KR20120132208A - Method of preparing fermented liquor using mushroom mycelial rice and seaweeds - Google Patents

Abstract

PURPOSE: A method for preparing fermentation wine using mushroom mycelium rice is provided to ensure various nutrients and to prevent or treat adult diseases such as diabetes or obesity. CONSTITUTION: A method for preparing fermentation wine of mushroom mycelium rice comprises: a step of dipping unpolished rice in warm water of 20-30 Deg.C. and fresh water of 20-40 Deg.C., and repeating the steps for 10-36 hours; a step of adjusting moisture content of rice by 25-50% and sterilizing at 100-140 Deg.C.; a step of sterilizing the rice and inoculating mushroom mycelium to the rice; a step of culturing the rice at 25-30 Deg.C. and drying to prepare mushroom mycelium rice; and a step of dipping and steaming the rice and fermenting. The method further comprises a step of mixing 1-100 parts by weight of dry marine algae based on the 100 parts by weight of the mushroom mycelium rice.

Description

Method of preparing fermented liquor using mushroom mycelial rice and seaweeds}

The present invention relates to a method for producing fermented wine using mushroom mycelium rice, which has excellent flavor and is effective in treating adult diseases such as diabetes or obesity.

Germination of the grain relaxes the binding structure of organic matter, making it soft and easy to digest in the body when eaten. In other words, when germinated, the grains loosen the bond structure that was hard to nourish new life, and as a result, the grain becomes soft. Soaking brown rice in water for more than a day and then softening it is because of the subtle differentiation caused by germination.

In addition, grains germinate to increase the amount of enzymes that break down the nutrients of the seeds themselves to nourish new life, and produce nutrients like never before. Fermentation has been widely used as a natural processing method of food, but germination was not so easy, and it was only enough to germinate barley into malt. However, as in the case of malt, the germination effect of starch is much increased because germination of barley increases the glycation enzymes by activating vitamin B1 in the barley with germination than when barley is used as it is. This malt is called malt in one shot and is used as a medicine to strengthen the stomach and spleen and to digest well.

When sprouting representative germination foods such as bean sprouts and mung bean sprouts, a large amount of ingredients that did not exist before germination are increased.For example, germinated brown rice ingredients sprouted with brown rice are well known such as gamma-originol, vitamins, amino acids, fatty acids, dietary fiber, and SOD. In addition to the ingredients, there are also new ingredients such as arabinoxy, which are attracting the attention of the academic community.

Therefore, germinated grains can be easily digested like fermented foods, increase the activity of enzymes in the body, and obtain various substances that have beneficial effects on the human body. After all, germinating grain is another breakthrough that can maximize the efficacy of food along with fermentation. In addition, by fully utilizing the usefulness of grain, it is an innovative way to heal disease and achieve disease-free longevity.

In general, a mushroom is a fungus that refers to a group of umbrella-shaped lampshades and sacks. These mushrooms are composed of fruit bodies with mycelium (Mycelium) and spores (spore) for reproduction. Compared to general plants, mycelium is root, stem and leaf, and fruit body is flower.

We may think that the freshly shaped fruiting bodies are all mushrooms, but in fact fruiting occurs in a very short period of time, and for the most part of the year, the fluffy, thin thread-like mycelium is white in organic matter such as humus and trees. They are parasitic or by-productive with fungi. The spores are hard and coated so that it is difficult to absorb digestion even if you eat mushrooms, spores germinate in the medium to become the first mycelium, and the compatible hyphae are cellular fusion with each other to become the second mycelia, while ingesting sufficient nutrients. Spread out.

The mycelium contains four times more nutrients than fruiting bodies, proteins, amino acids, vitamins, minerals, and various enzymes, so that the nutrition can be balanced enough to be reported. Especially, mycelium has 50 medicinal ingredients such as anticancer component and immune enhancing component than fruiting body. Because it contains about 60 times more, the mystery of the mushrooms will be hidden in the mycelium.

