WO2019131274A1

WO2019131274A1 - Method for producing fermentation product derived from green tea extract, and koji fermentation product derived from green tea extract

Info

Publication number: WO2019131274A1
Application number: PCT/JP2018/046312
Authority: WO
Inventors: 滋樹吉田; 均宮崎; 小嶋　誠
Original assignee: 国立大学法人筑波大学; 合同会社森のお茶工房
Priority date: 2017-12-26
Filing date: 2018-12-17
Publication date: 2019-07-04
Also published as: JPWO2019131274A1; TW201927326A

Abstract

To provide a novel product derived from green tea and a novel use thereof. Provided are: a method for producing a fermentation product derived from a green tea extract, said method being characterized by comprising fermenting the green tea extract using a koji mold; a koji fermentation product derived from a green tea extract; and a composition which comprises, as an active ingredient, the koji fermentation product derived from a green tea extract, preferably said composition being a medicinal composition or a food or beverage composition. Still preferably, the aforesaid composition is a composition for promoting adiponectin production, a composition for preventing, ameliorating or treating obesity, a skin care composition or a composition for preventing, ameliorating or treating arteriosclerosis.

Description

Process for producing fermented product derived from green tea extract and fermented product derived from green tea extract

The present invention provides a method for producing a fermented product derived from a green tea extract, a fermented product derived from a novel green tea extract, and a composition for promoting adiponectin production comprising the fermented product as an active ingredient, prevention, amelioration or treatment of obesity The present invention relates to a composition etc.

Green tea has long been used as a food and drink and is highly safe, and green tea contains green tea catechins, and it is known that this green tea catechins have an antibacterial action. Green tea extracts containing high concentrations of green tea catechins have been developed for industrial use of antibacterial activity, and are used as raw materials for oral hygiene products such as caries prevention and oral cleaning etc. and antibacterial masks etc. .
And in recent years, various physiological functions regarding green tea extract are further explored, and research and development of new applications utilizing this physiological function are made.
For example, Patent Document 1 provides a Helicobacter pylori removal agent, food / drink or food additive comprising a tea polyphenol or a tea extract as an active ingredient, which is used in combination with a proton pump inhibitor (PPI). Is disclosed.

JP, 2002-068992, A

Therefore, the main object is to provide new products and new applications derived from green tea.

The present inventors tried trial and error to provide new products and new applications starting from green tea. At that time, the present inventors attempted to ferment by adding a seed meal regardless of the common sense even though the green tea extract has an antibacterial action. And when the present inventors performed HPLC analysis about the fermented matter after koji fermentation, the peak patterns of both differed before and after fermentation, and koji fermentation was proceeding well. It could be confirmed. Thus, it was quite unexpected that the koji fermentation proceeded favorably with the green tea extract containing catechins having an antibacterial action at a high concentration.

Moreover, when the peak pattern of the HPLC analysis of the fermented manure derived from the green tea extract obtained by the present inventors was examined, the peak of catechins known for its anti-obesity action had almost disappeared. And, surprisingly, the fermented persimmon product derived from this green tea extract had a low concentration for promoting adiponectin production as compared with the green tea extract before fermentation. The present inventors conducted various physiological activity tests, and as a result, the green tea extract-derived fermented persimmon obtained was used to prevent, improve or treat obesity; beautifying; preventing, improving or treating arteriosclerosis, etc. I found that I could use it.

Since the present inventors did not detect peaks of catechins and teadenols which are known for their anti-obesity action, they were not detected, so that some components in the fermented green tea extract derived from the obtained green tea extract exert their efficacy. Think. From this, the present inventors have found that using green tea extract, green tea fermented products which are unprecedented can be obtained.
As such, the present inventors have completed the present invention.
That is, the present invention is as follows.

[1] A method for producing a fermented product derived from green tea extract, comprising fermenting the green tea extract with a koji.
[2] A fermented koji product derived from green tea extract.
[3] A composition comprising, as an active ingredient, a fermented persimmon derived from green tea extract.
[4] The composition is a composition for promoting adiponectin production, a composition for preventing, ameliorating or treating obesity, a composition for skin care, or a composition for preventing, ameliorating or treating arteriosclerosis The composition as described in [3].

According to the present invention, new products and new applications derived from green tea can be provided.
According to the present invention, it is possible to provide a new fermented green tea extract derived from green tea extract. According to the present invention, a composition for promoting adiponectin production comprising a fermented green tea extract derived from new green tea extract as an active ingredient; a composition for preventing, ameliorating or treating obesity; a skin care composition; preventing arteriosclerosis, A composition for improvement or treatment can be provided.
In addition, the effect described here is not necessarily limited, and may be any effect described in the present specification.

1A: standard catechins (from left: EGC, EC, EGCG, GCG), FIG. 1B: green tea extract (before fermentation), FIG. 1C: standard teadenol A, FIG. 1D: fermented mulberry (Production example 3: white Fig. 1E shows the results of HPLC analysis of bean sprout fermented product (Production Example 5: Long-white fungus (rice bran / wheat malt system)). The figure which shows the effect which the green tea extract which fermented green tea extract (manufacturing example 3: white bean sprout (rice bran and wheat straw type)) and the green tea extract before the fermentation give to the expression of adiponectin of mature adipocytes. is there. The upper part is the result of Western blot analysis of adiponectin, and the lower part is the result of quantifying its expression level. Cont is a control. The results of adiponectin (top: Fig. 3A) and fatty acid synthetase (FAS) (bottom: Fig. 3B) are shown in the administration group (right side) and the non-administration group (left side: Cont) in which the rat fermented product was administered to mice. . The upper part of Fig. 3A and Fig. 3B is the result of Western blot analysis of adiponectin, and the lower part is the result of quantifying its expression level. Cont is a control. * P <0.05 vs cont

Hereinafter, preferred embodiments of the present invention will be described. However, the present invention is not limited to the following preferred embodiments, and can be freely modified within the scope of the present invention. In the present specification, percentages are indicated by mass unless otherwise specified.

1. Method for Producing Fermented Product Derived from Green Tea Extract This embodiment is a method for producing a fermented product using green tea extract as a starting material, and is characterized in that the green tea extract is fermented in a pot, and is derived from green tea extract It is a manufacturing method of fermented material. By this, a fermented persimmon derived from green tea extract can be obtained.

The method for producing a fermented product derived from green tea extract according to the present embodiment includes the step of fermenting the green tea extract with a koji.
The method for producing a fermented product according to the present embodiment may include, other than the koji fermentation step, a step performed before and after fermentation, as appropriate. As a pre-fermentation step, for example, a step of extracting green tea, a seed meal preparation step, a preparation step of a culture medium for persimmon fermentation; a post-fermentation step, for example, a step of processing persimmon (for example, removal step), a preparation of persimmon fermented product Steps (for example, separation and purification steps of fermentation components) and the like can be mentioned.
Moreover, the process of the said fermented material manufacturing method may be integrated in the process of the manufacturing method (for example, food-drinks manufacturing method etc.) of a composition.

