JP5053667B2 - Novel lactic acid bacteria and lactic acid bacteria fermentation products for promoting adipocyte differentiation - Google Patents

Description

  The present invention relates to a novel lactic acid bacterium and an adipocyte differentiation promoting agent comprising a fermentation product of a plant material by lactic acid bacterium as an active ingredient, and more specifically, an action of promoting differentiation from undifferentiated adipocytes to adipocytes using the fermented product as an active ingredient. Thus, the present invention relates to an adipocyte differentiation promoting agent having a property of increasing small adipocytes. In particular, by improving the abnormal secretion of adipocytokines related to lifestyle-related diseases, foods for preventing, improving or treating a wide range of diseases such as metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension and arteriosclerosis , Feeds, cosmetics and pharmaceutical compositions.

  In recent years, as a result of the enrichment of eating habits, the number of adults who become obese or diabetic is rapidly increasing due to excessive caloric intake and lack of exercise. Obesity, diabetes, hyperlipidemia and hypertension, also called “death quartet”, are the leading causes of cardiovascular diseases such as arteriosclerosis. Recent studies have revealed that visceral fat accumulation exists upstream of metabolic syndrome, the quartet of death. Obesity is a condition in which fat cells accumulate excessive fat, and is known to cause hyperlipidemia, arteriosclerosis, and diabetes as complications. Until now, fat cells have been thought to be a tissue that only accumulates fat, but in recent years, fat cells secrete various adipocytokines including leptin, adiponectin, TNF-α, resistin, and free fatty acids. However, it has been found to be the most active endocrine organ in the living body. Normal adipocytes secrete adiponectin that acquires insulin sensitivity, but when they fall into obesity and fat cells accumulate fat and become enlarged, adiponectin secretion decreases and TNF-α and resistin that cause insulin resistance Secretion increases. This causes insulin resistance and is thought to lead to diseases such as metabolic syndrome, diabetes, hypertension, hyperlipidemia, obesity and arteriosclerosis.

  Therefore, in order to prevent and ameliorate diseases such as metabolic syndrome, diabetes, hypertension, hyperlipidemia, obesity, arteriosclerosis, it is necessary to promote differentiation of undifferentiated adipocytes and increase normal small adipocytes is important. Antidiabetic drugs and antihyperlipidemic drugs, which are pharmaceuticals, have a strong adipocyte differentiation promoting action, but these pharmaceuticals have strong side effects and have problems in continuous use. Therefore, it is considered important to ingest daily and simply as a safe food a material that has an action of promoting differentiation from undifferentiated fat cells to small fat cells and increasing normal small fat cells. As an example studied from such a viewpoint as a food material, for example, Patent Document 1 reports that an extract of a plant martico native to South America belonging to the genus Pepperaceae has an adipocyte differentiation promoting action.

  Wheat has been used for food since ancient times, and its safety has been confirmed. For young wheat leaves, squeezing the juice or making it into a food by pulverizing. The hulls of seeds are called “Bran” and are rich in dietary fiber, so they are edible in the form of cereals, etc. with the expectation of intestinal regulation. As for seeds, barley is indispensable for today's eating habits. Barley is used for beer by alcoholic fermentation, wheat is crushed and used for bread, udon, cakes, etc.

  Studies on the inhibitory action of adipocyte differentiation of these daily eaten barley have also been conducted. For example, Patent Document 2 discloses that adipocyte differentiation inhibition using as an active ingredient a component extracted from the seeds of gramineous plants. Agents have been reported.

  In addition, various studies have been conducted in search of new functions for fermented products produced by fermenting food ingredients. For example, in Patent Document 3, the barley is fermented after fermented, and the separated bran is steamed, cooled or frozen and thawed, and the resistant starch produced by wet heat treatment is mixed with the wheat grains or pulverized product thereof, Dieting agents by inhibiting digestive enzymes have been reported. However, it has not been known so far that the fermented products of wheat show an adipocyte differentiation promoting action.

JP 2006-28049 A JP 2005-247695 A JP-A-2005-245393

  The present invention provides an adipocyte differentiation promoting agent that eliminates the above-described conventional problems and has a function of promoting adipocyte differentiation, and can be taken safely and continuously every day. Objective.

  Therefore, the present inventors screened using 3T3-L1 cells, which are undifferentiated adipocytes, as an index for the purpose of increasing adipocytes differentiation and increasing small adipocytes. As a result, it has been found that an extract extracted from a fermentation product of lactic acid bacteria using wheat as a raw material exhibits an adipocyte differentiation promoting action. Further investigations have led to the discovery that differentiation from undifferentiated adipocytes to adipocytes is very strongly promoted when specific lactic acid bacteria are used, and the present invention has been completed.

That is, the present invention includes the following inventions.
(1) An adipocyte differentiation promoter comprising a fermented product of wheat lactic acid bacteria as an active ingredient.
(2) Lactic acid bacteria are selected from Leuconostoc mesenteroides RIE strain (FERM P-21110), Lactobacillus plantarum AYA strain (FERM P-21106) and Lactobacillus plantarum AYB strain (FERM P-21107) The adipocyte differentiation promoting agent according to (1), which is one or more types.
(3) As a fermentation product of wheat lactic acid bacteria, wheat germ or wheat bran Leuconostoc mesenteroides RIE strain (FERM P-21110), Lactobacillus plantarum AYA strain (FERM P-21106) or Lactobacillus plantarum AYB The adipocyte differentiation promoting agent according to (1), comprising a fermentation product of a strain (FERM P-21107).
(4) A food, feed or cosmetic comprising the adipocyte differentiation promoter according to any one of (1) to (3).
(5) The food, feed or cosmetic according to (4), which is for preventing or improving metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension or arteriosclerosis.
(6) A pharmaceutical composition for preventing or treating metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension or arteriosclerosis, comprising the adipocyte differentiation promoting agent according to any one of (1) to (3). .
(7) Leuconostoc mesenteroides RIE strain (FERM P-21110).
(8) Lactobacillus plantarum AYA strain (FERM P-21106).
(9) Lactobacillus plantarum AYB strain (FERM P-21107).