Conventional method for producing germinated grains or using mushroom mycelium, a method for producing germinated brown rice in the state of rice seed simply (National Patent Publication No. 2003-43495), sterilization of grains inoculated to inoculate mushroom seedlings and cultured grains Method (Domestic Patent Publication No. 2002-69859), Method of producing a cultivated barley cultured by inoculating a mushroom mushroom spawn after mixing and increasing the citrus juice gourd to the grain (barley) No.), a method of immersing the situation mushroom-containing solution in the germination stage of grain (Korean Patent Publication No. 2006-95105), by immersing the rice in the hot water extract by hot water extraction of the situation mushroom containing the biologically active ingredients of the fruiting body of the situation mushroom A method of producing a functional rice (Domestic Patent Publication No. 2003-86174), a function characterized in that the coating liquid mixed with shiitake mushroom concentrate and cyclodextrin (CD) is applied to rice and dried A functional rice obtained by immersing clean rice in Korean rice (Korean Patent Publication No. 2005-0082346) and clean rice, and heat-cleaning rice called boiled rice, containing fruit body extract of roe deer mushroom and heat sterilizing and drying clean rice again. 2003-0044268).

The conventional techniques described above have the disadvantage that the components such as beta glucan are significantly lowered by culturing the mushroom mycelium by using only the nutrients in the grain by culturing the mushroom bacteria in grains that have only germinated or grown in the rice seed state. In addition, the method of immersing the mushroom extract can not extract the active ingredient completely during the extraction of the mushroom extract, the extract is not absorbed well in the grain, there is a big problem that the extract is washed in the washing process even if the grain is coated on the outer wall .

Accordingly, the present applicant controls sterilized grains having no water content in patent application No. 2008-0120516 and then sterilizes to kill off microorganisms that cause off-flavor, and when inoculated with mushroom mycelium, the mushroom mycelium is the outer shell or pores of the grain. Penetrating through the surface of the grains through the decomposition of lignin, cellulose, hemicellulose and other fibers to grow by using a lot of ingredients such as beta glucan was found and filed.

However, examples of preparing alcoholic beverages using rice inoculated with the mushroom mycelium are not yet known.

Accordingly, the present inventors have completed the present invention by discovering that fermented wine can be produced by dipping and increasing the mushroom mycelial rice inoculated with a specific mushroom mycelium as a result of research and efforts to prepare fermented wine using mushroom mycelial rice. .

Therefore, an object of the present invention is to provide a method for producing fermented wine using mushroom mycelium rice.

The present invention

1) immersing the uncooked rice in 20 ~ 30 ℃ hot water for 10 to 30 hours, and repeating the germination and waiting process for 10 to 36 hours in 20 to 40 ℃ fresh water;

2) controlling the water content of the rice to 25 to 50%, sterilizing at 100 to 140 ℃;

3) After sterilization, it is selected from the group consisting of shiitake mushrooms, situation mushrooms, ganoderma lucidum mushrooms, fueryeong, agaricus, roe deer, cordyceps, cloud fingering, spirits, zelkovas, tops, birds, flowers, leaf mushrooms, chaga, longevity mushrooms and willows Inoculating the mushroom mycelium to rice;

4) culturing the rice inoculated mushroom mycelium at 25 ~ 30 ℃ and dried to obtain the mushroom mycelium rice; And

5) immersing and increasing the mushroom mycelial rice fermentation

Characterized in that the manufacturing method of mushroom mycelium rice fermented wine containing.

The fermented wine of the present invention includes a variety of nutrients contained in the mushroom mycelium rice and algae, it is excellent in flavor can meet the preferences of consumers. It can also be widely used for the treatment or prevention of adult diseases such as diabetes and obesity.

1 is a photograph of fermented base liquor used in preparing the fermented liquor of Example 4.
Figure 2 is a photograph of the fermented wine fermented by mixing the base wine and sulcus in Example 4.
Figure 3 is a photograph of the fermented wine fermented by mixing the base wine and sulcus in Example 5.
Figure 4 is a graph measuring the electron donating ability of mushroom mycelial rice and seaweed.
5 is a graph measuring the content of polyphenol compounds in mushroom mycelium rice and seaweed.
6 is a graph confirming the PTP1B inhibitory activity of the situation mushroom mycelium rice.
7 is a graph confirming the PTP1B inhibitory activity of shiitake mushroom mycelium rice.
8 is a graph confirming the PTP1B inhibitory activity of seaweed.
9 is a graph confirming the fat accumulation inhibitory effect of the situation mushroom mycelium rice.
10 is a graph confirming the fat accumulation inhibitory effect of shiitake mushroom mycelium rice.
11 is a graph confirming the fat accumulation inhibitory effect of seaweed.
12 is a graph confirming the CETP inhibitory activity of the situation mushroom mycelium rice.
Figure 13 is a graph confirming the CETP inhibitory activity of shiitake mushroom mycelium rice.
14 is a graph confirming the CETP inhibitory activity of seaweed.

