Experimental method for researching influence of Na2SeO3 on phellinus igniarius mycelium fermentation

Abstract

The invention discloses an experimental method for studying the influence of Na2SeO3 on phellinus igniarius mycelium fermentation, and belongs to the technical field of phellinus igniarius mycelium fermentation. Preparing strains to be tested; preparing a main reagent; preparing a culture medium; s2, fermentation experiment: performing fermentation experiment by adopting phellinus igniarius mycelium liquid with different Na2SeO3 concentrations, and researching mycelium yield and antioxidant activity change in fermentation; preparing a phellinus igniarius mycelium water extract; measuring the pH value and the acidity value of the fermentation liquor; determining the fresh weight of the phellinus igniarius mycelium; determining the antioxidant activity of the phellinus igniarius mycelium water extract; s3, performing experimental analysis; the pH value and the acidity value of the fermentation liquor under different Na2SeO3 concentrations are researched; the fresh weight of the phellinus igniarius mycelium under different Na2SeO3 concentrations is researched; the DPPH of the phellinus igniarius mycelia treated by different Na2SeO3 concentrations is researched; the reducing power of phellinus igniarius mycelia treated by different Na2SeO3 concentrations is researched; the yield and quality analysis of phellinus igniarius mycelia in different periods under the fixed Na2SeO3 concentration optimizes the fermentation conditions, and improves the antioxidant activity and medicinal value of phellinus igniarius.

Description

Technical Field

[0001] The invention relates to the technical field of mulberry linterus mycelium fermentation, in particular to an experimental method for studying the influence of Na2SeO3 on mulberry linterus mycelium fermentation. Background Art

[0002] The material basis of mulberry yellow is mainly composed of polysaccharides, flavonoids and triterpenoid compounds. The antioxidant, anti-inflammatory and anti-tumor properties formed by the synergistic effect of these active ingredients show unique application value in the field of adjuvant treatment of malignant tumors. With the deepening understanding of the medicinal value of mulberry yellow, its demand in the consumer market is also increasing. Liquid fermentation technology has become an effective way to achieve large-scale production of mulberry yellow. By optimizing fermentation conditions, such as temperature, pH value, dissolved oxygen and nutrient composition, the growth rate and biomass of mulberry yellow can be significantly improved to meet market demand.

[0003] For example, the patent publication number CN112322572A discloses a liquid fermentation method for increasing the yield of Phellinus igniarius mycelium, comprising: S1: preparing a liquid culture medium for culturing Phellinus igniarius; S2, inoculating the Phellinus igniarius species into the liquid culture medium for activation culture, and then inoculating the seed culture medium at an inoculum volume percentage of 10-15%, and carrying out seed culture to obtain a seed liquid; S3, inoculating the seed liquid prepared in S2 into the liquid culture medium at an inoculum volume percentage of 10-15%, and carrying out liquid fermentation culture at a temperature of 28-35°C for 1-3 days, adding a certain amount of vegetable oil and magnesium acetate tetrahydrate on any day during the culture process, and then culturing the seed liquid at a temperature of 23-28°C. Continue to culture for 2-3 days under the conditions of , and obtain a mixture of mulberry ignia mycelium and fermentation liquid; selenium, as one of the essential trace elements for the human body, has an important influence on the growth and metabolism of organisms. It plays an important role in anti-oxidation, anti-inflammatory, anti-tumor and immune regulation. Na2SeO3, as a commonly used selenium source, is widely used in selenium enrichment research of microorganisms. An appropriate amount of Na2SeO3 can promote the growth and metabolism of mulberry ignia, increase its biomass and active ingredient content. At present, the research on the effect of Na2SeO3 on mulberry ignia fermentation is still in its infancy. Different scholars have differences in selenium concentration, fermentation time and fermentation conditions, resulting in inconsistent research results.

[0004] In view of the above problems, it is necessary to further study the effect of Na2SeO3 on the liquid fermentation of Phellinus igniarius. Summary of the Invention

[0005] The purpose of the present invention is to provide an experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus igniarius mycelium, optimizing the fermentation conditions, and improving the antioxidant activity and medicinal value of Phellinus igniarius, which will not only help promote the development of the Phellinus igniarius industry, but also provide new ideas and strategies for the prevention and treatment of related diseases.

[0006] To achieve the above object, the present invention provides the following technical solution: an experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus igniarius mycelium, comprising the following steps: S1. Material preparation.

[0007] Prepare the test strain: Phellinus igniarius strain No. 2.

[0008] Prepare the main reagents: 1-diphenyl-2-trinitrophenylhydrazine, distilled water, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride, 0.1% ferric chloride, 1% potassium ferricyanide solution, 10% trichloroacetic acid, phenolphthalein, glucose, 95% ethanol, anhydrous ethanol, agar, and sodium hydroxide.

[0009] Prepare culture medium: PDA medium, liquid medium containing Na2SeO3.

[0010] Main instruments: digital constant temperature water bath, electric blast drying oven, visible spectrophotometer, high-speed centrifuge, electronic balance, vertical pressure steam sterilizer.

[0011] S2. Fermentation experiment.

[0012] (1) Activation and cultivation of mother culture: Take the slant mother culture block of Phellinus igniarius No. 2 and inoculate it into solid culture medium. Culture it in a constant temperature incubator at 28℃ for 6 days.

[0013] (2) Liquid fermentation experiments of Phellinus igniarius mycelium with different Na2SeO3 concentrations were conducted to study the changes in mycelium yield and antioxidant activity during fermentation at Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL.

[0014] (3) Prepare the aqueous extract of Phellinus igniarius mycelium.

[0015] (4) Determine the pH and acidity of the fermentation broth.

[0016] (5) Determine the fresh weight of Phellinus igniarius mycelium.

[0017] (6) Determination of the antioxidant activity of the aqueous extract of Phellinus igniarius mycelium.

[0018] S3. Experimental analysis.

[0019] (1) Study the pH and acidity of the fermentation broth at different Na2SeO3 concentrations.

[0020] (2) Study the fresh weight of Phellinus igniarius mycelium under different Na2SeO3 concentrations.

[0021] (3) Study the DPPH of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations.

[0022] (4) Study the reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations.

