligosaccharide. As used herein, cottonseed is a perennial seed belonging to the mallow family, and as long as the object of the present invention can be achieved, the raw materials used may include cotton, cotton, petals, linters, oils, etc. . Among cotton seeds, it is preferable to use cottonseed husk as a raw material because the cottonseed husk has a high hemicellulose content as a raw material for the xylooligosaccharide-containing polysaccharide.

  In addition, the bagasse of the present invention is a generic name for components other than sugar among the sugar cane constituents, and is industrially a squeezed rice cake obtained by collecting sugar, and if the object of the present invention can be achieved, the raw material to be used May include skin, piss, sugar and the like. The corn of the present invention is not particularly limited in its cultivar, production area, site, etc., but it is preferable to use a core or hull with a high hemicellulose content.

  The xylo-oligosaccharide of the present invention is obtained by washing a plant raw material in a dilute alkaline medium, removing impurities, and further extracting with an alkaline medium, or increasing the reactivity with xylanase by dilute alkali treatment in a specific concentration range. The residue is treated with alkaline hydrogen peroxide and / or sodium hypochlorite as necessary to improve decolorization and further reactivity with xylanase. It can be obtained by allowing xylanase to act on these extracts and / or processed products to hydrolyze the polysaccharides contained in the composition. The xylanase referred to in the present invention is a general term for enzymes having an action of hydrolyzing xylan-based substances, and any xylanase referred to in the present invention is included in the xylanase as long as it has xylan decomposing activity.

Examples of xylanase enzyme sources include, for example, purified xylanase-producing cells themselves, purified enzymes that accumulate or secrete xylanase-producing bacteria, and formulated purified enzymes together with additives such as excipients and stabilizers. Can be mentioned. In the case of a xylanase preparation, the additive added thereto is not particularly limited, and the dosage form may be any of powder, granule, liquid and the like.

  The origin of xylanase is not particularly limited, and examples of microorganisms that produce known xylanases include Basilis polymyxa, Bassilus poumillius, Bassilus subtilis, Cellvibrio bulbtus, Clostridium, Rumen bacteris, Aspergillus niger, Aspergillus olizae, Cephalosporia sacchari, Ceratocytosis paradoxa, Chetomium globosam, Cysosporum lignorum, Coniophora ebella, Fomes igiarius, Fomesmaruginatus, Gloeophyllum saepiarium, Helminthoisporium, Humicola insolens, Irpex lacteus, Lentites saepiaria, Malbranchea pullchea, Manulius lacrymans, Marulius silvester, Myrothesium verrucaria, Penicillium funuculosum, Penicillium verrufulosom, Penicillium wortimunni, Polyporus betulinus, Shizpphyllum communue, Trametes Produced by bacteria described in "Cellulase" (published by Kodansha Scientific (1987)) and "Encyclopedia of Cellulose" (published by Asakura Shoten (2000)) such as ersicolor, Tricoderma konningii, Tricoderma violet, Tricoderma sp Although xylanase can be mentioned, as long as it has an action of degrading xylanase, it is not limited to those derived from the above-mentioned known bacteria, and enzymes derived from novel bacteria are also included in the xylanase of the present invention.

The method for partial hydrolysis may be a known method and is not particularly limited. For example, a xylan-based substance is dissolved or suspended in an aqueous medium as a substrate, xylanase is added, A method of heating while stirring or shaking is mentioned.
In the above method, the reaction conditions such as dissolution, suspension, stirring method, xylanase / substrate addition method / addition order, concentration thereof, and the like are appropriately adjusted so as to improve the yield of xylo-oligosaccharide. In this case, the pH, temperature, and treatment time of the reaction solution may be within the range where the enzyme is not inactivated. Generally, when the reaction is performed at normal pressure, the temperature is 5 to 95 ° C., and the pH is It may be in the range of 1 to 11 and may be decomposed under pressure to improve the reaction rate and selectivity. For example, when using a xylanase derived from Tricoderderm (for example, trade name Sumiteam X manufactured by Shin Nippon Chemical Industry Co., Ltd.), the optimum reaction conditions for partial hydrolysis are pH 3.0 to 7.0, 30 to 60 ° C. It is preferable to process for 1 to 24 hours.

Next, food / medicine using the H. pylori inhibitor or bacteriostatic agent of the present invention will be described.
The Helicobacter pylori suppressor or bacteriostatic agent of the present invention can be used as a food / pharmaceutical as it is, among bifidobacteria, lactic acid bacteria, bacteroides, eubacterium, anaerobic streptococci, enterococci, Escherichia coli, etc. It may be used in combination with bacteria useful for the human body. The useful bacteria here will be described in more detail.

