Wild phellinus igniarius rich in polyphenol and application of wild phellinus igniarius

Abstract

The invention relates to wild phellinus igniarius rich in polyphenol and application thereof, and belongs to the technical field of microorganisms. The wild phellinus igniarius SH-27 provided by the invention is inonotus hispidus, and is preserved in the Guangdong Microbial Culture Collection Center on February 10, 2025, and the preservation number is GDMCC No: 65865. The sporocarp and the fermentation liquor of the wild phellinus igniarius SH-27 provided by the invention both have polyphenol compounds with relatively high concentration levels and have various active ingredients, mycelium pellets cultured by the liquid have the polyphenol content of 29.74 mg / g, the triterpene content of 3.52 mg / g, the polysaccharide content of 88.61 mg / g and the flavone content of 5.14 mg / g, and the average yield of polyphenol metabolites in the fermentation liquor reaches 33.72 mg / L. By optimizing the liquid culture medium, the yield of the mycelium pellets can be improved, and the yield of the mycelium pellets can be improved. The polyphenols content of mycelium pellets can reach 46.98 mg / g, the polyphenols content of sporocarps cultivated by the strain can reach 77.32 mg / g, various active components of the strain are beneficial to research and development of anti-inflammatory and anti-tumor drugs and improvement of human immunity, and the strain has extremely high research and development values.

Description

Technical Field

[0001] The invention relates to wild mulberry linterus rich in polyphenols and application thereof, belonging to the technical field of microorganisms. Background Art

[0002] Phellinus igniarius is a rare wood-dwelling basidiomycete with great medicinal potential. It is often found on the trunks of mulberry trees. Its fruiting body is yellow-brown, hence its name. It is also known as mulberry, mulberry ear, etc. Basidiomycota ) Agaricus Agaricomycetes ) Dermatomales ( Hymenochaetales ) Hymenochaetaceae ) Porites ( Inonotus ). It contains a wide variety of polyphenols with different structures, which have high biological activity and have good effects in anti-oxidation, anti-inflammatory, and anti-tumor.

[0003] Polyphenols are an important component of the secondary metabolites of Phellinus igniarius. Based on their structural characteristics, these polyphenols can be divided into pyrones, furanones, flavonoids, phenolic acids, catechins, isocoumarins, and other phenolic components. Pyrones, including styrylpyrones, benzopyrones, and their derivatives, are the most prominent of these. Their pharmacological activity increases with the number of phenolic hydroxyl groups they contain. They exhibit significant antioxidant, anti-inflammatory, anti-tumor, and immune system modulating effects. This study aimed to identify strains of Phellinus igniarius that produce high levels of polyphenols.

[0004] The medicinal benefits of mulberry fungus were first documented in Compendium of Materia Medica and Treatise on the Nature of Materia Medica. Its cold nature and slightly bitter flavor have been linked to its various benefits, including anti-tumor, gastric ulcer treatment, and liver protection. In Traditional Chinese Medicine, mulberry fungus is widely believed to benefit the five internal organs, detoxify, and stop diarrhea and bleeding. It is used to treat metrorrhagia, dysentery, rectal prolapse, and amenorrhea. Modern medical research has shown that mulberry fungus has immune-regulating, anti-aging, and anti-tumor properties.

[0005] Porites crassifolia has important medicinal value. It is commonly used in Northeast China to treat stomach ailments such as indigestion. In Xinjiang, it is collected as "Mulberry Gross" and used as a medicinal herb to treat difficult-to-treat conditions such as cancer, diabetes, gout, and arthritis. Currently, Porites crassifolia strains are scarce, and new strains have the potential to discover new active compounds. Therefore, discovering new strains of Porites crassifolia not only enriches the strain pool but also serves as an important avenue for discovering novel active compounds. Summary of the Invention

[0006] In view of the deficiencies in the prior art, the present invention provides a wild mulberry fungus rich in polyphenols and applications thereof. The mulberry fungus has the characteristic of high production of polyphenol compounds and has extremely high development value.

[0007] The technical solutions of the present invention are as follows: A wild mulberry fungus SH-27 rich in polyphenols, wherein the wild mulberry fungus SH-27 is a mulberry fungus ( Inonotus hispidus ), deposited in Guangdong Provincial Microbial Culture Collection on February 10, 2025, the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 65865.

