Preparation method for accelerating double-fungus mycelium fermentation of cereals

Abstract

The invention discloses a preparation method for accelerating double-fungus mycelium fermentation of cereals, and belongs to the technical field of processing of nutritious foods. According to the method, lucid ganoderma, hericium erinaceus, sodium selenite and grains are utilized to form complementary advantages, more nutrient substances beneficial to the human body are generated through fermentation decomposition of edible fungus hyphae on the grains, meanwhile, the growth speed of the hyphae is effectively increased through sodium selenite, and efficient conversion from inorganic selenium to organic selenium is further achieved. The content of beneficial components such as protein, polysaccharide and total phenols in the mycelium is remarkably increased in the innovative process. And the cereal food which is more healthy for consumers is researched and developed. According to the method, the culture time is remarkably shortened, the production efficiency is improved, the industrial production cost is reduced, and the economic benefits of enterprises are improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of processing of nutritional foods, and particularly relates to a preparation method for accelerating the fermentation of double-mushroom mycelia on grains. Background Art

[0002] Ganoderma lucidum is a fungus of the Ganodermataceae family and the Ganoderma genus, and is also known as single Ganoderma, Pycnoporus cinnabarinus or old wood fungus, etc. Ganoderma lucidum is one of the oldest mushrooms, a large dark mushroom with a smooth appearance and a woody texture. Ancient Chinese medical books, such as "Compendium of Materia Medica" and "Shennong's Classic of Materia Medica", describe that Ganoderma lucidum can improve intelligence and memory, extend lifespan and relieve stress. Currently, more than 240 compounds have been isolated from Ganoderma lucidum. Triterpenoids are the main components of Ganoderma lucidum and have a wide range of biological and pharmacological activities.

[0003] Hericium erinaceus, also known as Hericium or Hericium erinaceus, is a fungus of the Hericiaceae family and the Hericium genus. Its rich bioactive components endow it with high nutritional and medicinal value. Every 100 g of dried Hericium erinaceus contains approximately 22.3 g of crude protein, 3.5 g of crude fat, 64.8 g of carbohydrates and 4.3 g of moisture. Its fruiting body is medium-sized, large or very large, with a diameter of 3.5 - 10 (30) cm, fleshy, and its shape is head-shaped or obovate, resembling a monkey's head, so it is named "Hericium erinaceus".

[0004] Oats, also known as Bromus japonicus, are plants of the Poaceae family. They are a major miscellaneous grain crop originated from China, an annual cereal crop for both food and forage use, and are one of the eight major food crops in China. Their sown area and total output rank sixth, second only to wheat, rice, corn, barley and sorghum. Oats have high nutritional value and are rich in various macronutrients and micronutrients unique to oats. The protein content is rich, reaching 11% - 20%, higher than common grains such as corn, wheat, rice and sorghum. Oats contain dietary fibers such as β-glucan and lignin, which are beneficial to promoting health.

[0005] Selenium-enriched edible fungi: Selenium is an essential trace element for the human body and has important biological functions. For example, it participates in the synthesis of various proteins in the form of selenocysteine in the human body. If the body lacks selenium, it will lead to reduced immunity, organ dysfunction, and even cause arthritis and Kashin-Beck disease. Selenium cannot be synthesized in the human body and can only be ingested from the outside.

[0006] In recent years, it has been found that edible fungi have strong selenium-enriching ability because the ionic groups on the cell walls of edible fungi have the ability to adsorb elements. Modern research shows that the emergence of selenium-enriched edible fungi not only enriches the ways for people to supplement selenium from the outside, but also enhances the medicinal effect of edible fungi themselves and increases their medicinal value.

[0007] At present, there are many ready-to-eat whole grain products on the market. These products are convenient to consume, and many people use them as breakfast or daily meal replacements. However, most of these products have problems such as a single variety and insufficient nutritional components. Summary of the Invention

[0008] The object of the present invention is to provide a preparation method for accelerating the fermentation of grains by dual-fungal mycelia. In this method, Ganoderma lucidum, Hericium erinaceus, sodium selenite, and grains form complementary advantages. Through the fermentation and decomposition of grains by edible mushroom mycelia, more nutrients beneficial to the human body are produced. At the same time, sodium selenite is used to effectively accelerate the growth rate of mycelia, and the efficient conversion of inorganic selenium to organic selenium is also achieved. This innovative process significantly increases the content of beneficial components such as protein, polysaccharides, and total phenols in the mycelia, and further increases the content of nutritional components in the fermented grains.