[0023] (5) Analysis of the yield and quality of Phellinus igniarius mycelium at different stages under a fixed Na2SeO3 concentration.

[0024] Preferably, the PDA culture medium in S1 is prepared as follows: 20.0 g / L glucose, 200 g / L potato, 20.0 g / L agar powder, natural pH, autoclaved at 121°C for 30 min, and set aside after sterilization; the Na2SeO3-containing liquid culture medium is prepared as follows: 20.0 g / L glucose, 200 g / mL potato, Na2SeO3, fully mixed, natural pH, autoclaved at 121°C for 30 min, and set aside after sterilization.

[0025] Preferably, the fermentation experiment in S2 uses PDA culture medium, and Na2SeO3 is added to the culture medium to prepare a selenium-rich culture medium with Na2SeO3 concentrations of 0 mg / mL, 0.02 mg / mL, 0.14 mg / mL, 0.17 mg / mL, 0.20 mg / mL, 0.23 mg / mL, 0.26 mg / mL and 0.38 mg / mL.

[0026] Take a 50 mL Erlenmeyer flask and fill it with 30 mL of selenium-enriched culture medium. Inoculate three 1 cm diameter bacterial cakes and culture them in a shaker at 28°C and 180 r / min for 15 days.

[0027] After 15 days of culture, samples were taken to determine the pH value, acidity, fresh weight of Phellinus linteus mycelium, DPPH clearance rate and reducing power index of the fermentation broth at different concentrations.

[0028] Preferably, the fermentation experiment is designed to carry out liquid fermentation culture of Phellinus igniarius mycelium at two Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL.

[0029] After inoculation, samples were taken every 3 days, for a total of 5 times, to determine the fresh weight of the mycelium and antioxidant activity indicators.

[0030] Preferably, the preparation method of the water extract of Phellinus igniarius mycelium in S2 is: (1) Take the mycelium from the fermentation bottle, rinse it with distilled water, absorb the free water, add distilled water in a stoppered test tube at a material-water ratio of 1:5, and place it in a 95℃ water bath and heat for 3 hours.

[0031] (2) Remove the test tube from the water bath and centrifuge at 3000 rpm for 10 minutes.

[0032] (3) Carefully remove the clear bacterial liquid from the upper layer and transfer it to a new test tube or container for later use.

[0033] Preferably, the method for determining the pH and acidity of the fermentation broth in S2 is: (1) Use pH test paper to directly measure the pH value of the fermentation broth.

[0034] (2) Determination of acidity of fermentation broth: Take 1 mL of bacterial solution and place it in a test tube. Add 2 mL of distilled water to dilute it. Then add 0.05 mL of 0.5% phenolphthalein ethanol solution. After shaking, use a 0.1 mol / L NaOH solution as a titrant to titrate until the solution turns slightly red. If the color does not fade within 1 minute, the end point is calculated. The acidity of the bacterial solution is: Acidity = (Vsample-Vblank) × CNaOH × 10 Where Vsample is the milliliters of NaOH standard solution consumed, Vblank is the milliliters of NaOH standard solution consumed by the blank solution, and CNaOH is the concentration of the NaOH standard solution.

[0035] Preferably, the method for determining the fresh weight of Phellinus igniarius mycelia in S2 is: After filtering the bacterial liquid, the obtained mycelial balls are rinsed with distilled water for 3-5 times to remove the culture medium and other impurities attached to the surface of the mycelium.

[0036] The mycelium was transferred into a culture dish of known weight and weighed directly. The difference between the two weights was the fresh weight of the mycelium.

[0037] Preferably, the antioxidant activity determination of the water extract of Phellinus linteus mycelia in S2 comprises: (1) Determination of DPPH free radical scavenging ability Accurately weigh 20.0 mg of DPPH, dissolve it in anhydrous ethanol and dilute to 250.0 mL to obtain a 2.0×10-4 mol / L DPPH solution. Mix 2.0 mL of Phellinus igniarius aqueous extract at different concentrations with 2.0 mL of DPPH solution. After reacting for 30 minutes in the dark, measure the absorbance at a wavelength of 517 nm (A sample). Measure the absorbance of the mixture of 2.0 mL of DPPH solution and 2.0 mL of anhydrous ethanol (A blank). According to: Clearance rate = (1 − A sample / A blank) × 100% The DPPH clearance rate was calculated.

[0038] (2) Reducing power determination Take 3 mL of Phellinus igniarius aqueous extract at different concentrations, add 2.5 mL of phosphate buffer solution with a pH value of 6.6 and 2.5 mL of 1% potassium ferricyanide, and place in a constant temperature water bath at 50°C for 20 minutes, then cool rapidly; add 2.5 mL of 10% trichloroacetic acid, let it stand for a while, take 2.5 mL of the supernatant, add 0.5 mL of 0.1% ferric chloride and 2.5 mL of water, shake well, and let it stand for 10 minutes; measure its absorbance A at a wavelength of 700 nm. 700The absorbance reflects the reducing power of the bacterial solution.

[0039] Preferably, the experimental analysis results in S3 include: (1) pH and acidity of fermentation broth at different Na2SeO3 concentrations.

[0040] At a concentration of 0.14 mg / mL, the pH value of the Phellinus igniarius fermentation broth was the lowest.

[0041] When the concentration of Na2SeO3 increased to 0.26 mg / mL, the pH value of the bacterial solution decreased.

[0042] At a concentration of 0.38 mg / mL, the pH value of the Phellinus igniarius fermentation broth was the highest.

[0043] (2) Fresh weight of Phellinus igniarius mycelium at different Na2SeO3 concentrations: In the concentration range of 0 mg / mL-0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium showed an upward trend.

[0044] At a Na2SeO3 concentration of 0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium reached a maximum value of 7.02 g, which was 1.25 times that of the control group.

[0045] At a Na2SeO3 concentration of 0.14 mg / mL, the fresh weight of Phellinus igniarius mycelium was 6.45 g.

[0046] At a concentration of 0.2-0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium showed a downward trend, and the rate of decline increased.

[0047] When the concentration of Na2SeO3 increased to 0.26 mg / mL and 0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium decreased by 3.85 g and 5.03 g, respectively, compared with the control group.