  Examples of useful bacteria include Bifidobacterium adlescentis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium infantium Bifidobacterium longum, Bifidobacterium parvalorum, Lactobacillus acidophilus, Lactobacillus casei llus casei), Lactobacillus Sarubariusu (Lactobacillus salibarius), Lactobacillus gasseri (Lactobacillus gasseri), Lactobacillus fermentum (Lactobacillus fermentum), Streptococcus faecalis (Streptococcus faecalis), Streptococcus Pyro Jens (Streptococcus pyogenes), Pseudomonas Aerajinosa (Psudomonas aeruginosa) , Bacteroides desstasonis, Bacteroides ovatus, Bacteroides cetaeotamic Emissions (Bacteroides thetaiotaomicron), Bacteroides Barufantasu (Bacteroides vulfatus), Bacteroides Yuniforumisu (Bacteroides uniformis), Bacteroides melanin Roh Genis dregs (Bacteroides meraninogenicus), Fusobacterium barium (Fusobacterium varium), Fusobacterium Nekuroforamu (Fusobacterium necrophorum), Maegamonasu hyper mega-scan ( Maegamonas hypermegas, Mitsokeella multiacidus, Eubacterium limosum), Eubacterium nitritogenes, Eubacterium rectum (Eubacterium lentum), Eubacterium lentum, Enterobacter aerogenes (Enterobacter aerogenes) Clostridium bigermentans), Clostridium butyricum, Clostridium Clostridium form, Clostridium Um Kokkoidesu (Clostridium coccoides), Clostridium difficile (Clostridium difficile), Clostridium para Putri Fi cam (Clostridium paraputrificum), Clostridium Ramosamu (Clostridium ramosum), Clostridium Inokyumu (Clostridium innocuum), Clostridium sporogenes (Clostridium sporogenes), Propionibacterium acnes (Propionibacterium acnes), Peptococcus prevocii, Peptococcus anaerobias (Peptostrept) occus anaerobius), Peptostreptococcus Parubarasu (Peptostreptcoccus parvulus), Peptostreptococcus Purodakutasu (Peptostreptcoccus productus), Peptostreptococcus Asa Carolyn lithium dregs (Peptostreptcoccus asaccharolyticus), Peptostreptococcus Magnus (Peptostreptcoccus magnus), Peptostreptococcus Purebori (Peptostreptcoccus prevolli ), Beilonella alcaes, Beilonella parunla, Klebsiella Pumoniae (Klebsiella pneumoniae), Megasphaela elsdenii (Megaphaera elsdenii), etc. are mentioned, These useful bacteria may be used individually by 1 type, or may use 2 or more types together.

  The cellooligosaccharide and / or xylooligosaccharide of the present invention has an assimilability against bacteria belonging to the genus Lactobacillus having antibacterial activity of H. pylori. It is preferable because it is enhanced. In particular, it is preferable to use in combination with Lactobacillus gasseri OLL2716 strain. The useful bacteria referred to here are those that bring some physiological usefulness to humans when ingested. The useful bacteria of the present invention have improved the known enterobacteria and known enterobacteria listed above. In addition to those, newly discovered species are also included. Moreover, when using as an intestinal bacteria activator of this invention, even if it uses a living microbe itself, it is also free to use what was formulated by well-known methods, such as freeze-drying and compounding.

  Further, the cellooligosaccharide and / or xylo-oligosaccharide of the present invention is contained in one or more components selected from food materials, pharmaceutical medicinal ingredients, additives used in them, or the above-mentioned useful bacteria, It is also free to use as food / medicine.

  As described above, when the cellooligosaccharide and / or xylooligosaccharide of the present invention is used as a food / pharmaceutical, the addition amount of the cellooligosaccharide is 0.1% by mass or more. When the amount of the cellooligosaccharide of the present invention is less than 0.1% by mass, a sufficient effect of inhibiting Helicobacter pylori can not be obtained. The higher the amount of the cellooligosaccharide and / or xylooligosaccharide of the present invention, the more the above-mentioned effects can be obtained. However, when other components are added for reasons such as improving the dosing property, the cellooligosaccharide and / or xylooligosaccharide is added. The amount of sugar added is 99.99% by mass or less.

  Examples of the components used here include the following. The component used in the present invention is a food material, a pharmaceutical medicinal ingredient, or an excipient, a disintegrant, a binder, a fluidizing agent, a lubricant, a flavoring agent, a flavoring agent, a coloring agent, a sweetening agent, a solvent, an oil and fat, It is an additive such as a surfactant, thickener, gelling agent, and may be in any form such as powder, crystal, oil, liquid, semi-solid, such as `` Food Additives Official Document '', It is possible to use those described in “Japanese Pharmacopoeia” (all published by Yodogawa Shoten) and “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo). In addition, they may be coated for various purposes. These constituent components may be used alone or in combination. As addition amount of a structural component, it is 0.01 mass%-99.9 mass%.

  For example, anti-pyretic analgesics, antihypnotics, drowsiness preventives, antipruritics, pediatric analgesics, stomachic drugs, antacids, digestives, cardiotonic drugs, arrhythmic drugs, antihypertensives, vasodilators , Diuretics, anti-ulcer drugs, intestinal drugs, osteoporosis treatments, antitussive expectorants, anti-asthma drugs, antibacterial agents, frequent urination drugs, nourishing tonics, vitamins, etc. . The medicinal component can be used alone or in combination of two or more.

