JP5340528B2 - Lipid reducing agent in internal organs - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To obtain an agent for preventing or ameliorating lifestyle-related diseases, which reduces concentration of neutral fats and total cholesterol in the liver by oral intake in the field of food and medicine. <P>SOLUTION: The agent for preventing or ameliorating lifestyle-related diseases comprises a cellooligosaccharide as an active ingredient containing &ge;70 mass% of cellobiose, 0.1-30 mass% of one or more kinds selected from cellotriose, cellotetraose, cellopentaose and cellohexaose and &le;9 mass% of glucose. <P>COPYRIGHT: (C)2008,JPO&amp;INPIT

Description

  The present invention contains a cellooligosaccharide having a controlled sugar composition and a suppressed decrease in blood adiponectin concentration in an internal preparation in the food and pharmaceutical fields, thereby reducing neutral fat and total cholesterol concentration in the liver. The present invention relates to an agent for preventing or improving lifestyle-related diseases.

Cellooligosaccharide is a generic name for cellobiose, cellotriose, cellotetraose, cellopentaose and cellohexaose, and is a oligosaccharide in which 1 to 6 glucopyranose units are β-1,4 linked in one molecule.
Cellooligosaccharides are difficult to digest and absorb by humans and are useful as low-calorie sweeteners, as well as general foods, functional foods, cosmetics, pharmaceuticals and their additives, other chemical conversion materials, and fermentation materials. Yes (Non-Patent Document 1).

  The present invention focuses on intrahepatic lipids that can cause adult pathological conditions such as diabetes, arteriosclerosis, and cirrhosis, and has a specific sugar composition that suppresses a decrease in blood adiponectin concentration. By taking a preventive or ameliorating agent for lifestyle-related diseases containing cellooligosaccharide, it has the effect of reducing lipids in the liver.

The following reports have been made on attempts to lipid metabolism by cellooligosaccharides so far.
Non-Patent Documents 1 to 3 use a cellooligosaccharide having a cellobiose of 85.7% by mass, a cellotriose mass of 3.7%, and a glucose content of 9.3% by mass as a sugar composition. 7 parts by weight, 25 parts by weight casein, 5 parts by weight corn oil, 4 parts by weight mineral, 1 part by weight vitamin, 0.2 part by weight choline hydrochloride and 0.05 part by weight vitamin E) , The results of replacement with the cellooligosaccharide and breeding SD male rats for 4 weeks are described. According to this document, ingestion of cellooligosaccharide having the above-mentioned saccharide composition reduces serum total cholesterol, HDL-cholesterol, triglyceride, and body fat percentage relative to the additive-free system. However, although the cellooligosaccharides of this document have an effect of reducing serum lipids, there is no change in the visceral weight of the liver and the like, and no reduction effect has been observed for visceral lipids. On the other hand, the present invention is a preventive or ameliorating agent for lifestyle-related diseases that has a specific sugar composition, suppresses a decrease in blood adiponectin concentration, contains cellooligosaccharides, and is excellent in lipid reduction effect in the liver. Therefore, it is completely different from the serum lipid-lowering cellooligosaccharide of this document.

Patent Document 1 discloses a food and feed additive for promoting intestinal regulation and lipid metabolism characterized by containing lactic acid bacteria (Lactobacillus rahamnosus) and cellobiose in a mixing ratio of 1: 2.5-8. Have been described. In this document, 10% by mass of the basic food (casein 20.0 parts, DL-meotiin 0.3 parts by mass, minerals 3.5 parts by mass, vitamins 1.0 parts by mass, soybean oil 7.0 parts by mass) is obtained. And the results of breeding Wister male rats for 14 days are described. In this document, the cellobiose-only addition system has no effect on serum and visceral lipid metabolism compared to the addition of cellobiose, and the amount of lipid is reduced only when cellobiose and the lactic acid bacterium are used in combination. Therefore, as in the present invention, it contains a cellooligosaccharide whose sugar composition is highly controlled, and the cellooligosaccharide alone is essentially different from a lifestyle-related disease prevention or amelioration agent that reduces the amount of fat in the liver. In addition, the lipid metabolism-promoting food of this document requires the simultaneous intake of cellobiose and viable bacteria, so that the stability of viable bacteria when prepared as a preparation is poor, and there is a problem that the effect is reduced to some extent by taking. It was.
Therefore, non-insulin-elevating cellooligosaccharides with a specific sugar composition and reduced blood adiponectin concentration are used as active ingredients, and by taking these, lifestyle-related disease prevention that reduces lipids in the liver Or the improving agent was not known.

The latest technology of Wood Chemicals, published by CMC Publishing, 66-72 (2000) Journal of Japanese Society of Nutrition and Food, 49, No 3,143-148 (1998) Cellulose Communications, 5, No 2, 91-97 (1998) JP 2004-321068 A

  An object of this invention is to provide the lifestyle-related disease prevention or improvement agent which has the lipid reduction property in a liver.

The present inventors have found that cellooligosaccharides whose sugar composition is controlled within a specific range and in which the decrease in blood adiponectin concentration is suppressed have lipid lowering properties in the liver, have led to the present invention.
That is, the present invention is as follows.
(1) Cellobiose content is 95% by mass or more, 0.1 to 5% by mass of one or more selected from cellotriose, cellotetraose, cellopentaose and cellohexaose, and glucose content of 2% by mass or less An agent for reducing lipids in viscera, comprising cellooligosaccharide as an active ingredient, wherein the oligosaccharide has a reduction rate of neutral fat concentration in the liver of 15% or more.
(2) Lipid reduction in viscera according to (1) above, wherein the lipid reducing agent in the viscera is a reducing agent of neutral fat concentration in the liver and / or cholesterol concentration in the liver and / or fat mass in the abdominal cavity. Agent.
(3) The agent for reducing lipids in viscera according to (1) or (2) above, wherein the oligosaccharide is an oligosaccharide having a reduction rate of blood adiponectin concentration by oral intake of 30% or less.

