KR20010046517A - Functional beverage containing submerged culture broth of Phellinus sp. and process for preparation thereof - Google Patents

Abstract

PURPOSE: A process for preparing a functional beverage by liquid-culturing Phellinus linteus, mass-producing a mycelium and polysaccharides and mixing an additives is provided. Whereby, the beverage has excellent effect on increasing physiological activity such as reinforcement of immune activity, reduction of the cholesterol level in blood and reduction of the blood sugar level. CONSTITUTION: This functional beverage is prepared by liquid-culturing Phellinus linteus under optimum conditions, mass-producing a mycelium and polysaccharides, mixing a sweetening agent, a sour agent, raw drug preparations, vitamins and inorganic salts and sterilizing at 121deg.C for 30min. The beverage contains 0.01 to 30% by weight of a mycelium and polysaccharides and 0.28 to 55% by weight of food additives.

Description

Functional beverages containing liquid cultures of situational mushrooms and a method of manufacturing the same [Functional beverage containing submerged culture broth of Phellinus sp. and process for preparation eg}

The present invention relates to a functional beverage containing Phellinus sp. Liquid culture and a method for producing the same. More specifically, the present invention relates to a functional drink for enhancing human physiological activity, and a method for producing the same, in which an appropriate amount of mycelium and polymer polysaccharide obtained by liquid culture of mushrooms is mixed with various food additives.

Situation mushroom mycelium, also known as woody mud mushroom, has been used as a herbal medicine since various physiologically active functions have been demonstrated. However, in order to cultivate a large amount of situation mushrooms to use such medicinal functionality widely, mass culture technology is required, and research on mycelial solid culture technology has been actively conducted.

When the liquid culture of the situation mushroom mycelium, the present invention produces a polysaccharide having excellent physiological activity during the cultivation period, and when using this material together with the situation mushroom mycelium, human body physiology comparable to the case of using the mycelium or polysaccharide alone The present invention has been completed by discovering the activity enhancing effect and preparing the functional beverage.

Accordingly, an object of the present invention is to provide a functional beverage for enhancing the biological activity of the human body containing the situation mushroom mycelium obtained by liquid culture and the polymer polysaccharide. Another object of the present invention to provide a method for producing a functional drink for enhancing the human body physiological activity.

The above object of the present invention is a liquid culture under the optimum conditions, the mycelium and the polysaccharide produced in large quantities to produce a mycelium and a polysaccharide produced by the sweetener, acidulant, herbal medicines, vitamins and inorganic salts, and appropriately blended functional drinks After preparation, this function was achieved by investigating the immune activity, blood cholesterol lowering effect, hypoglycemic effect and exercise enhancing effect.

Hereinafter, the configuration and operation of the present invention.

Figure 1 shows the liquid culture process of the situation mushrooms added to the functional mushroom liquid culture-containing functional beverage of the present invention.

The invention edible industrial medium such as a pre-sale Phellinus received (Phellinus linteus) flow a synthetic medium, waste molasses, corn raffinate (corn steep liquor), starch hydrolysates (syrup), potato extract, sugar in Gyeongsangbuk Rural Development Liquid culture step of producing a mycelia or polysaccharides in maximum yield using; Functional beverages are mixed with the mycelium and polysaccharides with sweeteners, acidulants, herbal preparations, vitamins and salts, amino acids, minerals, iron salts, dietary fiber, natural juices, flavoring agents, stabilizers, caffeine, preservatives and nutrients. Manufacturing; Comprising the steps of investigating the effects of the human immune enhancement, blood glucose and blood lipid lowering, antihypertensive, exercise strength of the functional beverage prepared.

The strain used in the present invention was distributed at Gyeongbuk Rural Development Administration, and preserved by subcultured every 2 months at 4 ° C.