  The adipocyte differentiation promoter of the present invention can increase normal small adipocytes by promoting differentiation of undifferentiated preadipocytes. Therefore, it shows preventive, ameliorating, or therapeutic effects on lifestyle-related diseases such as metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, and arteriosclerosis caused by an increase in hypertrophic fat cells. Moreover, since the adipocyte differentiation promoter of the present invention has strong activity even at a low concentration and has high safety, it can be used by adding to various foods, cosmetics, feeds, pharmaceutical compositions and the like. It can be effectively used for the prevention, improvement or treatment of lifestyle-related diseases as described above.

  Hereinafter, an adipocyte differentiation promoting agent and a novel lactic acid bacterium that contain as an active ingredient a fermentation product obtained by fermenting the wheat of the present invention with lactic acid bacteria (hereinafter also referred to as the fermentation product of the present invention) will be described in detail.

(Wheat lactic acid bacteria fermentation product)
The barley used in the present invention is a plant belonging to the family Gramineae, and barley, wheat, rye, oats and the like can be used. These may be used alone or in combination of two or more. In addition, the whole plant may be used as wheat, and each part such as flower, ear, stem, stem, leaf, seed, root, embryo, endosperm, embryo, cocoon, bran, rice husk is used as wheat. May be. Each of these sites may be used alone or in combination of two or more. Among them, it is preferable to use wheat germ and wheat bran, particularly wheat germ and wheat bran alone or in combination, because a high effect is obtained. As wheat bran and wheat germ, those generally obtained industrially as a by-product of the edible milling process can be used. The wheat bran and the wheat germ may be used as they are obtained from the milling step, or may be used after a treatment such as sterilization of the attached bacteria by a steaming treatment or the like.

  The wheat used in the present invention can be subjected to a fermentation treatment as it is, but it is preferable to increase the surface area by previously crushing or crushing the wheat before the treatment because the fermentation can be performed efficiently. As a method of pulverizing or crushing, for example, a method of cutting with a slicer or a cutter and then crushing with a blender, a mixer or a grinding mill can be used. At that time, a small amount of water or ethanol may be added. In the present invention, wheat includes not only whole plants and whole grains, but also pulverized products thereof, parts of wheat and pulverized products thereof, and mixtures thereof. In the case of using wheat bran and wheat germ, a mixture containing embryo, endosperm crushed material and seed coat crushed material is usually obtained by pulverizing wheat using a known pulverizer, and a known mixture is obtained from the obtained mixture. A bran fraction or embryo fraction can be obtained using a classifier, for example, a sieve.

  The fermentation product of the present invention is produced by fermenting wheat with lactic acid bacteria. Examples of the lactic acid bacteria used at this time include strains belonging to the genus Leuconostoc, the genus Lactobacillus, the genus Lactococcus, or the genus Pediococcus and acting on wheat. For example, as lactic acid bacteria, Leuconostoc mesenteroides (Leuconostoc mesenteroides), Lactobacillus rhamnosus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus casei, Lactobacillus brelus (Lactobacillus brelus) ), Lactobacillus fermentum, Lactobacillus delbruckii, Lactobacillus sakei, Lactobacillus acidophilus, Lactobacillus bucheri, Lactobacillus buchli,・ Lactobacillus helveticus, Lactobacillus pentosus, Lactococcus lactis, Pediococcus acide And the like Rakutishi (Pediococcus acidilactici). These may be used alone or in combination of two or more.

  Among the above lactic acid bacteria, Leuconostoc mesenteroides RIE strain, Lactobacillus plantarum AYA strain or Lactobacillus plantarum AYB strain, which is a novel lactic acid bacterium found by the present inventors, is used. It is preferable because a fermentation product having a high effect can be obtained.

  Leuconostoc mesenteroides RIE strain is a strain isolated from pickles, and has the accession number FERM P-21110 on November 29, 2006, National Institute of Advanced Industrial Science and Technology, Patent Biological Deposit Center (East Tsukuba City, Ibaraki Prefecture) 1-chome 1-address 1 center 6) Lactobacillus plantarum AYA strain is a strain isolated from bread dough, and under the accession number FERM P-21106 on November 29, 2006, National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center (East Tsukuba City, Ibaraki Prefecture) 1-chome 1-address 1 center 6) Lactobacillus plantarum AYB strain is a strain isolated from bread dough, and under the accession number FERM P-21107 on November 29, 2006, National Institute of Advanced Industrial Science and Technology, Patent Biological Depositary Center (Higashi Tsukuba City, Ibaraki Prefecture) 1-chome 1-address 1 center 6)

The mycological properties of the novel lactic acid bacterium Leuconostoc mesenteroides RIE are as follows.
Bacterial morphology when cultured for 18 hours at 30 ° C. using MRS liquid medium (DIFCO) (1) Bacterial morphology Cocci (2) Gram staining Positive (3) Motility None (4) Spore None (5) Catalase None (6) facultative anaerobic (7) metabolism of glucose Convert to 50% or more lactic acid (8) Growth temperature range Growth is observed at 15 ° C to 45 ° C (9) Lactic acid fermentation Heterotype (10) Application of lactic acid Lightness D
(11) Carbohydrate fermentability Glycerol is negative, D-arabinose is negative, L-arabinose is positive, ribose is positive, D-xylose is positive, galactose is positive, glucose is positive, fructose is positive, mannose is positive, rhamnose Is negative, mannitol is positive, sorbitol is negative, α-methyl D glucoside is positive, amygdalin is positive, esculin is positive, salicin is positive, cellobiose is positive, maltose is positive, lactose is positive, melibiose is positive, sucrose is positive Positive for trehalose, negative for inulin, negative for melezitose, positive for raffinose, negative for starch, positive for gluconic acid.