[0008] The above-mentioned polyphenol-rich wild Morigenia igniarius was collected from the century-old ancient mulberry garden in Xiajin, Shandong Province, China. It was identified as a strain of Porites fusca. It was isolated and cultured from the wild fruiting body to obtain the mother strain, which was named SH-27.

[0009] The above-mentioned polyphenol-rich wild Morigensis SH-27 contains rich active ingredients. The polyphenol content of its liquid-cultured mycelium pellets is 29.74 mg / g, the triterpenoid content is 3.52 mg / g, the polysaccharide content is 88.61 mg / g, and the flavonoid content is 5.14 mg / g. The average yield of polyphenol metabolites in the fermentation broth is 33.72 mg / L. By optimizing the liquid culture medium, the polyphenol content of its mycelium pellets can reach 46.98 mg / g, and the polyphenol content of its cultivated fruiting bodies can reach 77.32 mg / g.

[0010] The method for culturing the mycelium of the wild Phellinus igniarius SH-27 comprises the following steps: Take the wild Phellinus igniarius SH-27 strain and inoculate it onto a PDA solid culture medium plate, culture it at 24-30℃ until a circular fungus lawn grows, take a circular fungus lawn with a diameter of 4-6 mm and inoculate it onto a PDA solid culture medium plate, culture it at 24-30℃ for 5-7 days until mycelium grows.

[0011] The liquid culture method of the wild Phellinus igniarius SH-27 comprises the following steps: The mycelium of wild Phellinus igniarius SH-27 was inoculated into liquid culture medium and cultured at 24-30°C and 100-200 rpm for 10-14 days until mycelial balls grew.

[0012] According to the present invention, the PDA solid culture medium preferably comprises the following components: 12 g / L potato extract powder, 20 g / L glucose, 15 g / L agar, the solvent is water, and the pH is natural.

[0013] Preferably, according to the present invention, the components of the liquid culture medium are: glucose 15~35g / L, yeast extract 0.5~3.5g / L, MgSO40.5~3.5g / L, KH2PO40.5~3.5g / L, vitamin B10.01~0.2g / L, the solvent is water, and the pH is 6.5~7.5.

[0014] Further preferably, the components of the liquid culture medium are: glucose 30.1 g / L, yeast extract 3.15 g / L, MgSO4 3.1 g / L, KH2PO4 1.1 g / L, vitamin B1 0.1 g / L, the solvent is water, and the pH is 7.2.

[0015] The method for culturing the fruiting body of the wild Phellinus igniarius SH-27 comprises the following steps: The cultivation medium was placed in a spawn bag, which was tied tightly and sterilized. The mycelial balls of wild Phellinus igniarius SH-27 were inoculated into the spawn bag. After culturing for 10 to 14 days, the fruiting bodies of wild Phellinus igniarius SH-27 were obtained by continuing to culture under the same conditions for 30 to 40 days.

[0016] Preferably, according to the present invention, the cultivation culture medium is composed of: 50~80 parts by mass of poplar wood chips, 10~30 parts by mass of wheat bran, 0.5~1 parts by mass of yeast extract, 0.5~1 parts by mass of calcium oxide, 1~2 parts by mass of calcium sulfate, and 0.5~1 parts by mass of baumazipi powder; the moisture content of the cultivation culture medium is 55~65%.

[0017] Further preferably, the poplar wood chips are sieved through a 20-30 mesh sieve, and the jujube peel powder is sieved through a 60-80 mesh sieve.

[0018] According to the preferred embodiment of the present invention, the sterilization conditions are: sterilization at 121° C. for 1.5 to 2 hours.

[0019] According to the preferred embodiment of the present invention, the spawn culture conditions are: 24-30° C., relative air humidity 60-70%, and light-shielding culture.

[0020] The method for preparing the polyphenol-rich wild Phellinus igniarius SH-27 fermentation broth comprises the following steps: 1) Take a 4-6 mm diameter mycelium from wild Phellinus igniarius SH-27 and inoculate it onto a PDA solid medium plate. Incubate for 7-8 days to activate the strain. 2) Take fresh bacterial lawns from the edge of the activated strain and inoculate them into PDB liquid medium. Cultivate for 7-10 days to obtain seed liquid. 3) The seed liquid is inoculated into a fermentation medium at a volume ratio of 5-15%, and fermentation is performed to obtain a polyphenol-rich wild phellinus igniarius SH-27 fermentation liquid.

