METHOD FOR CO-CULTURING INONOTUS OBLIQUUS, GANODERMA LUCIDUM, AND PHELLINUS LINTEUS MYCELIA

Abstract

The present invention relates to a method of co-culturing Inonotus obliquus, Ganoderma lucidum, and Phellinus linteus. The co-cultured mycelia prepared through the method of the present invention have high beta-glucan content and thus can exhibit superior health functionality, and can be used as an additive or a cooking seasoning in various foods. In addition, the use of the co-cultured mycelia in curing raw meat enables easy preparation of a meat-based food product that has a good taste and flavor.

Claims

1. A method of co-culturing Inonotus obliquus, Ganoderma lucidum and Phellinus linteus mycelia, comprising the following steps in order: (1) inoculating a fruit body tissue of each of Inonotus obliquus, Ganoderma lucidum and Phellinus linteus in Potato Dextrose Agar (PDA) and then separately culturing mycelia of each mushroom; (2) co-inoculating mycelia of three species of the Inonotus obliquus, Ganoderma lucidum and Phellinus linteus separately cultured in the step (1) in Potato Dextrose Broth (PDB); (3) culturing the PDB inoculated with the mycelia of three species for 4-6 weeks; (4) inoculating mycelia obtained through the culturing in the step (3) in a rice barley medium; and (5) further culturing the mycelia inoculated in the rice barley medium in the step (4) for 4-7 weeks to afford co-cultured mycelia.

2. The method of claim 1, wherein the culturing the mycelia in the step (3) is performed at 25-30° C.

3. The method of claim 1, wherein the rice barley medium in the step (4) is obtained by subjecting rice barley to soaking for 4-8 hr and then dehydration, adding calcium carbonate in an amount of 0.5-2 parts by weight based on 100 parts by weight of the dehydrated rice barley, and then performing sterilization at 120-125° C. for 30 min to 2 hr.

4. The method of claim 1, wherein the culturing in the step (5) is performed at 25-30° C.

5. A method of curing raw meat using co-cultured mycelia of Inonotus obliquus, Ganoderma lucidum and Phellinus linteus, comprising: obtaining co-cultured mycelia through the method of claim 1; and finely cutting the co-cultured mycelia isolated from a rice barley medium, adding meat with the finely cut co-cultured mycelia, a cumin powder, a basil powder, a dill weed powder, a dill seed powder, and a salt, and performing curing.

6. Co-cultured mycelia obtained through the method of claim 1.

7. Food additive comprising the co-cultured mycelia of claim 6.

8. A curing-agent composition for meat containing the co-cultured mycelia of claim 6, a cumin powder, a basil powder, a dill weed powder, a dill seed powder, and a salt.

Description

MODE FOR INVENTION

[0045] A better understanding of the present invention will be obtained through the following examples. However, the present invention is not limited to these examples, and may be embodied in other forms. These examples are provided to thoroughly explain the invention and to sufficiently transfer the spirit of the present invention to those skilled in the art.

Example 1. Co-Culture of Inonotus obliquus, Ganoderma lucidum and Phellinus linteus Mycelia

[0046] The fruit body tissues of Inonotus obliquus, Ganoderma lucidum, and Phellinus linteus were isolated and then inoculated in PDA, after which the mycelia of each mushroom were cultured at 27-29° C. for 2 weeks. PDB medium subdivided into 100 mt units was prepared, individual mushroom mycelia cultured in PDA were cut to 1 mm.sup.2 with a scalpel, and 5 pieces of each of the three species of strains that were cut were co-inoculated in each Erlenmeyer flask containing the PDB medium.

[0047] The co-inoculated medium was subjected to stationary culture in a BOD incubator (Bio-Oxygen Demand incubator, low-temperature incubator) at 27-28° C. and a humidity of 20% for 1 week. Here, stirring was performed for about 1 min every day during the culture. After 1 week, the flask that was cultured after co-inoculation was transferred to a shaking incubator and then culture was performed at 27° C. and 100 rpm for 4 weeks, thus preparing a co-cultured mycelial broth.