<Green tea extract>
The green tea extract is obtained by extraction from tea leaves, and can be obtained by a known method. Examples of tea leaves include tea leaves, fresh tea leaves and the like. As an example of the production method, it can be obtained, for example, by extracting from tea leaves and / or fresh tea leaves with hot water (0 to 100 ° C.) or a water-soluble organic solvent. Examples of the water-soluble organic solvent include lower alcohols having 1 to 4 carbon atoms (ethanol, 1,3-butylene glycol and the like) and the like, and one or more of them can be combined, and further water may be used. It is good also as a mixed water solvent. Alternatively, supercritical extraction means may be used. It is also possible to extract by adding an extraction aid (for example, an organic acid such as sodium ascorbate or an organic acid salt thereof).
The tea leaves are tea leaves obtained from tea leaves obtained from Camellia (eg, C. sinensis, C. assamica) or hybrids thereof. The tea leaves used in this embodiment are, for example, green teas (for example, Sencha, Bancha, Gyokuro, Tencha, Kettle-roasted tea, etc.), and non-fermented green tea leaves obtain the target fermented matter efficiently. It is preferable in point.
Moreover, as a commercially available green tea extract, for example, green tea extract BL (Access One Co., Ltd.), Polyphenon (Mitsui Norin Co., Ltd.), Theafran (Itoen Co., Ltd.), Sanphenone (Sun Chemical Co., Ltd.), etc. It can be mentioned.

It is preferable that the green tea extract contains tea catechins from the viewpoint that it can be fermented to a good degree and that the desired fermented product is obtained.
The main components of tea catechins are epicatechin (epicatechin, EC) and its hydroxy form, epigallocatechin (epigallocatechin, EGC), and their gallic acid esters, epicatechin gallate (epicatechin gallate, epicatechin gallate, ECG) And epigallocatechin gallate (epigallocatechin gallate, gallic acid epigallocatechin, EGCG), which are known to have contents in the order of EGCG>EGC>ECG> EC.

The amount of tea polyphenol in the green tea extract in the present embodiment is not particularly limited, but the lower limit is preferably 30% by mass or more, more preferably 40% by mass or more in dry matter conversion, and the upper limit is preferably Is 60 mass% or less, more preferably 50 mass% or less.
The amount of tea caffeine in the green tea extract according to the present embodiment is not particularly limited, but the upper limit thereof is preferably 10% by mass or less, more preferably 8% by mass or less, more preferably 6% by dry matter. The lower limit is preferably 1% by mass or more, more preferably 3% by mass or more.

The total amount of catechin in the green tea extract according to the present embodiment is not particularly limited, but the lower limit is preferably 20% by mass or more, more preferably 30% by mass or more in terms of dry matter, and the upper limit is preferably Is 60 mass% or less. Further, the total amount of catechin in the green tea extract is not particularly limited, but is preferably 20 to 60% by mass, more preferably 30 to 50% by mass in terms of dry matter, and the range can be excellent fermented The point and purpose are preferable in terms of efficiently obtaining a fermented product.
The amount of “EGCG” in the green tea extract is not particularly limited, but the lower limit is preferably 5% by mass or more, more preferably 10% by mass or more in terms of dry matter, and the upper limit is Preferably it is 30 mass% or less, More preferably, it is 20 mass% or less. In addition, the amount of "EGCG" in the green tea extract is preferably 10 to 30% by mass, more preferably 10 to 20% by mass, and the range can be good point fermentation and the target fermented product can be efficiently obtained. It is suitable in point.
The ratio of EGCG to the total catechin amount is preferably 20 to 60% by mass, and generally said to be about 30 to 60% by mass.
The form of the green tea extract used in the present embodiment is not particularly limited, and may be any of solid (powdery, granular and the like), semi-solid and liquid.

<麹>
The koji used in the present embodiment is not particularly limited, but koji containing koji mold (e.g., rice bran system, bean curd system, wheat straw system, etc.) is preferable in that the target fermented product can be efficiently obtained.
The koji can be used as a seed koji for producing a product such as sake, miso, soy sauce, shochu and the like by fermenting food using grains such as rice, soybean and wheat as raw materials. In general, Aspergillus niger is used in rice bran, which decomposes starch and proteins contained in cereals and grows using glucose and amino acids produced as nutrient sources.
The said koji is a point which a target fermented matter tends to exhibit various effects by what contains the koji mold (rice koji-type koji mold and / or the wheat koji-type koji fungus) which can be used as a rice koji and / or a wheat koji And is preferred.
Commercially available seeds can be used as the koji. For example, as a seed of commercial products, white birch snow komachi (8043) (Akita Inno Shoten), small yellow granular (8143) (Akita Inno Shoten), a seed of white persimmon (white bean sprout) (Hiroguchi Matsunosuke Shoten), Seeds of persimmon (yellow bean sprouts) (Hiroguchi Matsunosuke Shoten), long white fungus (manufactured by Ryohoku Co., Ltd.), improved long white fungus (manufactured by Ryohoku Co., Ltd.), seed oil for soy sauce (Biok Co., Ltd.), etc. And one or more selected from these.

The cocoons include microorganisms belonging to the genus Aspergillus (Aspergillus) (so-called Aspergillus spp.), Which are, for example, Aspergillus awamori, Aspergillus awamori ver. Aspergillus sojae, Aspergillus luchuensis, Aspergillus saitoi, Aspergillus usami and the like can be mentioned, and one or more species can be selected from these.
Among them, preferably, the Aspergillus fermentable Aspergillus oryzae can be used as a seed koji from the viewpoint of efficiently obtaining a target fermented product, and examples of Koji fungus used for the seed koji include, for example, Kawachi koji (Aspergillus kawachii), Oryzae (Aspergillus) oryzae), Aspergillus sojae, and Awamori (Aspergillus awamori), etc., and one or more species can be selected from these.
Persimmons have long been used in the traditional manufacturing industry of Japan, but there are liquid persimmons and solid persimmons in persimmon, and it is common to use solid persimmon when processing cereals. In the present embodiment, as shown in the after-mentioned Examples (examples), a solid koji seed meal such as a commercial product can be used for liquid culture, and furthermore, the koji fermented product of the present embodiment is obtained by this liquid culture. What I was able to do was quite surprising. Thereby, the mass production of the fermented mulberry product of this embodiment also becomes easier by liquid culture using a seed meal.

[Koji fermentation process]
The persimmon fermentation step of the present embodiment is preferably carried out by culturing persimmon in a medium containing a green tea extract, from the viewpoint that good persimmon fermentation can be performed and a target fermented substance can be efficiently obtained.
The medium containing the green tea extract used in the present embodiment can be prepared by mixing the basic medium composition and the green tea extract.
Further, the medium containing the green tea extract of the present embodiment is preferably a liquid medium, and is preferable in that it can be fermented in a favorable manner and can efficiently produce the target fermented product, and can easily recover the fermented product. .

