Novel hypsizygus tessulatus strain and method for producing biomass using same

Abstract

The present invention provides a Hypsizygus tessulatus SMESH1234 strain deposited as a novel mushroom strain under Accession No. KCCM13526P, a method for producing high-protein fungal biomass using the mycelium thereof, and biomass produced by the method. The biomass produced according to the production method of the present invention exhibits maximized durability and elasticity, thereby enabling use as a food substitute material for various functional foods and alternative meat products, and suggests a high value-added utilization strategy of fungal biomass as a raw material for sustainable food development.

Description

Novel Managak mushroom strain and method for producing biomass using the same

[0001] The present invention relates to a Hypsizygus tessulatus strain and a method for producing biomass using the same, and specifically to a novel Hypsizygus tessulatus strain (Hypsizygus tessulatus SMESH1234) and a method for producing biomass using the mycelium of the same.

[0002] As health and environmental sustainability emerge as major issues in modern society, the food industry is continuously exploring innovative materials to meet these demands. There is growing interest in alternative food ingredients that can promote health while simultaneously reducing carbon emissions and ecosystem destruction; in particular, there is an increasing demand for plant-based and eco-friendly protein materials to replace animal-based ingredients. However, traditional plant-based protein sources have limitations in terms of nutritional composition, taste, and texture, making it necessary to develop new alternative materials to complement them.

[0003] Mushroom mycelium is a material attracting attention as an alternative food ingredient capable of meeting these demands. Mycelium contains components beneficial to health, can be mass-produced with minimal resources, and is environmentally friendly due to its low carbon emissions. Furthermore, mycelium is evaluated as a suitable material for food applications because it grows rapidly and offers a variety of flavors and textures.

[0004] In utilizing such mushroom mycelium, it is necessary to improve productivity and quality stability, and to optimize culture conditions and environments to develop high-quality biomass suitable for various functional foods and alternative meat products.

[0005] The present invention aims to provide the SMESH1234 strain of the Managak mushroom of accession number KCCM13526P.

[0006] The present invention aims to provide a method for producing high-quality and high-functional fungal biomass using the mycelium of a Pleurotus ostreatus strain.

[0007] According to the first aspect of the present invention, the strain of *Hypsizygus tessulatus* SMESH1234, deposited under accession number KCCM13526P, may be provided.

[0008] According to a second aspect of the present invention, a method for producing biomass based on the mycelium of a Pleurotus ostreatus strain may be provided, comprising the steps of: liquid culturing the mycelium of the strain to obtain biomass; filtering the obtained biomass; adjusting the moisture content of the filtered biomass; secondary culturing the biomass with adjusted moisture content; and pressurizing the secondary cultured biomass.

[0009] According to an exemplary aspect of the present invention, the liquid culture step may be performed at 20 to 30°C for 10 to 20 days or at 22 to 27°C for 10 to 18 days.

[0010] According to an exemplary aspect of the present invention, the moisture content of the biomass can be controlled to 80 to 90% (v / w) or to 85 to 90% (v / w).

[0011] According to an exemplary aspect of the present invention, the secondary culture step may be performed at 20 to 30°C or at 22 to 27°C.

[0012] According to an exemplary aspect of the present invention, the pressurization step may be performed at a pressure of 10 to 30 kgf, or at a pressure of 15 to 20 kgf.

[0013] According to a third aspect of the present invention, a mycelium-based biomass of a Pleurotus ostreatus strain produced by the above method may be provided.

[0014] According to an exemplary aspect of the present invention, the biomass may have a network structure.

[0015] According to an exemplary aspect of the present invention, the biomass may have a thickness of 18 to 25 mm.

[0016] According to an exemplary aspect of the present invention, biomass may have a hardness of 10 to 14 N.

[0017] According to an exemplary aspect of the present invention, the biomass is 0.1 to 0.2 or 0.1 to 0.15 N / mm 2 It can have shear strength.

[0018] According to a fourth aspect of the present invention, a food containing biomass may be provided.

[0019] Through the method for producing biomass based on the mycelium of the Managara mushroom strain according to the present invention, it is possible to provide biomass having excellent durability and elasticity and a high-density texture.

