A strain of Hericium erinaceus with high selenium enrichment capacity, its cultivation and application

Abstract

This disclosure pertains to the field of Hericium erinaceus strains, specifically relating to a Hericium erinaceus strain with high selenium enrichment capacity, its cultivation, and its application. A Hericium erinaceus strain with high selenium enrichment capacity is provided, classified as Hericium erinaceus (Rullex F.) Pers, strain number HC008, deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC40188, dated May 9, 2022. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing. Compared to existing Hericium erinaceus strains on the market, its selenium enrichment capacity is significantly improved.

Description

Technical Field

[0001] This disclosure pertains to the field of Hericium erinaceus strains, specifically relating to a Hericium erinaceus strain with high selenium enrichment capacity, its cultivation, and its application. Background Technology

[0002] Hericium erinaceus (Rull ex F.) Pers belongs to the phylum Basidiomycota, class Basidiomycetes, order Russula, family Hericaceae. In addition to its delicious taste and rich nutrition, it also has the effects of lowering blood sugar, anti-oxidation and anti-aging, promoting peripheral nerve regeneration, inhibiting tumor cell growth, and enhancing immune regulation. It is especially effective in treating gastric ulcers and gastrointestinal cancer.

[0003] Selenium is an essential trace element for the human body. In the body, it mainly participates in the formation of glutathione peroxidase (GSH-Px), thioredoxin reductase (TrxR), and selenoprotein P (Selp), regulating oxidative stress, redox state, and key cellular activities. Selenium deficiency can lead to many diseases, such as Keshan disease and diseases caused by excessive oxidative stress, and can exacerbate the progression of these diseases. Most areas in my country are selenium-deficient, therefore, it is essential to supplement selenium through daily dietary intake.

[0004] Selenium exists in nature primarily as inorganic and organic selenium. Inorganic selenium is highly toxic, while edible fungi have a strong ability to enrich it, converting inorganic selenium into organic selenium that can be absorbed and utilized by the human body. Hericium erinaceus, as an edible fungus with excellent taste, nutrition, and health benefits, is naturally an important material chosen for selenium-enriched cultivation. Deep fermentation of edible fungi has advantages such as short cycle time, small footprint, low cost, suitability for industrial production, and controllable quality, thus showing broad application prospects in edible fungi production. Currently, the selenium enrichment capacity of commercially available Hericium erinaceus strains has not yet reached a very high level. Summary of the Invention

[0005] In view of the shortcomings of the existing technology, the purpose of this disclosure is to provide a strain, cultivation and application of Hericium erinaceus with high selenium enrichment capacity to improve selenium enrichment efficiency.

[0006] Firstly, the purpose of this disclosure is to provide a Hericium erinaceus fungus with high selenium enrichment capacity, which can improve selenium enrichment efficiency compared to existing Hericium erinaceus fungi.

[0007] The objective of this disclosure can be achieved through the following technical solutions:

[0008] A new Hericium erinaceus strain with high selenium enrichment, strain number HC008, is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC NO.40188 and deposit date of May 9, 2022. This new Hericium erinaceus strain is found to improve selenium enrichment efficiency compared to existing Hericium erinaceus strains.

[0009] Hericium erinaceus (Bull. ex Fr.) Pers., also known as monkey head mushroom, is a traditional Chinese medicine. It belongs to the kingdom Fungi, phylum Basidiomycota, class Basidiomycetes, order Polyporaceae, family Dentalceae, and is used medicinally for its fruiting body.

[0010] In some publications, the method for scaling up the Hericium erinaceus strain is described as follows:

[0011] The culture was carried out sequentially in test tubes, shake flasks, fermenters, and solid-state fermentation.

[0012] Test tube bacterial strains: The test tube culture medium is...

[0013] Glucose 5g / L~50g / L, wheat bran 50g / L~200g / L (boil for 30 min and filter the filtrate through gauze), pine sawdust 50g / L~200g / L (boil for 30 min and filter the filtrate through gauze), peptone: 1g / L~50g / L, yeast extract: 1g / L~50g / L, agar 20g / L~100g / L, potassium dihydrogen phosphate 0.5g / L~10g / L, magnesium sulfate 0.5g / L~10g / L, vitamin B1 0.5 mg / L~10 mg / L. Sterilize at 121℃ for 20 min. After cooling, inoculate at three points and incubate in a biochemical incubator at 25℃±1℃ for 5~7 days.