The basic medium composition is not particularly limited as long as it is a medium composition capable of fermenting koji, and inorganic salts, carbon sources (preferably sugars), nitrogen sources and the like used as nutrient sources of common koji are used. Good.
Examples of the saccharide include scroll, glucose, lactose, mannose, mannitol, sorbitol and the like, and one or more of them can be selected therefrom. The content of saccharides in the culture medium of the present embodiment is not particularly limited as saccharides, but is preferably 0.1 to 2% by mass in terms of good fermentability to obtain an objective fermented substance.
Examples of the inorganic salt include sodium nitrate, sodium nitrite, potassium phosphate, potassium phosphate, magnesium phosphate, magnesium sulfate, sodium chloride, potassium chloride, ferrous sulfate, manganese sulfate, copper sulfate And calcium carbonate, and one or more of them can be selected.

The content of the green tea extract in the medium used in the present embodiment is not particularly limited, but the lower limit thereof is preferably 0.1% by mass or more, more preferably 0.5% by mass or more, and the upper limit thereof is Preferably it is 40 mass% or less, More preferably, it is 30 mass% or less.
The content of the green tea extract in the culture medium of the present embodiment is preferably 1 to 20% by mass, more preferably 1 to 10% by mass, and the range is that it can be fermented to a good degree and the target fermented product is obtained. And is preferred.

The total amount of green tea catechins and the EGCG content in the medium of the present embodiment are not particularly limited, and can be adjusted with the green tea extract used in the medium and the amount used.
The total amount of green tea catechins in the culture medium of the present embodiment is preferably 0.1 to 5% by mass, and the range is preferable in that it can be fermented to a good degree and the target fermented product is obtained.
The EGCG content in the culture medium of the present embodiment is preferably 0.05 to 2.5% by mass, and the range is preferable in that it can be fermented to a good degree and the target fermented material is obtained.

The persimmon is added to the culture medium containing the green tea extract, and persimmon fermentation is carried out, but the amount of persimmon added at this time is not particularly limited, but the lower limit is preferably 0.001% by mass or more, the upper limit Is preferably added to the culture medium to 1% by mass or less. It is preferable to add koji to the culture medium to more preferably 0.005 to 0.5% by mass, and even more preferably 0.01 to 0.1% by mass.

As the fermentation conditions in the present embodiment, aerobic culture conditions are preferable in that they can be fermented in a favorable manner and can obtain a desired fermented substance. It is preferable to carry out aerobic culture using means such as shaking, aeration, stirring alone or in combination.
Furthermore, the culture temperature in the present embodiment is not particularly limited, and is preferably 10 to 40 ° C., more preferably 20 to 30 ° C.
The pH of the culture medium at the time of culture in this embodiment is preferably 4.0 to 8.0, more preferably 5.0 to 7.0, and still more preferably 5.5 to 6.5. .
The culture period in the present embodiment is not particularly limited, and is preferably 5 days or more, more preferably 10 to 30 days, and still more preferably 10 to 20 days.

The culture form in the present embodiment is not particularly limited, and may be continuous culture (perfusion culture etc.) or batch culture. The production state of the fermented mulberry during culture in this embodiment can be confirmed by HPLC analysis as shown in the following Example, whereby the culture period, addition of medium components, and recovery of fermented product can be performed. The timing or the like may be set.

As described above, the fermented persimmon product from the green tea extract of the present embodiment is produced in the culture medium.
In addition, the processing method of the koji of the koji fermented substance obtained by koji fermentation can be performed according to the treatment method of microorganisms, such as well-known fungi. For example, sterilization and sterilization of a rice cake by heating and / or filtration etc. are mentioned. When a seed meal or the like is used, the seed meal may be contained as it is because it has high safety, but in terms of changes in flavor and the like, it is desirable to sterilize or eliminate bacteria. As the sterilization, a filtration treatment for removing common fungi may be performed, and examples thereof include a treatment with a filter aid (activated clay, diatomaceous earth, etc.) or a membrane.
Moreover, the preparation method of the fermentation component in the obtained fermented persimmon can be performed according to the well-known isolation | separation purification method.

2. Green Tea Extract-Derived Fermented Persimmon Fermented Product According to the production method of the present embodiment, a fermented persimmon product derived from the green tea extract of the present embodiment can be obtained.
The fermented persimmon product derived from the green tea extract of the present embodiment is a novel persimmon fermented product in which catechins and teadenols are not detected as in the below-mentioned Examples and is not found in conventional green tea derived products. .
Although the component contained in the fermented green tea extract derived from the green tea extract of the present embodiment is currently under analysis, although the catechins and teadenol are not detected yet, as described later, It is considered that the koji fermentation produces and contains a novel component, since various effects are exhibited.

The fermented persimmon product manufactured by the manufacturing method of this embodiment has one peak with a relative retention time of 13 to 14 minutes in HPLC analysis of reverse phase column chromatography measured in the following Example. It is more preferred that this peak be the highest peak. By using this peak as an index, it is possible to determine the production state of the fermented koji product, and it is possible to enhance the recovery efficiency of the target peak component.

Furthermore, the content of catechins contained in the fermented green tea extract derived from green tea extract of the present embodiment is preferably 0.1% by mass or less, more preferably 0.05% by mass or less. It is preferable at the point which can exhibit the effect which does not originate from.
Furthermore, the content of teadenol A and / or B in the fermented green tea extract derived from the green tea extract of the present embodiment is preferably 0.1% by mass or less, more preferably 0.05% by mass or less, still more preferably 0 It is preferable that it is not more than .001% by mass because it can exert effects not derived from teadenol.

The fermented persimmon product derived from the green tea extract of the present embodiment is expected to further enhance various effects by combined use with known efficacy components (for example, catechins and / or teadenol etc.) and efficacy components to be found in the future. it can.

And, as shown in the following [Examples], the fermented persimmon product derived from the green tea extract of the present embodiment has an adiponectin production increasing action; adiponectin expression promoting action; adipocyte degrading enzyme ATGL expression promoting action; fat with little fat accumulation Inducing differentiation into cells; reducing the expression of transcription factor C / EBPα and PPARγ; suppressing the increase in fat accumulation due to fat cell hypertrophy; having a vascular endothelial wound healing effect; suppressing fatty acid synthetase (FAS) expression.