[0009] The present invention is achieved through the following technical solutions:

[0010] A preparation method for accelerating the fermentation of grains by dual-fungal mycelia, comprising the following steps:

[0011] S1. Select grains, rinse, remove dust and impurities, then place them in a container and boil until there is no white core and no blooming, then fish them out and drain the water for later use;

[0012] S2. Select an aqueous solution of sodium selenite, add it to the grains treated in S1 and mix well. After the aqueous solution of sodium selenite has fully penetrated, perform a micro-drying treatment on the grains until the grains do not stick to the hands;

[0013] S3. Add gypsum to the grains treated in S2, stir evenly, then place them in a test tube, seal, sterilize under high pressure, and after sterilization, cool and store in a laminar flow hood;

[0014] S4. Under aseptic conditions, separately pick Ganoderma lucidum mycelia and Hericium erinaceus mycelia and inoculate them into different regions of the same petri dish containing a solid activation medium, and culture them in the dark to obtain dual-fungal mycelia;

[0015] S5. Inoculate the dual-fungal mycelia obtained in S4 onto the test tube medium in S3 and perform constant-temperature and light-avoiding culture to obtain a preliminary product of selenium-rich dual-fungal mycelia grains;

[0016] S6. Dry the selenium-rich dual-fungal mycelia grains until the moisture content is not higher than 12-15%, and obtain the finished product of selenium-rich dual-fungal mycelia grains.

[0017] Ganoderma lucidum: It contains a variety of active ingredients such as Ganoderma lucidum polysaccharides, triterpenoids, nucleosides, purines, proteins, polypeptide amino acids, and trace elements such as organic selenium. As the main bioactive ingredient in Ganoderma lucidum, Ganoderma lucidum polysaccharides have functions such as regulating immunity, anti-tumor, and anti-aging.

[0018] Hericium erinaceus: In terms of functions, Hericium erinaceus has the functions of nourishing and strengthening the body, aiding digestion, and benefiting the five internal organs. Modern research shows that it contains active ingredients such as polypeptides, polysaccharides, fats, and proteins, and has certain curative effects on digestive tract tumors, gastric ulcers, duodenal ulcers, gastritis, abdominal distension, etc. The main effects of Hericium erinaceus polysaccharide: 1) Anti-ulcer and anti-inflammatory effects. 2) Anti-tumor effect. 3) Liver protection and liver nourishing effect. 4) Increasing immunity and anti-aging effect. 5) Improving the body's tolerance to hypoxia, increasing the cardiac output, and accelerating the body's blood circulation. 6) Lowering blood sugar and blood lipid levels

[0019] Oats: Rich and balanced in amino acid content; it has the effects of lowering blood sugar and blood lipid levels, enhancing immunity, inhibiting oxidation, anti-aging, and laxative, and is a high-quality coarse grain.

[0020] Finished product of selenium-rich double mushroom mycelium grains: The finished product of selenium-rich double mushroom mycelium grains has the effects of antioxidant, hypoglycemic, anti-tumor, anti-fatigue, antibacterial, etc., and has extremely high food and medicinal value.

[0021] Preferably, in the S1, the grains include any one of oats, corn, wheat, quinoa, rye, and highland barley;

[0022] The boiling duration is 10 - 15 min.

[0023] Preferably, in the S2, the mass concentration of the sodium selenite aqueous solution is 0.05%;

[0024] The mass ratio of the sodium selenite aqueous solution to the grains is 40 g:1000 g;

[0025] The temperature of the micro-drying is a constant temperature environment of 45 - 50 °C.

[0026] Preferably, in the S3, the mass ratio of gypsum to the grains is 10 g:1000 g.

[0027] The functions of gypsum: The action mechanism of gypsum includes directly supplementing sulfur and calcium, promoting the decomposition of organic matter, neutralizing oxalic acid, increasing the cellulose degradation rate and soluble sugar content, etc. At the same time, gypsum can also improve the buffering capacity of the culture medium, inhibit the side effects of too high pH value, prevent large fluctuations in the acidity and alkalinity of the culture medium, and enhance the coagulation effect on the bacterial cell interstitium, thereby reducing the damage to the culture structure.

[0028] Preferably, the solid activated culture medium is prepared by the following method:

[0029] S1. Cut 200 g of fresh peeled potatoes into pieces of 1 - 1.2 cm 3 Then add 1000 mL of distilled water and boil until mushy, filter with eight-layer gauze, and add distilled water to the filtrate to make up to 1000 mL to obtain the filtrate;

[0030] S2. Add glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder and vitamin B1 to the filtrate of S1, stir evenly, sterilize, cool and dispense into petri dishes. After cooling and solidifying, reserve for later use.