Hereinafter, the present invention will be described in detail.

The present invention is characterized in that the method for producing fermented wine using mushroom mycelium rice.

Mushrooms are ingested and grown by secreting enzymes from starch, glycosides, proteins, and minerals, as well as cell wall components such as cellulose, lignin, hemicellulose, which are the main components of wood and sawdust. In the present invention, by germinating rice that has not been milled, cellulose, lignin, hemicellulose contained in the shell and the shell, and mushroom nutrients can use many nutrients such as amino acids, sugars, vitamins and minerals during the germination process.

The manufacturing process of the mushroom mycelium rice is as follows.

In order to germinate rice which has not been milled, it is immersed in hot water at 20 to 30 ° C. for 10 to 30 hours. To germinate uncooked rice requires moisture and a constant temperature and must be immersed in water for a long time because it is surrounded by a hard shell. If the temperature of the hot water is less than 20 ℃, the germination of the rice is slow, if it exceeds 30 ℃, a variety of bacteria grow, delay the germination and odor occurs. After immersion in the hot water, the immersion and waiting process is repeated for 10 to 36 hours at intervals of 3 hours in fresh water. This is to dilute the contamination of various germs generated during germination.

In addition, it is preferable to apply a pressure of 10 to 20kg / cm ² for 1-2 seconds in order to generate larger voids between the shell and the grain in the germinated rice. This is due to the pressing of the grain, which results in larger grain and shell gaps. If the pores of the shell and the grain is large, the mycelia are easily infiltrated when inoculated with the mushroom mycelium, so that the grain can grow quickly. However, if the pressure of 20kg / cm ² or more, it is difficult to maintain the shape of the rice softened by germination.

When the rice is immersed in water, pores are generated, and in particular, when rice is germinated, the pores become much larger, and microorganisms that cause odors are generated. The microorganisms are killed through sterilization. At this time, after adjusting the rice to maintain the water content of 25 to 50%, it is preferable to sterilize 1-2 hours at 100 ~ 140 ℃. Due to the sterilization process, the physical properties of the rice is improved, water retention, breathability is good, and the nutrient source contained in the rice husk is easy to use for mycelial growth. If the moisture content of the grain is less than 25%, the water is evaporated during the culturing process, the mushroom mycelium is spread evenly as a whole can not grow. In addition, during the sterilization process, the grain containing water expands to push the outer shell and voids occur between the outer shell and the grain. If the moisture content exceeds 50%, the grain rapidly expands during sterilization and the outer shell bursts. The shape of the grains is deformed and hard to be milled, and the grains are entangled with each other, which causes a considerable difficulty in culturing the mycelium.

In addition, when sterilized after adding 1 to 10 parts by weight of rice bran, soy flour or corn flour with respect to 100 parts by weight of the rice, the ability to decompose cellulose, such as cellulose, hemicellulose, lignin, etc. contained in the shell is inferior Mushroom mycelia are supplied with insufficient nutrients through rice bran, soy flour or oak powder to help mycelia adhere to the skin.

After sterilization, mushroom mycelium is inoculated into sterile rice. The mushroom mycelium is selected from the group consisting of shiitake mushrooms, situation mushrooms, ganoderma lucidum mushrooms, fueryeong, agaricus, roe deer, cordyceps, cloud fingering, spirits, zelkova, spinning top, birds, flowers, leaf mushrooms and chaga, preferably shiitake The use of mushrooms, mushrooms or chaga is good for the flavor, fermentation, etc. of fermented wine produced by the later fermentation.

 The mushroom mycelium is preferably incubated for 10 to 20 days after inoculation into sterile liquid medium. When incubating the temperature is preferably in the range of 15 ~ 35 ℃, the growth of the mycelium is slowed out of the range.

After the culturing process, the mushroom mycelium rice is finally dried, preferably, the moisture content is adjusted to 15 to 17%. The drying process is made in a grain dryer attached to the automatic moisture meter, and adjusts the moisture content to 15 to 17%.