[0020] Through the method for producing biomass based on the mycelium of the Managara mushroom strain according to the present invention, it is possible to provide high-protein biomass as an alternative protein raw material.

[0021] The method for producing biomass based on the mycelium of a Pleurotus ostreatus strain according to the present invention can be usefully employed in the production of high-value-added alternative foods utilizing mushroom mycelium.

[0022] By providing a method for producing biomass based on the mycelium of a Pleurotus ostreatus strain according to the present invention, it is possible to contribute to the sustainable development of the food industry.

[0023] Figure 1 is a photograph showing the SMESH1234 strain according to the present invention being subcultured on a PDA plate medium.

[0024] Figure 2 is a photograph showing the network structure of a mycelium-based biomass of the SMESH1234 strain according to one embodiment of the present invention.

[0025] The present invention will be described in detail below with reference to the attached drawings. The following description should be understood as describing the present invention with specific examples, and the technical concept of the present invention is not limited to the description below. Furthermore, the attached drawings are provided to aid in understanding the present invention, and the technical concept of the present invention is not limited to the attached drawings. Additionally, the thickness or size of each component in the drawings may be exaggerated, omitted, or schematically depicted for convenience of explanation.

[0026] In the description of the structure of the present invention as described in this specification, positional relationships or directions are based on the drawings attached to this specification unless specifically stated otherwise.

[0027] In the description of the structure of the invention as described in this specification, descriptions of space or positional relationships refer to the relative positions between the components constituting the invention. Furthermore, unless specifically stated otherwise, another component may exist in the space between one component and another component. For example, when this specification refers to another component being located "above" or "on top" of one component, it includes not only the case where another component is located immediately above one component, but also the case where another component is located between one component and other components.

[0028] In this specification, singular expressions may be interpreted to include the plural unless specifically stated otherwise. In this specification, the expression "comprising" means that the configurations, parts, operations, features, numbers, etc. described in the description are present, and does not exclude the addition of one or more other configurations, parts, operations, features, numbers, etc.

[0029] In this specification, the term "mycelium" collectively refers to a state in which hyphae grow in a densely intertwined manner. It is observed in eukaryotic fungi and prokaryotic actinobacteria. Mushroom mycelium is an organ that absorbs nutrients for the survival of mushrooms and plays a role similar to that of plant roots. It is known that mushroom mycelium is much richer in nutrients and medicinal components compared to the fruiting body, which corresponds to the body.

[0030] In this specification, *Euonymus japonicus* mycelium refers to a form that can be produced by the germination of *Euonymus japonicus* spores in a natural state or by culturing *Euonymus japonicus* strains. Meanwhile, *Euonymus japonicus* mycelium contains various functional materials and therefore has excellent potential for use as a material for food, cosmetics, and medicine. Accordingly, the biomass based on the mycelium of the *Euonymus japonicus* strain according to the present invention can also be utilized as a material for cosmetics and medicine, as well as for food.

[0031] The present invention can be fully achieved by the following description. The following description should be understood as describing preferred embodiments of the present invention, but the present invention is not necessarily limited thereto. Furthermore, the attached drawings are for illustrative purposes only and do not limit the present invention; details regarding individual components may be appropriately understood in accordance with the specific intent of the relevant descriptions provided below.

[0032] The present invention provides the strain of *Hypsizygus tessulatus* SMESH1234, deposited under accession number KCCM13526P.

[0033] In the production of biomass through the cultivation of mycelium of the SMESH1234 strain according to the present invention, the biomass forms a network structure through liquid culture and secondary culture, and internal air bubbles of the biomass are removed through high-pressure treatment, thereby forming a dense texture while simultaneously maximizing the durability and elasticity of the biomass. The biomass produced by the above method can maintain texture stability compared to similar food materials such as soy meat or tofu even after cooking for a certain period of time. Furthermore, through the method of producing biomass according to the present invention, the biomass can be efficiently produced as a material suitable for various functional foods and alternative meat products.