  By taking the lifestyle-related disease prevention or ameliorating agent of the present invention, lipids in the liver can be reduced.

The present invention will be specifically described below, particularly focusing on preferred embodiments thereof.
The cellooligosaccharide in this invention needs to contain 70 mass% or more of cellobiose. A higher cellobiose content is preferable because the water solubility of the cellooligosaccharide is improved and the bioavailability at the time of taking is increased. The cellobiose content is preferably 86% by mass or more, more preferably 90% by mass or more, and particularly preferably 95% by mass or more.

In the cellooligosaccharide according to the present invention, the content of one or more selected from cellotriose, cellotetraose, cellopentaose and cellohexaose is 0.1 to 30% by mass. By containing these, the effect of reducing liver lipids is enhanced. However, if the content is too high, the solubility is lowered and the above-mentioned bioavailability is reduced. Therefore, this content needs to satisfy the above-mentioned range. A preferable range is 0.1 to 14% by mass, and more preferably 0.1 to 5% by mass.
The glucose content of the cellooligosaccharide in the present invention is 9% by mass or less. A low glucose content can suppress an increase in blood insulin concentration, and particularly increase lipid metabolism in the liver. The glucose content is preferably 3.5% by mass or less, and more preferably 2% by mass or less. The lower the glucose content, the greater the above-mentioned effect, so the lower limit is not particularly limited.

Below, the analysis method of the cellooligosaccharide and glucose content in the lifestyle-related disease prevention or improvement agent of this invention is described. The cellooligosaccharide 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: manufactured by Shimadzu Corporation, trade name: Asahipak NH ₂ P-50, mobile phase: acetonitrile or water = 75/25 (volume ratio)). The sugar composition of cellooligosaccharides is the mass percentage of the cell biose, cellotriose, cellotetraose, cellopentaose, cellohexaose peak area in the chromatogram obtained by the above method, and occupies the total mass. expressed. The content rate of glucose is also obtained by the same method, and is expressed as a percentage of the mass of glucose with respect to the total mass calculated from the peak areas of cellobiose and glucose.

  The cellooligosaccharide of the present invention is preferably non-insulin-elevating. This non-insulin-elevating property means that the blood insulin concentration after oral intake of cellooligosaccharide does not increase with respect to the insulin concentration before oral intake, and is measured by the following method. Seven-week-old SD rats were allowed free intake of AIN-93G (manufactured by Oriental Yeast Co., Ltd.) for 1 week, fasted for 1 week, fasted for 16 hours, and each cellooligosaccharide aqueous solution in Table 1 was 1500 mg / day using a sonde. kg was administered. Blood samples were collected from the extracorporeal vein under anesthesia before and 30 minutes after the administration, and the insulin concentration (ng / mL Shibayagi Levi insulin kit was used) was measured. The term “insulin non-rising property” as used herein means that the rate of increase in the insulin concentration is 30% or less. More preferably, the rate of increase in insulin concentration is 25% or less, and particularly preferably 20% or less.

  The cellooligosaccharide of the present invention preferably has a reduction rate of blood adiponectin concentration of 30% or less when taken orally. The blood adiponectin concentration is an index of abnormal lipid metabolism, and a low decrease rate is preferable because lifestyle-related diseases such as diabetes and arteriosclerosis do not occur. The blood adiponectin concentration referred to here is the adiponectin concentration (μg / mL-blood) measured by ELISA using blood (adiponectin ELISA kit manufactured by Otsuka Pharmaceutical Co., Ltd.). The rate of decrease can be determined by the following formula from the concentration of adiponectin measured using ingested and non-ingested cellooligosaccharides. Decrease rate (%) = ((concentration when cellooligosaccharide is not taken) − (concentration when cellooligosaccharide is taken)) / (concentration when cellooligosaccharide is not taken) (%). The smaller the adiponectin reduction rate, the greater the above-mentioned effect, so 25% or less is more preferable.

  The cellooligosaccharide of the present invention preferably has a reduction rate of neutral fat concentration in the liver of 15% or more. The higher the reduction rate of the neutral fat concentration in the liver, the higher the effect of preventing / ameliorating lifestyle-related diseases such as diabetes, arteriosclerosis, liver failure, and cirrhosis. The neutral fat concentration referred to here is the amount of neutral fat (mg / g-Wetliver) present per unit amount of the liver. The rate of decrease can be determined by the following formula from the intake of cellooligosaccharide and the neutral fat concentration in the non-ingested liver. Decrease rate (%) = ((concentration when cellooligosaccharide is not taken) − (concentration when cellooligosaccharide is taken)) / (concentration when cellooligosaccharide is not taken) (%). The above-mentioned effect becomes large, so that the fall rate of neutral fat density | concentration is large. Therefore, it is more preferably 20% or more.

  The cellooligosaccharide of the present invention preferably has a reduction rate of the total cholesterol concentration in the liver of 10% or more. The higher the reduction rate of total cholesterol in the liver, the higher the prevention / improvement effect of lifestyle-related diseases such as diabetes, arteriosclerosis, liver failure, and cirrhosis. The total cholesterol concentration here is also the total cholesterol amount (mg / g-Wetliver) present per unit amount of the liver, like the neutral fat concentration described above. The rate of decrease is determined by the following formula from the intake of cellooligosaccharide and the total cholesterol concentration in the non-intake liver. Decrease rate (%) = ((concentration when cellooligosaccharide is not taken) − (concentration when cellooligosaccharide is taken)) / (concentration when cellooligosaccharide is not taken) (%). The above-mentioned effect becomes large, so that the fall rate of total cholesterol concentration is large. Therefore, it is more preferably 15% or more.