[0048] Rice barley was soaked for 6 hr and then dehydrated for 8 hr, after which 1 g of calcium carbonate was added based on 100 g of the dehydrated rice barley, followed by homogeneous mixing and sterilization using a high-pressure sterilizer at 121° C. for 1 hr, thereby preparing a rice barley medium. After termination of sterilization, 5 mf of the co-cultured mycelial broth, obtained through culturing for 5 weeks, was aliquoted and inoculated per kg of the rice barley medium cooled to 25° C. After inoculation, culture was performed for 30 days in a culture room maintained at a temperature of 26-28° C. and a humidity of 45-50%.

[0049] After completion of culture, some of the co-cultured mycelia were dried at 57-60° C. for 24 hr using a dryer in the state in which the rice barley was included therein, and were pulverized using a pin mill, thus obtaining a powder.

[0050] The co-cultured mycelia that were not powdered were left behind for use as a raw-meat curing agent by removing the rice barley medium therefrom and finely cutting only the mycelia.

Example 2. Preparation of Cured Meat Using Co-Cultured Mycelia

[0051] 100 g of pork fillet, and appropriate amounts of the finely cut co-cultured mycelia remaining in Example 1, cumin powder, basil powder, dill weed powder, dill seed powder and salt were mixed to homogeneity, stored at room temperature of 25° C. for 4 hr, and then allowed to stand in a low-temperature warehouse at 7° C. for 24 hr, thereby preparing cured meat.

TABLE-US-00001 TABLE 1 Weight (g) Preparation Co-cultured Dill Dill conditions of Pork mycelia of Cumin Basil weed seed cured meat fillet Example 1 powder powder powder powder Salt Example 2-1 100 1.0 0.2 0.2 0.2 0.2 0.1 Example 2-2 100 1.0 0.1 0.3 0.1 0.3 0.1 Example 2-3 100 1.0 0.3 0.1 0.3 0.1 0.1 Example 2-4 100 5.0 0.1 0.1 0.1 0.1 0.1

Comparative Example 1. Preparation of Comparative Mycelia—i

[0052] The mycelia were prepared under respective conditions as shown in Table 2 below.

TABLE-US-00002 TABLE 2 Conditions Features Comparative In the method of Example 1, Inonotus ibliquus mycelia Example 1-1 were inoculated alone in PDB, rather than co-inoculating the mycelia of three species of mushrooms, and the subsequent culturing process was the same Comparative In the method of Example 1, Ganoderma lucidum mycelia Example 1-2 were inoculated alone in PDB, rather than co-inoculating the mycelia of three species of mushrooms, and the subsequent culturing process was the same Comparative In the method of Example 1, Phellinus linteus mycelia Example 1-3 were inoculated alone in PDB, rather than co-inoculating the mycelia of three species of mushrooms, and the subsequent culturing process was the same Comparative In the method of Example 1, the step of culturing PDB was Example 1-4 omitted, the mycelia of three species of mushrooms were directly inoculated in a rice barley medium, and the culture period was further increased by the time corresponding to the step of culturing PDB Comparative In the method of Example 1, the step of culturing PDB was Example 1-5 performed, followed by re-inoculation in a fresh PDB medium and additional liquid culture for 30 days (culturing in a rice barley medium was omitted)

Comparative Example 2. Preparation of Comparative Mycelia—ii

[0053] The co-cultured mycelia of Inonotus obliquus, Phellinus linteus and Sparassis crispa were prepared according to the method of Preparation Example 1 and Example 1 of Korean Patent No. 10-1652035.

Comparative Example 3. Preparation of Comparative Mycelia—iii

[0054] The co-cultured mycelia were prepared according to the method of Preparation Example 1 and Example 1 of Korean Patent No. 10-1652035 as in Comparative Example 2, with the exception that Ganoderma lucidum was used in lieu of Sparassis crispa.

Comparative Example 4. Preparation of Comparative Mycelia—iv

[0055] The co-cultured mycelia of Inonotus obliquus, Ganoderma lucidum and Phellinus linteus mycelia were prepared according to the method of Example 1 of Korean Patent No. 10-1358648.