  Excipients include starch acrylate, L-aspartic acid, aminoethylsulfonic acid, aminoacetic acid, candy (powder), gum arabic, gum arabic powder, alginic acid, sodium alginate, pregelatinized starch, inositol, ethylcellulose, ethylene Vinyl acetate copolymer, sodium chloride, olive oil, kaolin, cacao butter, casein, fructose, pumice grains, carmellose, carmellose sodium, hydrous silicon dioxide, dry yeast, dry aluminum hydroxide gel, dry sodium sulfate, dry magnesium sulfate, agar, Agar powder, xylitol, citric acid, sodium citrate, disodium citrate, glycerin, calcium glycerophosphate, sodium gluconate, L-glutamine, clay, clay granules, croscarmellose nato Um, cross polyvinyl pyrrolidone, magnesium aluminate silicate, calcium silicate, magnesium silicate, light anhydrous silicic acid, light liquid paraffin, cinnamon powder, crystalline cellulose, crystalline cellulose / carmellose sodium, crystalline cellulose (grain) , Synthetic aluminum silicate, synthetic hydrotalcite, sesame oil, wheat flour, wheat starch, wheat germ powder, rice, rice starch, potassium acetate, calcium acetate, cellulose acetate phthalate, safflower oil, white beeswax, zinc oxide, titanium oxide , Magnesium oxide, β-cyclodextrin, dihydroxyaluminum aminoacetate, 2,6-di-butyl-4-methylphenol, dimethylpolysiloxane, tartaric acid, potassium hydrogen tartrate, baked gypsum, sucrose fatty acid ester Tellurium Magnesium Hydroxide, Aluminum Hydroxide / Gel, Aluminum Hydroxide / Sodium Bicarbonate Coprecipitate, Magnesium Hydroxide, Suclan, Stearyl Alcohol, Stearic Acid, Calcium Stearate, Polyoxyl Stearate, Magnesium Stearate, Sterotex HM , Purified gelatin, purified shellac, purified sucrose, purified sucrose spherical granules, cetostearyl alcohol, cetopolyethylene glycol, gelatin, sorbitan fatty acid ester, D-sorbitol, tricalcium phosphate, soybean oil, soybean unsaponifiable matter, soybean lecithin, skim milk powder , Talc, ammonium carbonate, calcium carbonate, magnesium carbonate, neutral anhydrous sodium sulfate, low-substituted hydroxypropyl cellulose, dextran, dextrin, natural silicic acid Luminium, corn starch, tragacanth powder, silicon dioxide, calcium lactate, lactose, white petrolatum, sucrose, sucrose / starch spherical granules, powdered green leaf extract, naked malt green juice dry powder, honey, paraffin, potato starch, semi-digested body Starch, human serum albumin, hydroxypropyl starch, hydroxypropylcellulose, hydroxypropylmethylcellulose phthalate, phytic acid, glucose, glucose hydrate, partially pregelatinized starch, pullulan, propylene glycol, powdered reduced maltose starch syrup, powdered cellulose, pectin, bentonite , Sodium polyacrylate, polyoxyethylene alkyl ether, polyoxyethylene hydrogenated castor oil, polyoxyethylene polyoxypropylene glycol, polystyrene Sodium sulfonate, polyvinyl acetal diethylaminoacetate, polyethylene glycol, maltitol, maltose, D-mannitol, water candy, isopropyl myristate, anhydrous lactose, anhydrous calcium hydrogen phosphate, anhydrous calcium phosphate granule, magnesium aluminate metasilicate, methylcellulose , Cottonseed powder, cottonseed oil, owl, aluminum monostearate, glyceryl monostearate, sorbitan monostearate, medicinal charcoal, peanut oil, aluminum sulfate, calcium sulfate, granular corn starch, liquid paraffin, dl-malic acid, phosphorus Acid-hydrogen calcium, calcium hydrogen phosphate, calcium hydrogen phosphate granules, sodium hydrogen phosphate, potassium dihydrogen phosphate, calcium dihydrogen phosphate, phosphoric acid diwater Examples of excipients that are classified as excipients in “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo), “Japan Pharmacopoeia”, and “Food Additives Official Document” (published by Yodogawa Shoten) Even if it uses independently, it is also free to use 2 or more types together.

  Disintegrants include croscarmellose sodium, carmellose, carmellose calcium, carmellose sodium, celluloses such as low-substituted hydroxypropyl cellulose, carboxymethyl starch sodium, hydroxypropyl starch, rice starch, wheat starch, corn starch, potato “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo), starches such as starch, partially pregelatinized starch, synthetic polymers such as cross polyvinyl pyrrolidone and cross polyvinyl pyrrolidone copolymer Can be listed as a disintegrant in the book (published by Yodogawa Shoten). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  As binders, sugars such as sucrose, glucose, lactose, fructose, sugar alcohols such as mannitol, xylitol, maltitol, erythritol, sorbitol, gelatin, pullulan, carrageenan, locust bean gum, agar, gluconannan, xanthan gum, tamarind Water-soluble polysaccharides such as gum, pectin, sodium alginate, gum arabic, etc., celluloses such as crystalline cellulose, powdered cellulose, hydroxypropylcellulose, methylcellulose, starches such as pregelatinized starch, starch paste, polyvinylpyrrolidone, carboxyvinyl polymer, “Pharmaceutical additives” such as synthetic polymers such as polyvinyl alcohol, inorganic compounds such as calcium hydrogen phosphate, calcium carbonate, synthetic hydrotalcite, magnesium aluminate silicate, etc. Scripture "(Yakujinipposha issue)," the Japanese Pharmacopoeia ", mention may be made of what is classified as a binding agent in the" food additive official Manual "(Hirokawa Shoten). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  The fluidizers include “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo) such as silicon compounds such as hydrous silicon dioxide and light anhydrous silicic acid, “Japanese Pharmacopoeia”, “Food Additives Official Document” (Yodogawa Shoten) (Issued) can be classified as a fluidizing agent. Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

