METHOD FOR PREPARING BLOOD SUGAR CONTROL COMPOSITION CONTAINING CULTURED MUSHROOM MYCELIUM COMPLEX

Abstract

Proposed is a method for complex culture of Innotus obliquus, Phellinus linteus, and Ganoderma lucidum. The cultured mycelium complex prepared by the above method has a high content of beta-glucan, which has excellent health functionality and can be used as additives or cooking seasonings for various foods. In addition, when raw meat is salted using the cultured mycelium complex, meat dishes with good taste and flavor can be easily prepared.

Claims

1. A method of preparing a blood sugar control composition containing a cultured mushroom mycelium complex, the method comprising: (Step 1) inoculating the fruiting body tissues of three types of mushrooms including Innotus obliquus, Phellinus linteus, and Ganoderma lucidum into potato dextrose agars (PDA), respectively and culturing the mycelium of each of the mushrooms; (Step 2) inoculating a mycelium complex including the mycelium of each of the Innotus obliquus, Phellinus linteus, and Ganoderma lucidum cultured in Step 1 into potato dextrose broth (PDB); (Step 3) culturing the potato dextrose broth (PDB) inoculated with the mycelium of each of the three types of mushrooms for 4 to 6 weeks; (Step 4) inoculating the mycelium obtained through Step 3 into a rice barley medium; and (Step 5) obtaining a complex culture mycelium by culturing the mycelium inoculated into the rice barley medium for 4 to 7 weeks.

2. A composition for prevention or treatment of diabetes, the composition comprising a mushroom complex culture mycelium prepared by the method of claim 1.

3. A healthy functional food for prevention or treatment of diabetes, the food comprising a mushroom complex culture mycelium prepared by the method of claim 1.

4. A granule composition for controlling blood sugar and dietary, the composition comprising heat-treated products of Cucumis melo conomon, sweet pumpkin, Capsosiphon fulvescens, and Ecklonia cava in powder of a mushroom complex culture mycelium prepared by the method of claim 1.

5. A composition for prevention or treatment of diabetes, the composition comprising the granule composition of claim 4.

6. A healthy functional food for prevention or treatment of diabetes, the food comprising the granule composition of claim 4.

7. A method for preventing or treating diabetes, comprising administering the composition of claim 2 to a subject in need thereof.

Description

BEST MODE

[0045] Hereinafter, preferred embodiments of the present disclosure will be described in detail. However, the present disclosure is not limited to the embodiments described herein and may be embodied in other forms. Rather, it is provided so that this disclosure will be thorough and complete and will fully convey the spirit of the disclosure to those skilled in the art.

Preparation Example 1. Complex Culture of Innotus obliquus, Phellinus linteus, and Ganoderma lucidum

[0046] The fruiting bodies of Innotus obliquus, Phellinus linteus, and Ganoderma lucidum were isolated and inoculated into PDA, and then the mycelium of each mushroom was cultured at 27° C. to 29° C. for 2 weeks. After preparing the PDB medium subdivided into 100 ml units, each mushroom mycelium cultured in PDA was cut into 1 mm.sup.2 using a scalpel. The three strains were cut per one Erlenmeyer flask containing PDB medium were combined with 5 slices each.

[0047] A complexly inoculated medium was incubated in a stationary phase in a bio-oxygen demand incubator (BOD incubator, low-temperature incubator) for 1 week at 27° C. to 28° C. and 20% humidity, but was stirred for about 1 minute every day during incubation. After one week, the flask under the complex inoculation and culture was transferred to a shaking incubator and cultured at 27° C. and 100 rpm for 4 weeks to prepare a cultured mycelium complex culture solution.

[0048] After immersing rice barley for 6 hours, dehydrated for 8 hours, added 1 g of calcium carbonate based on 100 g of dehydrated rice barley, mixed evenly, and sterilized at 121° C. for 1 hour in an autoclave to prepare a rice barley medium. After completion of the sterilization, 5 ml of the cultured mycelium complex culture medium cultured for 5 weeks per 1 kg of the rice barley medium cooled to 25° C. was dispensed and inoculated. After inoculation, the cultured mycelium complex was cultured for 30 days in a culture room maintained at a temperature of 26° C. to 28° C. and a humidity of 45% to 50%.

[0049] The cultured mycelium complex was dried at 57° C. to 60° C. for 24 hours using a dryer while containing rice barley and pulverized using a pin mill grinder to prepare a powder.