The mycological properties of the novel lactic acid bacterium Lactobacillus plantarum AYA are as follows. Bacteria morphology when cultured for 18 hours at 30 ° C. using MRS liquid medium (DIFCO) (1) Bacteria morphology Bacilli (2) Gram staining Positive (3) Motility None (4) Spore None (5) Catalase None (6) Facultative anaerobic (7) Glucose metabolism Convert to 50% or more lactic acid (8) Growth temperature range Growth is observed at 15 ° C, 30 ° C and 35 ° C, but growth is not observed at 45 ° C ( 9) Lactic acid fermentation Homo type (10) Light application of lactic acid DL
(11) Fermentation of carbohydrates: glycerol is positive, D-arabinose is negative, L-arabinose is positive, ribose is positive, D-xylose is negative, galactose is positive, glucose is positive, fructose is positive, mannose is positive, rhamnose Is positive, mannitol is positive, sorbitol is positive, α-methyl D glucoside is negative, amygdalin is positive, esculin is positive, salicin is positive, cellobiose is positive, maltose is positive, lactose is positive, melibiose is positive, sucrose is positive Positive for trehalose, negative for inulin, positive for melezitose, positive for raffinose, negative for starch, positive for gluconic acid.

The mycological properties of the novel lactic acid bacterium Lactobacillus plantarum AYB are as follows. Bacteria morphology when cultured for 18 hours at 30 ° C. using MRS liquid medium (DIFCO) (1) Bacteria morphology Bacilli (2) Gram staining Positive (3) Motility None (4) Spore None (5) Catalase None (6) Facultative anaerobic (7) Glucose metabolism Convert to 50% or more lactic acid (8) Growth temperature range Growth is observed at 15 ° C, 30 ° C and 35 ° C, but growth is not observed at 45 ° C ( 9) Lactic acid fermentation Homo type (10) Light application of lactic acid DL
(11) Fermentation of carbohydrates: glycerol is positive, D-arabinose is negative, L-arabinose is positive, ribose is positive, D-xylose is negative, galactose is positive, glucose is positive, fructose is positive, mannose is positive, rhamnose Is negative, mannitol is positive, sorbitol is positive, α-methyl D glucoside is negative, amygdalin is positive, esculin is positive, salicin is positive, cellobiose is positive, maltose is positive, lactose is positive, melibiose is positive, sucrose is positive Positive for trehalose, negative for inulin, positive for melezitose, positive for raffinose, negative for starch, positive for gluconic acid.

  Fermentation products of wheat germ or wheat bran RIE strain, AYA strain or AYB strain are more preferable because they have a very high adipocyte differentiation promoting effect. Here, fermented products of wheat germ or wheat bran RIE strain, AYA strain or AYB strain include fermented wheat germ product of RIE strain, fermented wheat bran product, and fermented product of wheat germ and wheat bran mixture; Fermentation product of wheat germ, fermentation product of wheat bran, and fermentation product of wheat germ and wheat bran mixture by AYA strain; Fermentation product of wheat germ, fermentation product of wheat bran, and mixture of wheat germ and wheat bran by AYB strain Fermented products of wheat, fermented products of wheat germ, fermented products of wheat bran, and fermented products of wheat germ and wheat bran by two or more selected from RIE, AYA and AYB strains; and mixtures thereof Is done. In addition, the lactic acid bacteria fermentation products of wheat bran include those obtained by separating the bran fraction from the whole wheat fermentation product, and the lactic acid bacteria fermentation products of wheat germ separated the germ fraction from the whole wheat fermentation product. Also included.

  Fermentation treatment can be carried out by bringing lactic acid bacteria into contact with wheat, for example, whole wheat grains, each part of wheat and pulverized products thereof, and mixtures thereof. In terms of efficient treatment, it is preferable that lactic acid bacteria and wheat be dispersed in water in advance. The amount of water to be added is preferably equivalent to 5 times, more preferably 2 to 3 times the total mass of wheat. By adding water, the growth environment of lactic acid bacteria becomes suitable. In addition, you may sterilize wheat before a fermentation process, and you may heat-process in order to elute the component in wheat. The heat treatment can be performed by heating the wheat at a temperature in the range of 40 ° C to 120 ° C for 30 minutes to 24 hours. In that case, it is preferable to process at a high heating temperature for a short time. For example, the treatment is preferably performed at 40 ° C. to 60 ° C. for 3 hours to 24 hours, or at 60 ° C. to 120 ° C. for 30 minutes to 3 hours.

  In order to ferment the wheat in the present invention, examples of contacting the microorganism with the wheat include an embodiment in which the lactic acid bacterium adhering to the wheat as it is and an embodiment in which the lactic acid bacterium is added to the wheat. In order to obtain a fermentation product having a high adipocyte differentiation promoting effect, an embodiment in which lactic acid bacteria are added to wheat is preferable.

  The amount of lactic acid bacteria added to the wheat is not particularly limited, but is usually 0.005 to 10 parts by mass, preferably 0.05 to 5 parts by mass in terms of dry lactic acid bacteria per 100 parts by mass of wheat. As the lactic acid bacteria to be used, those obtained by separating the cultured lactic acid bacteria from the medium by filtration or centrifugation may be added, or those previously separated from the medium and freeze-dried may be used.

  In the case of lactic acid bacteria fermentation, an additive that promotes suitable fermentation of lactic acid bacteria may be added to promote fermentation. Such additives include glutamic acid or salts thereof, and sugars such as glucose, sucrose and maltose. The amount to be added is about 0.1 to 5% with respect to the total mass of wheat and lactic acid bacteria.

  During lactic acid bacteria fermentation, the pH may be adjusted to a value suitable for lactic acid bacteria in order to promote fermentation. The pH varies depending on the type of lactic acid bacterium used, but is generally in the range of 4.0 to 7.0. Fermentation at a low pH is preferable because the pH of the fermentation broth and fermentation product during fermentation can be kept low, and propagation of contaminating bacteria other than lactic acid bacteria can be suppressed.