[0021] According to the preferred embodiment of the present invention, the culture conditions in step 1) are: 24-30°C.

[0022] According to the preferred embodiment of the present invention, the culture conditions in step 2) are: 100-200 rpm, 24-30°C.

[0023] According to the preferred embodiment of the present invention, the culturing time of the seed solution in step 2) is 168 hours.

[0024] According to the preferred embodiment of the present invention, the PDB liquid culture medium in step 2) comprises the following components: 12 g / L potato extract powder, 20 g / L glucose, water as the solvent, and natural pH.

[0025] According to a preferred embodiment of the present invention, the inoculation amount of step 2) is: 3 to 4 pieces of bacterial moss with a diameter of 4 to 6 mm are inoculated per 150 mL of culture medium.

[0026] According to the preferred embodiment of the present invention, the fermentation culture conditions in step 3) are: pH 6.5-7.5, temperature 24-30° C., ventilation volume 2-4 L / h, and stirring speed 100-200 rpm.

[0027] Further preferably, the fermentation culture conditions in step 3) are: pH 7, temperature 30°C, ventilation volume 2 L / h, and stirring speed 100 rpm.

[0028] According to the preferred embodiment of the present invention, the fermentation culture cycle in step 3) is 5 to 12 days.

[0029] Further preferably, the fermentation culture cycle in step 3) is 8 days.

[0030] According to the present invention, preferably, the culture medium for the fermentation culture in step 3) is PDB liquid culture medium.

[0031] The application of the above-mentioned wild Morinda igniarius SH-27 in the preparation of antioxidant products.

[0032] The application of the wild Morinda igniarius SH-27 in the preparation of functional food or medicine for treating or preventing colitis.

[0033] Beneficial effects: (1) The wild Morinda igniarius SH-27 screened out by the present invention is rich in active ingredients. The polyphenol content of its liquid cultured mycelium pellets is 29.74 mg / g, the triterpenoid content is 3.52 mg / g, the polysaccharide content is 88.61 mg / g, and the flavonoid content is 5.14 mg / g. The average yield of polyphenol metabolites in the fermentation broth is 33.72 mg / L. By optimizing the liquid culture medium, the polyphenol content of its mycelium pellets can reach 46.98 mg / g, and the polyphenol content of its cultivated fruiting bodies can reach 77.32 mg / g. It has extremely high research and development value.

[0034] (2) The present invention has good application prospects in the preparation of functional foods or drugs for anti-oxidation, treatment or prevention of ulcerative colitis, and has great medicinal potential. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the mycelial morphology of wild Phellinus igniarius SH-27; Figure 2 This is a morphological diagram of the fruiting body of wild Phellinus igniarius SH-27. DETAILED DESCRIPTION

[0036] The technical solution of the present invention is further described below in conjunction with embodiments, but the protection scope of the present invention is not limited thereto.

[0037] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available materials and reagents unless otherwise specified.

[0038] Example 1 Isolation, Screening and Identification of Wild Phellinus igniarius (1) Wild Phellinus igniarius fruiting bodies were collected from the century-old ancient mulberry garden in Xiajin, Shandong Province, China. Under sterile conditions, the surface was disinfected by wiping with 75% (v / v) alcohol cotton. The alcohol cotton should not be too wet, and no alcohol or cotton wool should remain on the fruiting bodies after wiping. The blade was burned with an alcohol lamp to disinfect it, and then the blade was used to cut along the fiber of the fruiting body. After cutting, the blade was burned again, and a square was cut in the center of the cut surface with the blade to facilitate picking up with tweezers. The cut fruiting body tissue was picked up with pointed tweezers burned with an alcohol lamp; (2) Preparation of mother culture medium: After sterilizing the plate at 121°C under high temperature and high pressure, pour the prepared and sterilized PDA solid culture medium onto the clean bench and wait for it to solidify; The PDA solid culture medium comprises the following components: 12 g / L potato extract powder, 20 g / L glucose, 15 g / L agar, water as solvent, and natural pH.

[0039] (3) Preparation of mother culture: Transfer the fruiting body tissue separated in step (1) to the PDA solid culture medium in step (2) and culture in a 30°C constant temperature box in the dark; (4) Preparation of original seed: When the mycelium has grown but other bacteria have not yet grown, pick the tip of the mycelium and place it on the PDA solid culture medium, and culture it in a 30℃ constant temperature box in the dark.