[0048] (3) DPPH of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: At Na2SeO3 concentrations of 0-0.14 mg / mL, the DPPH radical scavenging rates of different samples increased with increasing sample concentrations.

[0049] When the concentration of Na2SeO3 was 0.14 mg / mL, the DPPH free radical scavenging rate was the highest, which was 2.44%.

[0050] When the concentration of Na2SeO3 was 0.02 mg / mL, the DPPH radical scavenging rate was 2.05%.

[0051] When the concentration of Na2SeO3 was 0.38 mg / mL, the DPPH radical scavenging rate was 1.75%.

[0052] When the concentration of Na2SeO3 was 0.26 mg / mL, the DPPH radical scavenging rate was 1.28%.

[0053] (4) Reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: When the concentration of Na2SeO3 was 0.14 mg / mL, the absorbance was 0.718, which was 46.83% higher than that of the control group.

[0054] When the concentration of Na2SeO3 was 0.17 mg / mL, the absorbance was 0.738, which was 60.12% higher than that of the control group.

[0055] Preferably, the fixed Na2SeO3 concentration in S3 is 0.14 mg / mL and 0.17 mg / mL, and the Phellinus igniarius is subjected to liquid fermentation culture at 28°C for 15 days, and the fresh weight, reducing power and DPPH clearance rate of the Phellinus igniarius at different stages of the fermentation process are measured.

[0056] The yield and quality analysis of Phellinus igniarius mycelium at different stages under fixed Na2SeO3 concentration includes: (1) Fresh weight of Phellinus igniarius at different stages At both concentrations, the changes in the fresh weight of Phellinus igniarius mycelia increased with the increase of fermentation time.

[0057] (2) DPPH of Phellinus igniarius mycelium at different stages The DPPH scavenging ability of Phellinus igniarius mycelia at both concentrations first increased and then decreased with the extension of fermentation time.

[0058] (3) Reducing power of Phellinus igniarius mycelium at different stages The reducing power of the Phellinus igniarius solution at both concentrations showed a trend of first increasing and then decreasing.

[0059] Compared with the prior art, the beneficial effects of the present invention are: the experimental method for studying the influence of Na2SeO3 on the fermentation of Phellinus igniarius mycelium uses Na2SeO3 as a selenium source, which promotes the growth of mycelium by participating in the metabolic process within Phellinus igniarius cells. During the entire stage of liquid fermentation of Phellinus igniarius, low concentrations of Na2SeO3 promote the growth of Phellinus igniarius mycelium and increase the antioxidant activity of Phellinus igniarius.

[0060] Experiments show that under the Na2SeO3 concentration environment of 0.17 mg / mL, the fresh weight of Phellinus igniarius reached the highest value, and indicators such as reducing power and DPPH clearance rate were also relatively high. Based on the setting of the Na2SeO3 concentration gradient in this experiment, not only the low concentration range was taken into consideration, but further concentration optimization experiments were also conducted to more accurately determine the optimal selenium concentration. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] Figure 1 Schematic diagram of pH value of fermentation broth treated with different Na2SeO3 concentrations in the present invention.

[0062] Figure 2 Schematic diagram of the acidity of fermentation broth treated with different Na2SeO3 concentrations in the present invention.

[0063] Figure 3 Schematic diagram of the fresh weight of bacterial pellets treated with different Na2SeO3 concentrations in the present invention.

[0064] Figure 4 This is a schematic diagram of the DPPH treatment of Phellinus igniarius mycelium with different Na2SeO3 concentrations in the present invention.

[0065] Figure 5 Schematic diagram of the reducing ability of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations in the present invention.

[0066] Figure 6 Schematic diagram of the fresh weight of the pellets at different stages of the present invention.

[0067] Figure 7 This is a schematic diagram of the DPPH of Phellinus igniarius mycelium at different stages of the present invention.

[0068] Figure 8 Schematic diagram of the reducing ability of Phellinus igniarius mycelium at different stages of the present invention. DETAILED DESCRIPTION

[0069] The following will clearly and completely describe the technical solutions in the embodiments of the present invention in conjunction with the accompanying drawings. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention. Example

[0070] See also Figures 1-8 The present invention provides the following technical solution: an experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus igniarius mycelium, comprising the following steps: S1. Material preparation.

[0071] Prepare the test strain: Phellinus igniarius strain No. 2.

[0072] Prepare the main reagents: 1-diphenyl-2-trinitrophenylhydrazine, distilled water, sodium dihydrogen phosphate, disodium hydrogen phosphate, sodium chloride, 0.1% ferric chloride, 1% potassium ferricyanide solution, 10% trichloroacetic acid, phenolphthalein, glucose, 95% ethanol, anhydrous ethanol, agar, and sodium hydroxide.

[0073] Prepare culture medium: PDA medium, liquid medium containing Na2SeO3.

[0074] Main instruments: digital constant temperature water bath, electric blast drying oven, visible spectrophotometer, high-speed centrifuge, electronic balance, vertical pressure steam sterilizer.

[0075] Instrument name model Manufacturer Digital constant temperature water bath HH-2 Shanghai Weicheng Instrument Co., Ltd. Electric blast drying oven DHG-9123A Shanghai Yiheng Scientific Instrument Co., Ltd. Visible spectrophotometer 722N Shanghai Jinghua Technology Instrument Co., Ltd. High-speed centrifuge H / T16 Jintan Hongke Instrument Factory electronic balance FA2204B Shanghai Tianmei Balance Instrument Co., Ltd. Vertical pressure steam sterilizer BXM-30R Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory S2. Fermentation experiment.

[0076] (1) Activation and cultivation of mother culture: Take the slant mother culture block of Phellinus igniarius No. 2 and inoculate it into solid culture medium. Culture it in a constant temperature incubator at 28℃ for 6 days.

[0077] (2) Liquid fermentation experiments of Phellinus igniarius mycelium with different Na2SeO3 concentrations were conducted to study the changes in mycelium yield and antioxidant activity during fermentation at Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL.

[0078] (3) Prepare the aqueous extract of Phellinus igniarius mycelium.

[0079] (4) Determine the pH and acidity of the fermentation broth.