Furthermore, the increase in adiponectin production has the following effects: (1) action to promote glucose uptake without insulin receptor mediated action; (2) action to decrease fatty acid in cells to sensitize insulin receptor; (3) liver Enhancement of insulin sensitivity by activating AMP kinase; (4) Burning action of fatty acid; (5) Atherosclerosis inhibitory action; (6) Anti-inflammatory action; (7) Hyaluronan synthesis promoting action in dermis is observed It is generally known.
Therefore, since the fermented product derived from the green tea extract of the present embodiment has an adiponectin production increasing action, it has a blood sugar level suppressing action, an antidiabetic action, a fat reducing action, an antiobesity action, a slimming action, an antihyperlipidemic action, It can exert anti-inflammatory and skin-refining effects.

In addition, since the fermented product derived from the green tea extract of the present embodiment has a lipolytic enzyme ATGL expression promoting action and a fatty acid synthetase (FAS) expression inhibiting action, lipolysis promoting action, fat reducing action, anti-obesity action ( In particular, it can exert the action of decomposing fat attached to the body.
The fermented product derived from the green tea extract of the present embodiment has an action to induce differentiation into fat cells with little fat accumulation; an action to reduce expression of transcription factors C / EBPα and PPARγ; and an action to suppress an increase in fat accumulation due to fat cell hypertrophy Therefore, it can exert a fat accumulation reducing action, a fat reducing action, an anti-obesity action (in particular, an action of suppressing fat accumulation on the body).

In addition, since the fermented product derived from the green tea extract of the present embodiment has a vascular endothelial wound healing action, it can exert antiarteriosclerosis.
In addition, the fermented product derived from the green tea extract according to the present embodiment has an action to suppress the decrease in expression of Catalase, which is an antioxidant enzyme, and Hemeoxygenase-1 (HO-1), which is an antioxidant enzyme, and an enzyme involved in active oxygen production. It has the effect of suppressing the expression level of NOX4 and the inflammatory factor TNFα. This can exert a reducing effect on bad oxidative stress in the body.

Thus, since the fermented persimmon product derived from the green tea extract according to the present embodiment has various actions as described above (hereinafter, also referred to as “action to increase adiponectin production” or “action of persimmon fermented product”), It can be used for prevention, improvement, or treatment of obesity, diabetes, arteriosclerosis, hyperlipidemia (hyperlipidemia), high blood pressure, skin care, skin care for skin care or maintenance, improvement of oxidative stress Or it can be used for reduction. Furthermore, the fermented persimmon product derived from the green tea extract of the present embodiment is effective for patients having these diseases or conditions or a preparation thereof.
In addition, as shown in Examples described later, the fermented persimmon product derived from the green tea extract of the present embodiment has an advantage that excellent effects can be obtained even with a simple dosage form such as oral intake and oral administration. Furthermore, since the fermented persimmon product derived from the green tea extract of the present embodiment can appropriately select the state of powder or liquid, etc., it can be conveniently used for ingestion or oral administration also in food and drink, medicine, feed and the like. It is also easy to provide food, drink, etc. containing the fermented persimmon product derived from the green tea extract of the present embodiment in the form of liquid, solid, tablet, jelly or the like. The persimmon fermented product derived from the green tea extract of the present embodiment has an advantage of being easy to exert an action on an adult or a mature animal.

In addition, this embodiment may be used for therapeutic purpose or non-therapeutic purpose.
"Non-therapeutic purpose" is a concept that does not include medical practice, that is, treatment of the human body by treatment. For example, health promotion, beauty practice, etc. may be mentioned.
"Amelioration" refers to the amelioration of a disease, condition or condition; prevention or delay of deterioration of a disease, condition or condition; reversal, prevention or delay of progression of a disease or condition.
"Prevention" refers to preventing or delaying the onset of a disease or condition in a subject of application, or reducing the risk of a disease or condition of interest.

Therefore, the fermented persimmon product derived from the green tea extract of the present embodiment is a preparation or composition intended to use the above-described action such as an adiponectin production increasing agent or a composition for increasing adiponectin production (hereinafter referred to as “formulation of the present embodiment Or (also referred to as “compositions”) can be used, and can further be used to produce these formulations or compositions.
The preparation or composition of the present embodiment itself exerts various effects such as the above-described adiponectin production-increasing effect and the like when it is ingested or administered to an animal (for example, a pet or the like) including a human. In addition, the preparation or composition may be for medicine, quasi-drug, for food or drink, or feed for human or non-human animals, or the medicine, quasi-drug, It may be a material or a preparation used by being mixed with food or drink or feed.

Moreover, the said food-drinks make the above-mentioned anti-obesity etc. a concept, and include the food-drinks which indicated that as needed, a functional food, the food for sick persons, and the food for specific health. These food and drink can be distinguished from general food and drink by the function indication.
Furthermore, it is also possible to distinguish it from the conventional food and drink by further blending the fermented persimmon product derived from the green tea extract of the present embodiment into a conventional food and drink, for example, persimmon derived from the green tea extract according to the present embodiment Tea-based products (for example, tea leaves, tea bags, instant teas, tea beverages, tea extract-containing products (confectionery, noodles, etc.) and the like to which a fermented substance is added can be mentioned.

In addition, the dosage form of the above-mentioned medicine (including quasi-drugs) containing the fermented persimmon derived from the green tea extract of the present embodiment is tablet, capsule, granule, powder, syrup, intravenous injection, Intramuscular injection, suppository, inhalant, transdermal absorption agent, eye drop, nasal drop, compress, bop, ointment, lotion, cream, preparation for oral cavity (dentifrice, liquid dentifrice, mouthwash, gum) It may be any of massage cream, ointment for oral cavity, pill tablets, troche, sore throat, and the like. The dosage form may also be oral administration (for internal use) or parenteral administration (for external use, injection).

Moreover, in order to prepare pharmaceutical preparations of such various dosage forms, the persimmon fermented matter derived from the green tea extract of the present embodiment is used alone or in combination with other pharmaceutically acceptable excipients, respectively. Agents, bulking agents, disintegrants, surfactants, lubricants, dispersants, buffers, preservatives, preservatives, flavoring agents, perfumes, coatings, carriers, diluents, medicinal ingredients other than the present invention, etc. It can be prepared. Moreover, among these administration forms, a preferable form is oral administration, and a liquid preparation for oral use can be prepared according to a conventional method by adding a flavoring agent, a buffer, a stabilizer and the like.

In general, the content of the fermented green tea extract-derived fermented substance in the preparation for oral administration is preferably 0.01% by mass or more, more preferably 0.1% by mass or more as a solid content concentration Moreover, preferably it is 20 mass% or less, More preferably, it is 10 mass% or less. Also, it is preferably 0.1 to 10% by mass, more preferably 0.5 to 5% by mass.