[0040] This strain grows slowly. After about three weeks of culture, the colony diameter is about 7~9cm. In the early stage of growth, white velvety hyphae grow. As the hyphae grow, they gradually change from white to yellow-brown, but the fresh hyphae at the edge of the colony are still white. Figure 1 As shown, this is the daily storage form of the strain. Mycelium or fungus moss can be picked from this strain in subsequent experiments.

[0041] (5) Expansion culture: Pick the hyphae from the edge of the colony of the original culture for liquid culture, culture in a shake flask at 24°C and 100 rpm for 10-14 days to obtain mycelial balls; The liquid culture medium is PDB liquid culture medium, which comprises 12 g / L potato extract powder, 20 g / L glucose, water as solvent, and natural pH.

[0042] (6) Identification of fungal species: The mycelial pellets obtained in step (5) were ground with liquid nitrogen, and the sample genome was extracted using a plant genomic DNA extraction kit. ITS-PCR amplification was performed using universal primers for the fungal internal transcribed spacer region, ITS1 / ITS4, synthesized by Shanghai Sangon Biotechnology Co., Ltd.; The primers are as follows: ITS1: 5'-TCCGTAGGTGAACCTGCGG-3', ITS4:5'-TCCTCCGCTTATTGATATGC-3', The ITS-PCR experiment was performed with a system of 50 μL, as shown in Table 1.

[0043] Table 1

[0044] The reaction conditions were as follows: initial denaturation at 94°C for 3 min, followed by 35 cycles of denaturation at 94°C for 30 s, annealing at 55°C for 30 s, and extension at 72°C for 1 min, with a final extension at 72°C for 5 min. The amplified products were sent to Shanghai Sangon Biotechnology Co., Ltd. for sequencing.

[0045] The above ITS sequences were BLASTed using the NCBI database and found to be Inonotus hispidus The clone SH1 similarity was as high as 99%, and it was identified as a strain of Porifera scalypifolia ( Inonotus hispidus ) and named SH-27. It was deposited in Guangdong Provincial Microbiological Culture Collection Center (GDMCC) on February 10, 2025. The address is: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 65865.

[0046] The wild mulberry fungus SH-27 screened by the present invention belongs to the family of Coriolus, the genus of Microporus, and has different shapes and colors, black or yellow-brown, with a cap in the shape of a horseshoe, and solitary fruiting bodies. The surface of some sample fruiting bodies is smooth, while the surface of some sample fruiting bodies has rough objects. The mycelium morphology is as follows: Figure 1 As shown, the fruiting body morphology is as follows Figure 2 The ITS sequence of the wild mulberry fungus SH-27 is shown in SEQ ID NO.1.

[0047] Example 2 Liquid Culture of Wild Phellinus Igneus SH-27 An alcohol lamp was lit on a sterile clean bench to create a local aseptic operating environment. Mycelial moss was picked from the mycelium of wild Phellinus igneus SH-27 and inoculated into the optimized culture medium for liquid culture. The culture was shaken at 24°C and 100 rpm for 10-14 days to obtain mycelial balls.

[0048] The optimized culture medium components are: glucose 30.1 g / L, yeast extract 3.15 g / L, MgSO4 3.1 g / L, KH2PO4 1.1 g / L, vitamin B1 0.1 g / L, solvent is water, pH 7.2.

[0049] Example 3 Determination method of active ingredient content of wild Phellinus igniarius SH-27 1. Determination of polyphenol content (1) Preparation of standard curve Accurately pipette 1 mL of the gallic acid standard stock solution (200 mg / L) into a 10 mL volumetric flask and dilute to volume with 70% ethanol to obtain the gallic acid working solution. Then, pipette 0, 0.2, 0.4, 0.6, 1.0, and 1.5 mL of the gallic acid working solution into separate 10 mL volumetric flasks. Add 2.5 mL of Folin-phenol reagent and shake well. Add 2.5 mL of 15% Na₂CO₃ solution, dilute to the mark with water, and shake well. Incubate in a 40°C water bath for 60 minutes, then allow to cool for 20 minutes. Measure the absorbance at 760 nm. Plot a standard curve with concentration as the horizontal axis and absorbance as the vertical axis.