The prevention of lifestyle-related diseases of the present invention means that the preventive agent comprising the cellooligosaccharide of the present invention as an active ingredient is taken orally, even when a high sugar, high fat, high calorie or other meal is ingested in the blood and It is possible to suppress the increase in visceral lipids and prevent lifestyle-related diseases such as diabetes, arteriosclerosis, obesity, liver failure, and cirrhosis.
On the other hand, the lifestyle-related disease improvement of the present invention is to reduce the blood and visceral lipids and improve the above-mentioned lifestyle-related diseases by ingesting the cellooligosaccharide of the present invention from the state in which the above-mentioned lifestyle-related diseases have occurred. It is to be. If the lifestyle-related disease prevention / amelioration agent comprising the cellooligosaccharide of the present invention as an active ingredient is taken orally, the above-mentioned prevention and improvement effects can be obtained.

The lifestyle-related disease preventive / ameliorating agent of the present invention has an action of lowering the neutral fat concentration and total cholesterol concentration in the liver. However, when taken orally, it also lowers blood lipids and visceral lipids other than the liver. It is what has.
The blood lipid as used in the present invention means blood neutral fat, HDL, LDL-cholesterol, and phospholipid, and is determined by a known measurement method using serum or plasma. The decrease in blood lipid means that the concentration of the above-mentioned blood lipid is measured, and is decreased from the value before administration of cellooligosaccharide or from the cellooligosaccharide non-administration state.

  Further, the visceral lipids of the present invention include the above-mentioned neutral fat and total cholesterol concentration in the liver, HDL, LDL-cholesterol, phospholipid concentration, epididymal lipids in the epididymis, and the peritoneal cavity around the kidney. It is the above-mentioned lipid existing in the inside. These are also expressed in lipid concentration or mass in the same way as blood lipids. The decrease in visceral lipid concentration means that the lipid concentration or mass has decreased before or after administration of cellooligosaccharide. Means that.

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 substance, one produced by condensation or sugar transfer of a monosaccharide such as glucose or a derivative thereof, What was obtained by the 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 enzyme secreted by cellulase-producing bacteria, and formulated purified enzyme together with excipients, stabilizers and other additives. . 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)) 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.

  As an example of the enzymatic decomposition method, for example, Japanese Patent Application No. 2004-218902, Japanese Patent Application No. 2004-323579, Japanese Patent Application No. 2005-125966, Japanese Patent Application No. 2005-125975, and Japanese Patent Application No. 2006-041149 (all applicants: Asahi Kasei Chemicals Corporation). It is preferable to use cellooligosaccharides produced by the method disclosed in.

Next, the lifestyle-related disease prevention / amelioration agent which uses the cellooligosaccharide of this invention as an active ingredient is demonstrated.
The preventive / ameliorating agent for lifestyle-related diseases comprising the cellooligosaccharide of the present invention as an active ingredient, whether the cellooligosaccharide of the present invention is used in powder form, alone or as an aqueous solution or dispersion, In addition to sugar, it is contained in one or more components selected from food materials, cosmetic materials, medicinal medicinal ingredients, or additives used in them, granules, molded products, aqueous solutions, aqueous dispersions, pastes It may be used as a gel food / medicine.

As for the compounding quantity of the cellooligosaccharide in the lifestyle-related disease prevention or improvement agent of this invention, 0.01-100 mass% is preferable. The amount of cellooligosaccharide here is the total amount of cellobiose, cellotriose, cellotetraose, cellopentaose and cellohexaose contained in the lifestyle-related disease prevention / amelioration agent of the present invention. If the amount is less than 0.01% by mass, a sufficient effect cannot be obtained in lipid metabolism in blood and / or viscera, which is not preferable. The higher the amount of cellooligosaccharide is, the more the above-described lipid metabolism effect is promoted. Therefore, the more preferable range is 0.6% by mass or more, and particularly preferably 0.75% by mass or more.
The lifestyle-related disease preventing / ameliorating agent of the present invention may contain a oligosaccharide other than glucose or a high-intensity sweetener in addition to the cellooligosaccharide of the present invention. The mixing ratio of the cellooligosaccharide and oligosaccharide or the high-intensity sweetener is not limited as long as the effects of the present invention are obtained. For example, the mass ratio of cellooligosaccharide or oligosaccharide is 0.1 / 99. 9-99.9 / 0.1.
In particular, the addition of indigestible oligosaccharides is preferable because in addition to lipid metabolism of cellooligosaccharides, intestinal regulation such as activation of useful intestinal bacteria can be obtained. Furthermore, it is preferable to add a high-intensity sweetener to the cellooligosaccharide of the present invention because the taste can be adjusted without increasing the calorie of the cellooligosaccharide.

  Examples of the oligosaccharide include saccharides such as galactose, fructose, mannose, arabinose, rhamnose, ribose, xylose, sorbose and their reduced products, sucrose, melibiose, trehalose, lactose, maltose, gentiobiose, laminaribiose, lactulose. Disaccharides such as xylobiose and their reduced products, lactosucrose, raffinose, maltototriose, isomaltose, palatinose, kestose, trisaccharides such as gentiosil cellobiose and their reduced products, maltotetraose, gentiosyl cellotri Tetrasaccharides such as ose and nystose and their reduced products, 5 or 6 saccharides such as maltopentaose and maltohexaose and their reduced products, β-cyclodextrin, γ-si Cyclodextrins, cyclic oligosaccharides such as dextran, indigestible dextrin, polydextrose, gum arabic, carboxymethylcellulose, guar gum, curdlan and other water-soluble polysaccharides and their reduced products, sorbitol, xylitol, multirole, mannitol, lactitol Such as sugar alcohols, etc. are classified as sweeteners in the “Food Additives Official Document” (published by Yodogawa Shoten). These oligosaccharides are subjected to chemical treatments such as carboxylation, ethylation, methylation, sulfate esterification, etc., for the purpose of improving solubility and the like even if the oligosaccharides are intact. A derivative may be used.