[0014] Shake flask culture medium: glucose 5-100 g / L, soluble starch 5-50 g / L, yeast extract 5-50 g / L, potassium dihydrogen phosphate 0.1-10 g / L, magnesium sulfate 0.1-10 g / L, sterilized at 121℃ for 20 min; after cooling, inoculate with test tube culture and place in a shaking incubator at 25℃±1℃ for 5-7 days;

[0015] Fermentation tank inoculum: The fermentation tank inoculum culture medium is: glucose 5-100g / L, soluble starch 5-50g / L, yeast extract 5-50g / L, potassium dihydrogen phosphate 0.1-10g / L, magnesium sulfate 0.1-10g / L, sterilized at 121℃ for 30min; after cooling, inoculate with shake flask inoculum, ambient temperature 26℃, and aerate during the culture process for 96 hours;

[0016] Solid-state fermentation: The inoculum in the fermentation tank is inoculated into a sterilized and cooled culture medium at an inoculation rate of 20%. The room temperature is maintained at 25℃±1℃ for mycelial culture. After the mycelium is dense and white and fills the bag, the mushroom production management is carried out. The humidity is 65%±5%. Maintaining a day-night temperature difference can stimulate its growth. When the spines grow to 1-1.5cm, they can be harvested.

[0017] The solid fermentation medium consists of 0-900 g / kg corn cobs, 0-900 g / kg cottonseed hulls, 0-900 g / kg sawdust, 100-500 g / kg wheat bran, 5-100 g / kg gypsum, and 5-100 g / kg white sugar. Add 1.2-1.5 times the amount of water to the mixture, pack it into culture bags, sterilize at 121°C for 120 minutes, and then cool after sterilization.

[0018] In some disclosures, the ability of the Hericium erinaceus strain to accumulate selenium increases with increasing inorganic selenium concentration in the culture medium, wherein the selenium content in the culture medium is ≤5000ppm.

[0019] In some disclosures, the Hericium erinaceus strain exhibits the strongest selenium accumulation capacity when cultured on a culture medium; the culture medium formulation consists of: 200–900 g / kg corn cobs, 200–900 g / kg cottonseed hulls, 100–500 g / kg wheat bran, 5–100 g / kg gypsum, and 5–100 g / kg white sugar.

[0020] In some publications, the described Hericium erinaceus strain showed the strongest ability to accumulate selenium when cultured at 25°C and 65% humidity.

[0021] Secondly, the purpose of this disclosure is to provide a food that enhances immunity, the food containing Hericium erinaceus powder, which is obtained by drying Hericium erinaceus fungus as described in the first aspect and then pulverizing it into a fine powder.

[0022] For example, Hericium erinaceus powder is a powder obtained by separating, extracting, and drying the culture of Hericium erinaceus through deep fermentation. This product contains polysaccharides, not less than 4.0%. Loss on drying: Take about 1g of this product and dry it at 105℃ to constant weight; the moisture content should not exceed 8.0% (GB5009.3 National Food Safety Standard - Determination of Moisture in Food). Ash content: Take 0.5-1g of this product and test it according to the method; the ash content should not exceed 8.0% (GB5009.4 National Food Safety Standard - Determination of Ash in Food). Heavy metals: Take 1g of this product and test it according to the method; the heavy metal content should not exceed 20 parts per million (Chinese Pharmacopoeia 2000 Edition, Part II, Appendix VIII H, Method II).

[0023] Hericium erinaceus powder is effective in strengthening the spleen and stomach, calming the nerves, and fighting cancer. It can be used to treat physical weakness, indigestion, insomnia, gastric ulcers, duodenal ulcers, and chronic gastritis.

[0024] Hericium erinaceus powder is typically made from the herb Hericium erinaceus, a common Chinese medicinal herb. Hericium erinaceus is the fruiting body of the fungus Hericium erinaceus (also known as hedgehog mushroom, spiky mushroom, or small spiky monkey head), belonging to the Hericium genus of the family Myricaceae. It is said to not only strengthen the spleen and stomach but also have a calming effect. For those experiencing indigestion, consuming a moderate amount of Hericium erinaceus powder can aid in recovery. Furthermore, it can improve weakness, fatigue, and insomnia. It can also be used as an adjunct treatment for duodenal ulcers, gastric ulcers, and chronic gastritis. Drinking a moderate amount of Hericium erinaceus powder daily can also help prevent digestive tract diseases.

[0025] Thirdly, the purpose of this disclosure is to provide the application of Hericium erinaceus (monkey head mushroom) in supplementing selenium in the human body, such as adding Hericium erinaceus powder as described in the first aspect to the food as a selenium supplement.

[0026] Hericium erinaceus (monkey head mushroom) has a high selenium content, which can be applied in food formulations. For example, Hericium erinaceus powder produced from Hericium erinaceus can be mixed into food. Hericium erinaceus has the effects of increasing appetite, enhancing gastric mucosal barrier function, improving lymphocyte transformation rate, and increasing white blood cell count. Therefore, it can improve the body's immunity to diseases. It is rich in selenium, so food can not only serve as a nutritional supplement to meet the body's daily nutritional needs, but also enhance the body's immunity.