[0050] (2) Detection of polyphenols in samples The mycelial pellets obtained in step (5) of Example 1 or the liquid culture of Example 2 were dried at 60°C and then ground into powder. 0.4 g of mycelial powder was accurately weighed and added with 4 mL of 70% ethanol. After ultrasonic extraction at 55°C for 1 h, the mixture was centrifuged at 6000 rpm for 10 min, and the supernatant was collected. The residue was extracted twice with 3 mL of 70% ethanol solution under the same extraction conditions as above. The supernatants from the three extractions were combined and diluted to 10 mL with 70% ethanol to obtain the ethanol extract sample solution for the determination of polyphenols, triterpenes, and flavonoids.

[0051] (3) Sample testing Pipette 1.0 mL of the alcohol extract into a 10 mL stoppered colorimetric tube. Add 2.5 mL of Folin-phenol reagent and shake well. Add 2.5 mL of 15% Na₂CO₃ solution, bring to volume with water, and shake well. Incubate in a 40°C water bath for 60 minutes, allow to cool for 20 minutes, and measure the absorbance at 760 nm. Calculate the total polyphenol concentration in each sample using the standard curve.

[0052] 2. Determination of polysaccharide content The extraction of polysaccharides was carried out according to the method of GB / T 15672-2009 for the determination of total sugar content in edible fungi. (1) Preparation of glucose standard curve To create a glucose standard curve: Pipette 0 mL, 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, and 1.0 mL of a glucose standard solution (100 mg / L) into a 10 mL stoppered colorimetric tube. Fill to 1.0 mL with distilled water. Add 1.0 mL of 5% (v / v) phenol solution to each solution. Quickly add 5.0 mL of concentrated sulfuric acid and allow to react for 10 minutes. Then, place the colorimetric tube in a 30°C water bath for 20 minutes. Measure the absorbance of a certain amount of the reaction solution at 490 nm. Based on the measured values, plot a standard curve with glucose concentration as the horizontal axis and absorbance as the vertical axis.

[0053] (2) Detection of polysaccharides in samples The mycelial pellets obtained from the liquid culture in step (5) of Example 1 were dried at 60°C and ground into powder. 0.25 g of the crushed mycelial powder was accurately weighed and added to 50 mL of distilled water. The mixture was then boiled in a water bath for 3 h. A group containing only distilled water and no mycelial powder was also boiled in a water bath for 3 h as a blank control. After the boiling water bath, the mixture was filtered, and the filtrate obtained was used as the sample for the intracellular polysaccharide content test.

[0054] (3) Sample testing Sample polysaccharide content determination: Accurately pipette 0.1 mL of each diluted sample (to ensure accurate experimental results, dilute the liquid to an absorbance between 0.2 and 0.8) into a 10 mL stoppered colorimetric tube. Fill to 1.0 mL with distilled water. Add 1.0 mL of 5% (v / v) phenol solution to the tube. Quickly add 5 mL of concentrated sulfuric acid, allow to react for 10 minutes, and place the tube in a 30°C water bath for 20 minutes. Measure absorbance using the same method as the standard curve. Perform three replicates for each sample. Use a blank control sample to zero the spectrophotometer. Calculate the polysaccharide content of each sample based on the standard curve.

[0055] 3. Determination of triterpenoid content (1) Standard curve: Weigh the oleanolic acid standard and add anhydrous ethanol to prepare a 0.2 mg / mL standard solution. Accurately measure 0, 0.1, 0.2, 0.3, 0.4, and 0.5 mL of the standard solution into centrifuge tubes. Evaporate the solvent. Accurately add 0.2 mL of freshly prepared 0.05 g / mL vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid. Shake well. Heat in a 70°C water bath for 15 minutes, cool in an ice bath for 5 minutes, add 4 mL of ethyl acetate, shake well, and use the vanillin-glacial acetic acid and ethyl acetate mixture as a blank. Measure the absorbance of the standard at 560 nm to construct a standard curve.

[0056] (2) Sample testing Take 1 mL of the ethanol extract sample, add 0.2 mL of freshly prepared 0.05 g / mL vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid, shake well, heat in a 70°C water bath for 15 minutes, cool in an ice bath for 5 minutes, add 4 mL of ethyl acetate, shake well, and measure the absorbance. Calculate the triterpenoid content in each sample using the standard curve.