Lubricants include: “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo) such as magnesium stearate, calcium stearate, stearic acid, sucrose fatty acid ester, talc, “Japan Pharmacopoeia”, “Food Additives Official Statement” (Made by Yodogawa Shoten) can be listed as a lubricant. Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  As a corrigent, "pharmaceutical additives encyclopedia" (published by Yakuji Nippo), such as glutamic acid, fumaric acid, succinic acid, citric acid, sodium citrate, tartaric acid, malic acid, ascorbic acid, sodium chloride, 1-menthol, Examples include those classified as flavoring agents in the “Japanese Pharmacopoeia” and “Food Additives Official Document” (published by Yodogawa Shoten). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  Perfumes include orange, vanilla, strawberry, yogurt, menthol, fennel oil, cinnamon oil, spruce oil, mint oil, and other “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo), “Japan Pharmacy” Can be listed as a flavoring agent and a fragrance in the “Method” and “Food Additives Official Document” (published by Yodogawa Shoten). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  Coloring agents include food colors such as Food Red No. 3, Food Yellow No. 5, Food Blue No. 1, etc., “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo), such as copper chlorofin sodium, titanium oxide, and riboflavin. Examples include those classified as colorants in “Pharmacopeia” and “Food Additives Official Document” (published by Yodogawa Shoten). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  Sweeteners include aspartame, saccharin, dipotassium glycyrrhizinate, stevia, maltose, maltitol, starch syrup, and amacha powder “Pharmaceutical Additive Encyclopedia” (published by Yakuji Nippo), “Japan Pharmacopoeia”, “Food Additives” What is classified as a sweetener in the "official document" (published by Yodogawa Shoten) can be mentioned. Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  The solvent is not particularly limited as long as it is used in pharmaceuticals, for example, “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo) such as alcohols such as methanol and ethanol, and ketones such as acetone, There are those classified as solvents in “Japanese Pharmacopoeia” and “Food Additives Official Document” (published by Yodogawa Shoten), and they can be used alone or in combination of two or more.

  Examples of fats and oils include stearic acid monoglyceride, stearic acid triglyceride, stearic acid sucrose ester, paraffins such as liquid paraffin, hard oils such as carnauba wax and hardened castor oil, castor oil, stearic acid, stearyl alcohol, polyethylene glycol The oils and fats listed in the “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo), “Japan Pharmacopoeia”, “Food Additives Official Document” (published by Yodogawa Shoten), etc. Two or more kinds can be used in combination.

  As a thickener, for example, “pharmaceutical additive encyclopedia” such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyacrylic acid, carboxyvinyl polymer, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, ethylcellulose, gum arabic, starch paste, etc. (Those published by Yakuji Nippo), “Japanese Pharmacopoeia”, “Food Additives Official Document” (published by Yodogawa Shoten) are listed. It is also free to do.

  Examples of the surfactant include phospholipid, glycerin fatty acid ester, polyethylene glycol fatty acid ester, sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene cetyl ether, polyoxyethylene stearyl ether, polyoxyethylene nonylphenyl ether, Polyoxyethylene polyoxypropylene glycol, polyoxyethylene sorbitan sun monolaurate, polysorbate, sorbitan monooleate, monostearate glyceride, monooxyethylene sorbitan monopalmitate, monooxyethylene sorbitan monostearate, polyoxymonooleate “Pharmaceutical Additives Encyclopedia” such as ethylene sorbitan, sorbitan monopalmitate, sodium lauryl sulfate, etc. ), "Japanese Pharmacopoeia", "Food Additives Official Document" (published by Yodogawa Shoten), those classified as surfactants are listed, and even if they are used alone, two or more may be used in combination Be free.

  Examples of the gelling agent include animal gelling agents such as gelatin, agar, xanthan gum, guar gum, gum arabic, curdlan, locust bean gum, carboxymethyl cellulose, hydroxyethyl cellulose, cellulose, microcrystalline cellulose, microcrystalline cellulose and the like. As a gelling agent for "Polymer additives encyclopedia" (published by Yakuji Nippo), "Japanese Pharmacopoeia", and "Food Additives Official Document" (published by Yodogawa Shoten) What is classified is mentioned, and it can be used alone or in combination of two or more.

  A food / pharmaceutical comprising the cellooligosaccharide and / or xylooligosaccharide of the present invention and one or more components selected from food materials, medicinal medicinal ingredients, useful bacteria, or additives used in the following. Although the manufacturing method will be described, the manufacturing method is not limited to the following method.

  The method for adding each component is not particularly limited as long as it is a commonly used method, but 1) Cellooligosaccharide and / or xylooligosaccharide and components are added simultaneously and mixed / dispersed. 2) Cellooligosaccharide And / or after mixing / dispersing the xylooligosaccharide and a specific component in advance, another component may be added and mixed / dispersed. 3) After mixing / dispersing two or more components in advance, cellooligo A method in which sugars and / or xylo-oligosaccharides are added and mixed / dispersed, or a combination of these addition methods may be used. When the components to be added are solutions, suspensions, and emulsions, it is preferable to employ a method of spraying them on cellooligosaccharide or other additives, since variation in component concentration in the final product is reduced.