  Lactic acid bacteria fermentation is preferably performed under anaerobic conditions in order to promote fermentation. In order to achieve anaerobic conditions, the fermenter may be simply sealed, or after sealing, the interior may be further filled with a gas such as nitrogen or carbon dioxide, or the pressure may be reduced.

  Although the temperature condition in the case of lactic acid bacteria fermentation is not restrict | limited in particular, Usually, it is 4-60 degreeC conditions. What is necessary is just to set temperature conditions suitably with the capacity | capacitance of fermentation, etc. The temperature condition can be adjusted by a thermostatic bath, a mantle heater, a jacket, or the like. In addition, the time required for fermentation may be appropriately set depending on the capacity of fermentation. The fermentation time is generally about 24 hours to 14 days when the fermentation temperature is 4 to 20 ° C, and about 6 to 72 hours when the temperature is 20 to 60 ° C.

  After completion of fermentation, fermentation can be stopped by adding sugar or sugar alcohol. Examples of such sugars include oligosaccharides (such as maltooligosaccharides and fructooligosaccharides) and sugar alcohols (such as sorbitol, xylitol, and maltitol). By adding these sugars, other functions can be imparted to the fermentation product of the present invention, such as intestinal regulation and immunostimulation. When stopping fermentation, the temperature of a fermented liquor can also be stopped by making high temperature, for example, 80-50 degreeC.

  After the fermentation is stopped, the fermented liquid may be used as it is as the fermentation product of the present invention, or may be further subjected to a concentration operation by centrifugation or the like and / or a solid-liquid separation or sterilization operation by filtration or the like as necessary. About sterilization, you may use together overheating type sterilization, pressure type sterilization, etc. separately. When the lactic acid bacterium is used as it is, the function of the lactic acid bacterium can be imparted to the fermentation product of the present invention.

  The fermented product of the present invention thus obtained may be used as it is, but it can also be used in various forms depending on the processing methods usually used by those skilled in the art, if necessary. As such an embodiment, for example, the fermentation product concentrate obtained by centrifuging and filtering the fermentation product as described above, solid-liquid separation and collecting the supernatant, the fermentation product or the Examples thereof include a fermentation product paste obtained by concentrating a fermented product concentrate, a powder obtained by drying and pulverizing the fermented product or the fermented product concentrated solution with an excipient as necessary, or a fermented product concentrated powder. These are all included in the fermentation product of the present invention.

  In the present invention, the fermented product of wheat lactic acid bacteria also includes an extract obtained by extracting the fermented product with water or an organic solvent. The active ingredient is concentrated by the extraction operation, and the adipocyte differentiation promoting activity can be further increased. The extraction method is not particularly limited as long as it is an extraction method using water or an organic solvent as an extraction solvent. However, the fermentation product, fermentation product extract, fermentation paste, powder or fermentation product extract powder (hereinafter referred to as a fermentation product or the like) is used. In some cases, a known method such as a method of immersing, stirring or refluxing in water or an organic solvent can be used. It can also be extracted by a supercritical fluid extraction method using carbon dioxide or the like. Regarding the lactic acid bacteria fermentation products of wheat, the above extract and those other than the extract may be referred to as “fermentation product extract” and “fermentation product before extraction”, respectively. Of lactic acid bacteria fermentation products.

  The organic solvent that can be used for extraction is not particularly limited, and specifically, lower alcohols such as methanol, ethanol, n-propanol, isopropanol, and n-butanol, and 1,3-butylene glycol, propylene glycol, glycerin. Alcohols that are liquid at room temperature such as polyhydric alcohols such as; ethers such as diethyl ether and propyl ether; esters such as butyl acetate and ethyl acetate; ketones such as acetone and ethyl methyl ketone; hexane; Can be mentioned. These may be used alone or in combination of two or more. Among the above organic solvents, alcohols that are liquid at room temperature, for example, lower alcohols having 1 to 4 carbon atoms, are preferably used from the viewpoint of operability and environmental influences, and safety due to the residual solvent. From the viewpoint, it is more preferable to use ethanol.

  In the present invention, the organic solvent includes a water-containing organic solvent in which an aqueous component is further contained in these organic solvents, and this may be used for extraction. From the viewpoint of maintaining high extraction efficiency, the content of the aqueous component in the water-containing organic solvent is usually 80% by volume or less, preferably 65% by volume or less, and more preferably 50% by volume or less.

  In this case, examples of the aqueous component include water and an aqueous solution of a water-soluble component. By mixing the organic solvent and an acidic aqueous solution, a neutral aqueous solution (or water), or a basic aqueous solution, the extraction conditions are set respectively. It can be adjusted to acidic, neutral, or basic conditions.

  As a specific extraction method, the above-mentioned fermentation products and the like are extracted in an extraction solvent heated at room temperature or under normal pressure or while being immersed or stirred, or extracted while refluxing in the extraction solvent. Etc. At that time, the extraction temperature is preferably 5 ° C. to the boiling point of the extraction solvent. Although extraction time can be suitably set with the kind of extraction solvent to be used, extraction conditions, and the content of aqueous components in the case of a water-containing organic solvent, it is usually about 30 minutes to 72 hours. When performing extraction by refluxing, it is preferable to use an organic solvent having a low boiling point so that the fermentation product or the like does not denature or undergo thermal decomposition.

  Next, the mixture containing the extract and the residue is subjected to filtration or centrifugation as necessary, and the solid component as a residue is removed to obtain an extract. The removed solid component can be subjected again to an extraction operation using an extraction solvent, and this operation may be repeated several times.