  High-intensity sweeteners include those classified as high-intensity sweeteners in the “Food Additive Official Document” (published by Yodogawa Shoten) such as saccharin, licorice (glycyrrhizin), stevia, aspartame, sucralose, and acesulfame potassium. It is. These high-intensity sweeteners may be used as they are, or derivatives obtained by substituting a part of the chemical structure with other substitution machines.

Constituents referred to in the present invention include oligosaccharides other than cellooligosaccharides, high-intensity sweeteners, food materials, cosmetic materials, pharmaceutical medicinal ingredients, pigments, fragrances, metals, ceramics or excipients, disintegrants, binders, It is an additive such as fluidizer, lubricant, flavoring agent, colorant, sweetener, solvent, fats and oils, surfactant, thickener, gelling agent, powder, crystal, oil, It can be in any form, such as liquid or semi-solid, for example, “Food Additives Official Document” (published by Yodogawa Shoten), “Cosmetic Raw Material Standards”, “Composition Standards by Cosmetic Variety” (both published by Yakuji Nippo), “Japan Those described in “Pharmacopeia” (published by Yodogawa Shoten) and “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo) can be used.
Further, they may be subjected to processing such as coating and liposome formation for various purposes. These constituent components may be used alone or in combination. The agent for preventing or improving lifestyle-related diseases of the present invention can be processed by known methods such as dissolution, mixing, dispersion, granulation, melting / solidification, compression, and drying.

  The lifestyle-related disease prevention or improvement agent which uses the cellooligosaccharide of this invention as an active ingredient below, and 1 or more types selected from a food material, a cosmetic material, a pharmaceutical active ingredient, or an additive used therewith Although the manufacturing method of the foodstuff or pharmaceutical containing a structural component is described, the manufacturing method of the lifestyle-related disease prevention / amelioration agent of this invention is not restrict | 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) Cello-oligosaccharide and component may be added simultaneously and mixed or dispersed. 2) Cellooligosaccharide and specific component 3) After mixing or dispersing in advance, another component is added and mixed or dispersed. 3) After mixing or dispersing two or more components in advance, cellooligosaccharide is added and mixed or dispersed. Or a combination of these addition methods.

  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 normal 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, and a rotation + up and down direction. 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, or a gelling agent is added as necessary to water or an organic solvent. Even if other components are added after the aqueous medium is added to the sugar in advance and dissolved or dispersed, 2) the cellooligosaccharide is added after the aqueous medium is added to the constituent in advance and dissolved or dispersed. Alternatively, 3) an aqueous medium may be added after the cellooligosaccharide and the constituent components are mixed or dispersed in advance, or a combination of these may be used. Each liquid, such as aqueous solution, dispersion, and emulsion obtained here, or each semi-solid food or medicine such as paste, gel, etc., is dried and subjected to processing such as granulation, coating, molding, etc. as necessary. Also good.

  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 an active ingredient solution or dispersion using a pressure nozzle, a two-fluid nozzle, a four-fluid nozzle, a rotating disk, an ultrasonic nozzle, etc., and an active ingredient solution or dispersion from a tubular nozzle are used. Any method of dropping may be used. When the active ingredient solution or dispersion is added, the active ingredient may be layered or coated so as to be laminated on the surface of the cellooligosaccharide particles, or may be supported on the cellooligosaccharide. As a binding solution, a mixture of cellooligosaccharide and other components may be granulated in a matrix. The effect is the same whether the layering or coating is wet or dry.

  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, rolling molding machines, etc., confectionery or cosmetic or pharmaceutical molding machines, cooked rice molding machines, compressed molding machines, wrapping 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. .

Next, an example of the component used in manufacture of the above-mentioned lifestyle-related disease prevention or improvement agent is described.
For example, food materials or additives used therein include, in addition to the cellooligosaccharides of the present invention, preservatives, shelf-life improvers, antioxidants, sweeteners, colorants, emulsifiers, thickeners as necessary. , Quality improvers, seasonings, acidulants, fortifiers, spices and herbs, food fragrances, enzymes and the like may be added. These food materials or additives can be used alone or in combination of two or more.

  Examples of preservatives and shelf-life improvers include sorbic acid, potassium sorbate, benzoic acid, sodium benzoate, p-hydroxybenzoate, sodium dehydroacetate, propionic acid, propionate, acetic acid, sodium acetate, glycine, ethyl Preservatives and longevity improvers in “Food Additive Official Documents” (published by Yodogawa Shoten) such as alcohol, polylysine, protamine, lysozyme, pectin degradation products, alanine, thiamine lauryl sulfate, yucca foam extract, chitosan, propylene glycol Can be classified.

  Antioxidants include, for example, “Food Additive Official Document” (published by Yodogawa Shoten) such as butylhydroxyanisole, butylhydroxytoluene, vitamin C, sodium ascorbate, erythorbic acid, sodium erythorbate, vitamin E, etc. Can be classified.

  Examples of the sweetener include saccharin, licorice, stevia, aspartame, sucralose, acesulfame potassium, fructose, panose, trehalose, xylitol, erythritol, sorbitol, lactitol, maltitol, reduced palatinose, reduced mizuame, mannitol, coupling sugar, Sweetened in "Food Additives Official Document" (published by Yodogawa Shoten) such as fructooligosaccharide, galactooligosaccharide, emulsifying oligosaccharide, nigerooligosaccharide, xylo-oligosaccharide, malto-oligosaccharide, isomalto-oligosaccharide, soybean oligosaccharide, palatinose, palatinose oligosaccharide, raffinose Those classified as fees are listed.