[0031] Preferably, in S2, the mixing ratio of glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder and vitamin B1 is 18 - 20 g / L: 1.8 - 2 g / L: 0.8 - 1 g / L: 2.8 - 3 g / L: 18 - 20 g / L: 0.008 - 0.01 g / L.

[0032] Preferably, in S2, sterilize at 120 - 121 °C for 15 - 20 min;

[0033] Cool to 45 - 50 °C and dispense into petri dishes.

[0034] Preferably, in S4, culture in the dark at 25 - 28 °C for 6 - 7 days.

[0035] Preferably, in S5, culture at a constant temperature of 25 - 28 °C for 4 - 5 days.

[0036] Preferably, in S6, dry at 50 - 52 °C for 10 - 12 h until the moisture content is not higher than 12 - 15%.

[0037] Compared with the prior art, the present invention has at least the following technical effects:

[0038] (1) The present invention provides a preparation method for accelerating the fermentation of double - fungus mycelia on grains. In this method, Ganoderma lucidum, Hericium erinaceus, sodium selenite and grains form complementary advantages. Through the fermentation and decomposition of grains by edible - fungus mycelia, more nutrients beneficial to the human body are produced. At the same time, sodium selenite is used to effectively accelerate the growth rate of mycelia, and the efficient conversion of inorganic selenium to organic selenium is also achieved. This innovative process significantly increases the content of beneficial components such as protein, polysaccharide and total phenol in the mycelia.

[0039] (2) In this preparation method, the mycelia of Ganoderma lucidum and Hericium erinaceus can co - exist on the same culture medium. The metabolites of Ganoderma lucidum and Hericium erinaceus promote each other. Ganoderma lucidum polysaccharide and Hericium erinaceus polysaccharide form a synergistic effect in co - fermentation, improving the content of overall active ingredients, and enabling higher yields of mycelia and polysaccharides. The polysaccharide obtained under co - culture has a better immune effect.

[0040] Ganoderma lucidum has the ability to decompose and utilize cellulose and lignin. Cereals (oats) are not only rich in nutrients such as protein, minerals, and vitamins, but also contain lignin in cereal (oat bran), making them excellent raw materials for cultivating Ganoderma lucidum. Inoculating Ganoderma lucidum strains into oats for solid-state fermentation to cultivate the cereal Ganoderma lucidum mycelium complex, while combining the nutrients and beneficial substances of both, produces a unique flavor, which is conducive to the development of new foods.

[0041] During the growth of microorganisms, they need to decompose and utilize the components in the culture medium. At the same time, the microorganisms themselves can also synthesize many nutrients during growth, causing significant changes in the nutrient components of the substrate. After fermenting oats with Ganoderma lucidum and Hericium erinaceus, the protein content of oats increases significantly, the fat content decreases, and the contents of various vitamins and amino acids all increase to varying degrees. The score of limiting amino acids in whole wheat increases, and the amino acid ratio becomes more balanced. Moreover, it also adds a strong flavor of edible fungi, with excellent color, aroma, and taste.

[0042] (3) In this preparation method, adding an appropriate amount of selenium to the culture medium has a promoting effect on the growth of Ganoderma lucidum and Hericium erinaceus mycelia. Research shows that sodium selenite has a greater impact on the growth of Ganoderma lucidum mycelia than carbon sources and nitrogen sources. Adding selenium during the fermentation process can effectively shorten the fermentation time by 20%.

[0043] Edible fungi have a strong ability to enrich selenium through the biotransformation method, enabling selenium to complete the material metabolism transformation in the edible fungi, converting the more toxic inorganic selenium into biologically active organic selenium such as selenium proteins and selenium polysaccharides that are easily absorbed and utilized by the human body. The synthesis of selenium polysaccharides not only converts inorganic selenium into organic selenium, but also changes the chemical structure and biological activity of natural polysaccharides, making them more easily absorbed and utilized by humans. Moreover, the scavenging rate of hydroxyl radicals and reducing ability of Ganoderma lucidum selenium polysaccharides are also significantly better than those of ordinary Ganoderma lucidum polysaccharides. At the same time, other selenium-free bioactive metabolites, including amino acids, polyphenols, flavonoids, cordycepin, and adenosine, are all improved to varying degrees under the influence of selenium enrichment culture. Therefore, selenium-enriched culture is an effective way to improve edible and medicinal fungi rich in trace elements and nutrients, providing a new way to supplement selenium for the human body.