[0080] (5) Determine the fresh weight of Phellinus igniarius mycelium.

[0081] (6) Determination of the antioxidant activity of the aqueous extract of Phellinus igniarius mycelium.

[0082] S3. Experimental analysis.

[0083] (1) Study the pH and acidity of the fermentation broth at different Na2SeO3 concentrations.

[0084] (2) Study the fresh weight of Phellinus igniarius mycelium under different Na2SeO3 concentrations.

[0085] (3) Study the DPPH of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations.

[0086] (4) Study the reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations.

[0087] (5) Analysis of the yield and quality of Phellinus igniarius mycelium at different stages under a fixed Na2SeO3 concentration.

[0088] The preparation method of PDA culture medium in S1 is as follows: 20.0 g / L glucose, 200 g / L potato, 20.0 g / L agar powder, natural pH, high pressure sterilization at 121°C for 30 min, and set aside after sterilization; the preparation method of liquid culture medium containing Na2SeO3 is as follows: 20.0 g / L glucose, 200 g / mL potato, Na2SeO3, fully mixed, natural pH, high pressure sterilization at 121°C for 30 min, and set aside after sterilization.

[0089] In the fermentation experiment in S2, PDA medium was used, and Na2SeO3 was added to the medium to prepare selenium-rich medium with Na2SeO3 concentrations of 0 mg / mL, 0.02 mg / mL, 0.14 mg / mL, 0.17 mg / mL, 0.20 mg / mL, 0.23 mg / mL, 0.26 mg / mL and 0.38 mg / mL.

[0090] Take a 50 mL Erlenmeyer flask and fill it with 30 mL of selenium-enriched culture medium. Inoculate three 1 cm diameter bacterial cakes and culture them in a shaker at 28°C and 180 r / min for 15 days.

[0091] After 15 days of culture, samples were taken to determine the pH value, acidity, fresh weight of Phellinus linteus mycelium, DPPH clearance rate and reducing power index of the fermentation broth at different concentrations.

[0092] The fermentation experiment was designed to carry out liquid fermentation culture of Phellinus igniarius mycelium at two Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL.

[0093] After inoculation, samples were taken every 3 days, for a total of 5 times, to determine the fresh weight of the mycelium and antioxidant activity indicators.

[0094] The preparation method of the water extract of Phellinus linteus mycelium in S2 is: (1) Take the mycelium from the fermentation bottle, rinse it with distilled water, absorb the free water, add distilled water in a stoppered test tube at a material-water ratio of 1:5, and place it in a 95℃ water bath and heat for 3 hours.

[0095] (2) Remove the test tube from the water bath and centrifuge at 3000 rpm for 10 minutes.

[0096] (3) Carefully remove the clear bacterial liquid from the upper layer and transfer it to a new test tube or container for later use.

[0097] The method for determining the pH and acidity of the fermentation broth in S2 is: (1) Use pH test paper to directly measure the pH value of the fermentation broth.

[0098] (2) Determination of acidity of fermentation broth: Take 1 mL of bacterial solution and place it in a test tube. Add 2 mL of distilled water to dilute it. Then add 0.05 mL of 0.5% phenolphthalein ethanol solution. After shaking, use a 0.1 mol / L NaOH solution as a titrant to titrate until the solution turns slightly red. If the color does not fade within 1 minute, the end point is calculated. The acidity of the bacterial solution is: Acidity = (Vsample-Vblank) × CNaOH × 10 Where Vsample is the milliliters of NaOH standard solution consumed, Vblank is the milliliters of NaOH standard solution consumed by the blank solution, and CNaOH is the concentration of the NaOH standard solution.

[0099] The method for determining the fresh weight of Phellinus linteus mycelium in S2 is: After filtering the bacterial liquid, the obtained mycelial balls are rinsed with distilled water for 3-5 times to remove the culture medium and other impurities attached to the surface of the mycelium.

[0100] The mycelium was transferred into a culture dish of known weight and weighed directly. The difference between the two weights was the fresh weight of the mycelium.

[0101] The antioxidant activity assay of the aqueous extract of Phellinus linteus mycelia in S2 includes: (1) Determination of DPPH free radical scavenging ability Accurately weigh 20.0 mg of DPPH, dissolve it in anhydrous ethanol and dilute to 250.0 mL to obtain a 2.0×10-4 mol / L DPPH solution. Mix 2.0 mL of Phellinus igniarius aqueous extract at different concentrations with 2.0 mL of DPPH solution. After reacting for 30 minutes in the dark, measure the absorbance at a wavelength of 517 nm (A sample). Measure the absorbance of the mixture of 2.0 mL of DPPH solution and 2.0 mL of anhydrous ethanol (A blank). According to: Clearance rate = (1 − A sample / A blank) × 100% The DPPH clearance rate was calculated.

[0102] (2) Reducing power determination Take 3 mL of Phellinus igniarius aqueous extract at different concentrations, add 2.5 mL of phosphate buffer solution with a pH value of 6.6 and 2.5 mL of 1% potassium ferricyanide, and place in a constant temperature water bath at 50°C for 20 minutes, then cool rapidly; add 2.5 mL of 10% trichloroacetic acid, let it stand for a while, take 2.5 mL of the supernatant, add 0.5 mL of 0.1% ferric chloride and 2.5 mL of water, shake well, and let it stand for 10 minutes; measure its absorbance A at a wavelength of 700 nm. 700 The absorbance reflects the reducing power of the bacterial solution.

[0103] The experimental analysis results in S3 include: (1) pH and acidity of fermentation broth at different Na2SeO3 concentrations.

[0104] like Figure 1 As shown in the figure, the pH value of the mulberry linterus fermentation broth treated with different Na2SeO3 concentrations was different. At a concentration of 0.14 mg / mL, the pH value of the mulberry linterus fermentation broth was the lowest, indicating that the addition of selenium promoted the metabolic activity of the mulberry linterus mycelium and produced more acidic substances.

[0105] When the concentration of Na2SeO3 increased to 0.26 mg / mL, the pH value of the bacterial solution decreased.

[0106] At a concentration of 0.38 mg / mL, the pH value of the mulberry ignia fermentation broth was the highest, indicating that high concentrations of selenium had a certain inhibitory effect on the growth of mulberry ignia, resulting in a slowdown in metabolic activity and a reduction in the production of acidic substances.