[0050] Meanwhile, this method is the same as the method disclosed in Example 1 of Korea Patent No. 10-1923408.

Comparative Preparation Example 1. Preparation of Each Mushroom Mycelium

[0051] Using the method of Example 1, but as a comparative condition, the single mycelium culture of Innotus obliquus, Phellinus linteus, and Ganoderma lucidum and the complex culture of Phellinus linteus and Innotus obliquus were performed in the same process to obtain a mushroom mycelium.

TABLE-US-00001 TABLE 1 Mycelium Comparative Preparation Innotus obliquus mycelium Example 1-1 Comparative Preparation Ganoderma lucidum mycelium Example 1-2 Comparative Preparation Phellinus linteus mycelium Example 1-3 Comparative Preparation Complex mycelium of Innotus Example 1-4 obliquus and Phellinus linteus

Experimental Example 1. Check the Blood Sugar Control Effect

Experimental Example 1-1. Blood Sugar Control Effect in Animal Model of Type 1 Diabetes

[0052] The animal model of type 1 diabetes animal model was induced by intravenous injection of 50 mg/kg of alloxan (Alloxan, Sigma, MA, USA) to ICR mice (male, 7 weeks old, 7 animals per group). In the experimental group, the mushroom mycelium was orally administered for 3 days before the administration of alloxan once a day at 2 mg/mouse by suspension of dry powder in phosphate-buffered saline (PBS). In addition, blood was collected up to the 9th day at 3-day intervals after alloxan administration, and the blood glucose content was measured and shown in Table 2 below.

TABLE-US-00002 TABLE 2 Blood glucose (mg/dL) Alloxan + Alloxan + Alloxan + Alloxan + Comparative Comparative Comparative Comparative Positive Alloxan + Preparation Preparation Preparation Preparation control Example Example Example Example Example Group 1 1-1 1-2 1-3 1-4 Alloxan (alloxan mycelium mycelium mycelium mycelium mycelium Untreated alone administration administration administration administration administration Condition group group) group group group group group 3 days 99 496 125 434 421 421 425 6 days 100 562 112 352 352 312 321 9 days 98 582 105 312 314 321 294

[0053] Referring to Table 2, it can be confirmed that blood sugar levels are maintained almost similar to those of non-diabetic induction mice, an alloxan-free group, throughout the administration period in the animals of the cultured mycelium complex administration group.

Experimental Example 1-2. Blood Sugar Control Effect in Animal Model of Type 2 Diabetes

[0054] The animal model of type 1 diabetes was used by purchasing db/db diabetic mice (5 weeks old, male). In the experimental group, the mushroom mycelium was suspended in PBS with dry powder and orally administered at 2 mg/mouse once a day for 8 weeks, and then the blood glucose content was measured and shown in Table 3 below.

TABLE-US-00003 TABLE 3 Blood glucose (mg/dL) Comparative Comparative Comparative Comparative Preparation Preparation Preparation Preparation Preparation Example Example Example Example Example 1 1-1 1-2 1-3 1-4 mycelium mycelium mycelium mycelium mycelium Untreated administration administration administration administration administration Condition group group group group group group 8 week 412 251 381 372 353 298

[0055] The db/db diabetic mouse is characterized by maintaining the blood glucose concentration at 390 to 420 mg/dL, but the results in Table 3 show that the cultured mycelium complex administration group in Preparation Example 1 is reduced by about 50% compared to the untreated group.

[0056] Through this animal experiment, it was confirmed that the cultured mycelium complex prepared in the present disclosure had very good blood sugar control ability.

[0057] Next, a formulation experiment was conducted to prepare a mushroom processed product in a state suitable for diabetic patients to eat and carry the cultured mycelium complex directly.

[0058] At this time, for the formulation, it was considered whether it is possible to control blood sugar quickly while easily relieving hunger during intake, whether it is possible to supply protein that may be insufficient for diabetic patients with insufficient muscle strength, and whether there is a good taste and aroma to continue taking the formulation.

[0059] In addition, the final form was decided to be prepared in the form of granules that are easy to take without water while being well packaged in disposable packaging paper.

[0060] Through preliminary experiments, in the powder mixture state, the powder mixture form was difficult to take it without water, so the powder mixture form was avoided. Since the hard pill formulation takes time to digest in the stomach after taking pill, fast blood sugar control is not good, so the pill formulation was also excluded from the experiment.