  The extract thus obtained may be used as a fat cell differentiation promoter of the present invention in a liquid state, and further, if necessary, dried or powdered by a method such as concentration or freeze-drying or spray-drying. May be used. Specific concentration and drying methods are not particularly limited, and examples thereof include filtration, centrifugation, centrifugal filtration, spray drying, spray cool, drum drying, vacuum drying, freeze drying, and the like alone or in combination. Can be adopted. In that case, you may add the excipient | filler normally used as needed.

  In the present invention, promotion of adipocyte differentiation refers to the action of differentiating undifferentiated adipocytes (preadipocytes) into adipocytes. Here, a fat cell refers to a cell containing a large amount of fat scattered between tissues or forming a fat tissue, and is classified into a brown fat cell and a white fat cell. In particular, white adipocytes are cells constituting subcutaneous fat and visceral fat. In this white adipose tissue, it is possible to increase small adipocytes from undifferentiated adipocytes, metabolic syndrome, obesity, diabetes, hyperlipidemia It is important for prevention and treatment of hypertension, arteriosclerosis, etc.

  The adipocyte differentiation promoting action can be evaluated by measuring GPDH (glycerol-3-phosphate dehydrogenase) activity which is a cell differentiation marker.

  The wheat lactic acid bacteria fermentation product obtained above is used alone as an adipocyte differentiation promoter, or as an adipocyte differentiation promoter containing other ingredients, or in foods, feeds, cosmetics or pharmaceutical compositions. Ingested continuously, the prevention, improvement and treatment effects of diseases such as metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension and arteriosclerosis caused by enlarged fat cells in accumulated visceral fat are expected.

  The adipocyte differentiation promoter of the present invention is usually taken in the range of 0.01 to 50 g per day for adults, preferably 0.1 to 5 g per day for adults, as the dry mass of the fermentation product before extraction. However, since the adipocyte differentiation promoter is highly safe, the amount of intake can be further increased. The daily intake may be taken once, but may be taken in several divided doses. The dry mass of the fermented product extract is ingested in the range of 0.001 to 10 g per day for an adult, preferably in the range of 0.01 to 5 g per day for an adult.

(Adipocyte differentiation promoter)
The adipocyte differentiation promoting agent of the present invention includes additives described later, other known adipocyte differentiation promoting substances, fat accumulation, as long as the effect of the present invention is not inhibited, in addition to the fermented product of wheat lactic acid bacteria as an active ingredient An inhibitory substance, a lipolysis promoting substance, a fat metabolism improving substance, or the like may be added singly or in combination.

  The form of the adipocyte differentiation promoter of the present invention is not particularly limited, and examples thereof include tablets, capsules, granules, powders, powders, syrups, dry syrups, liquids, suspensions, inhalants, and other oral agents, suppositories, and the like. The dosage form may be an enteral preparation such as an ointment, an external preparation for skin such as an ointment, cream, gel, or patch, a drip, an injection. Of these, oral preparations are preferred.

  Such a form of adipocyte differentiation promoter includes the above-mentioned ingredients, commonly used excipients, disintegrants, binders, lubricants, surfactants, alcohol, water, water-soluble polymers, sweeteners. Additives such as a corrigent and acidulant can be blended according to the dosage form, and can be produced according to conventional methods. Liquid preparations such as liquids and suspensions may be dissolved or suspended in water or other suitable medium immediately before taking. In the case of tablets and granules, the preparations may be prepared by a well-known method. The surface may be coated.

  When the adipocyte differentiation-promoting agent of the present invention contains the above-mentioned additives and other adipocyte differentiation-promoting substances, fat accumulation-inhibiting substances, lipolysis promoting substances, fat metabolism improving substances, etc., a fermentation product of wheat lactic acid bacteria that is an active ingredient The content of is different depending on the dosage form, but is usually 0.01 to 99% by mass, preferably 0.1 to 80% by mass, more preferably 1 to 75% by mass as the dry mass of the fermentation product before extraction. The dry mass of the fermentation product extract is usually from 0.0001 to 80 mass%, preferably from 0.001 to 50 mass%, more preferably from 0.1 to 40 mass%. It is desirable that the dosage per day can be managed so that the daily dosage of the active ingredient can be taken.

  Furthermore, the adipocyte differentiation promoter of the present invention may be allowed to coexist with various additives and other various substances used in the production of pharmaceuticals, foods, and feeds. Such substances and additives include various fats and oils, crude drugs, amino acids, polyhydric alcohols, natural polymers, vitamins, minerals, dietary fibers, surfactants, purified water, excipients, stabilizers, pH adjusters, Antioxidants, sweeteners, flavoring ingredients, acidulants, colorants, flavors and the like can be mentioned.

  Examples of the various oils and fats include vegetable oils such as soybean oil, corn oil, safflower oil, and olive oil, and animal oils such as beef tallow and sardine oil.

  Examples of the herbal medicine include cow yellow, ground yellow, eggplant, royal jelly, carrot and deer.

  Examples of the amino acid include glutamine, cysteine, leucine, arginine and the like.

  Examples of the polyhydric alcohol include ethylene glycol, polyethylene glycol, propylene glycol, glycerin, and sugar alcohol. Examples of the sugar alcohol include sorbitol, erythritol, xylitol, maltitol, mannitol and the like.

  Examples of the natural polymer include gum arabic, agar, water-soluble corn fiber, gelatin, xanthan gum, casein, gluten or gluten hydrolyzate, lecithin, starch, and dextrin.

  Examples of the various vitamins include vitamin C (ascorbic acid), vitamin B group, vitamin E (tocopherol), vitamin A, D, K, riboflavin butyrate, and the like. The vitamin B group includes vitamin B1 derivatives, vitamin B2, vitamin B6, vitamin B12, and various vitamin B complexes such as biotin, pantothenic acid, nicotinic acid, and folic acid. Examples of vitamin B1 derivatives include thiamine or a salt thereof, thiamine disulfide, fursultiamine or a salt thereof, dicetiamine, bisbuthiamine, bisbenchamine, benfotiamine, thiamine monophosphate disulfide, chicotiamine, octothiamine, prosultiamine, etc. All the compounds having the physiological activity of vitamin B1 are included.