  Examples of the colorant include Red No. 2, Red No. 3, Red No. 40, Red No. 102, Red No. 105, Red No. 106, Yellow No. 4, Yellow No. 5, Blue No. 1, Blue No. 2, Red No. 3 Rake, red 40 rake, yellow 4 rake, yellow 5 rake, blue 1 rake, blue 2 rake and other edible tars, carotene dye, anato dye, paprika dye, cochineal dye, gardenia dye, safflower dye, Examples include those classified as colorants in “Food Additive Official Documents” (published by Yodogawa Shoten) such as Benikouji dye, beet red, akane dye, spirulina dye, anthocyanin dye, and caramel.

  Examples of the emulsifier include those classified as emulsifiers in “official food additives” (published by Yodogawa Shoten) such as glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, saponin and lecithin. It is done.

  Examples of thickeners include guar gum, locust bean gum, carrageenan, agar, pectin, xanthan gum, gellan gum, gum arabic, glucomannan, alginic acid, psyllium seed gum, gelatin, methylcellulose, curdlan, crystalline cellulose, carboxymethylcellulose, tamarind Examples include those classified as thickeners in the “Food Additives Official Document” (published by Yodogawa Shoten) such as gums and indigestible dextrins.

  As quality improvers, those classified as quality improvers in the “Food Additives Official Statement” (published by Yodogawa Shoten) such as soy protein, wheat protein, casein, caseinate, milk protein concentrate, lactose, phosphate, etc. Can be mentioned.

  Examples of seasonings include sodium glutamate, nucleic acid seasonings, livestock extracts, marine extracts, vegetable extracts, yeast extracts, amino acid seasonings, fish sauce seasonings, kokumi seasonings, seasoning oils, etc. Those that are classified as seasonings in the “Kokusho Kosyo” (published by Yodogawa Shoten) are listed.

  As acidulants, for example, those classified as acidulants in the “Food Additive Official Document” (published by Yodogawa Shoten) such as citric acid, lactic acid, malic acid, tartaric acid, fumaric acid, succinic acid, adipic acid, gluconic acid, etc. Is mentioned.

  Examples of the reinforcing agent include vitamin A, vitamin B group, vitamin D, vitamin K, carotenoid, beef bone calcium, eggshell calcium, calcium carbonate, pearl shell, pearl powder, rice husk, scallop shell calcium, powder powder, fish bone Powders, dolomite, magnesium chloride, yeast minerals, heme iron, deep water, saltwater lake minerals, seaweed / plant-derived minerals, etc. classified as fortifiers (issued by Yodogawa Shoten) .

  Examples of spices and herbs include pepper, chili, pimenta, vanilla beans, cinnamon bark, clove, meat scraps, meat scrap flowers, cardamom, anise and large fennel, coriander, cumin, caraway, seaweed, juniper, ginger, saffron , Ukon, Laurel Time, Curry, etc., which are classified as spices and herbs in the “Food Additives Official Document” (published by Yodogawa Shoten).

  Examples of edible fragrances include fruit flavors such as peach flavor, orange flavor and lemon flavor, aroma flavors, sugar flavors such as maltol and furaneol, flavor enhancers such as sotron, polyphenols such as flavonoids and cacao mass, Examples include those classified as edible fragrances in the “Food Additive Official Document” (published by Yodogawa Shoten), such as precursor flavors, meat flavors, coffee flavors, milk flavors, menthols, and decalactones.

  Examples of the enzyme include α-amylase, β-amylase, glucose isomerase, protease, rennet, pancreatin, papain, lipase, cellulase, bromelain, pectinase, lysozyme, hesperidinase, pullulanase, transglutaminase, hemicellulase, β-galacta Examples include those classified as enzymes in the “Food Additives Official Document” (published by Yodogawa Shoten), such as ase, lactase, dextranase, invertase, catalase, deaminase, urease, tannase, lipoxygenase, and adenylaminase.

Next, an example of a medicinal medicinal ingredient or an additive used in them is shown.
Examples of medicinal medicinal ingredients include antipyretic analgesic / anti-inflammatory drugs, hypnotic sedatives, drowsiness preventives, antipruritics, pediatric analgesics, stomachic drugs, antacids, digestives, cardiotonic drugs, arrhythmic drugs, antihypertensive drugs, vasodilators Drugs, diuretics, anti-ulcer drugs, intestinal adjusters, osteoporosis drugs, antitussive expectorants, anti-asthma drugs, antibacterial agents, frequent urination improvers, nourishing tonics, vitamins, etc. Is the target. The medicinal component can be used alone or in combination of two or more.
Examples of additives used in pharmaceuticals include excipients, disintegrants, binders, fluidizers, lubricants, corrigents, fragrances, colorants, sweeteners, solvents, fats and oils, and thickeners. , Surfactants, gelling agents, and the like. These 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 polyvinylpyrrolidone, 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, green leaf extract powder, 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, mole, 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 granulated product, sodium hydrogen phosphate, potassium dihydrogen phosphate, calcium dihydrogen phosphate, phosphoric acid diwater Examples include sodium categorized as excipients in Japanese Pharmacopoeia (published by Yodogawa Shoten) and Pharmaceutical Additives Encyclopedia (published by Yakuji Nippo Co., Ltd.). In addition, two or more kinds can be used in combination.