In the present invention, vitamins were used alone or in combination of two or more kinds of nicotinic acid amide, vitamins A, B, C, D, E, folic acid. As amino acids, L-glycine, L-lysine, DL-methionine, L-valine, biotin, L-cysteine, L-cystine, L-tryptophan, L-threonine, and L-phenylalanine were used alone or in combination. . As the inorganic salt, calcium citrate, calcium carbonate, calcium pantothenate, calcium gluconate, vegetable calcium, animal calcium, and the like were used alone or in combination of two or more thereof. Iron salts were used alone or in combination of two or more kinds of iron lactate and iron citrate. Nutritional sources used honey or royal jelly. The dietary fiber was used alone or in combination of two or more kinds of extracts of various useful plants such as aloe and chicory, various gums such as xanthan gum, dextrin, soluble fiber and the like. The herbal preparations were used alone or in combination of two or more of Gugija, Ogapi, Baekchul, Baekjak, Green, Red Ginseng, Ginseng, Yin-yang Kwak, Uhwang, Hwangjeong. Natural juices were used alone or in combination of pineapple, lemon, citrus, orange, apple, pear, grapefruit, apricot, strawberry, peach, melon, guava, lemon, plum. Flavoring agents were used alone or in combination of two or more mixed fruit flavor, apple flavor, yogurt flavor, drink flavor. Sweeteners include white sugar, glucose, fructose, fructooligosaccharide, isomaltooligosaccharide, malto oligosaccharide, chitosan oligosaccharide, chito oligosaccharide, soy oligosaccharide, xylo oligosaccharide, isosugar (high fructose), invert sugar, aspartame, stevioside, sorbitol, It was used individually or in mixture of 2 or more types of mannitol. Acidifiers were used alone or in combination of two or more citric acid, DL- apple acid, succinic acid. Stabilizers were used alone or in combination of two or more polysorbate 20, polysorbate 40, polysorbate 80 in the sorbate, and preservatives are benzoic acid and its derivatives, dehydroacetic acid and its salts, sorbic acid and salts thereof It used individually or in mixture of 2 or more types.

Hereinafter, the specific method of the present invention will be described in detail with reference to Examples, but the scope of the present invention is not limited only to these Examples.

Example 1: Production of Mycelia and Polymer Polysaccharides by Situary Mushroom Liquid Culture

In this embodiment, the synthetic medium of the initial pH 4.5 composed of potato dextrose 24g / L, malt extract 10g / L, bactopeptone 1g / L for mycelial growth and polymer polysaccharide production; Edible industrial medium containing 1-10% W / V of molasses, corn steep liquor 1-10% W / V; Starch hydrolyzate (syrup) 1-10% W / V; Potato extract 1-10% W / V; After liquid culture of mycelium mushroom mycelium in a medium containing 1-10% W / V of sugar alone or two or more kinds, the liquid culture solution was put into a 500 mL flask containing 200 mL of medium in a stirred culture tank, and the aeration rate was 200 mL / min. , Aseptically inoculated into a 5 liter fermenter with a temperature of 20 ° C. and pH 4.5 and incubated for 12 days under the same conditions as above (FIG. 1). The culture was centrifuged at 10447 g for 20 minutes to obtain 14 g / L of mycelium. The supernatant was ethanol precipitated again (ethanol: culture solution = 4: 1), and then centrifuged at 10447 g for 20 minutes to give 6 g / of polysaccharide except the supernatant. L was obtained.

Example 2: Preparation of Functional Mushroom-Containing Mycelium and Polysaccharide Polysaccharides

1 to 100 g of the situation mushroom liquid culture solution containing both the situation mushroom mycelium and the polymer polysaccharide obtained in Example 1 was dissolved in 1 L of passion water or membrane filtration water, and then 0.001 to 5% by weight of nicotinic acid amide as a food additive, and 0.01 to 5 folic acid. Weight%, Aspartic acid 0.01 to 5%, Citrate 0.01 to 1.0%, Honey 0.01 to 3%, Dextrin 0.01 to 3%, Ginseng extract 0.01 to 10%, Apple 0.01 to 5%, Apple flavor 0.05 to 1% by weight, fructooligosaccharide 0.1 to 10% by weight, citric acid 0.05 to 5% by weight, polysorbate 0.01 to 3.0% by weight, benzoic acid 0.05 to 1% by weight, food additives 0.28 ~ 55% by weight Range. At this time, the situation mushroom mycelium and the polysaccharide is 0.01 to 30% by weight relative to the total composition ratio, preferably 0.1 to 10% by weight.