<Preparation Example 2. Manufacture of Heat-Treated Products of Cucumis melo Conomon, Sweet Pumpkin, Capsosiphon Fulvescens, and Ecklonia cava>

[0061] Cucumis melo conomon fruit, sweet pumpkin fruit were purchased and Cucumis melo conomon, sweet pumpkin uses only the pulp from which the seeds are removed. Capsosiphon fulvescens, and Ecklonia cava in their raw state were purchased and used.

[0062] Using this prepared material, Cucumis melo conomon, sweet pumpkin, Capsosiphon fulvescens, and Ecklonia cava, and water was put in an autoclave as shown in Table 2 and heated at 121° C., 1.5 atm for 1 hour, and then cooled to room temperature. Next, the cooled contents were put into a food grinder and pulverized to prepare a heat-treated product in the form of porridge.

TABLE-US-00004 TABLE 4 Cucumis melo Sweet Capsosiphon conomon Pumpkin fulvescens Ecklonia Water Condition (g) (g) (g) cava (g) (g) Preparation 100 100 50 50 300 Example 2-1 Preparation 100 100 30 70 300 Example 2-2 Preparation 100 140 30 30 300 Example 2-3 Preparation 100 60 70 70 300 Example 2-4

Comparative Preparation Example 2. Preparation of Comparative Conditions Heat-Treated Product

[0063] A heat-treated product was prepared under the conditions of Preparation Example 2, but the content of each component was prepared under the conditions of Table 5 below.

TABLE-US-00005 TABLE 5 Cucumis melo Sweet Capsosiphon conomon Pumpkin fulvescens Ecklonia Water Condition (g) (g) (g) cava (g) (g) Comparative 0 250 0 50 300 Preparation Example 2-1 Comparative 250 0 0 50 300 Preparation Example 2-2 Comparative 0 0 250 50 300 Preparation Example 2-3 Comparative 100 100 100 0 300 Preparation Example 2-4

Example 1 and Comparative Example 1. Preparation of Granular Compositions—I

[0064] Under the conditions of Table 6 below, each powder and heat-treated product were mixed, kneaded to make a dough, and granulated using a granulator. At this time, the granulation process is as follows. The dough was molded into granules using a reverse-rotating granulator (Garyeo Industrial Co.), and the molded ones were dried in a drying room at 50° C. for 5 hours so that the moisture content was 3 wt % to 4 wt %, and then granules molded with a diameter of 0.5 mm or more were recovered by sieving.

[0065] Among the granule raw materials, the powder of locust adult and silkworm pupa was prepared and used as follows, adult locust adult and silkworm pupa was purchased from an insect breeding farm, steamed for 30 minutes, dried, and then baked in an oven at 180° C. for 3 hours.

TABLE-US-00006 TABLE 6 Cultured Preparation mycelium Example 2-1 complex of Heat- Locust Silkworm Preparation treated adult pupa Example 1 product Water powder powder Condition Powder (g) (g) (g) (g) (g) Example 1-1 100 100 0 50 50 Example 1-2 100 100 0 30 70 Example 1-3 100 100 0 70 30 Example 1-4 100 80 0 60 60 Comparative 100 100 0 100 0 Example 1-1 Comparative 100 0 100 50 50 Example 1-2 Comparative 100 50 50 50 50 Example 1-3 Comparative 200 100 0 0 0 Example 1-4

Example 2 and Comparative Example 3. Preparation of Granular Composition—II

[0066] Granules were prepared by applying the granule preparing process of Example 1 and Comparative Example 1, but by changing the conditions of the heat-treated product to the conditions of Table 7 below.

TABLE-US-00007 TABLE 7 Condition Characteristic Example 2-1 Granules prepared by heat- treated product of Preparation Example 2-2 Example 2-2 Granules prepared by heat- treated product of Preparation Example 2-3 Example 2-3 Granules prepared by heat- treated product of Preparation Example 2-4 Comparative Granules prepared by heat- Example 2-1 treated product of Comparative Preparation Example 2-1 Comparative Granules prepared by heat- Example 2-2 treated product of Comparative Preparation Example 2-2 Comparative Granules prepared by heat- Example 2-3 treated product of Comparative Preparation Example 2-3 Comparative Granules prepared by heat- Example 2-4 treated product of Comparative Preparation Example 2-4

Experimental Example 2. Check the Degree of Granulation

[0067] Table 8 below shows whether granules are well produced according to the preparation conditions of the heat-treated product. It was determined that the granulation was good when it was 90% or more to maintain an intact form after the final preparation, and the condition was described when no granulation molding was performed or more than 5% of broken granulation was found during the drying process even after molding.