[0044] (4) In this preparation method, sodium selenite is specifically selected to enrich selenium in Ganoderma lucidum, and the organic selenium conversion rate of Ganoderma lucidum reaches 91%-100%. Cultivating selenium-enriched Ganoderma lucidum and Hericium erinaceus with an appropriate concentration of sodium selenite can not only effectively supplement selenium, but also synergistically promote the absorption of various trace elements by the mycelia, and can antagonize heavy metal elements harmful to the human body, which has important guiding significance for supplementing selenium elements in the human body.

[0045] Developing cereal foods that are healthier for consumers. This method significantly shortens the cultivation time, improves production efficiency, reduces industrial production costs, and increases the economic benefits of enterprises. Description of the Drawings

[0046] Figure 1 It is a comparison chart of variance analysis for sodium selenite concentration / mg / kg and hyphal length;

[0047] Figure 2 It is a comparison chart of post hoc multiple comparisons for sodium selenite concentration / mg / kg and hyphal length;

[0048] Figure 3 It is a comparison chart of variance analysis for sodium selenite concentration / mg / kg and hyphal length of Ganoderma lucidum strains;

[0049] Figure 4 It is a comparison chart of variance analysis for sodium selenite concentration / mg / kg and hyphal length of Hericium erinaceus beads;

[0050] Figure 5 It is a growth chart of hyphae and oats. Detailed implementation manners

[0051] The following will describe the implementation scheme of the present invention in detail in conjunction with embodiments. However, those skilled in the art will understand that the following embodiments are only used to illustrate the present invention and should not be regarded as limiting the scope of the present invention. For the specific conditions not specified in the embodiments, they are carried out according to the conventional conditions or the conditions recommended by the manufacturer. For the reagents or instruments not specified for the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0052] Example 1

[0053] A preparation method for accelerating the fermentation of double-fungus mycelium in grains, comprising the following steps:

[0054] S1. Select grains, rinse, remove dust and impurities, then place them in a container and boil until there is no white core and no blooming, then fish them out and drain the water for later use;

[0055] S2. Select an aqueous solution of sodium selenite, add it to the grains treated in S1 and mix well. After the aqueous solution of sodium selenite fully penetrates, perform a micro-drying treatment on the grains until the grains do not stick to the hands;

[0056] S3. Add gypsum to the grains treated in S2, stir evenly, then place them in a test tube, seal, sterilize under high pressure, and after sterilization, cool and store in a super clean bench;

[0057] S4. Under sterile conditions, respectively pick Ganoderma lucidum mycelium and Hericium erinaceus mycelium and inoculate them into different regions of the same culture dish containing a solid activation medium, and culture them in the dark to obtain double-fungus mycelium;

[0058] S5. Inoculate the double-fungus mycelium obtained in S4 onto the test tube medium in S3 and perform constant temperature and dark culture to obtain a preliminary product of selenium-rich double-fungus mycelium grains;

[0059] S6. Dry the selenium-rich dual-fungus mycelium grains until the moisture content is not higher than 12-15% to obtain the finished selenium-rich dual-fungus mycelium grains.

[0060] In S1, the grains are oats;

[0061] The boiling duration is 15 min.

[0062] In S2, the mass concentration of the sodium selenite aqueous solution is 0.05%;

[0063] The mass ratio of the sodium selenite aqueous solution to the grains is 40 g:1000 g;

[0064] The temperature of the micro-drying is a constant temperature environment of 50 °C.

[0065] In S3, the mass ratio of gypsum to grains is 10 g:1000 g.

[0066] The solid activated medium is prepared by the following method:

[0067] S1. Cut 200 g of fresh peeled potatoes into pieces of 1-1.2 cm 3 Then add 1000 mL of distilled water and boil until mushy, filter with eight-layer gauze, and make up the filtrate to 1000 mL with distilled water to obtain the filtrate;

[0068] S2. Add glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder, and vitamin B1 to the S1 filtrate, stir evenly, sterilize, cool, and dispense into petri dishes. After cooling and solidifying, reserve for use.

[0069] In S2, the mixing ratio of glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder, and vitamin B1 is 18 g / L:1.8 g / L:0.8 g / L:2.8 g / L:18 g / L:0.008 g / L.

[0070] In S2, sterilize at 121 °C for 20 min;

[0071] Cool to 50 °C and dispense into petri dishes.