  Examples of the mineral include calcium, magnesium, zinc, and iron.

  Examples of the dietary fiber include gums, mannan, pectin, hemicellulose, lignin, β-glucan, xylan, and arabinoxylan.

  Examples of the surfactant include glycerin fatty acid ester, sorbitan fatty acid ester, and sucrose fatty acid ester.

  Examples of the excipient include sucrose, glucose, corn starch, calcium phosphate, lactose, dextrin, starch, crystalline cellulose, and cyclodextrin.

  In addition to the above, other known adipocyte differentiation-promoting substances, fat accumulation-inhibiting substances, lipolysis promoting substances, fat metabolism improving substances, or various functional ingredients or additives, other than the above, for example, taurine, glutathione, carnitine , Creatine, coenzyme Q, glucuronic acid, glucuronolactone, red pepper extract, ginger extract, cacao extract, guarana extract, garcinia extract, theanine, γ-aminobutyric acid, capsaicin, capsiate, various organic acids, flavonoids, polyphenols, catechin , Xanthine derivatives, indigestible oligosaccharides such as fructooligosaccharides, polyvinylpyrrolidone and the like.

  The amount of these additives is appropriately determined according to the type of additive and the amount of intake to be desired, but is generally in the range of 0.01 to 90% by mass with respect to the total amount of the adipocyte differentiation promoter. Preferably, it is the range of 0.1-50 mass%.

  The adipocyte differentiation-promoting agent of the present invention has an action of increasing normal small adipocytes to normalize adipocytokine secretion by promoting adipocyte differentiation, and metabolic syndrome, diabetes, hyperlipidemia It exhibits excellent preventive, ameliorating and therapeutic effects on diseases such as hypertension, obesity and arteriosclerosis. In addition, it is safe and easy to take for a long time. Therefore, the adipocyte differentiation promoter of the present invention can also be used in foods, feeds and cosmetics for prevention and improvement of lifestyle-related diseases such as metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteriosclerosis and the like. . Furthermore, by ingesting the adipocyte differentiation promoting agent, food, cosmetics or feed of the present invention in a form in which the above intake can be controlled, metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteriosclerosis, etc. The present invention provides a preventive method and a therapeutic method for diseases related to hypertrophy of adipocytes, in other words, lifestyle-related diseases related to abnormal secretion of adipocytokines.

(Food and feed)
As described above, the adipocyte differentiation-promoting agent of the present invention has an adipocyte differentiation-promoting action, and is made from wheat having a dietary experience, and has high safety and good flavor. Furthermore, even if added to various foods, it does not inhibit the flavor of the food itself, so it can be added to various foods and taken continuously, metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteries It is expected to prevent and improve lifestyle-related diseases such as sclerosis.

  The food of the present invention contains the aforementioned adipocyte differentiation promoter. In the present invention, the food includes beverages. The food containing the adipocyte differentiation promoting agent of the present invention can be blended with the above adipocyte differentiation promoting agent in addition to health foods, functional foods, foods for specified health use, etc. that promote health by promoting adipocyte differentiation, All foods are included.

  Specific examples of food include liquid foods such as tube enteral nutrition, tablet foods, tablets, chewable tablets, tablets, powders, powders, capsules, granules, drinks, and other health foods and nutritional supplements. Foods: Green tea, oolong tea, and other tea drinks, soft drinks, jelly drinks, sports drinks, milk drinks, carbonated drinks, fruit juice drinks, lactic acid bacteria drinks, fermented milk drinks, powdered drinks, cocoa drinks, milk and purified water Spreads such as butter, jam, sprinkles and margarine; mayonnaise, shortening, custard cream, dressings, breads, cooked rice, noodles, pasta, miso soup, tofu, yogurt, soup or sauces, confectionery (e.g. biscuit or Cookies, chocolate, candy, cake, ice cream, chewing gum, tablet ), And the like.

  In addition to the above-mentioned adipocyte differentiation promoter, the food of the present invention includes other food materials, various nutrients, various vitamins, minerals, dietary fiber, various additives (for example, taste ingredients, sweet tastes) used in the production of the food. And sour agents such as organic acids, stabilizers, flavors) and the like, and can be produced according to a conventional method.

  In the food of the present invention, the content of the adipocyte differentiation promoter varies depending on the form of the food, but is usually 0.001 to 80% by mass, preferably 0.01 to 50% as the dry mass of the fermentation product before extraction. The dry mass of the fermentation product extract is usually from 0.0001 to 80% by mass, preferably from 0.001 to 50% by mass, more preferably from 0.1 to 20% by mass. More preferably, it is the range of 0.01-20 mass%. Since the adipocyte differentiation promoter of the present invention is highly safe, its content in food can be further increased. It is preferable that the daily intake can be controlled so that the intake of the above-described adipocyte differentiation promoting agent (as a dry mass of the fermentation product) per day for adults can be consumed.

  As described above, the food of the present invention has an action of differentiating undifferentiated adipocytes into adipocytes, and normalizes secretion of adipocytokines such as adiponectin, so that metabolic syndrome, diabetes, hyperlipidemia, hypertension In addition to excellent preventive and ameliorating effects on lifestyle-related diseases such as obesity and arteriosclerosis, it is safe and has no side effects. In addition, since the adipocyte differentiation promoter of the present invention has a good flavor and does not inhibit the flavor of the food even when added to various foods, the resulting food can be easily ingested for a long period of time and has excellent metabolic properties. Expected to prevent and improve syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteriosclerosis and the like. In addition, by eating and drinking the food of the present invention in a form in which the intake of the above-mentioned adult adipocyte differentiation promoting agent (as a dry mass of the fermentation product) can be controlled, metabolic syndrome, diabetes, Methods for preventing and improving lifestyle-related diseases such as hyperlipidemia, hypertension, obesity, and arteriosclerosis are provided.