[0057] 4. Determination of flavonoid content (1) Standard curve: Accurately weigh 10 mg of rutin standard, dried to a constant weight, dissolve in anhydrous ethanol, and adjust to volume in a 50 mL volumetric flask. Pipette 0.1 mL, 0.2 mL, 0.3 mL, 0.4 mL, and 0.5 mL of the 0.2 g / L rutin standard solution into a test tube and add anhydrous ethanol to make the volume 1.5 mL. Sequentially add 0.1 mL of 100 g / L aluminum nitrate solution and 0.1 mL of 98 g / L potassium acetate solution, and make up to 5 mL with water. Let the solution stand for 1 hour. Measure the absorbance at 420 nm using 30% (v / v) ethanol as a blank to construct a standard curve.

[0058] (2) Sample testing Take 1.5 mL of the alcohol extract sample and add 0.1 mL of 100 g / L aluminum nitrate solution and 0.1 mL of 98 g / L potassium acetate solution, respectively. Make up to 5 mL with water. Prepare three replicates for each sample and let it stand for 1 hour. Measure the absorbance using the same method as the standard curve. Calculate the flavonoid content in each sample using the regression equation of the standard curve.

[0059] The wild mulberry fungus SH-27 screened by the present invention has a liquid culture mycelial pellet with a polyphenol content of 29.74 mg / g, a polysaccharide content of 88.61 mg / g, a triterpene content of 3.52 mg / g, and a flavonoid content of 5.14 mg / g. By optimizing the liquid culture medium, the polyphenol content of the mycelial pellet can reach 46.98 mg / g.

[0060] Example 4 Preparation of wild Phellinus linteus SH-27 fruiting bodies 1. Preparation of cultivation medium The cultivation medium used is granular poplar wood chips. The cultivation medium formula is 77% poplar wood chips, 20% wheat bran, 1% yeast extract, 0.5% calcium oxide, 1% calcium sulfate, and 0.5% tamarind bark powder; the tamarind bark powder is sieved through a 60-mesh screen.

[0061] First, pick fresh, mold-free granular poplar chips, pass them through a 20-mesh sieve to remove wood blocks and other debris, weigh the poplar chips according to the formula, then put the weighed yeast extract, calcium oxide, calcium sulfate and baomazipi powder into water, add the poplar chips, stir evenly, adjust the moisture content of the cultivation medium to 60%, and then put it into the culture bag. The culture bag is a high-temperature resistant polypropylene bag (specifications 17cm×33cm×0.05cm). The bag should be moderately tight when filling, then tie the bag tightly, and use high-pressure steam sterilization at 121°C for 2 hours.

[0062] 2. Cultivation, inoculation and fungus culture On a clean bench, inoculate wild SH-27 mycelial pellets into spawn bags, inoculating 3-5 pellets per bag. Immediately move the inoculated bags to the incubation room and place them on a culture rack for growth. Incubate at 24°C, 70% relative humidity, and shaded from light. Regularly check for contamination and mycelial growth during incubation. Any contamination should be removed promptly to prevent spread. After 10-14 days of incubation, mycelium should cover the entire surface of the culture medium and penetrate deep into the cultivation medium. Continue incubation under the same conditions, and Phellinus igniarius fruiting bodies will emerge in approximately 30-40 days.

[0063] The cultured wild Phellinus igniarius SH-27 fruiting bodies were freeze-dried and ground into powder, and the polyphenol content of the fruiting bodies was measured to be 77.32 mg / g. Liquid Fermentation of Wild Phellinus Igneus SH-27 1) In a clean bench, use a 5 mm diameter, sterilized borer to inoculate a lawn of wild Phellinus igniarius SH-27 mycelium onto a PDA solid medium plate. Seal the plate with parafilm and place in a 30°C constant temperature incubator for 7 days to activate the strain. 2) Use a sterile 5 mm diameter borer to take three pieces of fresh moss from the edge of the activated strain obtained in step 1) and inoculate them into PDB liquid medium (150 mL of medium per 250 mL Erlenmeyer flask). Incubate in a constant temperature shaker at 30°C and 200 rpm for 168 h to obtain seed solution. The PDB liquid culture medium comprises the following components: 12 g / L potato extract powder, 20 g / L glucose, water as solvent, and natural pH.

[0064] 3) inoculating the seed solution obtained in step 2) at a 10% (v / v) inoculum into a 5-L fermentor containing 3 L of PDB liquid medium. Fermentation parameters were set at pH 7, temperature 30°C, aeration 2 L / h, and agitation 100 rpm. Fermentation was continued for 8 days to obtain a fermentation broth. 4) High performance liquid chromatography (HPLC) analysis determined the yield of polyphenol metabolites in the fermentation broth to be 33.72 mg / L.