[0072] In S4, culture in the dark at 28 °C for 7 days.

[0073] After the medium is sterilized, it must be placed at room temperature for more than 24 h. Only those without the growth of miscellaneous bacteria can be used. Before use, place it in a laminar flow hood and sterilize it by ultraviolet irradiation for 10 min.

[0074] In S5, culture at a constant temperature of 28 °C for 5 days.

[0075] In S6, dry at 50 °C for 10 h until the moisture content is not higher than 15%.

[0076] Example 2

[0077] A preparation method for accelerating the fermentation of cereal by dual - fungus mycelium, comprising the following steps:

[0078] S1. Select cereals, rinse, remove dust and impurities, then place them in a container and boil until there is no white core and no blooming, then take them out and drain the water, and reserve for later use;

[0079] S2. Select an aqueous sodium selenite solution, add it to the cereals treated in S1 and mix well. After the aqueous sodium selenite solution fully penetrates, perform micro - drying treatment on the cereals until the cereals do not stick to the hand;

[0080] S3. Add gypsum to the cereals treated in S2, stir evenly, then place them in a test tube, seal, sterilize under high pressure, and after sterilization, cool and store in a super - clean bench;

[0081] S4. Under aseptic conditions, separately pick Ganoderma lucidum mycelium and Hericium erinaceus mycelium and inoculate them into different regions of the same petri dish containing a solid activation medium, and culture in the dark to obtain dual - fungus mycelium;

[0082] S5. Inoculate the dual - fungus mycelium obtained in S4 onto the test - tube medium in S3 and culture at a constant temperature in the dark to obtain a preliminary product of selenium - rich dual - fungus mycelium cereal;

[0083] S6. Dry the selenium - rich dual - fungus mycelium cereal until the moisture content is not higher than 12 - 15%, to obtain the finished product of selenium - rich dual - fungus mycelium cereal.

[0084] In S1, the cereal is oats;

[0085] The boiling time is 10 min.

[0086] In S2, the mass concentration of the aqueous sodium selenite solution is 0.05%;

[0087] The mass ratio of the aqueous sodium selenite solution to the cereal is 40 g:1000 g;

[0088] The temperature of the micro - drying is a constant temperature environment of 45 °C.

[0089] In S3, the mass ratio of gypsum to the cereal is 10 g:1000 g.

[0090] The solid activation medium is prepared by the following method:

[0091] S1. Cut 200 g of fresh peeled potatoes into pieces of 1 - 1.2 cm 3 Then add 1000 mL of distilled water and boil until mushy, filter with eight - layer gauze, and make up the filtrate to 1000 mL with distilled water to obtain a filtrate;

[0092] S2. Add glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder and vitamin B1 to the filtrate of S1, stir evenly, sterilize, cool and dispense into petri dishes. After cooling and solidifying, reserve for later use.

[0093] In S2, the mixing ratio of glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder and vitamin B1 is 20 g / L: 2 g / L: 1 g / L: 3 g / L: 20 g / L: 0.01 g / L.

[0094] In S2, sterilize at 120 °C for 15 min;

[0095] Cool to 45 °C and dispense into petri dishes.

[0096] After the medium is sterilized, it must be placed at room temperature for more than 24 h. It can be used only when there is no growth of miscellaneous bacteria. Before use, place it in a laminar flow hood for ultraviolet irradiation sterilization for 10 min.

[0097] In S4, culture in the dark at 25 °C for 6 days.

[0098] In S5, culture at a constant temperature of 25 °C for 4 days.

[0099] In S6, dry at 52 °C for 12 h until the moisture content is not higher than 12%.

[0100] Experimental example:

[0101] I. Preparation of medium:

[0102] 1.1 Solid activation medium:

[0103] The medium formula is: 200 g / L of fresh peeled potato tubers, 20 g / L of glucose, 2 g / L of potassium dihydrogen phosphate, 1 g / L of magnesium sulfate, 3 g / L of peptone, 20 g / L of agar powder, 0.01 g / L of vitamin B1.

[0104] Cut the peeled potatoes into pieces about 1 cm 3 , add 1000 mL of distilled water and boil until soft and rotten, filter with eight-layer gauze to obtain a filtrate. Add distilled water to the filtrate to make up to 1000 mL. After adding other materials according to the formula, boil, adjust the natural pH, sterilize at 121 °C for 20 min, cool to 50 °C, dispense into petri dishes, and reserve after cooling and solidifying. After the medium is sterilized, it must be placed at room temperature for more than 24 h. It can be used only when there is no growth of miscellaneous bacteria. Before use, place it in a laminar flow hood for ultraviolet irradiation sterilization for 10 min.