The form of the food containing the fermented product derived from the green tea extract of the present embodiment includes soft drinks, tea drinks, tea leaves products, instant teas, coffee drinks, fruit drinks, carbonated drinks, juices, jellies, wafers, Other than foods and drinks such as biscuits, bread, noodles and sausages and various foods such as nutritional foods, there are further mentioned compositions for supplementing nutrition in the same form (tablets, capsules, syrups, etc.) as the orally administered preparation described above. Be

Various forms of food may be the above plants or their extracts alone, or other food materials, solvents, softeners, oils, emulsifiers, preservatives, perfumes, fragrances, stabilizers, coloring agents, antioxidants, moisturizing agents. The agent, the thickener, the active ingredient other than the present invention, etc. can be appropriately combined and prepared.

Generally, the content of the fermented product derived from the green tea extract of the present embodiment in the food or drink or feed is preferably 0.01% by mass or more, more preferably 0.1% by mass or more, as a solid content concentration. Moreover, preferably it is 10 mass% or less, More preferably, it is 5 mass% or less. Also, it is preferably 0.01 to 10% by mass, more preferably 0.5 to 5% by mass.

Further, examples of feeds include feeds for small animals used in rabbits, rats, mice and the like, feeds such as pet food used in dogs, cats and the like, and the like, and can be prepared in the same form as the above food.

The intake of the fermented product derived from the green tea extract of the present embodiment may vary according to the species, weight, sex, age, condition or other factors of the subject. The dose, route, interval of administration, and the amount and interval of intake can be determined as appropriate by those skilled in the art, but generally preferably 1 mg / 60 kg body weight or more, in terms of dry matter per adult, for adults. Preferably, it is 2 mg / 60 kg body weight or more, preferably 1000 mg / 60 kg body weight or less, more preferably 500 mg / 60 kg body weight or less, still more preferably 200 mg / 60 kg body weight or less. Also, it is preferably 1 to 2000 mg / 60 kg body weight, more preferably 2 to 1000 mg / 60 kg body weight, still more preferably 2 to 5000 mg / 60 kg body weight.

The present embodiment is not particularly limited as long as it is a person who needs or desires the administration or intake subject regardless of a sick person or a healthy person, but for example, a person with lack of exercise, obesity, diabetes etc. And middle-aged and elderly people, young and old men and women who care about their skin quality are preferable.
In addition, the present embodiment can be applied not only to obese patients but also to those who are prone to gain weight, those who desire appropriate weight maintenance, and the like.
Moreover, this embodiment is not only patients suffering from diabetes and insulin resistance, but also subjects suffering from diabetes and insulin resistance, for example, subjects having high postprandial hyperglycemia but no abnormality in fasting blood glucose, fasting The present invention can be applied to a subject who is not required to lower blood glucose but is desired to lower postprandial hyperglycemia.
In addition, the present embodiment can be applied to a hypertensive patient or a subject who wants to prevent hypertension or reduce the risk of the onset.
Furthermore, it can be applied to a subject in need of enhanced vascular protection or a subject in need of enhanced vascular protection.

In the present embodiment, the following aspects can also be adopted.
[1] A method for producing a fermented product derived from green tea extract, comprising fermenting the green tea extract with a koji.
Preferably, the koji contains a microorganism belonging to the genus Aspergillus, and more preferably, it is a rice bran-based yeast and / or a barley-based yeast.
Preferably, the green tea extract is 1 to 30% by mass in the liquid medium. More preferably, the total amount of green tea catechins is 0.1 to 5% by mass and / or the content of EGCG is 0.05 to 2.5% by mass in the liquid medium.
[2] A fermented koji product derived from green tea extract. Preferably, the fermented koji fermented product is a fermented product derived from a green tea extract obtained by the production method described in the above [1]. It is preferable that the subject of the fermented salmon product is a healthy person or a healthy animal, an adult or a mature animal.
[3] A composition or preparation containing the fermented persimmon product according to [2] above as an active ingredient. Preferably, it is a pharmaceutical composition, a food and drink composition, a feed composition or a preparation.
[4] The composition is a composition for promoting adiponectin production, a composition for preventing, ameliorating or treating obesity, a skin care composition, or a composition for preventing, ameliorating or treating arteriosclerosis; The composition or preparation according to the above [3], which is a composition for suppressing fatty acid synthetase (FAS) expression.
[5] The composition is for increasing adiponectin production; for promoting adiponectin expression; for promoting adipocyte-degrading enzyme ATGL expression; for inducing differentiation into adipocytes with little fat accumulation; for reducing expression of transcription factors C / EBPα and PPARγ The composition or the preparation according to the above [4], which is a composition having a fatty acid synthetase (FAS) expression suppressing action, for suppressing an increase in fat accumulation due to fat cell hypertrophy; for vascular endothelial wound healing;

[6] for promoting adiponectin production; for preventing, ameliorating or treating obesity; for skin care; or for preventing, ameliorating or treating arteriosclerosis; or derived from a green tea extract having a fatty acid synthetase (FAS) expression suppressing action Fermented salmon. Preferably, it is a fermented persimmon derived from a green tea extract obtained by the production method described in the above [1].
[7] Adiponectin production increase; Adiponectin expression promotion; Adipolytic enzyme ATGL expression promotion; For differentiation into adipocytes with little fat accumulation; For expression reduction of transcription factor C / EBPα and PPARγ; Adipocyte hypertrophy Or for suppressing endothelium fat accumulation increase; or for vascular endothelial wound healing; or for a fatty acid synthetase (FAS) expression inhibitory action, a persimmon fermented product derived from green tea extract.

[8] Use for medical purpose or non-medical purpose for any of the following applications of the fermented persimmon product derived from the green tea extract according to the above [2].
Adiponectin production promotion; prevention, amelioration or treatment of obesity; beautiful skin; or prevention, amelioration or treatment of arteriosclerosis; adiponectin expression promotion; adipocyte decomposing enzyme ATGL expression promotion; Decreased expression of C / EBPα and PPARγ; suppression of fat accumulation increase due to adipocyte hypertrophy; vascular endothelial wound healing; or suppression of fatty acid synthetase (FAS) expression.
Preferably, it is a fermented persimmon derived from a green tea extract obtained by the production method described in the above [1].
[9] Use of a fermented persimmon derived from green tea extract for the production of the composition or preparation of the above [3] or [4].

Hereinafter, the present invention will be described in more detail based on examples. The embodiments described below show an example of a representative embodiment of the present invention, and the present invention is not limited to the following embodiment.

<Method of Producing Fermented Persimmon Fermented from Green Tea Extract>
1. Green tea extract As green tea extract, green tea extract BL (trade name: Access One Inc.) (green tea polyphenol 46.5% by mass, total amount of green tea catechins 40.11% by mass, EGCG content 13.6% by mass, green tea caffeine 5.2 The thing of mass%) was used.
Moreover, in the specification of green tea extract BL, the water content is 5% or less by the Karl Fischer method. Tea polyphenols can be measured by iron tartrate colorimetric method, and total amount of green tea catechins, EGCG content and green tea caffeine can be measured by HPLC method.