Next, immersed and cooked mushroom mycelial rice prepared above to fermentation to produce a fermented wine.

At this time, the fermented liquor can be prepared through a conventional casting method for producing a base liquor by using rice (scattered yeast) prepared by increasing the rice, sowing seed (종), the mushroom mycelium Rice can be used in the manufacture of scoops.

In more detail, first, the semi-process of washing white rice or non-rice in clean water, and the immersion process of sinking the semi-finished rice in order to manufacture the base liquor are used. The purpose is to remove foreign substances such as dust on the surface of the rice and to absorb the proper amount of water.

After going through the immersion as described above will be cooked (증) in a steaming pot. The cooker is a process of making gourd rice, and the purpose of cooker is to make the starch with the strong water vapor of 100 ℃ or higher on the water absorbed rice to facilitate the action of various enzymes. After passing through the semi-mime- steaming process, the rice is cooled at room temperature and is cooled and stored.

On the other hand, the final state refers to the fungus bacteria used as spawn (種 菌) when manufacturing the soup (麴) and usually to grow a specific mold in millet or rapeseed to contain a lot of spores. This final state is to enter the starch raw material such as sweet potato, potato, barley and multiply into artificially increased starch raw material.

Yeast is a group of single-celled organisms that are fungi or mushrooms but do not have mycelia and do not have photosynthesis or motility. They are used to make sake.

Jumo (酒 확대) refers to an expanded culture of yeast for fermentation of base liquor. Base liquor should contain a lot of healthy enzymes and acids (citric acid, lactic acid, etc.). The presence of acid serves to prevent the production of bacteria in the base liquor.

Base liquor can be obtained by mixing fermented rice, sake, and immigration fermented in a jar, etc., which are prepared through the above process, and preferably fermented at 20 to 30 ° C. for 30 to 50 hours. It is good to get it.

On the other hand, the mushroom mycelium rice after the semi-process, the soaking process and the steaming process as described above to produce a sake by cooling, and then mixed with the base sake can be fermented to obtain a final fermented wine. At this time, it is preferable that the fermentation made after mixing with the base liquor proceeds for 20 to 80 hours at 20 to 30 ℃.

In the case of preparing the sulcus using the mushroom mycelial rice, it is more preferable in terms of flavor and functional fermentation of fermented wine by mixing 1 to 100 parts by weight of dried seaweed with respect to 100 parts by weight of mushroom mycelial rice, and more preferably 10 It is preferable to add 50 parts by weight. In addition, the seaweed may be mixed with mushroom mycelial rice powder after drying.

By the seaweed is preferred to use a steam (Porphyra Tenera), seaweed in (Laminaria), wakame (Undaria pinnatifida), Sargassum (Sargassum fulvellum), kkosiraegi (Gracilaria verrucosa), fusiformis (Hizikia fusiforme) or agar (Gelidium amansii) Most preferably, laver ( Porphyra Tenera ) is used.

In addition, the preparation of the sulcus can be fermented by mixing normal white rice with mushroom mycelium rice, fermenting by mixing 10 to 200 parts by weight, more preferably 50 to 150 parts by weight of white rice with respect to 100 parts by weight of mushroom mycelial rice. good.

The fermented liquor finally obtained by the above process is excellent in flavor, but may exhibit antidiabetic and anti-obesity effects due to raw materials.

Hereinafter, the present invention will be described in detail with reference to the following examples, but the present invention is not limited thereto.

Example 1: Preparation of Mushroom Mycelia Grain

After immersing the uncooked rice in water at 25 ℃ for 1 hour, drain it by putting it in a tray, drain it, and adjust the water content to 35% using a grain moisture meter, and then put it in 18 sterilizable 1 liter glass bottles. Put in. The germinated rice was sterilized by maintaining the glass bottle at 121 ° C. for 90 minutes. Then, inoculated each mushroom mycelium of Table 1 in a glass bottle containing sterilized rice and incubated for 20 days at the temperature shown in the table. The mushroom mycelium is obtained by subcultured on the PDA (Potato dextrose agar) medium, the mushroom spawn is distributed from the Applied Microorganisms Department of Agricultural Science and Technology Research Institute, National Institute of Agricultural Science and Technology, Jeonbuk Agricultural Research Institute, Spawn Association, Agricultural Development Research Institute, etc. And it was inoculated into sterile liquid medium containing 0.3% by weight of soybean meal.