[0105] 1.2 Selenium-promoted oat medium:

[0106] 1.2.1 Verification of the effect of different concentrations of sodium selenite on the growth of Ganoderma lucidum mycelium

[0107] The selenium enrichment experiment was carried out on the mycelium growth stage of the same Ganoderma lucidum strain by combining with the selenium concentration gradient method. The sodium selenite concentrations in the oat medium were set at 0 mg / kg (CK), 5 mg / kg, 10 mg / kg, 15 mg / kg, and 20 mg / kg respectively, and 8 groups of parallels were set for each experimental treatment group.

[0108] The activated Ganoderma lucidum strain was inoculated into the oat test tube media with 5 different sodium selenite concentrations respectively according to the aseptic operation procedure, and cultured at a constant temperature of 28 °C. Observe the growth of the mycelium in the oat test tube medium. When the length of the mycelium in the test tube reached 1 cm, draw a line as the starting length, record and calculate the mycelium growth rate every 24 h, and determine whether there is a growth-promoting effect on the Ganoderma lucidum strain when adding sodium selenite by comparison.

[0109] Taking 15 d as the end point, the average length of the mycelium of the Ganoderma lucidum strain when the mycelium grew for 15 d is shown in Table 1.

[0110] Table 1 Comparison of the average length of mycelium of different Ganoderma lucidum strains when growing for 15 d / cm

[0111]

[0112]

[0113] As can be seen from Table 1 above, using one-way ANOVA to study different sodium selenite concentrations / mg / kg samples all showed significance (p < 0.05) for the mycelium length, which means that different sodium selenite concentrations / mg / kg samples have differences in the mycelium length.

[0114] Specific analysis shows that:

[0115] The sodium selenite concentration / mg / kg showed significance at the 0.01 level for the mycelium length (F = 119.269, p = 0.000).

[0116] And specifically comparing the differences, the comparison results of the average scores of the groups with obvious differences are "20.0 > 0.0; 30.0 > 0.0; 40.0 > 0.0; 20.0 > 10.0; 30.0 > 10.0; 40.0 > 10.0; 30.0 > 20.0; 40.0 > 20.0; 40.0 > 30.0".

[0117] In summary, different sodium selenite concentrations / mg / kg samples all showed significant differences in the mycelium length.

[0118] As Figure 1 shown, it is the comparison chart of the analysis of variance of sodium selenite concentration / mg / kg and mycelium length.

[0119] Figure 1 The results showed that through the selenium concentration gradient method for selenium-enriched cultivation of the mycelia of the strain, it was found that adding sodium selenite to the culture medium could promote the growth of the mycelia of Ganoderma lucidum strains.

[0120] 1.2.2 Verify the effects of sodium selenite at different concentrations on the growth of Hericium erinaceus mycelia

[0121] Combined with the selenium concentration gradient method, a selenium-enriched experiment was carried out on the mycelial growth stage of the same Hericium erinaceus strain. Sodium selenite concentrations of 0 mg / kg (CK), 10 mg / kg, 20 mg / kg, and 30 mg / kg were set in the oat medium, and 8 replicates were set for each experimental treatment group.

[0122] The activated Hericium erinaceus strain was inoculated into the oat test tube media with 4 different sodium selenite concentrations respectively according to the aseptic operation procedure, and cultured at a constant temperature of 28°C. Observe the growth of the mycelia in the oat test tube media. When the length of the mycelia in the test tube reached 1 cm, mark it as the starting length, record and calculate the mycelial growth rate every 24 h, and determine that the addition of sodium selenite has a growth-promoting effect on the Hericium erinaceus strain by comparison.

[0123] Taking 15 d as the end point, the average length of the mycelia of the Hericium erinaceus strain when the mycelia grew for 15 d is shown in Table 2.

[0124]

[0125] It can be seen from Table 2 above that all samples with different sodium selenite concentrations / mg / kg showed significant differences (p < 0.05) in the growth length of Hericium erinaceus mycelia, which means that samples with different sodium selenite concentrations / mg / kg have differences in the growth length of Hericium erinaceus mycelia.

[0126] As Figure 2 shown, it is a post hoc multiple comparison contrast diagram of sodium selenite concentration / mg / kg and mycelial length.