[0107] like Figure 2 As shown in the data, at a Na2SeO3 concentration of 0.14 mg / mL, the acidity of the Phellinus igniarius liquid was the highest, and the acidity was second at a Na2SeO3 concentration of 0.02 mg / mL, indicating that acidic substances would be accumulated during the liquid fermentation of the Phellinus igniarius mycelium. In the concentration range of 0.20 mg / mL-0.38 mg / mL, the acidity of the Phellinus igniarius fermentation liquid tended to decrease. The acidity of the 0.23 mg / mL Na2SeO3 concentration treatment group was equivalent to that of the 0.38 mg / mL Na2SeO3 concentration treatment group. The acidity was lowest at a Na2SeO3 concentration of 0.38 mg / mL. This indicates that an appropriate amount of selenium can promote the growth of Phellinus igniarius mycelium, while excessive or insufficient amounts will have an adverse effect on the growth, further verifying the inhibitory effect of high concentration selenium on the growth of Phellinus igniarius. Except for the 0.14 mg / mL and 0.02 mg / mL concentration treatment groups, the acidity of the other treatment groups was lower than that of the control group, but the difference in acidity among the treatment groups was not significant.

[0108] (2) Fresh weight of Phellinus igniarius mycelium at different Na2SeO3 concentrations: like Figure 3 As shown in the figure, with the increase of Na2SeO33 concentration, the fresh weight of Phellinus igniarius mycelium showed a trend of first increasing and then decreasing.

[0109] In the concentration range of 0 mg / mL-0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium showed an upward trend.

[0110] At a Na2SeO3 concentration of 0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium reached a maximum value of 7.02 g, which was 1.25 times that of the control group.

[0111] At a Na2SeO3 concentration of 0.14 mg / mL, the fresh weight of Phellinus igniarius mycelium was 6.45 g.

[0112] At a concentration of 0.2-0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium showed a downward trend, and the rate of decline increased.

[0113] When the concentration of Na2SeO3 increased to 0.26 mg / mL and 0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium decreased by 3.85 g and 5.03 g, respectively, compared with the control group.

[0114] This shows that the growth rate of Phellinus igniarius is fastest at a concentration of 0.17 mg / mL, at which point the biomass accumulates the most. As the concentration continues to increase, the degree of inhibition on the growth of Phellinus igniarius increases.

[0115] (3) DPPH of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: like Figure 4 As shown in the figure, at the Na2SeO3 concentration of 0-0.14 mg / mL, the DPPH radical scavenging rate of different samples increased with the increase of sample concentration.

[0116] When the concentration of Na2SeO3 was 0.14 mg / mL, the DPPH free radical scavenging rate was the highest, which was 2.44%.

[0117] When the concentration of Na2SeO3 was 0.02 mg / mL, the DPPH radical scavenging rate was 2.05%.

[0118] When the concentration of Na2SeO3 was 0.38 mg / mL, the DPPH radical scavenging rate was 1.75%.

[0119] When the concentration of Na2SeO3 was 0.26 mg / mL, the DPPH radical scavenging rate was 1.28%.

[0120] Except for the 0.26 mg / mL Na2SeO3 treatment group, the DPPH radical scavenging rates of the other three treatment groups were greater than that of the control group, which indicates that an appropriate amount of selenium can not only promote the growth of Phellinus igniarius, but also significantly improve its antioxidant activity. When the Na2SeO3 concentration is too high, the antioxidant activity of Phellinus igniarius will be inhibited.

[0121] The experiment found that a selenium concentration of 0.14 mg / mL is a more suitable condition, which can promote the growth of Phellinus igniarius and improve its antioxidant activity. The research results provide a scientific basis and data support for the optimization of the liquid fermentation process of Phellinus igniarius mycelium, and also lay the foundation for further exploring the effects of Na2SeO3 on the growth and metabolism of Phellinus igniarius.

[0122] (4) Reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: like Figure 5 As shown in the figure, the reducing power of Phellinus igniarius mycelium under different Na2SeO3 concentrations was determined. The reducing power is one of the important indicators to measure the antioxidant activity. The reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations was different. With the increase of Na2SeO3 concentration, the reducing power of Phellinus igniarius mycelium first increased and then decreased.

[0123] When the concentration of Na2SeO3 was 0.14 mg / mL, the absorbance was 0.718, which was 46.83% higher than that of the control group.

[0124] When the concentration of Na2SeO3 was 0.17 mg / mL, the absorbance was 0.738, which was 60.12% higher than that of the control group.

[0125] The reducing power of Phellinus igniarius mycelium in the Na2SeO3 treatment groups of 0.02 mg / mL, 0.20 mg / mL, 0.23 mg / mL and 0.26 mg / mL was stronger than that in the control group. The reducing power of the Na2SeO3 treatment group of 0.38 mg / mL was the lowest. The results showed that the Na2SeO3 treatment group of 0.17 mg / mL showed stronger reducing ability than other treatment groups. This may be because the Phellinus igniarius mycelium produced higher polyphenol compounds under this concentration treatment. At the selenium concentration of 0.17 mg / mL, the reducing power of the Phellinus igniarius liquid was the strongest, indicating that the antioxidant activity of Phellinus igniarius was the highest at this concentration. This result was consistent with the trend of fresh weight change of Phellinus igniarius, further proving that the selenium concentration of 0.17 mg / mL is the optimal concentration for liquid fermentation of Phellinus igniarius.

[0126] In summary, the experiment found that the selenium concentration of 0.17 mg / mL is the optimal concentration for liquid fermentation of Phellinus igniarius. At this concentration, the biomass accumulation of Phellinus igniarius is the largest and the antioxidant activity is the highest.

[0127] The Na2SeO3 concentrations in S3 were fixed at 0.14 mg / mL and 0.17 mg / mL, and Phellinus igniarius was fermented in liquid at 28°C for 15 days. The fresh weight, reducing power and DPPH scavenging rate of Phellinus igniarius at different stages of the fermentation process were determined.