2. Culturing method 100 mL of a liquid medium having the medium composition shown in Table 1 was dispensed in advance into a 500 mL shouldered flask which was dry heat-sterilized and subjected to dry heat sterilization, and autoclave sterilization was performed according to a conventional method. After cooling to room temperature, 1 cup of various types of seed meal (Table 2, 20 to 60 mg) was inoculated directly with a spatula. After inoculation, the cells were cultured at 25 ° C. and shaking at 120 oscillations / min for 17 days.
After the culture, cells were removed by suction filtration with filter paper (Advantec No. 2), and used as a culture supernatant in subsequent experiments of the physiological activity test.

3. HPLC Analysis Conditions The culture supernatant was subjected to component analysis by HPLC analysis of reverse phase column chromatography after filtration through a 0.22 μm disk filter. The analysis conditions are as follows.
<Analytical conditions>
Column: Reversed phase column: TSK gel ODS-80Ts (5 μm. 4.6 × 250 mm)
Mobile phase: (A) 1.0% acetic acid
(B) Acetonitril
Gradient: A: B = 90: 10 (0 min) → 20: 80 (45 min)
Flow rate: 0.6 ml / min
Detection: UV 280 nm
Injection volume: 20 μL
Standard product 1: catechin standard product mixture Epigallocatechin (EGC), epicatechin (EC), epigallocatechin gallate (EGCG), epicatechin gallate (ECG) each 0.025 mg / mL
Standard product 2: Teadenol A 0.025 mg / mL

Regardless of which type of seed meal is used, catechins and other green tea components found in the culture medium at the early stage of culture are almost completely abolished by the culture, and only compounds showing a peak at a relative retention time of 13 to 14 minutes are detected. The It is unclear whether this peak is a single compound or a plurality of compounds, and is under analysis. The peak of catechins and teadenol can not be detected at 0.01 mg / mL or less in the culture solution after koji fermentation by using any seed koji, and the height of this 13 to 14 minute peak is 1.00. And the catechins and teadenols were less than 0.005.
Also, the polyphenol content of the uninoculated medium and the supernatant of each culture was measured as the polyphenol content by the Folin-Ciocalteu method using gallic acid as a standard. In each supernatant after cultivation, the amount of polyphenols was significantly reduced as in HPLC analysis, and the amount of polyphenols was significantly reduced, and was at least 13% or less when the amount of polyphenols in the uninoculated medium was 100%.
In addition, FIG. 1 shows the analysis results of Production Example 3 (white bean sprout) (FIG. 1D) and Production Example 5 (long white bacteria) (FIG. 1E) of representative culture supernatants (the vertical axis of the chromatogram is identical). scale).
Production Example 3 (White Sprout) and Production Example 5 (Long White Fungus) are used as rice bran and wheat straw-based seed meal.
Further, FIG. 1A shows the analysis results of the catechins of standard product 1, FIG. 1B shows the green tea extract (before fermentation), and FIG. 1C shows the analysis results of teadenol A of standard product 2.

<Experimental method of physiological activity>
1. Wound healing experiment Pig-derived vascular endothelial cells were seeded at 3 × 10 ⁵ cells / dish in a 3.5 cm dish and allowed to adhere for 7 hours. After synchronized culture for 17 hours in a medium containing 1% FBS, Hydroxytyrosol was added at a final concentration of 50 μM and a fermentation product at a final concentration of 1/1000 dilution. After compound addition, the bottom of the dish was artificially scratched using a tip of a 200 μL tip, and observed and photographed under a microscope. After 24 hours, the photographs were taken again, and the degree of wound repair was assessed by counting the number of cells present within the range of the wound created initially. Photographing performed three views per dish.

2.3T3-L1 cell culture and addition of compound High-glucose DMEM was used to culture 3T3-L1 cells, and the cells were cultured at 37 ° C. in a 5% CO ₂ environment. The cells were cultured until reaching confluence with DMEM containing 10% calf serum, and after 2 days, they were cultured for 2 days by changing to differentiation induction medium (Insulin, DEX, IBMX, DMEM containing 10% FBS). Then, the culture was continued in the presence of insulin and 10% FBS, repeating the medium exchange every two days. The cells around day 12 after induction of differentiation were used as mature adipocytes, and the cells around day 18 as a model for hypertrophic fat cells. In the experiments to evaluate the effect of compounds on the differentiation of 3T3-L1 cells into mature adipocytes, the compounds were added with insulin two days after conversion to differentiation-inducing medium. In the experiment to observe the effect of the compound on mature adipocytes, the compound was added to the mature adipocytes after 12 days and cultured for 24 hours. Furthermore, in the experiment to examine the effect on the change to hypertrophied adipocytes, the compound was added to the cells after 12 days and cultured for 6 days while repeating the medium exchange in the presence of the compound. After the above treatment, Oil Red O staining and Western blot analysis were performed.

3. Oil Red O Stain Oil Red O was dissolved at a concentration of 1 g / 200 mL (99% isopropanol) to make a stock solution and diluted to 60% with water at the time of use. The cells after each stimulation were washed 3 times with PBS (−), fixed with a 10% formalin solution for 30 minutes. After washing three times with water, 60% isopropanol was added and allowed to stand for 5 minutes, and then oil red O solution was added to stain the lipid. After 20 minutes, it was again washed three times with water, and DMSO was added to extract the oil red O dye, which was collected in a 1.5 mL tube. The supernatant was collected after centrifugation for 10 minutes at 10,000 rpm and quantified by measuring the absorbance at 540 nm.

4. Preparation of total lysate After each cell culture, the cells were washed twice with PBS (-) and immediately stored at -80 ° C. The cells are then thawed on ice and Lysis buffer (50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM EDTA, 50% Glycerol, 100 mM NaF, 10 mM Sodium pyrophosphate, 1% Triton X-100, 1 mM NaVO4, 150 μL of 1 mM PMSF, 10 μg / mL Antipain, 10 μg / mL Leupeptin, 10 μg / mL Aprotinin) was added and allowed to stand for 5 minutes, and the cells were scraped with a scraper and collected in a 1.5 mL tube. These were allowed to stand on ice for 15 minutes, centrifuged at 14,000 rpm for 15 minutes, and the supernatant was recovered.

5. Electrophoresis and antigen-antibody reaction The protein concentration of the collected sample was measured using BCA Protein assay kit (Pierce), and proteins corresponding to 20 μg were subjected to 7.5%, 10%, 12.5%, 15% acrylamide gel SDS-PAGE. did. After electrophoresis, it was transferred to a PVDF membrane (Millipore), blocked with 5% BSA / TBS-T (10 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20), and reacted with various primary antibodies. After washing the membrane for 15 minutes with TBS-T, an antigen-antibody reaction was carried out using a secondary antibody that recognizes each primary antibody. The membrane is again washed with TBS-T for 15 minutes, and Chemi-lumione (Nacalai Tesque) or Clarity Max Western ECL Substrate (BIO-RAD) is used for band detection, and chemiluminescence by Hose radish peroxidase (HRP) is C -Detected by DiGit Blot Scanner (LI-COR). 5% BSA / TBS-T was used for antibody dilution. The dilution ratio, reaction temperature and time were in accordance with the attached protocol of each antibody.