[0127] Figure 2 The results showed that this experiment proved that selenium has a promoting effect on the growth of Ganoderma lucidum and Hericium erinaceus mycelia.

[0128] 1.2.3 Oat pretreatment

[0129] Take 1 kg of oats, rinse the wheat grains twice with clean water to remove impurities such as dust and wheat bran, and at the same time reduce the base number of miscellaneous bacteria in the culture medium. Boil the oats in a pot, and continue to boil for 15 min after the water boils. When the wheat grains reach the standard of "no white core and no blooming", take them out in time and drain the excess water.

[0130] 1.2.4 Precise addition and mixing of selenium element (sodium selenite)

[0131] Measure 40 g of 0.05% sodium selenite aqueous solution with a graduated cylinder and add it to the oats, ensuring thorough mixing to allow the sodium selenite to fully penetrate.

[0132] The precise addition of selenium was particularly emphasized, aiming to ensure that an appropriate concentration of selenium could not only effectively promote mycelial growth, cell proliferation and differentiation, and improve the conversion rate of inorganic selenium to organic selenium, but also not cause toxicity to organisms.

[0133] As Figure 3 shown, it is a comparative diagram of the analysis of variance of the sodium selenite concentration / mg / kg and the mycelial length of Ganoderma lucidum strains.

[0134] As Figure 4 shown, it is a comparative diagram of the analysis of variance of the sodium selenite concentration / mg / kg and the mycelial length of Hericium erinaceus strains.

[0135] Combined with the above Figure 3 and Figure 4 , due to the mixed growth of Ganoderma lucidum strains and Hericium erinaceus strains, when adding sodium selenite, a concentration of 20 mg / kg that has a promoting effect on both Ganoderma lucidum strains and Hericium erinaceus strains was selected as the optimal concentration.

[0136] 1.2.5 Addition of gypsum and encapsulation of the culture medium

[0137] Place the oats obtained in 1.2.4 (oats added with sodium selenite) in a constant temperature environment of 50 °C and slightly dry it until the wheat grains are not sticky to the touch, then add 10 g of gypsum and stir evenly. Pack it into test tubes with a specification of 18 × 180 mm, with 30 g in each tube (containing 0.3 g of gypsum and 30 g of oats in combination), with a height of about 13 cm, and the tightness should be appropriate, sealed with cotton plugs.

[0138] 1.2.6 Sterilization

[0139] Place the sealed test tubes in an autoclave for sterilization, sterilize at 121 °C for 40 minutes. After sterilization, cool naturally, and take them out when the temperature of the culture medium drops to about 40 °C and put them into a laminar flow hood.

[0140] II. Preparation of the mixed strain of Ganoderma lucidum and Hericium erinaceus (in this technical solution, Ganoderma lucidum and Hericium erinaceus are selected as the mother strains):

[0141] 2.1 Under aseptic conditions, use an inoculation spatula to pick a piece of Ganoderma lucidum mycelium and a piece of Hericium erinaceus mycelium respectively and transfer them to two different positions of the same solid activation medium, and culture them in the dark at 28 °C for 7 days; thus, the mixed mycelium of Ganoderma lucidum and Hericium erinaceus is obtained.

[0142] 2.2 Preparation of selenium-accelerated double-strain fermentation mycelium oats of Ganoderma lucidum and Hericium erinaceus:

[0143] During inoculation, first repeatedly burn the inoculation spatula with the outer flame of an alcohol lamp. After cooling, use the inoculation spatula to pick up the mycelium at the mixed growth site of Ganoderma lucidum and Hericium erinaceus, and quickly inoculate it onto the selenium-enriched oat test tube medium. Then quickly burn the mouth of the test tube with the outer flame of the alcohol lamp and cover the test tube with a stopper to complete the inoculation.

[0144] 2.3 Place the inoculated selenium-accelerated dual-fungus mycelium oat medium in a sterile incubator at 28 °C for constant temperature and light-free cultivation. After cultivating for about 5 days, it can be seen that the mycelia of Ganoderma lucidum and Hericium erinaceus grow all over the surface and inside of the oats, and selenium-enriched Ganoderma lucidum and Hericium erinaceus mycelium oats are obtained.

[0145] As Figure 5 shown, it is the growth diagram of mycelium and oats. In the figure, A is the diagram of oat mycelium taken out of the test tube; B is the growth diagram of mycelium and oats.

[0146] 2.4 Take out the selenium-accelerated dual-fungus mycelium oats and place them in a drying oven at 52 °C for about 10 hours until the moisture content is 15%, and the production of mycelium oats is completed.