  Furthermore, the adipocyte differentiation promoting agent of the present invention can be blended not only in human foods but also in livestock, racehorses, pets and other feeds. Since feed is almost equal to food except that the subject is other than humans, the above description about food can be applied to feed as well.

(Cosmetics)
As described above, the adipocyte differentiation-promoting agent of the present invention has a fat cell differentiation-promoting action and is made from wheat that has been eaten as a raw material, and since it is highly safe, it can also be used as a cosmetic material. . Since it can be applied continuously, it is expected to prevent and improve obesity and cellulitis.

  Although it does not specifically limit as cosmetics, From a functional surface, the emulsion for face or body, a cosmetic liquid, a cream, a lotion, an essence, a pack, a sheet | seat etc. are preferable, for example. The addition amount is not particularly limited, but as an example, the dry mass of the fermentation product before extraction is usually 0.001 to 80% by mass, preferably 0.01 to the weight of the cosmetic base material. It is 50 mass%, More preferably, it is the range of 0.1-20 mass%, As dry mass of the extract of a fermentation product, it is usually 0.0001-60 mass%, Preferably 0.001-40 mass% More preferably, the range of 0.01 to 20% by mass is appropriate.

(Pharmaceutical composition)
The adipocyte differentiation-promoting agent of the present invention has excellent preventive, ameliorating and therapeutic effects on diseases such as metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteriosclerosis, etc. Alternatively, it can be a pharmaceutical composition for treatment. That is, the pharmaceutical composition of the present invention contains the above-described adipocyte differentiation promoter of the present invention. In other words, the pharmaceutical composition of the present invention is a fermentation obtained by subjecting the above wheat to lactic acid bacteria fermentation. Product (wheat lactic acid bacteria fermentation product) is included as an active ingredient.

  Regarding the pharmaceutical composition of the present invention, the compounding amount of the fermented product of wheat lactic acid bacteria, its intake (dosage), other active ingredients, functional ingredients and additives, their compounding quantity, dosage form, etc., which are active ingredients Is the same as described for the adipocyte differentiation promoter.

  By administering the pharmaceutical composition of the present invention, undifferentiated adipocytes differentiate and normal small adipocytes increase, so that adipocytokine secretion is normalized, metabolic syndrome, diabetes, hyperlipidemia, hypertension It is possible to treat or prevent diseases associated with fat cell hypertrophy such as obesity and arteriosclerosis, in other words, diseases associated with abnormal secretion of adipocytokines. In addition, since the pharmaceutical composition of the present invention is safe and has no side effects, it can be continuously administered for a long period of time, and has excellent metabolic syndrome, diabetes, hyperlipidemia, hypertension, obesity, arteriosclerosis. The prevention and treatment effects are expected.

  EXAMPLES Hereinafter, although this invention is demonstrated further more concretely based on an Example, this invention is not limited to these Examples.

(Wheat processing)
Wheat was used as the wheat, and the dried products of the bran portion and the germ portion were each pulverized using a mixer. Each pulverized product was sterilized using an autoclave and then used for the fermentation treatment described below.

(Cultivation of lactic acid bacteria)
Culture of the novel lactic acid bacteria of the present invention, Leuconostoc mesenteroides RIE strain (FERM P-21110), Lactobacillus plantarum AYA strain (FERM P-21106) and Lactobacillus plantarum AYB strain (FERM P-21107) The following method was used.

  The RIE strain, AYA strain, and AYB strain were separately inoculated with a platinum loop in 5 mL of GYP medium and cultured at 30 ° C. for 48 hours (preculture). Next, 1 mL of the preculture was inoculated into 30 mL of GYP medium, and precultured at 30 ° C. for 24 hours (main culture). The obtained main culture was centrifuged (5000 rpm, 15 minutes), and the cells were collected from the precipitate. The cells were washed twice with sterilized water and then centrifuged (5000 rpm, 15 minutes), and the cells were made up to 3 mL with sterilized water to obtain a cell solution.

  The composition of the GYP medium is shown below.

Example 1
A raw material solution having the composition shown in Table 2 was prepared for each part of wheat, and 0.4 mL of the RIE bacterial cell solution was inoculated thereto, followed by culturing at 30 ° C. for 48 hours. After completion of the culture, heat treatment (121 ° C., 15 minutes) was performed.

  After completion of the culture, the culture solution was centrifuged (7000 rpm, 5 minutes, 4 ° C.), and the precipitate was separated and recovered. Subsequently, this precipitate was freeze-dried, suspended in 10 mL of ethanol, and extracted by shaking at room temperature for 24 hours. Centrifugation (7000 rpm, 5 minutes, 4 ° C.) was performed, and the supernatant was filtered with a 0.45 μm filter and freeze-dried to obtain an extract.

(Example 2)
An AYA strain was used instead of the RIE strain in the same manner as in Example 1, and a fermented treatment was performed using a pulverized product of wheat bran and wheat germ to prepare an extract.

(Example 3)
Using the AYB strain instead of the RIE strain, an extract was prepared in the same manner as in Example 1 using a wheat bran and wheat germ pulverized product.

(Comparative Example 1)
In Example 1, in place of the pulverized product of each part of wheat, fermentation was performed using the RIE strain in the same manner as in Example 1 except that a pulverized maitake portion was prepared according to Table 3. An extract of the fermentation product was prepared.

(Comparative Example 2)
In Example 2, in place of the pulverized product of each part of wheat, fermentation was performed using the AYA strain in the same manner as in Example 2 except that the pulverized maitake portion was prepared and used according to Table 3. An extract of the fermentation product was prepared.

(Comparative Example 3)
In Example 1, without using the pulverized product of each part of wheat, an extract was prepared by inoculating 0.4 g each of RIE strain and AYA strain into 10 g of sterile water and culturing at 30 ° C. for 48 hours. Comparative Example 3 was obtained.