[0065] The wild Morinus igniarius SH-27 provided by the present invention has polyphenol compounds at relatively high concentrations in its fruiting bodies and fermentation broth, and has multiple active ingredients. The liquid-cultured mycelial pellets have a polyphenol content of 29.74 mg / g, a triterpene content of 3.52 mg / g, a polysaccharide content of 88.61 mg / g, and a flavonoid content of 5.14 mg / g. The average yield of polyphenol metabolites in the fermentation broth reaches 33.72 mg / L. By optimizing the liquid culture medium, the polyphenol content of the mycelial pellets can reach 46.98 mg / g, and the polyphenol content of the cultivated fruiting bodies can reach 77.32 mg / g. The various active ingredients of the strain are helpful for the research and development of anti-inflammatory and anti-tumor drugs and for improving human immunity. The strain is a strain with development prospects.

Claims

1. A wild mulberry fungus SH-27 rich in polyphenols, wherein the wild mulberry fungus SH-27 is a thick hairy fibrous fungus ( Inonotus hispidus ), deposited in Guangdong Provincial Microbial Culture Collection on February 10, 2025, the deposit address is 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou, and the deposit number is GDMCC No: 65865.

2. The method for culturing mycelium of wild Phellinus igniarius SH-27 according to claim 1, characterized in that: The steps include: Take the wild Phellinus igniarius SH-27 strain and inoculate it onto a PDA solid culture medium plate, culture it at 24-30℃ until a circular fungus lawn grows, take a circular fungus lawn with a diameter of 4-6 mm and inoculate it onto a PDA solid culture medium plate, culture it at 24-30℃ for 5-7 days until mycelium grows.

3. The liquid culture method of wild Phellinus igniarius SH-27 according to claim 1, comprising the following steps: The mycelium of the wild Phellinus igniarius SH-27 according to claim 2 is picked and inoculated into a liquid culture medium, and cultured at 24-30° C. and 100-200 rpm for 10-14 days until a mycelial ball is grown.

4. The culture method according to claim 3, wherein The components of the liquid culture medium are: glucose 15-35 g / L, yeast extract 0.5-3.5 g / L, MgSO4 0.5-3.5 g / L, KH2PO4 0.5-3.5 g / L, vitamin B1 0.01-0.2 g / L, the solvent is water, and the pH is 6.5-7.5; Further preferably, the components of the liquid culture medium are: glucose 30.1 g / L, yeast extract 3.15 g / L, MgSO4 3.1 g / L, KH2PO4 1.1 g / L, vitamin B1 0.1 g / L, the solvent is water, and the pH is 7.

2.

5. The method for culturing the fruiting body of the wild Phellinus igniarius SH-27 according to claim 1, comprising the following steps: The cultivation medium is placed in a spawn bag, the bag is tied tightly and then sterilized, the mycelial pellets of the wild mulberry fungus SH-27 according to claim 3 are inoculated into the spawn bag, and after culturing for 10 to 14 days, the wild mulberry fungus SH-27 fruiting bodies are obtained by continuing to culture under the same conditions for 30 to 40 days.

6. The culture method according to claim 5, wherein The cultivation medium is composed of: 50-80 parts by mass of poplar wood chips, 10-30 parts by mass of wheat bran, 0.5-1 part by mass of yeast extract, 0.5-1 part by mass of calcium oxide, 1-2 parts by mass of calcium sulfate, and 0.5-1 part by mass of baomazizi powder; the moisture content of the cultivation medium is 55-65%; Further preferably, the poplar wood chips are sieved through a 20-30 mesh sieve, and the baomazi peel powder is sieved through a 60-80 mesh sieve.

7. The culture method according to claim 5, wherein The sterilization conditions are: sterilization at 121° C. for 1.5 to 2 hours.

8. The culture method according to claim 5, wherein The spawn culture conditions are: 24-30° C., relative air humidity 60-70%, and light-shielding culture.

9. Use of the wild Phellinus igniarius SH-27 according to claim 1 in the preparation of antioxidant products.

10. Use of the wild Morinda igniarius SH-27 according to claim 1 in the preparation of functional foods or medicines for treating or preventing colitis.

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