[0147] Except for oats, the same method is applicable to various grains such as corn, quinoa, wheat, rye, and hulless barley.

[0148] 2.5 Detection report of the components of the final product oats

[0149]

[0150]

[0151] 2.6 Selenium content table of the final product

[0152] Category Corn Quinoa Wheat Rye Highland barley Selenium mg / kg 10.2 19.1 5.6 9.8 0.7

[0153] Finally, it should be noted that the above are only the preferred embodiments of the present invention and are not used to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention shall be included within the protection scope of the present invention.

Claims

1. A preparation method for accelerating the fermentation of grains by dual fungal mycelia, characterized in that, It includes the following steps: S1. Select grains, rinse them, remove dust and impurities, then place them in a container and boil until there is no white core and no blooming, then fish them out and drain the water for later use; S2. Select sodium selenite aqueous solution, add it to the grains treated in S1 and mix well. After the sodium selenite aqueous solution fully penetrates, perform micro-drying treatment on the grains until the grains do not stick to hands; S3. Add gypsum to the grains treated in S2 and stir evenly, then place them in a test tube, seal it, sterilize under high pressure, and after sterilization, cool and place it in a super clean bench for storage; S4. Under aseptic conditions, respectively pick Ganoderma lucidum mycelium and Hericium erinaceus mycelium and inoculate them into different areas of the same petri dish containing solid activated medium, and culture them in the dark to obtain dual-fungus mycelium; S5. Inoculate the dual-fungus mycelium obtained in S4 onto the test tube medium in S3 and culture it at a constant temperature in the dark to obtain the initial product of selenium-enriched dual-fungus mycelium grains; S6. Dry the selenium-enriched dual-fungus mycelium grains until the moisture content is not higher than 12 - 15%, to obtain the finished product of selenium-enriched dual-fungus mycelium grains.

2. The preparation method of a kind of fermented cereal for accelerating the mycelium of two bacteria according to claim 1, wherein, In S1, the grains include any one of oats, corn, wheat, quinoa, rye, and highland barley; The boiling duration is 10 - 15 min.

3. The preparation method of a kind of fermented cereal for accelerating the fermentation of dual mycelium according to claim 1, characterized in that, In S2, the mass concentration of the sodium selenite aqueous solution is 0.05%; The mass ratio of the sodium selenite aqueous solution to the grains is 40 g:1000 g; The temperature of the micro-drying is a constant temperature environment of 45 - 50 °C.

4. A preparation method for accelerating the fermentation of grains by dual mycelia according to claim 1, characterized in that, In S3, the mass ratio of gypsum to the grains is 10 g:1000 g.

5. The preparation method of a kind of preparation for accelerating the fermentation of double-fungus mycelium of grains according to claim 4, characterized in that, The solid activated medium is prepared by the following method: S1. Cut 200 g of fresh peeled potatoes into pieces of 1 - 1.2 cm 3 Then add 1000 mL of distilled water and cook until mushy. Filter with eight - layer gauze. Add distilled water to the filtrate to make up to 1000 mL to obtain the filtrate; S2. Add glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder, and vitamin B1 to the filtrate in S1, stir evenly, then sterilize, cool, and dispense it into petri dishes, and leave it for use after cooling and solidifying.

6. A preparation method for accelerating the fermentation of grains by dual mycelia according to claim 5, characterized in that In S2, the mixing ratio of glucose, potassium dihydrogen phosphate, magnesium sulfate, peptone, agar powder, and vitamin B1 is 18 - 20 g / L:1.8 - 2 g / L:0.8 - 1 g / L:2.8 - 3 g / L:18 - 20 g / L:0.008 - 0.01 g / L.

7. A preparation method for accelerating the fermentation of grains by dual mycelia according to claim 5, characterized in that, In S2, sterilize at 120 - 121 °C for 15 - 20 min; Cool to 45 - 50 °C and dispense it into petri dishes.

8. A preparation method for accelerating the fermentation of grains by dual mycelium according to claim 1, characterized in that, In S4, culture in the dark at 25 - 28 °C for 6 - 7 days.

9. A preparation method for accelerating the fermentation of grains by dual mycelia according to claim 1, characterized in that, In S5, culture at a constant temperature of 25 - 28 °C for 4 - 5 days.

10. A preparation method for accelerating the fermentation of grains by dual mycelia according to claim 1, characterized in that, In S6, dry at 50 - 52 °C for 10 - 12 h until the moisture content is not higher than 12 - 15%.

Images