The devices used here are small suction transport devices, pneumatic transport devices, bucket conveyors, pressure-feed transport devices, vacuum conveyors, vibratory quantitative feeders, sprays, funnels, etc. May be. The mixing method of each component is not particularly limited as long as it is a normal method, but a container rotary mixer such as a V type, W type, double cone type, container tack type mixer, or high-speed stirring is used. A stirring mixer such as a mold, a universal stirring type, a ribbon type, a pug type, and a Nauta type mixer, a high-speed fluid mixer, a drum mixer, and a fluidized bed mixer may be used. A shaker mixer such as a shaker can also be used.

  The dispersion method is not particularly limited as long as it is a commonly performed dispersion method, but it is a one-way rotation type such as a portable mixer, a three-dimensional mixer, a side mixer, a multi-axis rotation type, a reciprocating reversal type, a vertical movement type, a rotation + up and down Mixing method using stirring blades such as mobile type, pipe type, jet type stirring and mixing method such as line mixer, gas blowing type stirring and mixing method, high shear homogenizer, high pressure homogenizer, ultrasonic homogenizer, etc. The method may also be a container-shaking mixing method using a shaker, or a combination of these methods.

  Further, in the above mixing and dispersion, there is no particular limitation on the order of adding an aqueous medium to which a surfactant, a thickener, and a gelling agent are added to water or a water / organic solvent as necessary. Even if other constituent components are added to cellooligosaccharide and / or xylooligosaccharide in advance after the aqueous medium is dissolved and dispersed, 2) after the aqueous medium is added to the constituent components in advance and dissolved / dispersed 3) Cello-oligosaccharide and / or xylooligosaccharide may be added, 3) Cellulo-oligosaccharide and / or xylooligosaccharide and components may be mixed / dispersed in advance, and then an aqueous medium may be added, or a combination of these may be used. . Each liquid or semi-solid food / cosmetic product / pharmaceutical such as aqueous solution, dispersion, paste, and emulsion obtained here is dried as necessary and subjected to processing such as granulation, coating, and molding.

  The granulation / coating method is not particularly limited as long as it is a known method, but either a stirring method or a fluidized bed method may be used, or a method combining them may be used. Examples of the agitation granulator include a one-way rotary type such as a portable mixer, a three-dimensional mixer, and a side mixer, a multi-axis rotary type, a reciprocating reversal type, a vertical movement type, a rotation + vertical movement type agitator, and a fluidized bed type. An upper spray type, a central spray type, a lower spray type, a combined stirring type, a central can jet type, a Wurster type and the like can be mentioned. Moreover, you may give the dry granulation which uses a roller compactor.

  Regarding the coating, a granulated product may be obtained in advance, and a known coating may be applied thereto, or after coating, another coating may be applied to form a multilayer. As a spraying method of the coating agent, a method of spraying a component solution / dispersion using a pressure nozzle, a two-fluid nozzle, a four-fluid nozzle, a rotating disk, an ultrasonic nozzle, etc., and a component solution / dispersion from a tubular nozzle are used. Any method of dropping may be used. When the component solution / dispersion is added, the component solution / dispersion may be applied to the cellooligosaccharide particles, such as layering or coating, where the component is laminated on the surface of the cellooligosaccharide particles. Cellooligosaccharide particles or a mixture of cellooligosaccharide and other components may be granulated in a matrix using the liquid as a binding liquid. The effect is the same whether the layering or coating is wet or dry.

The cellooligosaccharide and / or xylooligosaccharide of the present invention is excellent in the retention of liquid components, and when it is used as a core particle, aggregation of particles with layering and coating can be suppressed.
In addition, when the component is a solution, suspension, or emulsion, the component solution, suspension, or emulsion can be converted into a component solution, suspension, or emulsion such as dipping using cellooligosaccharide particles or a mixture of cellooligosaccharide and other additives as a carrier. A method of immersing and holding the components can be used. Although depending on the conditions such as the component type and concentration, even the liquid dipping method such as dipping is excellent in that the uniformity of the component concentration is practically maintained and the process is simple compared to the above spraying.

Further, when the constituent component is a solution, suspension, or emulsion, the constituent solution is obtained by using cellooligosaccharide and / or xylooligosaccharide particles, or a mixture of cellooligosaccharide and / or xylooligosaccharide particles and other additives as a carrier. Alternatively, after immersing in a suspension or emulsion, the dispersion may be spray-dried to form a composite.
The cellooligosaccharide and / or xylooligosaccharide particles or the mixture of cellooligosaccharide and / or xylooligosaccharide particles and other additives before and after the addition of the component solution / dispersion are in a state in which the respective unit particles are individually dispersed. Alternatively, it may take the form of an agglomerated granulated product.

  The molding method is not particularly limited as long as it is a commonly performed method, but a mold may be used, and a known molding method such as compression, melting, injection, rolling, etc. can be applied, and a method combining these But you can. Examples of molding machines used here include compression molding machines, melt molding machines, injection molding machines, and rolling molding machines. Confectionery / cosmetics / pharmaceutical molding machines, cooked rice molding machines, compressed molding machines, packaging machines Well-known molding machines, such as a rice cake manufacturing apparatus, a dumpling / wrapping molding machine, and a foundation substrate compression molding machine, can be used.