  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 “Nippon Pharmacopoeia” (published by Yodogawa Shoten) such as starches, starches such as partially pregelatinized starch, synthetic polymers such as cross polyvinyl pyrrolidone and cross polyvinyl pyrrolidone copolymer, “Pharmaceutical additives encyclopedia” (Pharmaceutical Daily Inc.) Issued) is classified as a disintegrant. 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, "Japanese Pharmacopoeia" such as synthetic polymers such as polyvinyl alcohol, inorganic compounds such as calcium hydrogen phosphate, calcium carbonate, synthetic hydrotalcite, magnesium aluminate silicate, etc. Hirokawa Shoten) include those classified as a binder "Pharmaceutical additives Dictionary" (Yakujinipposha Corporation published). Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  Examples of the fluidizing agent include those classified as fluidizing agents in the “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.) such as silicon compounds such as hydrous silicon dioxide and light anhydrous silicic acid. . 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 magnesium stearate, calcium stearate, stearic acid, sucrose fatty acid ester, talc, etc. “Japan Pharmacopoeia” (published by Yodogawa Shoten), “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.) And those classified as lubricants. 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, glutamic acid, fumaric acid, succinic acid, citric acid, sodium citrate, tartaric acid, malic acid, ascorbic acid, sodium chloride, 1-menthol and other "Japanese Pharmacopoeia" (published by Yodogawa Shoten), What is classified as a corrigent in “Pharmaceuticals” (published by Yakuji Nippo Co., Ltd.) 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.

  Perfumes include orange, vanilla, strawberry, yogurt, menthol, fennel oil, cinnamon oil, spruce oil, mint oil, and other Japanese pharmacopoeia (published by Yodogawa Shoten) "(Published by Yakuji Nippo Co., Ltd.)" can be listed as flavoring agents and fragrances. 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., “Japanese Pharmacopoeia” (published by Yodogawa Shoten) such as copper chlorofin sodium, titanium oxide, and riboflavin, “Pharmaceutical Additives” What is classified as a colorant in “Encyclopedia” (published by Yakuji Nippo Co., Ltd.) 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.

  Sweeteners include aspartame, saccharin, dipotassium glycyrrhizinate, stevia, maltose, maltitol, starch syrup, and amacha powder, etc., “Pharmaceutical Additives Encyclopedia” (Yakuji Nippo Inc.) ) Issue)) can be listed as sweeteners. Even if it uses individually by 1 type chosen from the above, it is also free to use 2 or more types together.

  Solvents are not particularly limited as long as they are used in pharmaceuticals. For example, “Japan Pharmacopoeia” (published by Yodogawa Shoten) such as alcohols such as methanol and ethanol, and ketones such as acetone, Examples include those classified as solvents in the “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.), 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 Such as “Japanese Pharmacopoeia” (published by Yodogawa Shoten) and “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.). They can also be used together.

  As the thickener, for example, “Japanese Pharmacopoeia” such as hydroxypropylcellulose, hydroxypropylmethylcellulose, polyacrylic acid, carboxyvinyl polymer, polyethylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, methylcellulose, ethylcellulose, gum arabic, starch paste, etc. (Ayukawa Shoten)), “Thickeners” listed in “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.), which can be used alone or in combination of two or more. is there.

  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, glyceride monostearate, monooxyethylene sorbitan monopalmitate, monooxyethylene sorbitan monostearate, polyoxymonooleate "Japanese Pharmacopoeia" (published by Yodogawa Shoten), such as ethylene sorbitan, sorbitan monopalmitate, sodium lauryl sulfate, Those classified as surfactants listed chemicals additives Dictionary "(Yakujinipposha Corporation issued), the use of it alone, it is free to combination of two or more.

  Examples of the gelling agent include animal gelling agents such as gelatin, vegetable polysaccharides such as agar, xanthan gum, guar gum, gum arabic, curdlan, locust bean gum, carboxymethylcellulose, hydroxyethylcellulose, cellulose, microcrystalline cellulose, Examples of chemically synthesized polymers such as polyvinylpyrrolidone that are classified as gelling agents in the “Japanese Pharmacopoeia” (published by Yodogawa Shoten) and “Pharmaceutical Additives Encyclopedia” (published by Yakuji Nippo Co., Ltd.) Even if it uses singly, it is also free to use 2 or more types together.

  Examples of the cellooligosaccharide-containing food of the present invention include, for example, gels such as jelly, pudding, and yogurt, mayonnaise, dressings, sauces, sauces, soups, processed vegetables and other seasonings, curry, hayashi, meat sauce, stew , Retort food such as soup, chilled food, hamburger, bacon, sausage, salami sausage, livestock processed products such as hams, water paste products such as salmon, chikuwa, fish ham and sausage, fried rice cake, bread, raw noodles, dry noodles, Processed wheat foods such as macaroni, spaghetti, Chinese bun skin, cake mix, premix, white sauce, gyoza and spring rolls, canned curry, sauce, soup, boiled, jam, bottling, candy, Lozenges, tablet confectionery, chocolate, biscuits, cookies, rice confectionery, Japanese and Western confectionery, Western confectionery, snack confectionery Confectionery and sugar confectionery, puddings, 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. It may be used for tea beverages such as lactic acid or dairy beverages, sencha, oolong tea, matcha tea, black tea and the like.

  Examples of the cellooligosaccharide-containing pharmaceutical of the present invention include, for example, solid preparations of tablets, powders, fine granules, granules, extracts, and pills. Other than the above solid preparations, confectionery, health food, food What is used for foodstuffs, such as a feeling improvement agent and a dietary fiber reinforcement | strengthening agent, solid foundation, a bath preparation, an animal medicine, a diagnostic agent, an agrochemical, a fertilizer, a ceramic catalyst, etc. is also contained in this invention.

  Cellooligosaccharides used in the present invention are improved intestinal flora, activation of lactic acid bacteria, lactobacilli, etc., blood sugar concentration, blood insulin concentration reduction, blood cholesterol reduction, body fat percentage reduction, lipid Various physiological activities such as carbohydrate metabolism-promoting function, bowel movement / stool odor improvement, anti-tactile properties, etc. can be expected. In addition to the above normal food applications, functional foods, health foods, diet foods, etc. May be used.