Comparative Example 5. Preparation of Comparative Cured Meat—i

[0056] The cured meat was prepared in the same manner as in Example 2, with the exception that the co-cultured mycelia of Example 1, cumin powder, basil powder, dill weed powder, dill seed powder and salt were mixed under the conditions of Table 3 below.

TABLE-US-00003 TABLE 3 Weight (g) Preparation Co-cultured Dill Dill conditions of Pork mycelia of Cumin Basil weed seed cured meat fillet Example 1 powder powder powder powder Salt Comparative 100 1.0 0.0 0.0 0.4 0.4 0.1 Example 5-1 Comparative 100 1.0 0.0 0.4 0.4 0.0 0.1 Example 5-2 Comparative 100 1.0 0.4 0.0 0.4 0.0 0.1 Example 5-3 Comparative 100 1.0 0.0 0.8 0.0 0.0 0.1 Example 5-4 Comparative 100 1.8 0.0 0.0 0.0 0.0 0.1 Example 5-5

Comparative Example 6. Preparation of Comparative Cured Meat—ii

[0057] The cured meat was prepared in the same manner as in Example 2, with the exception that the mushroom mycelia C were used under the conditions of Table 4 below, in lieu of using the co-cultured mycelia of Example 1.

TABLE-US-00004 TABLE 4 Preparation conditions of Type of mushroom mycelia used for cured meat preparation of cured meat Comparative Example 6-1 Mushroom mycelia of Comparative Example 1-1 Comparative Example 6-2 Mushroom mycelia of Comparative Example 1-2 Comparative Example 6-3 Mushroom mycelia of Comparative Example 1-3 Comparative Example 6-4 Mushroom mycelia of Comparative Example 1-4 Comparative Example 6-5 Mushroom mycelia of Comparative Example 1-5 Comparative Example 6-6 Mushroom mycelia of Comparative Example 2 Comparative Example 6-7 Mushroom mycelia of Comparative Example 3 Comparative Example 6-8 Mushroom mycelia of Comparative Example 4

Comparative Example 7. Preparation of Comparative Cured Meat—iii

[0058] The cured meat was prepared in the same manner as in Example 2, with the exception that a curing composition and pork fillet were mixed and then allowed to stand in a low-temperature warehouse at 7° C. for 28 hr.

Experimental Example 1. Evaluation of Beta-Glucan Content

[0059] The beta-glucan content in the mushroom mycelia was evaluated by the Korea Institute of Analysis and Technology upon request. The results thereof are shown in Table 5 below. It was confirmed that the co-cultured mycelia obtained through the method of Example 1 had the highest beta-glucan content.

TABLE-US-00005 TABLE 5 Conditions Beta-glucan content (mg/g) Example 1 196.1 Comparative Example 1-1 100.6 Comparative Example 1-2 102.3 Comparative Example 1-3 102.5 Comparative Example 1-4 112.4 Comparative Example 1-5 124.4 Comparative Example 2 149.2 Comparative Example 3 131.4 Comparative Example 4 136.2

Experimental Example 2. Sensory Evaluation of Food Using Mushroom Mycelial Powder as Cooking Seasoning

[0060] 50 people of all ages and both genders were made to taste bean sprout soup seasoned with salt and various powdery mushroom mycelia, and the evaluation thereof was marked on a 5-point scale. Here, no seasonings other than the salt and the mushroom mycelial powder were added to the bean sprout soup, so only the effect of the mushroom mycelial powder on enhancing the taste was observed. Each soup was evaluated as 5 points (very good), 4 points (good), 3 points (normal), 2 points (slightly bad) or 1 point (bad), and the average value thereof is shown in Table 6 below.

[0061] As is apparent f rom the results of Table 6, it was confirmed that the preference for flavor and taste of bean sprout soup cooked with the co-cultured mycelial powder obtained through the method of Example 1 was the highest.