  In particular, regarding compression molding, a method of compression molding into a desired shape using a mold, or a method of pre-compressing into a sheet shape and then cleaving into a desired shape may be used. As the compression molding machine, for example, a roller press such as a hydrostatic press, a briquetting roller press, a smooth roller press, a compressor such as a single punch tablet press, or a rotary tablet press can be used. . The molded product obtained by the above-described method may be coated by a known method. When the molded product is a tablet, it may be coated with sugar to form a sugar-coated tablet. In particular, the cellooligosaccharide of the present invention may be blended in the sugar coating layer in the sugar-coated tablet.

  Examples of the cellooligosaccharide and / or xylooligosaccharide-containing food of the present invention include, for example, gels such as jelly, pudding, yogurt, mayonnaise, dressings, sauces, sauces, soups, processed vegetables, seasonings, curry, Hayashi, meat sauce, stew, soup and other retort foods, chilled food, hamburger, bacon, sausage, salami sausage, hams and other livestock products, salmon, chikuwa, fish meat ham and sausage, fried rice cakes, bread, Wheat processed foods such as raw noodles, dry noodles, macaroni, spaghetti, Chinese bun skin, cake mix, premix, white sauce, gyoza and spring rolls, canned curry, sauce, soup, boiled, jam, etc. Bottled products, candy, troches, tablet confectionery, chocolate, biscuits, cookies, rice confectionery, Japanese and Western confectionery Confectionery such as Western confectionery, snacks, candy, pudding, fries, croquettes, dumplings, cooking processed products such as Chinese bun, vegetable paste, minced meat, fruit paste, a paste such as seafood paste. In addition, there are dairy products such as ice cream, ice milk, lact ice, whipped cream, condensed milk, butter, yogurt, cheese and white sauce, and processed oils and fats such as margarine, fat spread and shortening. In addition, carbonated drinks such as cola, carbonated drinks, alcohol drinks, fruit drinks mixed with dairy products, fruit juice or drinks containing fruits, milk drinks, coffee, milk, soy milk, cocoa milk, fruit milk, yogurt, etc. May be used for tea beverages such as lactic acid / milky beverages, sencha, oolong tea, matcha tea, black tea, and the like.

  Examples of the cellooligosaccharide and / or xylooligosaccharide-containing pharmaceutical of the present invention include solid preparations such as tablets, powders, fine granules, granules, extracts, and pills.

The present invention will be described based on examples, but the present invention is not limited thereto.
[Production Example 1]
Trichoderma reesei GL-1 strain (Tricoderderma reesei GL-1 strain (Incorporated Administrative Agency, National Institute of Advanced Industrial Science and Technology, Patent Biodeposition Center, receipt number FERM BP-10323)) was inoculated on a normal agar medium, and cultured at 37 ° C. for 7 days , 1 platinum spore from the surface of the medium, 1 g polypeptone, 0.5 g yeast extract, 2 g potassium phosphate, 1.5 g ammonium sulfate, 0.3 g magnesium sulfate, 0.3 g calcium chloride, 1 mL trace element (6 mg boric acid) Ammonium molybdate tetrahydrate 26 mg, iron chloride (3) hexahydrate 100 mg, copper sulfate pentahydrate 40 mg, manganese sulfate tetrahydrate 8 mg, zinc sulfate heptahydrate 200 mg in a total amount of 100 mL of purified water 1 mL of Adecanol, crystalline cellulose (Asahi Kasei Chemica) Was inoculated's trade name PH-101) 10 g in a medium suspended and dissolved in purified water total amount 1L, for 5 days under aeration-agitation culture at 28 ° C.. During the culture, the pH of the medium was adjusted to 2.8-4.7 using an aqueous sodium hydroxide solution. The cultured liquid is centrifuged, the supernatant is sterilized with a microfiltration membrane having an opening of 0.46 μm, and the filtrate is ultrafiltered with a molecular weight cut off of 13,000 (trade name: Microza Pencil type module ACP manufactured by Asahi Kasei Chemicals). -0013) was concentrated 10 times by volume to obtain a crude enzyme.

  Next, use a commercially available softwood-derived dissolving pulp, and hydrolyze it with a hydrochloric acid concentration of 0.4% aqueous hydrochloric acid at 120 ° C. for 1 hour to wash the acid-insoluble residue and filter to obtain a wet cake. It was. Using this wet cake as an aqueous dispersion with a concentration of 10% cellulose, an ultra-high performance disperser / wet pulverizer (manufactured by Ashizawa Co., Ltd., trade name: Pearl Mill RL φ1mm zirconia bead filling rate 80%) A grinding treatment was performed to obtain a cellulose fine particle dispersion (average polymerization degree 220, diethyl ether soluble matter content 0.7%, average particle diameter 0.7 μm, colloidal component content 51.5%).

The ground cellulose was suspended and dissolved in a 50 mM acetic acid-sodium acetate buffer (pH 4.5) so that the crude cellulose was 2% by mass and the protein concentration was 0.25%, and the total volume was 1000 mL. . The glass flask was placed in a 55 ° C. water bath and reacted for 4 hours while stirring the interior. After completion of the reaction, 300 μL of the reaction solution was dispensed in a suspended state, the enzyme and undegraded cellulose were removed using an ultrafiltration module (fractional molecular weight 10,000), and then the sugar concentration was analyzed by high performance liquid chromatography. . The sugar concentration of the reaction solution was cellotriose to cellohexaose 0.2% by mass, cellobiose 1.5% by mass, and glucose 0.3% by mass.
The reaction solution was filtered through an ultrafiltration membrane having a molecular weight cut off of 13000 (trade name Micro-The Pencil type module ACP-0013 manufactured by Asahi Kasei Chemicals), and the resulting filtrate was deionized with a cation / anion exchange resin. Distillation was performed at 70 ° C. under reduced pressure to obtain an aqueous solution having a sugar concentration of 20 times.