[0027] Of course, the Hericium erinaceus fungus described in this application can also be used in the following ways, such as:

[0028] Enhancing Immune Function: Fifty-five days after syngeneic bone marrow transplantation, recipient mice exhibited severely impaired immune function. Cellular immune functions, such as the ability of spleen cells to produce interleukin-2 (IL-2), were significantly reduced. After 15 days of continuous intraperitoneal injection of Hericium erinaceus polysaccharide and thymosin, the ability of mouse spleen cells to produce IL-2, as well as their proliferative response to concanavalin A (ConA) stimulation and mixed lymphocyte culture response, were significantly enhanced. This indicates that the combined use of Hericium erinaceus polysaccharide and thymosin significantly promotes the early recovery of immune function in syngeneic bone marrow transplanted mice compared to thymosin alone. Intraperitoneal injection of 2 mg / mouse of Hericium erinaceus polysaccharide for 7 consecutive days significantly improved the phagocytic function of peritoneal macrophages in mice; it also promoted hemolysin production and increased humoral immunity. Intraperitoneal injection of 2.88 mg / mouse for 8 consecutive days significantly counteracted the leukopenia induced by cyclophosphamide poisoning, with the leukopenia rate being only half that of the control group. Hericium erinaceus polysaccharide has a strong promoting effect on the proliferation of mouse thymocytes activated by ConA in vitro, and can also promote the proliferation of splenic lymphocytes. It also has a synergistic effect on B cells stimulated by lipopolysaccharide (LPS).

[0029] Antitumor effect: Sarcoma S180 cells were inoculated subcutaneously in the left anterior axilla of male Swiss mice, followed by oral administration of Hericium erinaceus polysaccharide at doses of 50 mg / kg, 100 mg / kg, and 200 mg / kg once daily for 7 consecutive days. The results showed that all three dose groups inhibited tumor growth and significantly activated natural killer (NK) cell activity. Tumor weight was negatively correlated with the corresponding splenic NK cell activity. Hericium erinaceus also inhibited the hepatocarcinogenic effect of aflatoxin in rats and reduced the number of lesions on liver sections.

[0030] Anti-ulcer and hypoglycemic effects: Pepsin inhibition and adsorption experiments demonstrated that the mechanism of action of Hericium erinaceus tablets in treating gastric ulcers may be due to the inhibition of pepsin activity, thereby promoting ulcer healing. Hericium erinaceus polysaccharides can lower blood glucose levels in normal mice and tetrahydropyrimidine-induced diabetic mice.

[0031] Anti-aging effects: Hericium erinaceus mycelium polysaccharides and fruiting body polysaccharides can significantly increase the flight ability of fruit flies, reduce the content of lipofuscin in the myocardial tissue of newly hatched fruit flies and mice, and increase the specific activity of superoxide dismutase (SOD) in the brain and liver of mice.

[0032] Fourthly, the application of Hericium erinaceus, as in the first aspect, in increasing the activity of glutathione peroxidase in the liver and blood.

[0033] Fifthly, the application of Hericium erinaceus, as described in the first aspect, in enhancing the activity of superoxide dismutase in the liver and blood.

[0034] Sixthly, the application of Hericium erinaceus, as described in the first aspect, in reducing malondialdehyde levels in the liver and blood.

[0035] The beneficial effects of this disclosure are:

[0036] The Hericium erinaceus strain of this application can improve selenium enrichment efficiency compared to existing Hericium erinaceus strains. Attached Figure Description

[0037] To more clearly illustrate the technical solutions in the embodiments of this disclosure or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, those skilled in the art can obtain other drawings based on these drawings without creative effort.

[0038] Figure 1 electron micrograph Detailed Implementation

[0039] The technical solutions of the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. Based on the embodiments of this disclosure, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this disclosure.

[0040] A type of Hericium erinaceus fungus with high selenium enrichment capacity, classified as Hericium erinaceus (strain number HC008), has been deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 40188 on May 9, 2022. The deposit address is No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing.

[0041] The Hericium erinaceus strain obtained in this application was isolated from the fruiting bodies of Hericium erinaceus collected from trees in Qimen County, Huangshan City, Anhui Province. 16S rDNA identification was performed. This Hericium erinaceus strain is used to enhance selenium enrichment capacity. Microscopic observation revealed its physical morphology as follows: nearly spherical, white, blocky, and soft in appearance, with a narrow basal attachment point. Spines approximately 1-1.5 cm long cover the entire fruiting body. Under the microscope, the hyphae consist of closely packed tubular cells with thin walls, septa, and branches. Due to its high selenium enrichment capacity, Hericium erinaceus can be applied to… For example, in food formulations, Hericium erinaceus powder produced from Hericium erinaceus fungi can be mixed into food. Hericium erinaceus has effects such as increasing appetite, strengthening the gastric mucosal barrier, improving lymphocyte transformation rate, and increasing white blood cell count, thus enhancing the body's immunity against diseases. It is rich in selenium, making the food not only a nutritional supplement to meet the body's daily nutritional needs but also to improve immunity. Adding selenium-enriched Hericium erinaceus fruiting body product (selenium-enriched Hericium erinaceus powder) to food can supplement the body with selenium. This selenium-enriched Hericium erinaceus powder can also be added to other food ingredients, such as sorbitol and magnesium stearate.