Finally, the rice mycelium cultured mushroom mycelium was dried to obtain a mushroom mycelium grain. Table 1 shows the culture state of the mycelia according to the mushroom mycelium.

Different kinds of mushrooms Culture temperature (℃) Outer shell culture Wheat culture state Shiitake mushrooms 15 to 25  + +++ Situation Mushroom 25-30  +++ +++ Gongji mushroom 25-30  +++ +++ Ghost 25-30 - +++ Agaricus 25-30 - +++ Spindle 20 to 28  +++ +++ Cordyceps 25-30 - +++ Fingering 25-30  +++ +++ Nerd 25-30  + +++ top 18-20  + +++ Birds 20 to 25  + +++ Blossom Mushroom 20 to 25 - + Leaf mushroom 20 to 25 - +++ Chaga Mushroom 25-30 - +

※ + + + Sleeping + + Growing + Normal-Not growing

Example 2 Preparation of Germinated Mushroom Mycelium Grains

After immersing the uncooked rice in 25 ℃ water for 24 hours, soaked in 30 ℃ fresh water every 3 hours and withdrawn for 24 hours to allow the rice to germinate.

 The germinated rice was put into a net or net, and the water was removed, and then adjusted to a moisture content of 35% using a grain moisture meter, and then put into a sterilizable 1 liter 18 glass bottle. The germinated rice was sterilized by maintaining the glass bottle at 121 ° C. for 90 minutes. Thereafter, inoculated with the mushroom mycelium cultured in Example 1 in a glass bottle containing sterilized rice and incubated for 20 days. Finally, the rice mycelium cultured mushroom mycelium was dried to obtain a mushroom mycelium grain. Table 2 shows the culture state of the mycelia according to the mushroom mycelium.

Different kinds of mushrooms Culture temperature (℃) Outer shell culture Wheat culture state Shiitake mushrooms 15 to 25  +++ +++ Situation Mushroom 25-30  +++ +++ Gongji mushroom 25-30  +++ +++ Ghost 25-30 - +++ Agaricus 25-30 - +++ Spindle 20 to 28  +++ +++ Cordyceps 25-30 - +++ Fingering 25-30  +++ +++ Nerd 25-30  +++ +++ top 18-20  +++ +++ Birds 20 to 25  +++ +++ Blossom Mushroom 20 to 25 - + Leaf mushroom 20 to 25 + +++ Chaga Mushroom 25-30 + +

※ + + + Sleeping + + Growing + Normal-Not growing

Experimental Example 1 beta glucan content of mushroom mycelium grains

The content of the beta glucan in the mushroom mycelium grains obtained by the method of the above example and the 9-minute sea bream and pottery by-products was measured using the situation mushroom mycelium. In addition, after immersing brown rice in water for 8 hours, sterilizing for 30 minutes at 121 ℃, inoculated with the situation mushroom mycelium and incubated for 10 to 20 days at 25 ℃ cultured brown rice and the content of beta glucan in the situation mushroom cultured in oak Was measured and compared, and the results are shown in Table 3 below.

division Beta Glucan Content (% by weight) Brown rice cultured for 10 days 0 Brown rice cultured for 20 days 2.7 Situation Mushroom (Oak Culture) 13.5 Situation mushroom mycelium grains (Example 1) 17.8 Situation mushroom mycelium grains (Example 2) 18.3 Mill By-Products (Example 1) 25.2 Mill By-Products (Example 2) 27.5

Example 3 Influence of Mycelial Growth with Additives

When sterilized in Example 2, 5 mg by weight of rice bran, soy flour or oak powder was added to 100 parts by weight of the germinated grains, and the mycelium culture state of the rice hull was shown in Table 4 below.