[0034] According to one embodiment of the present invention, the present invention provides a method for producing biomass through mycelial culture of the Hypsizygus tessulatus SMESH1234 strain, comprising the following steps.

[0035] A step of obtaining biomass by liquid culturing the mycelium of *Hypsizygus tessulatus* strain SMESH1234;

[0036] A step of filtering the obtained biomass;

[0037] A step of controlling the moisture content of filtered biomass;

[0038] A step of secondary culturing of moisture-controlled biomass; and

[0039] Step of pressurizing the secondary cultured biomass.

[0040] According to one embodiment of the present invention, liquid culture may mean culturing a strain or mycelium in a liquid culture medium. By liquid culturing the strain or mycelium, a pure mycelial biomass, which is the culture product, can be obtained. The liquid culture medium in the liquid culture step may be any one selected from the group consisting of Potato Dextrose Broth, Yeast-Malt medium, Czapek-Dox Broth, MCM medium (mushroom complete medium), Macaya-Lizano medium, etc., but is not limited thereto, and preferably may be a cellulose or glucose-based medium. The culture temperature is a temperature suitable for culturing the mycelium of the Pleurotus ostreatus strain, and may be 20 to 30°C, 20 to 29°C, or 21 to 28°C, and preferably may be 22 to 27°C. The culture period may be 10 to 20 days, 10 to 18 days, 12 to 18 days, 12 to 16 days, or 13 to 17 days, but preferably 10 to 18 days. The pH of the medium may be 4 to 7, 4 to 6, 4.5 to 6.5, 5 to 7, 5 to 6.5, 4.2 to 6.8, or 4.8 to 6.5, and preferably 5.5 to 6.5. If the pH of the medium has a value below or above the above range, the growth amount of the mycelium may decrease rapidly.The above medium may further include a carbon source, and the carbon source of the medium may be at least one selected from the group consisting of cellulose, hemicellulose, lignin, polysaccharides, starch, trehalose, maltose, sucrose, fructose, galactose, mannose, glucose, yeast extract, dextrin, glycerol, or organic acids, but is not limited thereto. Meanwhile, the concentration of the carbon source may be 1 to 10%, 1.5 to 8.5%, 2 to 9%, 2.5 to 9.5%, 1.5 to 4.5%, 2 to 5%, 2.5 to 5.5%, 1.5 to 2.5%, 2.5 to 3.5%, or 2 to 3% of the liquid medium concentration, but is not limited thereto. The medium may further include a nitrogen source, and the nitrogen source of the medium may be at least one selected from inorganic or organic forms such as yeast extract, peptone, soytone, nitrate, and ammonia, but is not limited thereto. The nitrogen concentration may be 0.01 to 0.6%, 0.02 to 0.7%, 0.02 to 0.06%, 0.03 to 0.07%, 0.05 to 0.4%, 0.1 to 0.5%, 0.15 to 0.35%, or 0.2 to 0.4%, but is not limited thereto. The above medium may further contain inorganic elements, inorganic dyes, etc., and the inorganic element used in the medium may be at least one selected from the group consisting of phosphorus (P), potassium (K), magnesium (Mg), silicon (Si), sodium (Na), iron (Fe), aluminum (Al), zinc (Zn), boron (B), manganese (Mn), molybdenum (Mo), etc., but is not limited thereto. Inorganic salts may include KH2PO4, KH2PO4, (NH4)2HPO4, NH4H2PO4, etc. as phosphates, and MgSO4, etc. as sulfates, but are not limited thereto.

[0041] According to one embodiment of the present invention, filtration may be performed using a filter paper, a filter cloth filter, a filtration device, etc., and the filtration device may be, for example, a rotary drum filtration device (rotary drum vacuum filtration device), but is not limited thereto, and any known filtration method may be used.

[0042] According to one embodiment of the present invention, moisture content control may be achieved using methods such as drying, but is not limited thereto, and any known drying method may be used. Meanwhile, according to an exemplary embodiment of the present invention, the moisture content of the biomass may be 70 to 95% (v / w), 75 to 90% (v / w), 80 to 90% (v / w), 80 to 93% (v / w), 83 to 90% (v / w), or 85 to 90% (v / w), but is not limited thereto, and preferably may have a moisture content of 80 to 90% (v / w).