[Production Example 2]
100 mL of the cellooligosaccharide aqueous solution obtained in Production Example 1 was introduced into a 200 mL glass flask and cooled from 70 ° C. to 5 ° C. at a rate of 10 ° C. per hour while stirring. The crystallization was carried out at a rate of 10 g / min so as to be%. The cellooligosaccharide crystallized in the aqueous solution was filtered under reduced pressure, dried, pulverized and sieved to obtain cellooligosaccharide powder CE-1. The sugar composition of the obtained cellooligosaccharide powder is shown in Table 1.

[Production Example 3]
The strain of Production Example 1 was replaced with cellobibrio gilbus ATCC13127, the pH during culture was changed to 4-10, and the buffer during enzyme reaction was changed to a phosphate buffer with pH 6.5. A cellooligosaccharide aqueous solution was prepared in the same manner as in Production Example 1.
The obtained cellooligosaccharide aqueous solution is passed through a column filled with activated carbon, and a glucose-rich fraction is collected, dried in a 60 ° C. aerated oven for 16 hours, and powdered by the same operation as in Production Example 2, and cellooligosaccharide is obtained. Powder CE-2 was obtained. The sugar composition of the obtained cellooligosaccharide powder is shown in Table 1.

[Example 1]
Using CE-1 as the cellooligosaccharide, the following test solution was prepared, and the increase inhibition rate of H. pylori was measured. The results are shown in Table 2.
<Test solution>
(0) Celluligosaccharide-free 5% horse serum-added 1/10 Brucella broth (1) 2% CE-1 dissolved 5% horse serum-added 1/10 Brucella broth (2) CE-1 dissolved 5% 1/10 Brucella broth supplemented with 5% horse serum (3) CE-1 dissolved in 10% 5% horse serum <culture method>
A test strain (Helicobacter pylori ATCC 43504) was cultured in sheep blood agar medium K (BBL) at 35 ° C. for 3 days by microaerobic culture, and then subjected to McFarland No. 2 (about 10 ⁷ to 10 ⁸ CFU / mL). This is diluted 100 times with a sterilized physiological saline solution to obtain a bacterial solution for addition. 1 mL of the bacterial solution for addition is added to 9 mL of each test solution, and cultured under shaking at 35 ° C. under microaerobic conditions. Each test solution is collected after 72 hours of culture, and a 10-fold diluted series solution is prepared with a sterile physiological saline solution. 50 μL of the stock solution and each dilution series solution are smeared on sheep blood agar medium K and cultured under microaerobic conditions at 35 ° C. for 4-5 days. After 0 hour, only the test solution is quantified. The number of bacteria that have grown after culturing is counted and the number of viable bacteria per mL is obtained. The increase suppression rate was calculated | required with the following formula | equation.
Increase suppression rate (%) = (viable cell count 2-viable cell count 1) / viable cell count 2 × 100

[Example 2]
From Example 1, the cellooligosaccharide was changed to CE-2, and the amount of cellooligosaccharide added in the test solution was 2% to prepare the following test solution (4). The increase inhibition rate was calculated. The results are shown in Table 2.
<Test solution>
(4) 1/10 Brucella broth with 5% horse serum and 2% CE-2 dissolved

[Comparative Example 1]
From Example 1, instead of cellooligosaccharide, D-glucose (manufactured by Wako Pure Chemical Industries, Ltd.) was used to prepare the following test solution (5) with the addition amount in the test solution being 2%. In the same manner as above, the increase inhibition rate of H. pylori was determined. The results are shown in Table 2.
<Test solution>
(5) 1/10 Brucella broth with 5% horse serum in which 2% D-glucose is dissolved

From Examples 1 and 2, cellooligosaccharide had an effect of suppressing Helicobacter pylori. Furthermore, Example 1 shows that the proliferation inhibitory rate of Helicobacter pylori improves and the inhibitory effect of Helicobacter pylori is high, so that the addition amount of cellooligosaccharide increases.
In contrast to Example 2, Example 1 has a high cellobiose content, a low cellotriose-cellohexaose content, and a low glucose content, but with the same addition amount, the viable number of Helicobacter pylori is small, and the cellobiose content in the cellooligosaccharide The higher the value, the higher the suppression effect.
Furthermore, from Comparative Example 1, glucose did not have an effect of suppressing Helicobacter pylori.

[Example 3]
CE-1 was used as the cellooligosaccharide, and the addition amount was 5% and 10%, and the cells were cultured in the same manner as in Example 1. The culture time was shaken from 48 to 96 hours, and the number of viable bacteria was compared with no addition. The results are shown in FIG.
FIG. 1 shows that when cellooligosaccharide is added at a high concentration, not only suppression or bacteriostasis but also H. pylori can be killed in a short time.
Below, the prescription example of the foodstuff and the pharmaceutical containing the inhibitor of H. pylori or bacteriostatic agent of this invention is illustrated as a reference example below.

[Example 4]
The cellooligosaccharides of Examples 1 to 3 were changed to xylooligosaccharides, and the increase inhibition rate of H. pylori was determined in the same manner. The sugar composition of the xylo-oligosaccharide used was as follows.
Xylobiose 36.4%, Xylooligoose or higher xylo-oligosaccharides 7.11%, and other xylose.