Different kinds of mushrooms Not added Rice bran 5% 5% soy flour Soybean flour 5% Shiitake mushrooms +++ ++++ ++++ + Situation Mushroom +++ ++++ ++++ +++ Gongji mushroom +++ ++++ ++++ +++ Spindle +++ ++++ ++++ +++ Fingering +++ ++++ ++++ +++ Nerd +++ ++++ ++++ +++ top +++ ++++ ++++ +++ Birds +++ ++++ ++++ +++ Ghost - +++ +++ +++ Agaricus - +++ +++ +++ Cordyceps - + +++ + Blossom Mushroom - + + + Leaf mushroom + +++ +++ + Chaga Mushroom + + + +

※ + + + Sleeping + + Growing + Normal-Not growing

Example 4 Preparation of Fermented Wine Using Mushroom Mycelial Rice

First, 16 kg of non-glutinous rice was washed in clean water, soaked for 8 hours, and then submerged. Then, steam was absorbed in a steamer and steamed with strong steam of 100 ° C. or more. Thereafter, cooling was carried out at room temperature to obtain a rice cake.

The starch raw material was increased and propagated to artificially increased starch raw material to obtain immigration, and the yeast was expanded and cultured to obtain a hair seed.

Thereafter, the jar was fertilized with the main hair, fermented at 25 ° C. for 36 hours, and cooled to prepare a base liquor.

Meanwhile, 16 kg of the situation mushroom mycelial rice prepared in Example 2 was washed in clean water, soaked for 8 hours, and then submerged, and then steamed with strong steam of 100 ° C. or more in rice absorbed with water in a steaming pot. It was prepared by cooling at room temperature. Thereafter, the prepared base liquor and the sulcus were mixed, and then put in a jar and fermented at 25 ° C. for 48 hours to obtain a fermented liquor.

Example 5 Preparation of Fermented Wine Using Mushroom Mycelial Rice and Seaweed Powder

Fermented liquor was prepared in the same manner as in Example 4, except that 300 g of dried laver powder was added to the situation mushroom mycelium rice at the same time to prepare the sulcus in Example 4.

Experimental Example 2 Analysis of Components of Components Used in Fermented Wine

2-1. General Ingredient Analysis

The general components of the situation mushroom mycelium rice, shiitake mushroom mycelium rice and seaweed of Example 2 used in the production of fermented wine were measured according to the A.O.A.C. method, and are shown in Table 5 below. In other words, moisture is the atmospheric pressure drying method, crude protein is the Kjeldahl method using the automatic nitrogen distillation unit (KJELTEC 2200 SYSTEM, Foss, Sweden), and crude fat is the Soxhlet method using the automatic fat extraction unit (SOXTEC AVANTI 2055 SYSTEM, Foss, Sweden). Inquiry was measured by the dry painting method. Crude carbohydrate was obtained by subtracting 100 from the sum of moisture, crude fat, crude protein and crude ash.

sample Component Analysis (%) moisture Crude fat Crude protein Views min Crude carbohydrate Situation Mushroom
Mycelium Rice 13.2 0.5 8.8 1.1 76.4 Shiitake mushrooms
Mycelium Rice 15.6 1.0 15.1 2.8 65.5 Kim 12.1 4.5 30.2 15.9 37.3

2-2. Electron donating ability

Electron-donating activity (EDA) was measured by modifying the Blois method. That is, add 3 mL of a 1 × 10 ^-4 M 1,1-di-phenyl-2-picrylhydrazyl (DPPH) solution (dissolved in 99.9% methanol) to 0.2 mL of the sample, shake for 10 seconds, and react for 10 minutes. The decrease in absorbance at nm was measured, and the electron donating ability was expressed as a percentage of the absorbance difference between the sample addition and the non-addition as shown in Equation 1, and the results are shown in FIG. 4.

[Equation 1]

2-3. Polyphenol Compound Content

900 μL of distilled water was added to 100 μL of the sample, and 0.2N Folin-Ciocalteu's phenolic reagent was added thereto and mixed. After standing at room temperature for 3 minutes, 1 mL of 1% sodium carbonate solution was added thereto, followed by mixing and shaking. The absorbance was measured at 760 nm after standing at room temperature for 1 hour. In this case, the standard material is treated in the same way as the sample using tannic acid to prepare a standard curve, and then the concentration of the phenolic compound in the sample is determined in consideration of the dilution factor by reading the concentration of the standard substance for absorbance of the sample. It was calculated and the results are shown in FIG.