[0128] The yield and quality analysis of Phellinus igniarius mycelium at different stages under fixed Na2SeO3 concentration includes: (1) Fresh weight of Phellinus igniarius at different stages like Figure 6 As shown in the figure, at two concentrations, the changes in the fresh weight of Phellinus linteus mycelia increased with the increase of fermentation time.

[0129] In the early stage of fermentation, due to sufficient nutrition, the bacteria grew rapidly and the fresh weight increased rapidly. As the fermentation progressed, the nutrition was gradually consumed, the growth rate of the bacteria slowed down, and the rate of increase in fresh weight also slowed down accordingly. At the end of fermentation, the fresh weight of Phellinus igniarius at a concentration of 0.17 mg / mL was slightly higher than that at a concentration of 0.14 mg / mL, but the difference was not significant. This may be because under the stimulation of higher concentrations of Na2SeO3, the bacteria can produce more biomass, but too high a concentration may also have a certain inhibitory effect on the growth of the bacteria.

[0130] (2) DPPH of Phellinus igniarius mycelium at different stages like Figure 7 As shown in the figure, the DPPH scavenging ability of Phellinus igniarius mycelia at two concentrations first increased and then decreased with the extension of fermentation time.

[0131] The DPPH radical scavenging ability of Phellinus igniarius mycelium was significantly enhanced in the early stage of culture (3-6 days). The DPPH radical scavenging ability of Phellinus igniarius mycelium reached a peak on the 6th day, and then the clearance rate showed a downward trend. At 9-12 days, the downward trend slowed down; then at 12-15 days, the downward trend began to rise again.

[0132] The DPPH scavenging ability of Phellinus igniarius mycelium at a concentration of 0.17 mg / mL was higher than that at a concentration of 0.14 mg / mL overall. At the peak, the DPPH scavenging ability of Phellinus igniarius mycelium at a concentration of 0.17 mg / mL was 34.29% higher than that at a concentration of 0.14 mg / mL.

[0133] (3) Reducing power of Phellinus igniarius mycelium at different stages like Figure 8 As shown in the figure, the reducing power of the Phellinus igniarius solution at the two concentrations showed a trend of first increasing and then decreasing.

[0134] In the early stage of fermentation, since the bacteria are in a rapid growth stage and have vigorous metabolism, the reducing substances produced gradually increase, resulting in an increase in reducing power. When the fermentation enters the middle and late stages, the growth rate of the bacteria slows down, the metabolism tends to be stable, and its reducing power is correspondingly inhibited. At concentrations of 0.14 mg / mL and 0.17 mg / mL, the reducing power reaches a peak value on the 9th day of fermentation, and at a concentration of 0.17 mg / mL, the reducing power of the mulberry ignia mycelium is higher than that of 0.14 mg / mL. Experiments show that on the ninth day of culture, the reducing power of the mulberry ignia mycelium treated with 0.17 mg / mL of Na2SeO3 is better.

[0135] As a form of sodium selenate, Na2SeO3 mainly participates in various biochemical reactions in the body in the form of selenium. Selenium is one of the essential trace elements for organisms. It plays a key role in anti-oxidation, detoxification and metabolic regulation. During the entire stage of liquid fermentation of Phellinus igniarius, the addition of Na2SeO3 significantly affected the growth characteristics and antioxidant capacity of Phellinus igniarius.

[0136] Using Na2SeO3 as a selenium source promotes mycelial growth by participating in metabolic processes within Phellinus igniarius cells. Selenium is a key component of numerous antioxidant enzymes, such as glutathione peroxidase, which scavenge free radicals within cells, protecting cell membranes and DNA from oxidative damage. Appropriate doses of selenium supplementation enhance the activity of these enzymes, thereby improving the antioxidant capacity of Phellinus igniarius cells and reducing cell damage caused by oxidative stress. This protective effect contributes to the normal growth and reproduction of Phellinus igniarius mycelium, significantly increasing its growth rate and biomass in a culture medium containing an appropriate concentration of Na2SeO3.

[0137] The effect of Na2SeO3 on the antioxidant capacity of Phellinus igniarius is very significant. By measuring the reducing power of Phellinus igniarius mycelium at Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL, it was found that with the increase of Na2SeO3 concentration, the antioxidant capacity of Phellinus igniarius first increased and then weakened; under the appropriate Na2SeO3 concentration conditions, the reducing power and DPPH clearance rate of Phellinus igniarius liquid reached the maximum value, indicating that the antioxidant capacity of Phellinus igniarius was the strongest at this time. This may be related to the metabolism and distribution of selenium in Phellinus igniarius cells. An appropriate amount of selenium can significantly increase the activity of antioxidant enzymes in Phellinus igniarius cells, thereby increasing the antioxidant capacity of cells.

[0138] The results obtained in this experiment have certain similarities and differences with other experiments on the effect of Na2SeO3 on the liquid fermentation of Phellinus igniarius.

[0139] From the aspect of consistency, this experiment found that low concentration of Na2SeO3 has the effect of promoting the growth of Phellinus igniarius mycelium and increasing the antioxidant activity of Phellinus igniarius, which is consistent with the results of earlier studies. Studies have shown that the appropriate addition of selenium can significantly increase the biomass and polysaccharide content of Phellinus igniarius, thereby enhancing the antioxidant activity of Phellinus igniarius. In terms of differences, the optimal Na2SeO3 concentration found in this experiment is different from that of other studies. The experiment shows that under the Na2SeO3 concentration environment of 0.17 mg / mL, the fresh weight of Phellinus igniarius reaches the highest value, and indicators such as reducing power and DPPH clearance rate are also relatively high. This difference may be due to inconsistent experimental conditions, such as the formula composition of the fermentation medium, fermentation time, temperature, humidity and the genetic properties of the strain itself. Different studies may have different methods and indicators used in determining antioxidant activity, which will also cause different results.

[0140] In setting up the Na2SeO3 concentration gradient in this experiment, not only the low concentration range was taken into consideration, but further concentration optimization experiments were also conducted to more accurately determine the optimal selenium concentration. This experimental design may also be one of the reasons why the results of this experiment are different from those of other experiments.