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 (Incorporated Administrative Agency, National Institute of Advanced Industrial Science and Technology, Patent Biodeposition Center, receipt number FERM ABP-10323) was inoculated on a normal agar medium, cultured at 37 ° C for 7 days, and then the surface of the medium 1 spore ears, 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) 26 mg of tetrahydrate, 100 mg of iron chloride (3) hexahydrate, 40 mg of copper sulfate pentahydrate, 8 mg of manganese sulfate tetrahydrate, and 200 mg of zinc sulfate heptahydrate were dissolved in 100 mL of purified water in total. ), Adecanol 1 mL, crystalline cellulose (product of Asahi Kasei Chemicals) No. PH-101) was inoculated into a medium suspended and dissolved in 1 L of purified water in a total amount, and cultured with aeration and stirring at 28 ° C. for 5 days. During the culture, the pH of the medium was adjusted to 2.8 to 4.7 using an aqueous sodium hydroxide solution. The culture solution 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. Cellooligosaccharide crystallized in an aqueous solution at 25 ° C. is filtered under reduced pressure, dried with a ventilating dryer at 40 ° C., pulverized in a mortar, and then sieved with a sieve having an opening of 150 μm, and the powder under the sieve is 45 μm. The fine powder was removed using a sieve to obtain cellooligosaccharide powder CE-1. The sugar composition of the obtained cellooligosaccharide powder is shown in Table 1.

[Production Example 3]
Manufacture except that the strain of Production Example 1 is replaced with Cellobibrio gilvus, the pH during culture is changed to 4-10, and the buffer during enzyme reaction is changed to a phosphate buffer of pH 6.5. A cellooligosaccharide aqueous solution was prepared in the same manner as in Example 1.
The obtained cellooligosaccharide aqueous solution was passed through a column filled with activated carbon to remove the cellobiose-rich fraction, and cellooligosaccharide powder CE-2 was obtained in the same manner as in Production Example 2. The sugar composition of the obtained cellooligosaccharide powder is shown in Table 1.

[Production Example 4]
In the method for producing cellooligosaccharide of Production Example 2, ethanol was added 2.5 times by weight to the cellooligosaccharide aqueous solution at room temperature and cooled and crystallized in the same manner as in Production Example 1 to obtain cellooligosaccharide powder CE-3.
[Production Example 5]
The cellooligosaccharide aqueous solution obtained in Production Example 3 was directly spray-dried (Tokyo Rika spray dryer outlet temperature 72 ° C.) to obtain cellooligosaccharide powder CE-4.

<Confirmation test for non-insulin elevation>
Seven-week-old SD rats were allowed free intake of AIN-93G (manufactured by Oriental Yeast Co., Ltd.) for 1 week, fasted for 1 week, fasted for 16 hours, and each cellooligosaccharide aqueous solution in Table 1 was 1500 mg / day using a sonde. kg was administered. Blood samples were collected from the extracorporeal vein under anesthesia before and 30 minutes after the administration, and the insulin concentration (ng / mL Shibayagi Levi insulin kit was used) was measured. The increase rate of insulin concentration was less than 30% in CE-1 to 3, and it was confirmed that these cellooligosaccharides are non-insulin increasing.

<Improvement of lifestyle-related diseases in spontaneous type 2 diabetic mice>
[Example 1 , 2, Reference Example 1 ]
As an experimental animal, a 5-week-old male KK- ^Ay mouse (Ta / Jcl manufactured by CLEA Japan) was used. Mice are placed in individual stainless steel cages, room temperature 23 ± 5 ° C., humidity 40-70%, light / dark 12 hours each (lighting 7 am-7pm), ventilation 12 times / hour (fresh sterilized by filter) Animals were raised in a breeding room maintained in air.
First, pre-breeding for 5 days was performed, and during the pre-breeding, powdered feed (CRF-1 Oriental Yeast Co., Ltd.) was placed in a feeder and allowed to freely take. In addition, tap water was used as drinking water and allowed to freely ingest during the breeding period.
After the preliminary breeding, the grouping was performed in a group of 10 animals so that the blood glucose level and the body weight before grouping of each group were almost equal by a random method using a computer. After grouping, the powdered feed was replaced with a standard diet (AIN-93G manufactured by Oriental Yeast Co., Ltd.) and reared in the same manner. The effect of adding cellooligosaccharide is to fix sucrose in the above-mentioned standard food, and 2.5 or 5% by mass of the components other than sucrose is added to each cellooligosaccharide powder of CE-1 to 3 shown in Table 1. Instead, comparison was made. Table 2 shows the results of 30-day breeding and 60-day breeding.

[Comparative Examples 1 and 2]
Breeding was performed under the same conditions as in Example 2 except that CE-3 and CE-4 were used as cellooligosaccharides. The results are shown in Table 2.

[Comparative Example 3]
As a control, no cellooligosaccharide was added and the standard diet alone was raised. The results are shown in Table 2. Food intake and body weight were not significantly different between Examples and Comparative Examples throughout the study.

<Feed composition>
Casein 20.0 mass%
L-cysteine 0.3% by mass
Corn starch 39.7% by mass
Pregelatinized corn starch 13.2% by mass
Sucrose 10.0% by mass
Soybean oil 7.0% by mass
Cellulose powder 5.0% by mass
AIN-93 mineral mixture 3.5% by mass
AIN-93 vitamin mix 1.0% by mass
Choline bitartrate 0.25 mass%
Tertiary butyl hydroquinone 0.0014% by mass
* In the cellooligosaccharide addition system, the above-mentioned sucrose amount of 10.0% by mass was fixed, and the other components were mixed to replace 5.0% by mass with respect to the total amount of feed.