<Results of physiological activity test>
<Adiponectin expression level increase confirmation test>
As described above, the results obtained using each fermented koji product derived from green tea extract fermented by various koji in Table 2 are shown below.
Fermented fish products of Preparation Examples 1 to 6 were added at a final concentration of 1/1000 dilution to mature adipocytes, cultured for 24 hours, then cell lysate was prepared, and the protein expression level of adiponectin was quantified by Western blot method . Cont indicates a control without addition of compound. Α-Tubulin was used as an internal standard for Western blot analysis.
The green tea extract-derived fermented rattan fermented products (Production Examples 1 to 6) in which the green tea extract was fermented with Koji type all increased the expression level of adiponectin in mature adipocytes.

<Wound healing promotion effect confirmation test of vascular endothelial cells>
Production Example 5: Pig-derived vascular endothelial cells were diluted with a final concentration of 1/1000 of a koji fermentation product of long white fungus (rice bran / wheat bran system), and the wound healing effect was evaluated at 24 hours. Cont shows the control which does not add a compound.
A fermented mulberry product (Production Example 5: A long white fungus (rice bran / wheat bran system)) exhibited a wound healing promoting effect of vascular endothelial cells.

<Test to confirm adiponectin expression level increase: Before and after fermentation>
A fermented product (Production Example 3: white bean sprout (rice bran and wheat bran system)) and its pre-fermented product are added to mature adipocytes at a final concentration of 1/1000 dilution, and after culturing for 24 hours, cell lysate is prepared The protein expression level of adiponectin was quantified by the Western blot method. Cont shows the control which does not add a compound. Α-Tubulin was used as an internal standard for Western blot analysis. The results are shown in FIG.
The fermented rattan product (Production example 3: white bean sprout (rice bran and wheat bran system)) increased the expression level of adiponectin, while the tea leaf extract before fermentation is adiponectin under the same conditions (1/1000 dilution). There was no effect of increasing the expression level of This means that a compound newly produced from catechins by microorganism-controlled fermentation has a stronger adiponectin producing action than catechins.

<Enzyme ATGL expression increase confirmation test>
To the mature adipocytes, fermented koji fermentation products (Production Example 3: white bean sprouts (rice bran and wheat bran), Production Example 5: long-white fungus (rice bran and wheat straw) at a final concentration of 1/1000 dilution, After culturing for 24 hours, cell lysate was prepared, and the protein expression amount of enzyme ATGL involved in lipolysis was quantified by Western blot method. Cont shows the control which does not add a compound. Α-Tubulin was used as an internal standard for Western blot analysis.
Fermented fermented koji (Production example 3: White bean sprout (rice bran and wheat straw type), Production example 5: Long white fungus (rice bran, wheat straw type) increases expression of the enzyme ATGL involved in lipolysis of fat cells Teadenol A (0.3 μM) did not show its effect while showing a trend.

<Confirmation induction test to fat cells with little fat accumulation>
8 days after induction of differentiation into fat precursor cells, fermented rattan products (Production example 3: white bean sprout (rice bran and wheat straw type), production example 5: long white fungus (rice bran and wheat straw type)) final concentration 1 / A 1000 dilution, Teadenol A was added to a final concentration of 0.3 μM, and the fat accumulated in the cells was quantified by staining with Oil Red O. Cont shows the control which does not add a compound.
Fermented fermented persimmon (Production example 3: white bean sprout (rice bran and wheat straw type), Production example 5: Long white fungus (rice bran and wheat straw type)) is used to differentiate 3T3-L1 cells into mature adipocytes. It suppressed fat accumulation. This result means that, when fat precursor cells differentiate into fat cells in the body, the fermentation product may produce fat cells with less fat accumulation.

<Test for increased expression of transcription factor C / EBPα and PPARγ>
Two days after the induction of differentiation induction, fermented fish products (Production Example 3: White bean sprouts (rice bran and wheat straw type), Production Example 5: long white fungus (rice bran and wheat straw type)) were added to fat precursor cells at a final concentration of 1 The cell lysate was prepared after 8 days of culture, and protein expression levels of transcription factors C / EBPα and PPARγ involved in fat differentiation were quantified by Western blot method. Cont shows the control which does not add a compound. Α-Tubulin was used as an internal standard for Western blot analysis.
Fermented product of persimmon (Production example 3: White bean sprout (rice bran and wheat straw type), Production example 5: Long white fungus (rice bran and wheat straw type)) is involved in differentiation from 3T3-L1 cells to mature adipocytes Decreases the expression of factor C / EBPα and PPARγ. This result supports the result that the fermentation product partially suppresses fat accumulation in the process of differentiation from 3T3-L1 cells to mature adipocytes.

<Confirmation test of fat accumulation increase by adipocyte hypertrophy>
Fermented product of koji (Production example 3: white bean sprout (rice bran / wheat straw type), production example 5: long white fungus (rice bran / wheat rod type)) at a final concentration of 1/1000 in mature adipocytes 12 days after the initiation of differentiation Fat added at dilution and accumulated after 6 days of culture was quantified by staining with oil red O. Each Cont on Day 12 and Day 18 indicates a control without adding a compound.
Continued culture of mature adipocytes greatly increased the amount of fat in the cells. The increase was partially suppressed by the koji fermented product (Production Example 3: white bean sprout (rice bran and wheat straw type), Production Example 5: long white fungus (rice bran and wheat straw type)). This result indicates that the fermented product may suppress further increase of fat accumulation due to fat cell hypertrophy in the body.

<Test for expression level of adiponectin and lipolytic enzyme ATGL>
Fermented product of koji (Production example 3: white bean sprout (rice bran / wheat straw type), production example 5: long white fungus (rice bran / wheat rod type)) at a final concentration of 1/1000 in mature adipocytes 12 days after the initiation of differentiation The dilution was added, and after 6 days of culture, the intracellularly accumulated fat was stained and quantified using Oil Red O. Each Cont on Day 12 and Day 18 indicates a control without adding a compound.
By continuing the culture of mature adipocytes, the expression levels of adiponectin and lipolytic enzyme ATGL in cells were greatly reduced. The decrease was partially suppressed by the koji fermented product (Production Example 3: White bean sprouts (rice bran and wheat straw type), Production Example 5: Long-white fungus (rice bran and wheat straw type)). This result indicates that the fermented salmon product may improve the decrease in the production of adiponectin, which is an extremely good hormone, by fat cell hypertrophy in conjunction with obesity, and the decrease in lipolytic ability by the decrease in ATGL production.