[0042] Example 1: Isolation of Hericium erinaceus strains (tissue isolation method)

[0043] Separation method: In a sterilized laminar flow hood, wipe the surface of the collected Hericium erinaceus fruiting bodies with 75% alcohol for disinfection. Then, tear open the Hericium erinaceus fruiting bodies and, using a sterilized inoculation needle, pick up a 1cm section at the junction of the cap and stipe or at the gills. 2 Prepare inoculation blocks by taking three small pieces and placing them evenly on the slant of a test tube. Incubate in an incubator at 25℃±1℃. Maintain aseptic technique during inoculation. After 5–7 days of incubation, dense white mycelia and aerial hyphae will grow on the culture medium, yielding the slant test tube culture.

[0044] The culture medium consists of: glucose 5g / L–50g / L, wheat bran 50g / L–200g / L (boiled for 30 minutes and then filtered through gauze), pine sawdust 50g / L–200g / L (boiled for 30 minutes and then filtered through gauze), peptone 1g / L–50g / L, yeast extract 1g / L–50g / L, agar 20g / L–100g / L, potassium dihydrogen phosphate 0.5g / L–10g / L, magnesium sulfate 0.5g / L–10g / L, and vitamin B1 0.5mg / L–10mg / L.

[0045] Example 2: Activation of Hericium erinaceus strain (activation method)

[0046] Activation method: Using a sterilized inoculation needle, pick up 1cm of the bacterial culture from the slant test tube. 2 Prepare inoculation blocks by taking three small pieces and placing them evenly on a test tube slant. Incubate at 25℃±1℃ for 5-7 days until the mycelium covers the slant and grows white. Then, repeat the above procedure to transfer the culture again for activation. The activation medium consists of: glucose 5g / L~50g / L, wheat bran 50g / L~200g / L (boil for 30 minutes and filter the filtrate through gauze), pine sawdust 50g / L~200g / L (boil for 30 minutes and filter the filtrate through gauze), peptone: 1g / L~50g / L, yeast extract: 1g / L~50g / L, agar 20g / L~100g / L, potassium dihydrogen phosphate 0.5g / L~10g / L, magnesium sulfate 0.5g / L~10g / L, and vitamin B1 0.5mg / L~10mg / L.

[0047] In this embodiment, the mycelium of Hericium erinaceus strain is transferred to a suitable activation medium and cultured at 25℃±1℃ for 5-7 days until the mycelium is dense. Then, the strain is transferred again according to the above operation method to activate the strain.

[0048] Example 3: Method for scaling up the culture of Hericium erinaceus strain (scale-up culture method)

[0049] Expanded culture process: in vitro culture → shake flask culture → fermenter culture → solid-state fermentation culture

[0050] Test tube bacterial cultures: glucose 5g / L–50g / L, wheat bran 50g / L–200g / L (boil for 30 minutes, then filter the filtrate through gauze), pine sawdust 50g / L–200g / L (boil for 30 minutes, then filter the filtrate through gauze), peptone 1g / L–50g / L, yeast extract 1g / L–50g / L, agar 20g / L–100g / L, potassium dihydrogen phosphate 0.5g / L–10g / L, magnesium sulfate 0.5g / L–10g / L, vitamin B1 0.5mg / L–10mg / L. Sterilize at 121℃ for 20 minutes. After cooling, inoculate at three points and incubate in a biochemical incubator at 25℃±1℃ for 5–7 days.

[0051] Shake flask culture medium: glucose 5-100 g / L, soluble starch 5-50 g / L, yeast extract 5-50 g / L, potassium dihydrogen phosphate 0.1-10 g / L, magnesium sulfate 0.1-10 g / L, sterilized at 121℃ for 20 min. After cooling, inoculate with test tube culture and incubate in a shaking incubator at 25℃±1℃ for 5-7 days.

[0052] Fermentation tank starter culture: The fermentation tank starter culture medium consists of: glucose 5-100 g / L, soluble starch 5-50 g / L, yeast extract 5-50 g / L, potassium dihydrogen phosphate 0.1-10 g / L, and magnesium sulfate 0.1-10 g / L. The fermentation tank volume is 600 L. Sterilize at 121℃ for 30 min. After cooling, inoculate with 200 ml of starter culture in a shake flask. Incubate at an ambient temperature of 26℃ for 96 hours with aeration.