TABLE-US-00006 TABLE 6 Conditions Flavor Savory taste Delicate taste Example 1 4.4 4.5 4.2 Comparative Example 1-1 3.1 2.9 3.0 Comparative Example 1-2 3.0 2.8 3.1 Comparative Example 1-3 2.5 3.2 3.2 Comparative Example 1-4 3.2 3.4 2.7 Comparative Example 1-5 3.1 2.9 3.3 Comparative Example 2 3.4 3.1 3.1 Comparative Example 3 3.5 3.0 3.2 Comparative Example 4 3.2 3.4 3.3

Experimental Example 3. Sensory Evaluation of Food Cooked with Cured Meat

[0062] The cured meat prepared in each of Example 2, Comparative Example 5 and Comparative Example 6 was coated with a batter made from Bekul frying powder and breadcrumbs and then fried.

[0063] For the meat fries thus prepared, sensory evaluation was performed using 100 people of ages 20-50 and both genders. Table 7 below shows the results according to a 5-point scale (5 points: very good, 4 points: good, 3 points: normal, 2 points: slightly bad, 1 point: very bad).

TABLE-US-00007 TABLE 7 Delicate Conditions Flavor Chewiness Softness taste Example 2-1 4.4 4.1 4.2 4.0 Example 2-2 4.2 4.2 4.3 4.1 Example 3-3 4.0 4.4 4.1 4.3 Example 2-4 4.1 4.2 4.0 4.2 Comparative Example 5-1 2.6 4.3 4.1 3.4 Comparative Example 5-2 3.1 4.2 4.2 3.3 Comparative Example 5-3 3.1 4.1 4.2 3.6 Comparative Example 5-4 2.5 4.4 4.3 2.2 Comparative Example 5-5 1.9 4.3 4.2 2.2 Comparative Example 6-1 2.9 2.7 2.7 3.2 Comparative Example 6-2 2.5 3.2 2.8 2.9 Comparative Example 6-3 2.9 3.0 2.8 3.2 Comparative Example 6-4 2.8 3.2 3.1 2.8 Comparative Example 6-5 2.8 2.7 3.2 3.1 Comparative Example 6-6 3.0 3.2 3.0 3.3 Comparative Example 6-7 2.2 3.3 3.2 3.5 Comparative Example 6-8 2.2 3.2 3.0 2.8 Comparative Example 7 2.1 2.9 2.9 2.5

[0064] As is apparent from the results of Table 7, the meat fries prepared from the raw meat cured with the co-cultured mycelia obtained through the method of the present invention exhibited the highest scores in flavor, chewiness, softness and delicate taste. Meanwhile, the cumin powder, basil powder, dill weed powder, and dill seed powder contained in the curing composition are very effective at removing the odor of meat, but the scent thereof is strong, so they may be rejected by those who ingest them, but the strong scent thereof was deemed to be alleviated because the co-cultured mycelia of the present invention was used therewith in the curing process.

Experimental Example 4. Evaluation of Low-Temperature Storage Period of Cured Meat

[0065] The cured meat prepared in Example 2 and Comparative Example 6 was stored at 2° C. for a long period of time, and the storage stability in the cured state was compared on the 3.sup.rd day, 7.sup.th day, and 14.sup.th day. In Table 8 below, a good state is indicated by and decay initiation or progression is indicated by x. With reference to Table 8, it was confirmed that only the cured meat of Examples 2-1 to 2-4, cured with the co-cultured mycelia obtained through the method of the present invention, exhibited good low-temperature storage stability until the 14.sup.th day.

TABLE-US-00008 TABLE 8 Conditions 3.sup.rd day 7.sup.th day 14.sup.th day Example 2-1 ⊚ ⊚ ⊚ Example 2-2 ⊚ ⊚ ⊚ Example 2-3 ⊚ ⊚ ⊚ Example 2-4 ⊚ ⊚ ⊚ Comparative Example 6-1 ⊚ X X Comparative Example 6-2 ⊚ X X Comperative Example 6-3 ⊚ X X Comparative Example 6-4 ⊚ X X Comparative Example 6-5 ⊚ X X Comparative Example 6-6 ⊚ ⊚ X Comparative Example 6-7 ⊚ ⊚ X Comparative Example 6-8 ⊚ ⊚ X