[Comparative Example 2]
The cellooligosaccharides of Examples 1 to 3 were changed to maltose (Malose manufactured by Wako Pure Chemical Industries), and the increase inhibition rate of H. pylori was determined by the same method.

[Reference Example 1]
CE-1 was used as a cellooligosaccharide, and an acidic sports drink was prepared according to the following formulation 1. As a production method, the raw materials of Formula 1 were mixed in advance, dissolved in water, and sterilized by heating in an autoclave at 121 ° C. for 20 minutes, and then produced as a prototype.
<Prescription 1: Acidic sports drink>
1) Sucrose 4.0% by mass
2) Citric acid (hydrate) 0.1% by mass
3) Ascorbic acid 0.1% by mass
4) Grapefruit straight juice 1.0% by mass
5) Cellooligosaccharide CE-1 10.0% by mass
6) Water 83.8% by mass

[Reference Example 2]
CE-1 was used as the cellooligosaccharide, and orange juice was prepared according to the following formulation 12. As a production method, the raw materials of prescription 2 were mixed in advance, dissolved in water, sterilized by heating in an autoclave at 121 ° C. for 20 minutes, and prototyped.
<Prescription 2: Acidic sports drink>
1) Sucrose 7.5% by mass
2) Citric acid (hydrate) 0.1% by mass
4) Orange 4 times concentrated fruit juice 5.0% by mass
5) Cellooligosaccharide CE-1 10.0% by mass
6) Water 76.9% by mass

[Reference Example 3]
To 98 parts of orange juice obtained in Example 5, 2.0 parts of commercially available gelatin was added and dissolved at 90 ° C. with stirring. After dissolution, it was sealed in a 100 mL plastic container and stored at 5 ° C. for 18 hours to prepare a cellooligosaccharide-containing jelly.

[Reference Example 4]
In the method of Example 6, gelatin was replaced with commercially available agar to prepare cellooligosaccharide-containing agar.

[Reference Example 5]
Using CE-1 as the cellooligosaccharide, 90 parts of cellooligosaccharide and 10 parts of corn starch were previously mixed in a plastic bag for 3 minutes, 0.5 parts of magnesium stearate was added, and further mixed for 30 seconds. A 200 mg cello-oligosaccharide-containing tablet was obtained with a clean press 12HUK, φ8mm, 12R mortar 12 stand, rotation speed 53rpm) and a striking pressure of 15kN.

  The cellooligosaccharide composition of the present invention has excellent handleability and suppresses or bacteriostatic increases in Helicobacter pylori, so that in addition to normal food materials, functional food materials, food and pharmaceutical additives It can be suitably used in the food / pharmaceutical field as an agent or pharmaceutical.

The viable H. pylori count for each culture time in Example 3 is shown.

Claims (11)

  An inhibitor or bacteriostatic agent for Helicobacter pylori containing a water-soluble saccharide having a structure in which a plurality of sugars are chemically bonded and having at least one β bond in the structure.   2. The inhibitor or bacteriostatic agent of H. pylori according to claim 1, wherein the β bond is a β1-4 bond.   The polysaccharide according to claim 1 or 2, wherein the polysaccharide contains at least one glucose and / or xylose as a constituent sugar, and the β1-4 bond is a β1-4 glucoside bond and / or a β1-4 xyloside bond. An inhibitor or bacteriostatic agent for Helicobacter pylori.   The cellobiose content is 50% by mass or more, the content of one or more selected from the group consisting of cellotriose, cellotetraose, cellopentaose and cellohexaose is 0 to 50% by mass, and the glucose content is 30% by mass or less. The suppressor or bacteriostatic agent of Helicobacter pylori according to any one of claims 1 to 3, comprising a cellooligosaccharide as an active ingredient.   The content of one or more selected from the group consisting of xylobiose, xylotriose, xylotetraose, xylopentaose, xylohexaose, xyloheptaose and water-soluble xylan having a degree of polymerization exceeding 7 is 30% by mass or more. The suppressor or bacteriostatic agent of Helicobacter pylori according to any one of claims 1 to 3, which contains xylooligosaccharide having a xylose content of 70% by mass or less as an active ingredient.   The inhibitor or bacteriostatic agent according to any one of claims 1 to 5, wherein the inhibitor or bacteriostatic agent and the food material or the medicinal medicinal component are dispersed or dissolved in water or a water / organic solvent mixture. An aqueous composition, wherein the content of is 0.1% by mass or more.   Furthermore, 1 or more types chosen from the group which consists of surfactant, a thickener, and a gelatinizer are contained, The aqueous composition of Claim 6 characterized by the above-mentioned. The aqueous composition according to claim 6 or 7, wherein the aqueous composition is in any one of an aqueous solution, a water dispersion, a paste form, and a gel form.   An inhibitor or bacteriostatic agent according to any one of claims 1 to 5 and at least one selected from food ingredients, pharmaceutical medicinal ingredients, food additives and pharmaceutical additives, Content is 0.1 mass% or more, The solid composition characterized by the above-mentioned.   The solid composition according to claim 9, wherein the solid composition is in the shape of any one of a powder, a granule, and a molded body.   The solid composition according to claim 10, wherein the surface is coated.

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