[0053] Solid-state fermentation: The inoculum in the fermenter is inoculated into a sterilized and cooled culture medium at a rate of 20%. The room temperature is maintained at 25℃±1℃ for mycelial cultivation. Once the mycelium has thickened and whited into the bag, fruiting management begins, with humidity at 65%±5%. Maintaining a day-night temperature difference can stimulate growth. Harvesting can begin when the spines have grown to 1–1.5 cm.

[0054] The solid fermentation medium consists of 0-900 g / kg corn cobs, 0-900 g / kg cottonseed hulls, 0-900 g / kg sawdust, 100-500 g / kg wheat bran, 5-100 g / kg gypsum, and 5-100 g / kg white sugar. 1.2-1.5 times the amount of water is added to the mixture, which is then placed into a special edible mushroom cultivation bag and sterilized at 121℃ for 120 minutes. After sterilization, the mixture is cooled.

[0055] Example 4: Verification of the selenium enrichment capacity of the Hericium erinaceus strain of this application and commercially available Hericium erinaceus varieties.

[0056] A solid fermentation medium with the same inorganic selenium concentration of 500 ppm was set up. Group 1 used a patented strain, Group 2 used a commercially available Hericium erinaceus strain (model 920), Group 3 used a commercially available Hericium erinaceus strain (model 4916), and Group 4 used a commercially available Hericium erinaceus strain (model 6). Environmental factors were kept constant to verify the selenium enrichment effect.

[0057] The groups are as follows (the solid fermentation medium is: corn cob 0-900g / kg, cottonseed hull 0-900g / kg, sawdust 0-900g / kg, wheat bran 100-500g / kg, gypsum 5g-100g / kg, white sugar 5g-100g / kg, with 1.2-1.5 times the amount of water added):

[0058] The dry weight of each material was 50 kg, and each batch contained approximately 120 bags. After sterilization and cooling, inoculation, mycelial culture, and fruiting culture were carried out. The inoculation ratio and culture method were kept consistent, and the culture period was approximately 60 days. After harvesting, the materials were dried and pulverized, and the total selenium content in each group of Hericium erinaceus powder was tested.

[0059] Selenium content detection method: Method 1, hydride atomic fluorescence spectrometry, as specified in GB5009.93 "National Food Safety Standard - Determination of Selenium in Food". The results for each group are as follows:

[0060]

[0061] The results above show that the patented Hericium erinaceus strain has a much higher capacity for selenium enrichment than ordinary commercially available Hericium erinaceus strains.

[0062] Example 5: Verification of the relationship between the selenium enrichment capacity of the Hericium erinaceus strain under this application and the concentration of inorganic selenium in the culture medium.

[0063] Solid-state fermentation media with different inorganic selenium concentrations were set up. Control group 1 had no added inorganic selenium; experimental group 2 had an inorganic selenium concentration of 500 ppm; experimental group 3 had an inorganic selenium concentration of 1000 ppm; experimental group 4 had an inorganic selenium concentration of 2000 ppm; experimental group 5 had an inorganic selenium concentration of 3000 ppm; and experimental group 6 had an inorganic selenium concentration of 5000 ppm. Other environmental factors remained the same. The selenium enrichment effect was verified as follows:

[0064] The groups are as follows (the solid fermentation medium is: corn cob 0-900g / kg, cottonseed hull 0-900g / kg, sawdust 0-900g / kg, wheat bran 100-500g / kg, gypsum 5g-100g / kg, white sugar 5g-100g / kg, with 1.2-1.5 times the amount of water added):

[0065]

[0066] The dry weight of each material was 50 kg, and each batch contained approximately 120 bags. After sterilization and cooling, inoculation, mycelial culture, and fruiting culture were carried out. The inoculation ratio and culture method were kept consistent, and the culture period was approximately 60 days. After harvesting, the materials were dried and pulverized, and the total selenium content in each group of Hericium erinaceus powder was tested.

[0067] Selenium content detection method: Method 1, hydride atomic fluorescence spectrometry, as specified in GB5009.93 "National Food Safety Standard - Determination of Selenium in Food". The results for each group are as follows:

[0068]

[0069] The results above show that the selenium enrichment capacity of this Hericium erinaceus strain increases with the increase of inorganic selenium concentration in the solid culture medium. The selenium enrichment capacity is very strong, but the increase slows down after reaching a certain level. When the selenium content reaches 5000 ppm, it completely inhibits the growth of Hericium erinaceus.

[0070] Example 6 verifies the best selenium enrichment effect of this application under specific conditions.