Example 3: Preparation of functional beverages containing mushroom mycelium

A functional beverage containing only mycelia obtained by liquid culture of the situation mushrooms in Example 1 was prepared in the same manner as in Example 2. That is, in Example 1, 1 to 100 g of the mycelium obtained by centrifuging the liquid mushroom culture medium for 10 minutes at 10447 g was dissolved in 1 L of passion water or membrane filtration water, and then, as a food additive, nicotinic acid amide, folic acid, aspartic acid, calcium citrate, Honey, dextrin, ginseng, apple, apple flavor, fructooligosaccharide, citric acid, polysorbate, benzoic acid were added in the same amount as in Example 2. At this time, the situation mushroom mycelium was 0.01 to 30% by weight and preferably 0.1 to 10% by weight relative to the total composition ratio as in Example 2.

Example 4: Preparation of Functional Beverage Containing Polysaccharides of Situation Mushrooms

In Example 1, a functional beverage containing the polymer polysaccharide obtained by centrifuging the liquid mushroom culture medium and centrifuging the supernatant was prepared in the same manner as in Example 2. That is, in Example 1, the supernatant obtained by centrifuging the liquid mushroom culture in 10447 g for 20 minutes was ethanol precipitated again (ethanol: culture solution = 4: 1), followed by centrifugation for 20 minutes at 10447 g. Or melt 1 to 100 g in 1 L membrane filtration water and then add nicotinic acid amide, folic acid, aspartic acid, calcium citrate, honey, dextrin, ginseng, apple, apple flavor, fructooligosaccharide, citric acid, polysorbate, benzoic acid as food additives Example 2 And added in the same amount. At this time, the situation mushroom polymer polysaccharide was 0.01 to 30% by weight and preferably 0.1 to 10% by weight relative to the total composition ratio as in Example 2.

Example 5: Functional Evaluation of Situary Mushroom Mycelium and Beverages Containing Polysaccharides

In this example, the main functionality of the functional beverage containing both the situation mushroom mycelium and the polymer polysaccharide prepared in Example 2 was evaluated. Immune activity was measured by the Mayer method. That is, 50 μl of NHS (normal human serum), GVB ⁺⁺ (gelatin veronal buffer saline pH 7.2) and the sample were mixed and reacted at 37 ° C. for 30 minutes, and then 350 μl of GVB ⁺⁺ was added to the reaction solution. This was serially diluted from 10 to 160 times. 250 mL of 750 μl GVB ⁺⁺ and positive sensitized blood cells (IgM haemolysin sensitized sheep erythrocyte, 10 ⁸ cell / mL) were added thereto and reacted for 1 hour, followed by 2.5 mL of PBS (phosphate buffer saline, pH7.4). After centrifugation, the absorbance of the supernatant was measured at 412 nm. Anti-complement activity was indicated by ITCH ₅₀ , the inhibition of 50% total complement hemolysis. In order to verify the exact activity of the sample, anti-complementary polysaccharide CAP-O (ITCH ₅₀ = 80%) with known titer was calculated as a standard. The calculation method is as follows.