[0043] According to one embodiment of the present invention, secondary cultivation may refer to culturing biomass with controlled moisture content in a medium. The medium may be any one selected from the group consisting of PDA (Potato Dextrose Agar) medium, MEA (Malt Extract Agar) medium, YMEA (Yeast Malt Extract Agar) medium, CMA (Corn Meal Agar) medium, Czapek-Dox Agar medium, MYA (Malt Yeast Agar) medium, Wheat Bran Agar medium, cellulose-based medium, sawdust-based medium, bran-based medium, rice-based medium, corn-based medium, sugarcane-based medium, or glucose medium, but is not limited thereto, and preferably may be a cellulose-based medium. During the secondary cultivation, a molding die may be used to shape the cultured biomass into a desired form, and the size and shape of the molding die may be determined according to the purpose of use of the biomass as the final product, and is not limited to a specific shape. The secondary culture may be performed, for example, at a temperature of 20 to 30°C, 21 to 29°C, or 20 to 28°C, but is not limited thereto, and preferably at a temperature of 22 to 27°C. The secondary culture may be performed, for example, for 1 to 7 days, 2 to 6 days, 3 to 8 days, or 1 to 4 days, but is not limited thereto, and preferably at 1 to 2 days. Through the liquid culture and secondary culture processes, the biomass according to the present invention may form a network tissue or structure, and accordingly, the elasticity of the material may be increased.

[0044] According to one embodiment of the present invention, the pressurization treatment may be performed by a pressurization means, which may be a compressor, an inert gas, etc., but is not limited thereto, and any known pressurization method may be used. According to an exemplary embodiment of the present invention, the pressurization treatment may be performed inside a mold. According to an exemplary embodiment of the present invention, the pressure applied during pressurization may be, for example, 10 to 30 kgf, 12 to 28 kgf, or 14 to 24 kgf, but is not limited thereto, and preferably, a pressure of 15 to 20 kgf may be applied. Through the pressurization treatment, air bubbles inside the biomass may be removed and a uniform texture may be formed, thereby increasing the density of the biomass according to the present invention and enabling it to have more robust physical properties.

[0045]

[0046] The present invention will be explained in more detail below through examples. These examples are merely illustrative of the invention, and therefore the scope of the invention should not be interpreted as being limited by these examples.

[0047]

[0048] Isolation of the *Hypsizygus tessulatus* strain

[0049] Hypsizygus tessulatus, a species of Hypsizygus tessulatus that grows naturally on Hwangbangsan Mountain in Jeonju, Jeollabuk-do, was collected to isolate a pure strain of the species. A portion of the fruiting body was taken and inoculated onto a PDA medium (containing 200g of potato, 20g of dextrose, and 20g of agar), and the grown strain was cultured in an incubator at 22 to 27°C. Subculture was repeated several times from the cultured Hypsizygus tessulatus until a pure strain was isolated, thereby isolating a pure strain of Hypsizygus tessulatus.

[0050]

[0051] Identification of strain *Hypsizygus tessulatus* SMESH1234

[0052] The Hypsizygus tessulatus strain according to the present invention was subcultured on PDA plates to confirm that it was a pure culture strain. Based on the results of morphological observations, such as fruiting bodies and spores, that appeared when the mushroom grew, the strain was named Hypsizygus tessulatus SMESH1234 and deposited with the Korean Culture Center of Microorganisms (KCCM) on November 5, 2024, receiving accession number KCCM13526P. The Hypsizygus tessulatus SMESH1234 strain is shown in Figure 1.