[0071] Different culture medium groups were set up, the main difference being the selection of several commonly used main ingredients, with the same concentration of inorganic selenium (500 mg / kg). After the fruiting bodies were harvested, the selenium content in the fruiting bodies of different groups was tested to determine the optimal culture medium ratio.

[0072] The solid fermentation medium from Example 4 can be formulated as follows:

[0073] Group 1: 360g / kg cottonseed hulls + 360g / kg corn cobs + 260g / kg wheat bran + 10g / kg gypsum + 10g / kg white sugar

[0074] Group 2: 360g / kg sawdust + 360g / kg cottonseed hulls + 260g / kg wheat bran + 10g / kg gypsum + 10g / kg white sugar

[0075] Group 3: 360g / kg sawdust + 360g / kg corn cob + 260g / kg wheat bran + 10g / kg gypsum + 10g / kg white sugar

[0076] The dry weight of each material was 50 kg, and each batch contained approximately 120 bags. After sterilization and cooling, inoculation, mycelial culture, and fruiting culture were carried out. The inoculation ratio and culture method were kept consistent, and the culture period was approximately 60 days. After harvesting, the materials were dried and pulverized, and the total selenium content in each group of Hericium erinaceus powder was tested.

[0077] Selenium content detection method: Method 1, hydride atomic fluorescence spectrometry, as specified in GB5009.93 "National Food Safety Standard - Determination of Selenium in Food". The results for each group are as follows:

[0078]

[0079] The results above show that different culture medium ratios have a significant impact on the selenium enrichment capacity of Hericium erinaceus. The culture medium formula combination of group 3 (cottonseed hull 360g / kg + corn cob 360g / kg + wheat bran 260g / kg + gypsum 10g / kg + white sugar 10g / kg) has the strongest selenium enrichment capacity.

[0080] Example 7: Specific implementation example of temperature and humidity combination:

[0081] Different cultivation temperatures were set, and the humidity was maintained at 65% ± 5%. After the fruiting bodies were harvested, the selenium content in the fruiting bodies of different groups was tested to determine the optimal combination of temperature and humidity.

[0082] The groups are as follows (the solid fermentation medium is: 360g / kg cottonseed hulls + 360g / kg corn cobs + 260g / kg wheat bran + 10g / kg gypsum + 10g / kg white sugar, with 1.2 to 1.5 times the amount of water added to the materials).

[0083] Group 1: Temperature 16℃

[0084] Group 2: Temperature 20℃

[0085] Group 3: Temperature 25℃

[0086] Group 4: Temperature 30℃

[0087] The dry weight of each material was 50 kg, and each batch contained approximately 120 bags. After sterilization and cooling, inoculation, mycelial culture, and fruiting culture were carried out. The inoculation ratio and culture method were kept consistent, and the culture period was approximately 60 days. After harvesting, the materials were dried and pulverized, and the total selenium content in each group of Hericium erinaceus powder was tested.

[0088] Selenium content detection method: Method 1, hydride atomic fluorescence spectrometry, as specified in GB5009.93 "National Food Safety Standard - Determination of Selenium in Food". The results for each group are as follows:

[0089]

[0090] The results above show that different temperature and humidity combinations have a significant impact on the selenium enrichment capacity of Hericium erinaceus, with group 3 (temperature 25℃ + humidity 65%) showing the strongest selenium enrichment capacity.

[0091] Example 8: Practical application of Hericium erinaceus strain

[0092] Practical Application Example 1: The selenium-enriched Hericium erinaceus fruiting body product (selenium-enriched Hericium erinaceus powder) enriched using this Hericium erinaceus strain can be used to produce selenium-enriched edible fungus powder or selenium-containing food raw materials for food fortification. It can be added to "rice and its products", "wheat and its products", "wheat flour and its products", bread, "biscuits", "modified milk powder (children and adults)", "dairy beverages", etc.; "selenium-enriched Hericium erinaceus dry powder" food raw material can be used as a raw material for the production of functional foods such as candy tablets and solid beverages.

[0093] Practical Application Example 2: Used to supplement the selenium needed by the human body and increase the selenium content in the blood.

[0094] Adding selenium-enriched Hericium erinaceus fruiting body products (selenium-enriched Hericium erinaceus powder) from this Hericium erinaceus strain to food can supplement the human body with selenium. For example, this selenium-enriched Hericium erinaceus powder can be mixed with other food ingredients, such as sorbitol and magnesium stearate, to make 0.5g / tablet compressed candies. Each tablet contains 50μg of selenium. Eight users were selected to compare their blood selenium levels before and after use. One tablet was consumed daily for two months. The data are as follows:

[0095]

[0096]

[0097] As can be seen from the table above, products (compressed candies) containing selenium-enriched Hericium erinaceus powder can indeed significantly increase the selenium content in the human blood and supplement the body with selenium.