(Test Example 1) Cell differentiation activity confirmation test (1) Differentiation and culture into adipocytes 3T3-L1 cells, which are undifferentiated adipocytes, are seeded in a 24-well plate so as to be 5 × 10 ⁴ cells per well (1 mL ), Differentiation induction medium (0.5 mM 1-methyl-3-isobutylxanthine, 0.25 μM dexamethasone, 10 μg / mL 10% FCS-added DMEM medium containing insulin) was added to 0.5 mL, and cultured for 2 days, and adipocytes Differentiation was induced. Thereafter, the culture was changed to 0.5 mL of a maturation promotion medium (DMEM medium supplemented with 10% FCS containing 5 μg / mL insulin) and cultured for 7 days.

  At this time, the extracts obtained in Examples 1, 2, 3, and Comparative Examples 1, 2, and 3 were added to the differentiation induction medium and the maturation promotion medium so that the dry weight was 100 or 200 μg / mL. After completion of the culture, GPDH (glycerol-3-phosphate dehydrogenase) activity, which is a cell differentiation marker, was measured using a GPDH measurement kit (Hokudo Co., Ltd.).

(2) Measurement of GPDH activity After completion of the culture, the well was washed with 0.5 mL of PBS (−) three times, and then 0.5 mL of the enzyme extract was added, and the cells were detached from the bottom of the well by pipetting. Next, the peeled cells were put together with the enzyme extract into a sampling tube, and the sampling tube was immersed in liquid nitrogen for about 10 minutes to freeze the contents. Thereafter, the sampling tube was immersed in a water bath set at 37 ° C. to melt the contents. This freeze-thaw was repeated 3 times in total to disrupt the cells, and the resulting disrupted solution was centrifuged at 15,000 rpm for 5 minutes at 4 ° C., and the supernatant was recovered. The collected supernatant was used as a specimen.

  100 μL of the reaction substrate solution was placed in a 96-well plate and heated at 25 ° C. for about 5 minutes. Next, 50 μL of the sample was placed in the 96-well plate and stirred well, and the decrease in absorbance at 340 nm was measured over time (10 minutes every other minute) to determine the amount of change in absorbance per minute. . The obtained change in absorbance was applied to the following formula to determine GPDH activity. The results are shown in Table 4 as relative values when the control group (wells to which neither extract nor lactic acid bacteria are added) is taken as 100%.

  GPDH activity (U / mL) = ΔOD (340 nm) /min×0.482

  As is apparent from Table 4, lactic acid bacteria alone did not show adipocyte differentiation promoting activity. However, it has been clarified that the fermented wheat product produced by the lactic acid bacterium of the present invention exhibits a high differentiation promoting activity, and exhibits a very high differentiation promoting effect when fermented using the RIE strain and the AYA strain. On the other hand, fermented maitake with lactic acid bacteria did not show fat differentiation promoting activity.

(Example 4) Manufacture of tablets 20 g of an extract from a fermentation product of wheat bran RIE obtained in the same manner as in Example 1, 73.8 g of crystalline cellulose (Asahi Kasei) and 5 g of polyvinylpyrrolidone (BASF) were mixed. Then, 30 mL of ethanol was added thereto, and granules were produced according to a conventional method by a wet method. After the granules were dried, 1.2 g of magnesium stearate was added to give granules for tableting, and tableting was performed using a tableting machine to produce 100 tablets each having 1 g (extracted per tablet). 0.2g of product).

(Example 5) Production of granules 10 g of an extract from a fermentation product of wheat bran AYA obtained in the same manner as in Example 2, 170 g of lactose (DMV) and 60 g of crystalline cellulose (Asahi Kasei) were mixed. Ethanol (130 mL) was added to the mixture and kneaded for 5 minutes by a conventional method using a kneader. After kneading, the mixture was sieved with 10 mesh and dried at 50 ° C. in a dryer. After drying, the particles were sized to obtain 240 g of granules. This was individually packaged so as to be 4 g per package (including 0.167 g of extract per package).

(Example 6) Manufacture of syrup agent While boiling 400 g of purified water, 750 g of sucrose and 1 g of an extract from a fermented lactic acid bacterium of AYA strain were added in the same manner as in Example 2. The solution was dissolved and heated, and purified water was added thereto to make a total volume of 1000 mL to produce a syrup (containing 0.02 g of extract per 20 mL).

(Example 7) Manufacture of cream The cream was manufactured with the following composition.

Claims (10)

Containing fermentation product by Leuconostoc mesenteroides RIE strains wheat germ or wheat bran (FERM P-21110) or Lactobacillus plantarum AYA strain (FERM P-21106), fat cell differentiation promoting agent. The adipocyte differentiation promoting agent according to claim 1, comprising a fermentation product of a wheat bran Leuconostoc mesenteroides RIE strain (FERM P-21110). The adipocyte differentiation promoter according to claim 1, comprising a fermentation product of wheat germ Leuconostoc mesenteroides RIE strain (FERM P-21110). The adipocyte differentiation promoting agent according to claim 1, comprising a fermentation product of Lactobacillus plantarum AYA strain (FERM P-21106) of wheat bran. The adipocyte differentiation promoting agent according to claim 1, comprising a fermentation product of Lactobacillus plantarum AYA strain (FERM P-21106) of wheat germ. A food, feed or cosmetic comprising the adipocyte differentiation promoter according to any one of claims 1 to 5 . The food, feed or cosmetic according to claim 6 , which is for preventing or improving metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension or arteriosclerosis. A pharmaceutical composition for preventing or treating metabolic syndrome, obesity, diabetes, hyperlipidemia, hypertension or arteriosclerosis, comprising the adipocyte differentiation promoting agent according to any one of claims 1 to 5 .   Leuconostoc mesenteroides RIE strain (FERM P-21110).   Lactobacillus plantarum AYA strain (FERM P-21106).