In order to measure the effect of lowering cholesterol in the blood, the rat was ingested with hyperlipidemia for 2 weeks after oral administration of hyperlipidemia, followed by fasting the experimental animals for 12 hours, anesthetizing with ether, and recovering the liver tissue. Was taken from. Blood was centrifuged and used for experiments after plasma separation. Total cholesterol content was measured using the kit using enzyme method (AM 202-K). To measure the hypoglycemic effect, 5 week-old male rats were purchased and adapted to solid feed (Samyang Feed) for 1 week, and then fed to AIN-76 diet for 4 weeks. The breeding room was lit and turned off every 12 hours while maintaining a temperature of 20 ° C. and a humidity of 50%. Distilled water was used for water, and distilled water and diet were taken at will. During the experiment, dietary intake was measured every 2 days and weight gain was measured every 3 days. Rats were divided into 10 groups in each group. First, groups with similar average weights were classified, and then randomly classified so that the average weight of each group was similar. Diabetes induced streptozotocin (STZ) (Sigma chemical Co., 50mg / kg BW / 0.01M citrate buffer, pH), which is known to cause hyperglycemia by specifically destroying β-cells in the pancreas and lowering insulin secretion. 4.5) was injected intramuscularly into experimental animals to experimentally induce diabetes in the form of IDDM. After fasting for 24 hours after STZ injection, urine glucose was measured, and the equivalent weight of 300 mg / dl or more was determined to be diabetes induced. The rats proved to be diabetic with STZ were orally administered to the rats at a dose of 50-100 mg / kg for two weeks, and the sample intake efficiency was increased by fasting before the diet. The normal control group that did not cause diabetes was injected with the same amount of saline to give the same stress as the diabetic group. Likewise, the same stress was given by oral administration of physiological saline to both the diabetic induction control group and the normal control group that did not orally administer the sample. After oral administration of the sample for 2 weeks, the animals were fasted for 12 hours, anesthetized with ether, and then opened, and blood was collected from the abdominal arteries. Blood glucose levels were measured using a Japanese Eiken kit (GLZYME) prepared by the glucose oxidase method using the hydrogen peroxide as a principle by the action of glucose oxidase from glucose. The improvement of exercise power was measured by the swimming limit measurement method. The swimming pool of the mouse is an acrylic product of 90 × 45 × 45 cm (L × W × H), filled with water up to 38 cm from the bottom of the pool, pump (C-P60H, 日本) and flow meter (FC). -A20, Japan Hitachi Institute) was used. The water temperature was adjusted to 34 degreeC by the heater installed in the bottom of a swimming pool. The water outlet made a precise, straight hole in the pipe of the vinyl product, affecting the entire water. And the pipe is installed 2 to 3 cm below the surface of the water to maintain a constant flow direction of water, the flow rate of the water was measured by the aberration type flow meter (SPC-type, Nippon Kogyo Co., Ltd.) at the surface of the pool. During the test period, the flow rate of the pool was set at 8 L / min (surface flow velocity at the center of the pool was about 166 cm / sec). The mice were introduced to the pool to start swimming, and when the mice became tired after the continuous swimming, they were confirmed to go to the rear of the pool. Therefore, the board is inclined at the rear of the swimming pool, and the water is installed downward so that the tired mouse descends into the water by the flow of water at the rear of the swimming pool, and if the mouse stays in the water for more than seven seconds, the athletic ability is achieved. The mouse was taken out of the pool and time was recorded. This swimming time showed the mouse's athletic ability. After swimming, the mice were moistened, dried and placed in a breeding case. After 48 hours of rest and swimming, the experiment was repeated. The experimental results are shown in Table 1, and the additive-free beverage was a control group without adding the mycelium and the polysaccharide of the liquid mushroom culture medium, and the added beverage A was the liquid mushroom mycelium and the polysaccharide polysaccharide solution 100g / L and the beverage B was added. 75 g / L and added drink C is 50 g / L added. In the immunological activity, the beverages A, B, and C were significantly higher than those of the non-added beverages, and in particular, the beverage A was high. In addition, the beverages A, B, and C were superior to the non-added beverages, and the added beverage A was the most excellent in the blood cholesterol lowering effect, the hypoglycemic effect, and the exercise enhancing effect.

Functional Evaluation of Beverages Containing Mycelium Liquid Culture Liquid Mycelia and Polysaccharides Prototype Group Functionality No-added drinks Additive Drink A Additive drink B Additive Drink C Immune activity ｛ITCH50 (%)｝ 23.5 ± 2.1 42.3 ± 3.2 35.8 ± 2.4 33.7 ± 2.8 Blood cholesterol lowering effect (mg / ㎗) 62.7 ± 3.8 45.1 ± 3.6 46.5 ± 3.8 52.4 ± 3.1 Hypoglycemic effect (mg / ㎗) 165.4 ± 20.5 150.6 ± 21.8 152.6 ± 20.3 155.1 ± 26.4 Exercise power enhancement effect (min) 30.2 ± 5.5 40.2 ± 3.7 47.5 ± 3.3 35.6 ± 2.8

Example 6: Functional Evaluation of Drinks Containing Situation Mushroom Mycelia

The main functionality of the functional beverage containing the mycelium produced from the situation mushroom liquid culture prepared in Example 3 was evaluated by the same method as in Example 5. Experimental results, as shown in Table 2, wherein the additive-free beverage is a control group without adding the mushrooms mycelium, the added beverage A is 6g / L lyophilized mushroom mushroom mycelium, 4g / L, added beverage C is 2g / L is contained. As shown in Table 2, Immune activity was higher in all of the added beverages A, B, and C compared to the non-added beverages. Especially, the added beverage A was higher, and the blood cholesterol lowering effect, blood sugar lowering effect, and exercise enhancing effect were all higher than the additive-free beverage. A, B and C were excellent, especially the added beverage A was the highest.