TABLE-US-00008 TABLE 8 Condition State Example 1-1 Good Example 1-2 Good Example 1-3 Good Example 1-4 Good Example 2-1 Good Example 2-2 Good Example 2-3 Good Comparative Poor granulation using a granulator Example 1-1 Comparative Poor granulation using a granulator Example 1-2 Comparative Poor granulation using a granulator Example 1-3 Comparative Poor granulation using a granulator Example 1-4 Comparative Crumbling during drying process after Example 2-1 granule preparing Comparative Crumbling during drying process after Example 2-2 granule preparing Comparative Crumbling during drying process after Example 2-3 granule preparing Comparative Crumbling during drying process after Example 2-4 granule preparing

[0068] As a result, it can be confirmed that the granule formulation is completely prepared in Examples 1 and 2, and the conditions of Comparative Examples 1 and 2 were not well kneaded in the granulator, resulting in poor granulation formation or crumbling while drying the produced granules.

[0069] Through this, it can be seen that the content of each raw material sample, which is a raw material for granules, or the conditions under which the heat-treated product was prepared in particular among the raw material samples are important.

Experimental Example 3. Check the Blood Sugar Lowering Effect by Taking Granules

[0070] Five people in their 40s and 60s who had more than 10 days of experience during 1 month measuring symptoms with an empty blood sugar of 110 to 130 mg/dl (pre-diabetes state, diabetes must exceed 130 mg/dl) were allowed to take the compositions of Examples 1 and 2 respectively for 1 month and the period during which the fasting blood sugar increased to 110 mg/dl or more was measured during the dosing period. Although they were not provided with a separate dietary table, they were instructed to take 5 g of each granule within 30 minutes after each meal. The measured values are shown in Table 9 below, and the number of occurrences of hyperglycemia symptoms for each group was recorded as an average value. The decimal point of the average value is rounded off. At this time, the group ingesting the cultured mycelium complex powder of Preparation Example 1 was also set as the comparative group.

[0071] As a result of the confirmation, it can be seen that the number of times of hyperglycemia was significantly reduced for 1 month in all the granules taking groups of Examples 1 and 2. In particular, the effect was similar to that of the complex cultured mushroom mycelium powder of Preparation Example 1, which is a result of proving that the continuous intake is more important than the intake amount of the complex cultured mushroom.

TABLE-US-00009 TABLE 9 Number of times of hyperglycemia in 1 Condition month while taking Example 1-1 2 Example 1-2 3 Example 1-3 2 Example 1-4 3 Example 2-1 2 Example 2-2 1 Example 2-3 3 Preparation Example 1 2

Experimental Example 4. Sensory Evaluation

[0072] Through the results of the previous blood sugar drop effect, the following sensory evaluation confirmed whether granular formulations are easy to take and whether they can be applied as health supplements that can relieve hunger, which usually occurs in people with high blood sugar symptoms. In this case, among the samples that were tried for granulation formulation, those that were not granulated well were mixed with each raw material sample, dried, and then powdered to be taken.

[0073] To this end, adults in their 20s and 60s, men and women of all ages were divided into two groups, and 5 g of the powder or granules were taken within 30 minutes after every meal in 20 people for 3 days. Take it on an empty stomach to relieve hunger or check the taste preference for 1 to 5 points (1 point: Very bad to 5 points: Very good) was scored. In addition, in this case, the cultured mycelium complex powder of Preparation Example 1 was presented as a comparative group.

TABLE-US-00010 TABLE 10 Condition Relieve hunger Taste preference Example 1-1 4.3 4.1 Example 1-2 4.2 4.5 Example 1-3 4.3 4.2 Example 1-4 4.1 4.1 Example 2-1 4.2 4.4 Example 2-2 3.9 4.2 Example 2-3 4.2 3.8 Comparative 2.4 2.9 Example 1-1 Comparative 2.5 2.8 Example 1-2 Comparative 2.9 2.1 Example 1-3 Comparative 2.1 Example 1-4 Preparation 2.3 Example 1

[0074] As a result, it can be confirmed that the granule formulations of Examples 1 and 2 are helpful in relieving hunger and have good taste, so they are good products for use as health supplements. In particular, it can be seen that the cultured mycelium complex itself is significantly superior to the powder, and it is also confirmed that the raw material of the granules and the preparing conditions of the heat-treated product affect both the feeling of hunger and the taste of the granule formulation.