<A test to confirm the reduction in the expression levels of the antioxidant enzymes Catalase and Hemeoxygenase-1 (HO-1)>
Fermented product of koji (Production example 3: white bean sprout (rice bran / wheat straw type), production example 5: long white fungus (rice bran / wheat rod type)) at a final concentration of 1/1000 in mature adipocytes 12 days after the initiation of differentiation After adding for dilution and culturing for 6 days, cell lysate was prepared, and protein expression levels of antioxidant enzymes Catalase and Hemeoxygenase-1 (HO-1) were quantified by Western blot method. Each Cont on Day 12 and Day 18 indicates a control without adding a compound. Α-Tubulin was used as an internal standard for Western blot analysis.
By continuing the culture of mature adipocytes, the expression levels of intracellular antioxidant enzymes Catalase and Hemeoxygenase-1 (HO-1) were greatly reduced. The decrease was remarkably improved by the fermented rattan product (Production Example 3: white bean sprout (rice bran and wheat straw type), Production Example 5: long-white fungus (rice bran and wheat straw type)). Adipocyte hypertrophy is an inflammatory response and generates reactive oxygen species. Decreased expression of antioxidant enzymes due to hypertrophy means further increase of reactive oxygen species generated. The ability of the koji fermentation product to improve the reduced expression of antioxidant enzymes leads to the ability to alleviate this bad oxidative stress cycle in the body.

<Confirmation test of increase in expression levels of enzyme NOX4 and inflammatory factor TNFα involved in active oxygen production>
Fermented product of koji (Production example 3: white bean sprout (rice bran / wheat straw type), production example 5: long white fungus (rice bran / wheat rod type)) at a final concentration of 1/1000 in mature adipocytes 12 days after the initiation of differentiation After adding for dilution, and culturing for 6 days, cell lysate was prepared, and the protein expression levels of reactive oxygen producing enzyme NOX4 and inflammatory factor TNFα were quantified by Western blot method. Each Cont on Day 12 and Day 18 indicates a control without adding a compound. Α-Tubulin was used as an internal standard for Western blot analysis.
Continued culture of mature adipocytes greatly increased the expression levels of the enzyme NOX4 and the inflammatory factor TNFα, which are involved in the production of reactive oxygen in cells. The increase in TNFα was suppressed by the koji fermentation product (Production Example 3: White bean sprouts (rice bran and wheat straw type), Production Example 5: Long-white fungus (rice bran and wheat straw type)). In addition, the increase in NOX4 was suppressed by the fermented product of Production Example 2: white bean sprout (rice bran), but the effect was observed for the fermented koji of Long White Fungus (rice bran / wheat straw). I could not find it. The increased expression of TNFα and NOX4 due to hypertrophy is directly linked to the increase of reactive oxygen species, and suppressing the expression means reducing the oxidative stress in the body caused by the fattening of fat cells in conjunction with obesity.

<Mouse administration of fermentation product: Adiponectin expression promoting action of fatty tissue and fatty acid synthetase (FAS) expression suppressing action>
Animal experimentation:
Mature female ICR mice (Japan SLC Co., Ltd.) were divided into a fermentation product administration group (6 animals) and a non-administration group (6 animals). To the administration group, a freeze-dried product (Production Example 5) of a fermentation culture supernatant prepared using a long-white fungus (Hyrodoku) was administered at 10 mg / kg body weight / day for 10 days. The administration was carried out by mixing the powder of the above-mentioned dried product with the powder feed MF powder (Nippon SLC Co., Ltd.) of a normal food and taking it freely. Ten days later, adipose tissue was excised and stored at -80.degree.

Protein preparation from tissue:
Adipose tissue stored at -80 ° C is thawed on ice, then partially excised and tissue Lysis buffer (50 mM HEPES pH 7.5, 50 mM NaCl, 1 mM EDTA, 50 mM NaF, 1 mM NaVO4, 25 mM β The supernatant was recovered by homogenization in -glycerophosphate, 200 .mu.M dithiothreitol, 0.02% Triton X-100) and centrifugation at 14,000 rpm for 30 minutes. The resultant was further centrifuged at 14,000 rpm for 15 minutes, and the collected supernatant was used as a final sample.

Electrophoresis and antigen-antibody reaction:
The protein concentration of the collected sample was measured using a BCA Protein assay kit (Pierce), and 20 μg of the protein was subjected to 10% acrylamide gel SDS-PAGE. After electrophoresis, it was transferred to a PVDF membrane (Millipore), blocked with 5% BSA / TBS-T (10 mM Tris-HCl, 150 mM NaCl, 0.1% Tween 20), and reacted with various primary antibodies. After washing the membrane for 15 minutes with TBS-T, an antigen-antibody reaction was carried out using a secondary antibody that recognizes each primary antibody. The membrane is again washed with TBS-T for 15 minutes, and Chemi-lumione (Nacalai Tesque) or Clarity Max Western ECL Substrate (BIO-RAD) is used for band detection, and chemiluminescence by Hose radish peroxidase (HRP) is C -Detected by DiGit Blot Scanner (LI-COR). 5% BSA / TBS-T was used for antibody dilution. The dilution ratio, reaction temperature and time were in accordance with the attached protocol of each antibody.

·result:
Western blot blots of adiponectin and fatty acid synthetase (FAS) and their quantification are shown in FIG. Β-actin was used as an internal standard for quantification. As evident from the intensity of the band, the expression of adiponectin in the fermentation product-administered group showed a clearly high level compared to the non-administered group. On the other hand, the expression of FAS showed a low value in the fermentation product administration group. From these results, it has become clear that the fermented product has an effect of increasing adiponectin expression not only in cultured adipocytes but also at the individual level. Furthermore, the fermented product is considered to be capable of suppressing lipid synthesis by reducing the expression of FAS.

The green tea extract-derived koji ferment has an action to promote the expression of adiponectin in mouse adipose tissue and an action to suppress the expression of fatty acid synthetase (FAS). And, the fermented persimmon product derived from green tea extract is capable of promoting adiponectin expression promoting action and expression suppressing action of fatty acid synthetase by oral administration, and is obese in intake of a beverage or food or drink, or even in oral administration of a pharmaceutical product or the like. Various symptoms, diseases or conditions such as diabetes, hyperlipidemia, hypertension and the like can be prevented, ameliorated or treated. In addition, the persimmon fermented product derived from green tea extract is very effective for adults with so-called adult diseases or their spares.

Claims

A method for producing a fermented product derived from green tea extract, comprising fermenting the green tea extract with a koji.
Persimmon fermented matter derived from green tea extract.
A composition comprising a fermented persimmon derived from green tea extract as an active ingredient.
The composition according to claim 3, which is a composition for promoting adiponectin production, a composition for preventing, ameliorating or treating obesity, a composition for skin care, or a composition for preventing, ameliorating or treating arteriosclerosis. Composition of