[0053]

[0054] Biomass production using the mycelium of *Hypsizygus tessulatus* SMESH1234 strain

[0055] Mycelia of the *Hypsizygus tessulatus* strain SMESH1234 cultured on PDA medium were cut into 4x4 mm pieces, and 3 to 5 pieces were inoculated into a 250 mL Erlenmeyer flask containing 10 g of PDA medium (potato extract 4 g / L, glucose 20 g / L, agar 15 g / L). The mixture was then liquid-cultured in an aerobic liquid incubator at 22 to 27°C for 10 to 18 days to form initial mycelia. The biomass, which is the liquid-cultured mycelial product, was filtered through filter paper and adjusted to have a moisture content of 80 to 90%. The moisture-controlled biomass was then secondarily cultured in a molding mold with a cellulose-based medium (containing glucose 20 g / L) at 22 to 27°C for 1 to 2 days to strengthen the high-density network structure of the biomass. Subsequently, a pressure of 15 to 20 kgf was applied inside the molding die to remove air bubbles inside the biomass and to form a uniform texture, and the manufactured biomass is shown in Fig. 2.

[0056]

[0057] Measurement of physical properties

[0058] The physical and structural characteristics of the mycelium-based biomass of the prepared *Hypsizygus tessulatus* SMESH1234 strain were compared with similar food materials and are shown in Table 1 below. Thickness was measured five times using 20 x 20 sections, and the average value was calculated. Hardness was measured using a texture analyzer (Zwick Roell 500N Zwicki) with a 3 mm diameter circular probe. Cutting strength was measured at a speed of 1.5 mm / sec and 400 mm 2 It was measured based on the cross-sectional area.

[0059]

[0060] Food Material Manchurian Mushroom Mycelium Biomass Tofu (Solid Type) Soy Meat (Textured Plant Protein) Shiitake Mushroom (After Drying) Sweet Potato Starch Jelly Thickness (mm) 20.0 18~20 22~25 18~22 20~25 Hardness (N) 11.1 10~12 12~14 10.5 11 Shear Strength (N / mm 2 )0.120.10~0.130.13~0.150.110.11~0.12

[0061] As described above, it was confirmed that the mycelium-based biomass of the *Hypsizygus tessulatus* SMESH1234 strain according to the present invention has physical properties similar to those of similar food materials such as tofu and soy meat. This suggests that the biomass according to the present invention can be utilized as a stable food material with enhanced elasticity.

[0062]

[0063] The embodiments of the present invention have been described above. However, those skilled in the art may make various modifications to the present invention within the scope of the technical concept of the present invention as described in the claims, such as simple design changes, omission of some components, and simple changes in use, depending on the specific application of the technology, and it is obvious that such modifications are also included within the scope of the rights of the present invention.

[0064]

[0065] Depository Name: Korean Culture Collection Center (KCCM)

[0066] Trustee Number: KCCM13526P

[0067] Date of Consignment: 20241105

Claims

1. Strain of *Hypsizygus tessulatus* SMESH1234 deposited under accession number KCCM13526P.

2. A step of obtaining biomass by liquid culturing the mycelium of the strain of claim 1; A step of filtering the obtained biomass; A step of controlling the moisture content of filtered biomass; A step of secondary culturing of biomass with controlled moisture content; and A step of pressurizing the secondary cultured biomass; comprising Method for producing biomass based on the mycelium of a Pleurotus ostreatus strain.

3. In Paragraph 2, The liquid culture step is characterized by being performed at 20 to 30℃ for 10 to 20 days. Method for producing biomass based on the mycelium of a Pleurotus ostreatus strain.

4. In Paragraph 2, The moisture content of the biomass is controlled to 80 to 90% (v / w). Method for producing biomass based on the mycelium of a Pleurotus ostreatus strain.

5. In Paragraph 2, The second culture step is characterized by being performed at 20 to 30℃. Method for producing biomass based on the mycelium of a Pleurotus ostreatus strain.

6. In Paragraph 2, The pressurization step is characterized by being performed at a pressure of 10 to 30 kgf. Method for producing biomass based on the mycelium of a Pleurotus ostreatus strain.

7. Manufactured by the method of any one of paragraphs 2 through 6 Biomass based on the mycelium of the Pleurotus ostreatus strain.

8. In Paragraph 7, Biomass is characterized by having a network structure. Biomass based on the mycelium of the Pleurotus ostreatus strain.

9. Food containing the biomass of Paragraph 7.

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