2-4. Free amino acid measurement

50 mL of 70% ethanol was added to 20 mL of the sample, which was connected to a reflux condenser and heated to reflux at 100 ° C for 1 hour, followed by suction filtration (Whatman No. 3). The filtrate was concentrated under reduced pressure to 2 to 3 mL under 40 ° C. The concentrate and the concentrated water were washed with a small amount of distilled water, transferred to a separatory funnel, and 20 mL of diethyl ether was added to remove the lower layer. I was. The dried sample solution was dissolved in lithium citrate buffer (pH 2.2), and 25 mL was analyzed by an amino acid autoanalyzer (Biochrom 30, Amersham Biosciences Ltd., England).

The analysis results are shown in Table 6 below.

Experimental Example  3: Anti-diabetic  Active test

Situary mushroom mycelium rice, shiitake mushroom mycelium rice and seaweed of Example 2 were each obtained by infiltrating into 80% ethanol aqueous solution for 24 hours to obtain an extract. Each extract was powdered by removing the solvent using an evaporator and then lyophilizing.

The powder was used to identify PTP1B inhibitory activity known as negative regulator of insulin signaling system.

PTP1B content was measured using a PTP1B fluorometric assay kit (ABNOVA, USA). First, 30 μL of distilled water, 30 μL of 10X PTP assay buffer and 5 μL of 10X flouro-phospho-substrate were added to each well of a 96 well plate, and then incubated for 15 minutes at room temperature by adding recombinant PTP1B. Next, 20 μL of developing buffer was added to each well, followed by incubation at room temperature for 15 minutes, and the reaction was stopped after adding 25 μL of stopsolution. The fluorescence intensity was measured at excitation wavelength 502 nm and measurement wavelength 530 nm, and is calculated as in Equation 2 below.

&Quot; (2) "

The measurement results for the situation mushroom mycelium rice, shiitake mushroom mycelium rice and seaweed samples are shown in Figures 6 to 8, respectively.

As shown in Figure 6, the situation mushroom mycelium rice showed a significant inhibitory effect at the p <0.05 level statistically at 1 ㎍ / mL, compared to the control, 3, 10, 30, 100, 300, 1,000 ㎍ / In mL, a significant inhibitory effect was found at the p <0.01 level.

As shown in FIG. 7, the shiitake mushroom mycelium rice showed a significant inhibitory effect at the p <0.01 level statistically at 1 ~ 1,000 ㎍ / mL compared to the control.

In addition, as shown in FIG. 8, laver showed a significant inhibitory effect at p <0.05 level at 10, 30 ㎍ / mL, and significant at p <0.01 level at 100 and 1,000 ㎍ / mL. Inhibitory effect was shown.

As a result, the fermented wine of the present invention was expected to be able to increase the action of insulin and delay the absorption of glucose by inhibiting the PTP1B enzyme involved in the dephosphorylation in the insulin signaling process of the body tissues, thereby preventing or improving diabetes.

Experimental Example 4: Anti-obesity Activity Test

4-1. Fat accumulation inhibitory effect

In order to measure anti-obesity activity, experiments were performed by culturing all adipocytes, 3T3-L1 cells.

10% bovine calf serum was added to Dulbecco's modified Eagle medium and 1% penicillin / threptomycin as an antibiotic was used as a culture medium of the cells, and cultured in a humidified 5% CO ₂ incubator at 37 ° C. When the cells were 80-90% fused, the cells were separated by treatment with 0.25% trypsin-EDTA, the cells were collected in a centrifuge, and a suspension solution was made and cultured in 24-well plates. Subsequently, differentiation was induced by treating differentiation medium in which 0.5 mM IBMX, 1 μM dexamethasone, and 5 μg / mL insulin were added to DMEM culture medium containing 10% fetal bovine serum. After 2 days, 4 days, and 6 days, the cells were replaced with a culture medium containing only 5 μg / mL of insulin.

Next, the cells were confirmed to inhibit fat accumulation of each sample by oil red O staining.

A 0.35 g Oil Red O reagent was dissolved in 100 mL isopropanol and filtered to obtain a stock solution, and a working solution was prepared by mixing 60% by weight of the stock solution with 40% by weight of distilled water. After 8 days from the start of the differentiation of the 3T3-L1 cells, the medium was removed, the cell culture was washed with PBS, and fixed with 5 μl of 10% formaldehyde and fixed for 5 minutes. Again 500 μL of 10% formaldehyde was added to each well and fixed for 1 hour. After removing formaldehyde and washing with distilled water, 500 μL of the working solution was placed in each well, dyed at room temperature for 30 minutes, and washed three times with distilled water. The stained cells were observed under a microscope and 100% isopropanol was added to elute lipids accumulated in the cells. The cells were transferred to 96 well plates and absorbed at 500 nm to evaluate the degree of lipid accumulation. The degree of lipid accumulation according to the treatment of each sample extract is shown in FIGS. 9 to 11.