Functional Evaluation of Drinks Containing Mycelium Produced from Liquid Mushroom Culture Prototype Group Functionality No-added drinks Additive Drink A Additive drink B Additive Drink C Immune activity ｛ITCH50 (%)｝ 23.5 ± 2.1 42.3 ± 2.8 40.4 ± 2.6 34.3 ± 2.2 Blood cholesterol lowering effect (mg / ㎗) 62.7 ± 3.8 52.4 ± 3.1 54.2 ± 3.7 56.4 ± 4.1 Hypoglycemic effect (mg / ㎗) 165.4 ± 20.5 138.3 ± 20.9 143.3 ± 22.5 148.3 ± 21.8 Exercise power enhancement effect (min) 30.2 ± 5.5 42.2 ± 3.5 39.7 ± 4.1 35.4 ± 3.8

Example 7: Functional Evaluation of Functional Beverage Containing Polysaccharides of Situation Mushrooms

The main functionality of the functional beverage prepared by adding the polymer polysaccharide produced from the situation mushroom liquid culture prepared in Example 4 was evaluated in the same manner as in Example 5, and the results are shown in Table 3. The additive-free beverage is a control group that does not add the lyophilized polymer polysaccharide contained in the liquid mushroom culture medium, and the additive beverage A is 6 g / L of the lyophilized polymer polysaccharide contained in the liquid mushroom liquid culture medium, and the beverage B is 4 g / L and the beverage added. C is a functional beverage prepared by adding 2 g / L, respectively. As shown in Table 3, Immune activity was higher in all of the added beverages A, B, and C than in the non-added beverages, and in particular, the added beverage A was high, and in addition, in the blood cholesterol lowering effect, hypoglycemic effect, and exercise enhancing effect, all the additive beverages were added. A, B and C were excellent, especially the added beverage A was the highest.

Functional Evaluation of Beverages Containing Lyophilized Polymer Polysaccharides Produced from Situation Mushroom Liquid Culture Prototype Group Functionality No-added drinks Additive Drink A Additive drink B Additive Drink C Immune activity ｛ITCH50 (%)｝ 23.5 ± 2.1 45.7 ± 3.1 42.4 ± 2.8 38.7 ± 3.5 Blood cholesterol lowering effect (mg / ㎗) 62.7 ± 3.8 46.2 ± 3.3 49.5 ± 3.5 53.9 ± 2.8 Hypoglycemic effect (mg / ㎗) 165.4 ± 20.5 143.5 ± 23.7 147.4 ± 25.3 150.2 ± 20.5 Exercise power enhancement effect (min) 30.2 ± 5.5 39.4 ± 2.8 35.2 ± 3.1 32.2 ± 3.0

As described through the above examples, the present invention situational mushroom functional beverage prepared by formulating a suitable amount of liquid culture and food additives containing all or any of the situation mushroom mycelium and the polymer polysaccharide in passion and membrane filtration water It is a very useful invention in the health food industry because it has an excellent effect of enhancing the body's physiological activities such as immune enhancement, blood cholesterol lowering, blood sugar lowering, exercise strengthening.

Claims (3)

Functional drink containing mycelium and polymer polysaccharide together or singly obtained by liquid culture of situation mushrooms. The situation mushroom mycelium and the polymer polysaccharide according to claim 1 are added to 0.01 or 30% by weight to all or each of the passion water or membrane filtration water, and 0.28 to 55% by weight of food additives are sterilized for 30 minutes at 121 ℃ Situation mushrooms containing functional beverage manufacturing method. According to claim 2, Food additives are vitamins and salts, amino acids, minerals, nutrients, dietary fiber, iron salts, natural juices, caffeine, flavoring agents, sweeteners, acidulants, preservatives, stabilizers, herbal preparations Method for manufacturing functional drinks containing mushrooms.