[0098] Practical Application Example 3: The selenium-enriched Hericium erinaceus powder produced by this strain helps maintain the normal physiological function of the immune system.

[0099] The effect on organ indices in mice was investigated using 40 Kunming mice (18–22 g) provided by Anhui Medical University, randomly divided into four groups. Group 1 was the control group, fed a low-selenium basal diet; Group 2 was the sodium selenite group, receiving a certain amount of sodium selenite daily by gavage in addition to the low-selenium basal diet (selenium content equivalent to the daily gavage dose of dry powder in Group 4); Group 3 was the common Hericium erinaceus group, receiving a certain amount of common Hericium erinaceus powder daily by gavage in addition to the low-selenium basal diet (equivalent to the daily gavage dose of dry powder in Group 4); and Group 4 was the selenium-enriched Hericium erinaceus fruiting body group, receiving the same dose daily by gavage at 30 times the recommended human dose. Mice were sacrificed after 30 days by bone marrow removal. The spleen, thymus, and liver were dissected, accurately weighed, and the organ weight index was calculated.

[0100] Organ body mass index = weight of immune organs (mg) / body weight (g) × 10.

[0101] Effects of each treatment group on organ indices in mice (mg / 10g)

[0102] blank Sodium selenite Common monkey head mushroom Selenium-enriched Hericium erinaceus spleen 42.5 43.6 43.3 44.1 thymus 14.9 15.8 15.3 16.7 liver 41.2 43.3 42.1 44.8

[0103] This shows that the non-specific immunity of mice was improved after gavage administration of selenium-enriched Hericium erinaceus fruiting bodies compared with other groups.

[0104] Practical application example 4: The antioxidant effect of selenium-enriched Hericium erinaceus powder produced by this strain;

[0105] Forty Kunming mice, weighing 18–22 g, provided by Anhui Medical University, were randomly divided into four groups. Group 1 was the control group, fed a low-selenium basal diet; Group 2 was the sodium selenite group, receiving a certain amount of sodium selenite daily by gavage in addition to the low-selenium basal diet (selenium content equivalent to the daily gavage dose of dried powder in Group 4); Group 3 was the common oyster mushroom group, receiving a certain amount of common oyster mushroom powder daily by gavage in addition to the low-selenium basal diet (equivalent to the daily gavage dose of dried powder in Group 4); and Group 4 was the selenium-enriched oyster mushroom fruiting body group, receiving the same dose daily by gavage at 30 times the recommended human dose. Mice were sacrificed after 30 days via medullary amputation. Blood was collected via retinal bleeding, anticoagulated with pre-prepared heparin sodium (50 U / ml whole blood), centrifuged at 3500 rpm for 10 min, and the plasma was separated and stored at 4°C for analysis.

[0106] Determination of GSH-Px activity: The activity of GSH-Px is expressed as the rate of reaction catalyzing GSH. The specific procedures were performed according to the kit instructions.

[0107] SOD activity was detected using the xanthine oxidase method, and the specific steps were performed according to the kit instructions.

[0108] MDA content detection: The TBA method was used, and the specific steps were performed according to the kit instructions.

[0109] Effects of selenium-enriched Hericium erinaceus fruiting bodies on glutathione peroxidase (GSH-Px) activity in mouse liver and blood:

[0110] blank Sodium selenite Common monkey head mushroom Selenium-enriched Hericium erinaceus plasma 367.534 443.156 371.088 521.341 liver 525.546 589.562 538.124 639.167

[0111] This shows that after oral administration of selenium-enriched Hericium erinaceus fruiting bodies, the activity of glutathione peroxidase in the blood and liver of mice increased. Sodium selenite and ordinary Hericium erinaceus also showed some improvement compared to the control group, but the effect was not as good as that of the selenium-enriched Hericium erinaceus powder group.

[0112] Effects of selenium-enriched Hericium erinaceus fruiting bodies on superoxide dismutase (SOD) activity in mouse liver and blood

[0113] Sound (U / ml):

[0114] blank Sodium selenite Common monkey head mushroom Selenium-enriched Hericium erinaceus plasma 267.135 289.435 271.132 328.173 liver 331.013 351.143 336.874 381.078

[0115] This indicates that the levels of superoxide dismutase in the blood and liver of mice after gavage administration of selenium-enriched Hericium erinaceus fruiting bodies are high.

[0116] Increased enzyme activity

[0117] Effects of selenium-enriched Hericium erinaceus fruiting bodies on malondialdehyde (MDA) levels in mouse liver and blood (U / ml):

[0118] blank Sodium selenite Common monkey head mushroom Selenium-enriched Hericium erinaceus plasma 7.3 6.8 6.6 4.7 liver 7.5 6.6 6.4 4.9

[0119] This indicates that the malondialdehyde (MDA) content in the blood and liver of mice decreased after gavage administration of selenium-enriched Hericium erinaceus fruiting bodies.