As shown in FIG. 9, the situation mushroom mycelial rice showed a significant inhibitory effect at a p <0.01 level in a concentration-dependent manner compared to the control group, and as shown in FIG. 10, the shiitake mushroom mycelium rice was 10 compared to the control group. In pg / mL, p <0.05, 30, 100, and 300 μg / mL showed significant inhibitory effects at p <0.01.

In addition, as shown in FIG. 11, laver showed a significant fat accumulation inhibitory effect at p <0.05 level at 100 μg / mL and p <0.01 level at 300 μg / mL compared to the control group.

4-2. CETP Inhibitory Activity

CETP content was measured using a CETP inhibitor assay kit (BioVision, USA). After 160 μL of the sample was put in a 96 well plate, 3 μL of Rabbit Serum was added and reacted. In each well, 10 μL of Donor Molecule, 10 μL of Acceptor Molecule, and 10 μL of 10X CETP Assay Buffer were incubated at 37 ° C. for 30 minutes, and the fluorescence intensity was measured at an excitation wavelength of 465 nm and a measurement wavelength of 535 nm. The results of measuring CETP inhibitory activity are shown in FIGS. 12 to 14, respectively.

As shown in FIG. 12, the situation mushroom mycelial rice had a concentration-dependent inhibitory effect, and showed a significant inhibitory effect at p <0.05 level at 100 μg / mL, p <0.01 level at 300 μg / mL Showed significant inhibitory effect.

As shown in FIG. 13, the shiitake mushroom mycelium rice had a concentration-dependent inhibitory effect, and showed a significant inhibitory effect at p <0.05 level at 30 μg / mL, and p <at 100 and 300 μg / mL. There was a significant inhibitory effect at the 0.01 level.

In addition, as shown in FIG. 14, laver showed a significant inhibitory effect at p <0.05 level at 30 μg / mL, and a significant inhibitory effect at p <0.01 level at 100 and 300 μg / mL.

As a result, it was confirmed that the fermented strain of the present invention can be effectively used for the treatment of obesity.

Claims (6)

1) immersing the uncooked rice in 20 ~ 30 ℃ hot water for 10 to 30 hours, and repeating the germination and waiting process for 10 to 36 hours in 20 to 40 ℃ fresh water;
2) controlling the water content of the rice to 25 to 50%, sterilizing at 100 to 140 ℃;
3) After sterilization, it is selected from the group consisting of shiitake mushrooms, situation mushrooms, ganoderma lucidum mushrooms, fueryeong, agaricus, roe deer, cordyceps, cloud fingering, spirits, zelkovas, tops, birds, flowers, leaf mushrooms, chaga, longevity mushrooms and willows Inoculating the mushroom mycelium to rice;
4) culturing the rice inoculated mushroom mycelium at 25 ~ 30 ℃ and dried to obtain the mushroom mycelium rice; And
5) immersing and increasing the mushroom mycelial rice fermentation
Mushroom mycelium rice fermented wine manufacturing method comprising a.
The method according to claim 1, wherein 1 to 100 parts by weight of dried seaweed is mixed with 100 parts by weight of the mushroom mycelial rice, followed by fermentation.
The method of claim 2, wherein the algae are Kim (Porphyra Tenera), seaweed in (Laminaria), wakame (Undaria pinnatifida), Sargassum (Sargassum fulvellum), kkosiraegi (Gracilaria verrucosa), fusiformis (Hizikia fusiforme) or agar (Gelidium amansii) Method for producing mushroom mycelial rice fermented wine, characterized in that.
The method according to claim 1, wherein 10 to 200 parts by weight of white rice is mixed and fermented with respect to 100 parts by weight of the mushroom mycelial rice.
According to claim 1, wherein the fermentation method of mushroom mycelium rice fermented wine, characterized in that made for 20 to 80 hours at 20 to 30 ℃.
The method according to claim 1, wherein 1 to 10 parts by weight of rice bran, soy flour or corn flour are added to 100 parts by weight of rice, followed by sterilization.

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