<Test items and methods>
(Weight measurement)
The body weight of each mouse was measured 30 days and 60 days after grouping.
(Feed consumption measurement)
After 8, 15, 30, and 60 days after grouping, the food intake of each mouse was measured on that day.
(Blood collection and blood analysis)
After the grouping, about 100 μL of blood was collected from the tail vein using heparinized capillaries on the 15th, 30th, and 60th days. The obtained blood was centrifuged with a centrifuge (trade name CF8DL, manufactured by Hitachi Koki Co., Ltd.) at 4 ° C., 1972 G, for 15 minutes, and the following items were measured for plasma.
・ Adiponectin concentration in blood (using Atsuka Pharmaceutical Adipo ELISA kit)
・ Plasma triglyceride (using Triglyceride E-Test Wako manufactured by Wako Pure Chemical Industries)
・ Plasma total cholesterol (Cholesterol-E-Test Wako manufactured by Wako Pure Chemical Industries, Ltd. is used)
(Measurement on isolated organs)
Sixty days after administration, the liver and epididymal fat were removed under intraperitoneal anesthesia with 50 mg / kg of ketamine hydrochloride and 2 mg / kg of xylazine, and then weighed and frozen in liquid nitrogen. Lipids in the liver were extracted from the liver with hexane and the total cholesterol concentration and neutral fat concentration in the liver were measured.

From the results shown in Table 2, the reduction rate of blood adiponectin was suppressed to 30% or less and the neutral fat concentration reduction rate in the liver was 15% or more in the same breeding period. On the other hand, Comparative Examples 1 and 2 had neither a blood adiponectin reduction rate nor a liver neutral fat concentration reduction rate within the scope of the present invention.
In addition, the cellooligosaccharide addition system of Examples 1 and 2 and Reference Example 1 showed a significant decrease in plasma and intraperitoneal lipids compared to Comparative Example 3. When Example 1 and Example 2 were compared, when the cellooligosaccharide content in the feed increased, the effect appeared in a short period, and the absolute value decreased in the long-term administration. According to the comparison between Example 2 and Reference Example 1 , when the content of cellotriose to cellohexaose was high, the expression of effects in blood was slow, but the decrease in liver lipid was promoted.
In addition, when Comparative Example 1 is compared with Examples 1 and 2 and Reference Example 1 , even if the glucose content in the feed is large, it has an effect of reducing the epididymis lipid, but the effect on blood lipid is delayed, and the liver There was no significant difference in lipids.
Furthermore, since the cellobiose content of Comparative Example 2 is less than 70% by mass and is not within the scope of the present invention, the effect on the amount of epididymis lipid and blood lipid was smaller than that of Example and Comparative Example 1.

<Improvement of lifestyle-related diseases in healthy rats>
[Examples 3 and 4 ]
As experimental animals, 5-week-old male SD rats (manufactured by CLEA Japan) were used. Rats are placed in individual stainless cages, room temperature 23 ± 5 ° C., humidity 40-70%, light and darkness 12 hours each (lighting 7 am to 7 pm), ventilation 12 times / hour (fresh sterilized by filter) Animals were raised in a breeding room maintained in air.
First, pre-breeding for 5 days was performed, and during the pre-breeding, powdered feed (CRF-1 Oriental Yeast Co., Ltd.) was placed in a feeder and allowed to freely take. In addition, tap water was used as drinking water and allowed to freely ingest during the breeding period.
After the preliminary breeding, the grouping was performed in a group of 10 animals so that the blood glucose level and the body weight before grouping of each group were almost equal by a random method using a computer. After grouping, the animals were reared in the same manner without changing the powdered feed.
The cellooligosaccharide was obtained by dissolving or suspending the above-mentioned CE-1 in purified water, and administering 160, 1000 mg / kg once / day for 13 weeks using a sonde. After administration, 30 mg / kg of pentobarbital sodium was intraperitoneally administered and anesthetized, and then 2.0 to 2.5 mL of blood was collected from the abdomen of the posterior vena cava, and 3.8 w / v% sodium citrate 0.1 mL 0.9 ml of blood was dispensed into the test tube containing the sample and centrifuged at 1870 G for 15 minutes. Using the plasma after centrifugation, the blood adiponectin concentration, the plasma and the neutral fat concentration in the liver were measured in the same manner as described above.

[Comparative Example 4]
The animals were reared in the same manner as in Example 3 except that no cellooligosaccharide was administered, and blood adiponectin concentration, plasma and liver triglyceride concentration were measured.
Food intake and body weight were not significantly different between Examples and Comparative Examples throughout the study.

  From Table 3, the Examples had a reduction rate of adiponectin concentration of 30% or less, and a reduction rate of neutral fat concentration in the liver of 15% or more, which was within the scope of the present invention. In the cellooligosaccharide administration system, blood and liver lipid concentrations decrease in a dose-dependent manner, and in a high-dose system, the liver weight tends to decrease. As a result of this test, in healthy rats, it was possible to reduce blood and hepatic lipid concentrations by ingesting cellooligosaccharide without changing food intake.

  The lifestyle-related disease preventive / ameliorating agent containing the cellooligosaccharide of the present invention as an active ingredient can be suitably used in general foods, functional foods, and pharmaceutical fields because it can reduce hepatic lipids.

Claims (3)

  A cellooligosaccharide having a cellobiose content of 95% by mass or more, 0.1 to 5% by mass of one or more selected from cellotriose, cellotetraose, cellopentaose and cellohexaose, and a glucose content of 2% by mass or less. An agent for reducing lipids in internal organs, which is contained as an active ingredient, and the oligosaccharide has a reduction rate of neutral fat concentration in the liver of 15% or more.   The agent for reducing lipids in viscera according to claim 1, wherein the agent for reducing lipids in the viscera is a agent for reducing neutral fat concentration in the liver and / or cholesterol concentration in the liver and / or fat mass in the abdominal cavity.   The agent for reducing lipids in viscera according to claim 1 or 2, wherein the oligosaccharide is an oligosaccharide having a reduction rate of blood adiponectin concentration by oral intake of 30% or less.