[0120] It can be used for the large-scale cultivation of Hericium erinaceus, for the cultivation of selenium-enriched Hericium erinaceus, with strong enrichment capacity, greatly saving costs, for strain research, and through the metabolism of Hericium erinaceus strains, inorganic selenium is converted into organic selenium that is more easily absorbed by the human body before the cultivation of selenium-enriched Hericium erinaceus mycelium. The resulting selenium-enriched Hericium erinaceus mycelium has a high selenium content, is more easily absorbed by the human body, and has better nutritional and health benefits.

[0121] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of this disclosure. In this specification, the illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0122] The foregoing has shown and described the basic principles, main features, and advantages of this disclosure. Those skilled in the art should understand that this disclosure is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of this disclosure. Various changes and modifications can be made to this disclosure without departing from its spirit and scope, and all such changes and modifications fall within the scope of this disclosure as claimed.

[0123]

Claims

1. A type of Hericium erinaceus fungus with high selenium enrichment capacity, characterized in that, The Hericium erinaceus strain mentioned is strain number HC008, deposited at the Collection Center of the Institute of Microbiology, Chinese Academy of Sciences, with accession number CGMCCNO.40188 and deposit date of May 9, 2022. The method for scaling up the Hericium erinaceus strain: The culture was carried out sequentially in test tubes, shake flasks, fermenters, and solid-state fermentation. Test tube culture medium: glucose 5g / L~50g / L, wheat bran 50g / L~200g / L, pine sawdust 50g / L~200g / L, peptone 1g / L~50g / L, yeast extract 1g / L~50g / L, agar 20g / L~100g / L, potassium dihydrogen phosphate 0.5g / L~10g / L, magnesium sulfate 0.5g / L~10g / L, vitamin B1 0.5mg / L~10mg / L. Sterilize at 121℃ for 20min, cool, and inoculate at three points. Place in a biochemical incubator at 25℃±1℃ for 5-7 days. After boiling wheat bran for 30 minutes, filter the filtrate through gauze; after boiling pine sawdust for 30 minutes, filter the filtrate through gauze. Shake flask culture medium: glucose 5-100 g / L, soluble starch 5-50 g / L, yeast extract 5-50 g / L, potassium dihydrogen phosphate 0.1-10 g / L, magnesium sulfate 0.1-10 g / L, sterilized at 121℃ for 20 min; after cooling, inoculate with test tube culture and place in a shaking incubator at 25℃±1℃ for 5-7 days; Fermentation tank inoculum: The fermentation tank inoculum culture medium is: glucose 5-100g / L, soluble starch 5-50g / L, yeast extract 5-50g / L, potassium dihydrogen phosphate 0.1-10g / L, magnesium sulfate 0.1-10g / L, sterilized at 121℃ for 30min; after cooling, inoculate with shake flask inoculum, ambient temperature 26℃, and aerate during the culture process for 96 hours; Solid-state fermentation: The inoculum in the fermentation tank is inoculated into a sterilized and cooled culture medium for cultivation. The inoculation amount is 5% to 20%. The room temperature is maintained at 25℃±1℃ for mycelial cultivation. After the mycelium is dense and white and fills the bag, the mushroom management is carried out. The humidity is 65%±5%. Maintaining the temperature difference between day and night can stimulate its growth. When the stingers grow to 1 to 1.5 cm, they can be harvested. The solid fermentation medium consists of 0-900 g / kg corn cobs, 0-900 g / kg cottonseed hulls, 0-900 g / kg sawdust, 100-500 g / kg wheat bran, 5-100 g / kg gypsum, and 5-100 g / kg white sugar. Add 1.2-1.5 times the amount of water to the mixture, pack it into culture bags, sterilize at 121°C for 120 minutes, and then cool after sterilization. The selenium enrichment capacity of the Hericium erinaceus strain increased with the increase of inorganic selenium concentration in the culture medium, and the selenium content in the culture medium was ≤5000ppm. The Hericium erinaceus strain exhibited the strongest selenium accumulation capacity when cultured on the culture medium. The culture medium formula consisted of: 200–900 g / kg corn cobs, 200–900 g / kg cottonseed hulls, 100–500 g / kg wheat bran, 5–100 g / kg gypsum, and 5–100 g / kg white sugar. The Hericium erinaceus strain exhibited the strongest selenium enrichment capacity when cultured at 25°C and 65% humidity.

2. A food product for enhancing immunity, characterized in that, The product contains selenium-enriched Hericium erinaceus powder obtained by drying Hericium erinaceus as described in claim 1 and then pulverizing it into a fine powder.

3. An application of Hericium erinaceus as described in claim 1 in supplementing selenium in the human body.

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