[0141] This experiment focused on exploring the effects of different concentrations of Na2SeO3 on the liquid fermentation of Phellinus igniarius. The Na2SeO3 concentration gradient values ​​of 0, 0.02, 0.14, 0.17, 0.20, 0.23, 0.26, and 0.38 mg / mL were set to deeply analyze the effects of Na2SeO3 on the growth and antioxidant activity of Phellinus igniarius mycelium. After 15 days of liquid fermentation, the pH value under each concentration condition, the fresh weight of the strain after Phellinus igniarius fermentation, the acidity value, and the reducing power and DPPH of Phellinus igniarius mycelium were measured. The experimental results show that within the gradient range of selenium concentration, the growth and antioxidant activity of Phellinus igniarius show a changing pattern.

[0142] When the Na2SeO3 concentration was 0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium reached its peak. This result confirmed that adding an appropriate amount of selenium can significantly promote the growth of Phellinus igniarius mycelium, while excessive or insufficient selenium will have a certain inhibitory effect on growth. By comparing the fermentation and culture conditions of Phellinus igniarius mycelium at different concentrations, under the conditions of Na2SeO3 concentration gradient consisting of 0, 0.02, 0.14, 0.20, 0.26, and 0.38 mg / mL, this article found that the mycelium at a concentration of 0.14 mg / mLNa2SeO3 had a higher DPPH scavenging ability, which showed that the antioxidant activity of Phellinus igniarius was significantly improved at this concentration; at a Na2SeO3 concentration of 0.17 mg / mL, the antioxidant activity of Phellinus igniarius mycelium reached the best level. This discovery provides a scientific basis for the addition of selenium in the fermentation stage of Phellinus igniarius liquid, and also provides data support for the subsequent use of a large amount of Phellinus igniarius liquid with high antioxidant activity in health products, medicines and other fields.

[0143] It is worth noting that under the Na2SeO3 concentration environment of 0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium was the largest and the antioxidant activity was better. This may be because selenium plays multiple functions in the liquid fermentation stage of Phellinus igniarius, which not only accelerates the development of mycelium but also enhances the antioxidant activity of the mycelium. The enhancement of antioxidant activity may be more complicated, involving many aspects such as the interaction between selenium and the internal metabolic pathways of Phellinus igniarius.

[0144] The experimental results show that under appropriate selenium concentration conditions, the growth state and antioxidant activity of Coriolus igniarius can be greatly improved. The experimental results expand the research scope of Coriolus igniarius liquid fermentation and also provide new ideas and methods for the development and utilization of Coriolus igniarius.

[0145] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "connected" and "connection" should be understood in a broad sense. For example, they can refer to fixed connection, detachable connection, or integral connection; mechanical connection, electrical connection; direct connection, or indirect connection through an intermediary. Those skilled in the art will understand the specific meanings of the above terms in the present invention based on the specific circumstances.

[0146] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. An experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium, comprising the following steps: S1. Material preparation; Prepare the test strain: Phellinus igniarius strain 2; Prepare the following reagents: 1-diphenyl-2-trinitrophenylhydrazine, distilled water, sodium dihydrogen phosphate, sodium hydrogen phosphate, sodium chloride, 0.1% ferric chloride, 1% potassium ferricyanide solution, 10% trichloroacetic acid, phenolphthalein, glucose, 95% ethanol, anhydrous ethanol, agar, and sodium hydroxide. Prepare culture medium: PDA medium, liquid medium containing Na2SeO3; Main instruments: digital constant temperature water bath, electric blast drying oven, visible spectrophotometer, high-speed centrifuge, electronic balance, vertical pressure steam sterilizer; S2, fermentation experiment; (1) Activation and cultivation of mother culture: Take the slant mother culture block of Phellinus igniarius No. 2 and inoculate it into solid culture medium, and culture it in a constant temperature incubator at 28°C for 6 days for use; (2) Using liquid fermentation experiments with Phellinus igniarius mycelium at different Na2SeO3 concentrations, the changes in mycelium yield and antioxidant activity during fermentation at Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL were studied; (3) Preparation of aqueous extract of Phellinus igniarius mycelium; (4) Determine the pH and acidity of the fermentation broth; (5) Determination of the fresh weight of Phellinus igniarius mycelium; (6) Determination of the antioxidant activity of the aqueous extract of Phellinus igniarius mycelium; S3, experimental analysis; The pH and acidity of the fermentation broth were studied at different Na2SeO3 concentrations; The fresh weight of mycelium of Phellinus linteus under different Na2SeO3 concentrations was studied; The DPPH of Phellinus linteus mycelium was studied with different Na2SeO3 concentrations. The reducing power of Phellinus linteus mycelium treated with different Na2SeO3 concentrations was studied; Analysis of the yield and quality of Phellinus igniarius mycelium at different stages under fixed Na2SeO3 concentration.

2. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The PDA culture medium in S1 is prepared as follows: 20.0 g / L glucose, 200 g / L potato, 20.0 g / L agar powder, natural pH, and autoclaved at 121° C. for 30 min. After sterilization, it is ready for use. The preparation method of the liquid culture medium containing Na2SeO3 is as follows: 20.0 g / L glucose, 200 g / mL potato, and Na2SeO3, fully mixed, natural pH, and autoclaved at 121°C for 30 minutes. After sterilization, it is set aside.

3. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The fermentation experiment in S2 used PDA medium, and Na2SeO3 was added to the medium to prepare selenium-enriched medium with Na2SeO3 concentrations of 0 mg / mL, 0.02 mg / mL, 0.14 mg / mL, 0.17 mg / mL, 0.20 mg / mL, 0.23 mg / mL, 0.26 mg / mL and 0.38 mg / mL; Take 50 mL Erlenmeyer flasks and fill them with 30 mL of selenium-enriched medium. Inoculate three 1 cm diameter bacterial cakes and culture them in a shaker at 28°C and 180 rpm for 15 days. After 15 days of culture, samples were taken to determine the pH value, acidity, fresh weight of Phellinus linteus mycelium, DPPH clearance rate and reducing power index of the fermentation broth at different concentrations.

4. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 3, characterized in that: The fermentation experiment was designed to carry out liquid fermentation culture of Phellinus igniarius mycelium at two Na2SeO3 concentrations of 0.14 mg / mL and 0.17 mg / mL; After inoculation, samples were taken every 3 days, for a total of 5 times, to determine the fresh weight of the mycelium and antioxidant activity indicators.

5. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The preparation method of the water extract of Phellinus igniarius mycelium in S2 is: (1) Take the mycelium from the fermentation bottle, rinse it with distilled water, absorb the free water, add distilled water at a material-water ratio of 1:5 in a stoppered test tube, and heat it in a 95℃ water bath for 3 hours; (2) Remove the test tube from the water bath and centrifuge at 3000 rpm for 10 min. (3) Carefully remove the clear bacterial liquid from the upper layer and transfer it to a new test tube or container for later use.

6. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The method for determining the pH and acidity of the fermentation liquid in S2 is: (1) Use pH test paper to directly measure the pH value of the fermentation broth; (2) Determination of acidity of fermentation broth: Take 1 mL of bacterial solution and place it in a test tube. Add 2 mL of distilled water to dilute it. Then add 0.05 mL of 0.5% phenolphthalein ethanol solution. After shaking, use a 0.1 mol / L NaOH solution as a titrant to titrate until the solution turns slightly red. If the color does not fade within 1 minute, the end point is calculated. The acidity of the bacterial solution is: Acidity = (Vsample-Vblank) × CNaOH × 10 Where Vsample is the milliliters of NaOH standard solution consumed, Vblank is the milliliters of NaOH standard solution consumed by the blank solution, and CNaOH is the concentration of the NaOH standard solution.

7. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The method for determining the fresh weight of Phellinus linteus mycelia in S2 is: After filtering the bacterial solution, the obtained mycelial pellets were rinsed with distilled water 3-5 times to remove the culture medium and other impurities attached to the surface of the mycelium; The mycelium was transferred into a culture dish of known weight and weighed directly. The difference between the two weights was the fresh weight of the mycelium.

8. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The antioxidant activity determination of the water extract of Phellinus linteus mycelia in S2 includes: (1) Determination of DPPH free radical scavenging ability Accurately weigh 20.0 mg of DPPH, dissolve it in anhydrous ethanol and dilute to 250.0 mL to obtain a 2.0×10-4 mol / L DPPH solution. Mix 2.0 mL of Phellinus igniarius aqueous extract at different concentrations with 2.0 mL of DPPH solution. After reacting for 30 minutes in the dark, measure the absorbance at a wavelength of 517 nm (A sample). Measure the absorbance of the mixture of 2.0 mL of DPPH solution and 2.0 mL of anhydrous ethanol (A blank). According to: Clearance rate = (1 − A sample / A blank) × 100% Calculate the DPPH clearance rate; (2) Reducing power determination Take 3 mL of Phellinus igniarius aqueous extract at different concentrations, add 2.5 mL of phosphate buffer solution with a pH value of 6.6 and 2.5 mL of 1% potassium ferricyanide, and place in a constant temperature water bath at 50°C for 20 minutes, then cool rapidly; add 2.5 mL of 10% trichloroacetic acid, let it stand for a while, take 2.5 mL of the supernatant, add 0.5 mL of 0.1% ferric chloride and 2.5 mL of water, shake well, and let it stand for 10 minutes; measure its absorbance A at a wavelength of 700 nm. 700 The absorbance reflects the reducing power of the bacterial solution.

9. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The experimental analysis results in S3 include: (1) pH and acidity of fermentation broth at different Na2SeO3 concentrations: At a concentration of 0.14 mg / mL, the pH value of the Phellinus igniarius fermentation broth was the lowest; When the concentration of Na2SeO3 increased to 0.26 mg / mL, the pH value of the bacterial solution decreased; At a concentration of 0.38 mg / mL, the pH value of the Phellinus igniarius fermentation broth was the highest; (2) Fresh weight of Phellinus igniarius mycelium at different Na2SeO3 concentrations: In the concentration range of 0 mg / mL-0.17 mg / mL, the fresh weight of Phellinus igniarius mycelium showed an increasing trend; At a concentration of 0.17 mg / mL Na2SeO3, the fresh weight of Phellinus igniarius mycelium reached a maximum of 7.02 g, which was 1.25 times that of the control group; At a Na2SeO3 concentration of 0.14 mg / mL, the fresh weight of Phellinus igniarius mycelium was 6.45 g; At a concentration of 0.2-0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium showed a downward trend, and the rate of decline increased; When the concentration of Na2SeO3 increased to 0.26 mg / mL and 0.38 mg / mL, the fresh weight of Phellinus igniarius mycelium decreased by 3.85 g and 5.03 g, respectively, compared with the control group; (3) DPPH of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: At Na2SeO3 concentrations of 0-0.14 mg / mL, the DPPH radical scavenging rates of different samples increased with increasing sample concentrations; When the concentration of Na2SeO3 was 0.14 mg / mL, the DPPH radical scavenging rate was the highest, which was 2.44%; When the concentration of Na2SeO3 was 0.02 mg / mL, the DPPH radical scavenging rate was 2.05%; When the concentration of Na2SeO3 was 0.38 mg / mL, the DPPH radical scavenging rate was 1.75%; When the concentration of Na2SeO3 was 0.26 mg / mL, the DPPH radical scavenging rate was 1.28%; (4) Reducing power of Phellinus igniarius mycelium treated with different Na2SeO3 concentrations: When the concentration of Na2SeO3 was 0.14 mg / mL, the absorbance was 0.718, which was 46.83% higher than that of the control group; When the concentration of Na2SeO3 was 0.17 mg / mL, the absorbance was 0.738, which was 60.12% higher than that of the control group.

10. The experimental method for studying the effect of Na2SeO3 on the fermentation of Phellinus linteus mycelium according to claim 1, characterized in that: The Na2SeO3 concentration in the S3 was fixed at 0.14 mg / mL and 0.17 mg / mL, and the Phellinus linteus was subjected to liquid fermentation at 28°C for 15 days, and the fresh weight, reducing power and DPPH clearance rate of the Phellinus linteus at different stages of the fermentation process were measured; The yield and quality analysis of Phellinus igniarius mycelium at different stages under fixed Na2SeO3 concentration includes: (1) Fresh weight of Phellinus igniarius at different stages At both concentrations, the changes in the fresh weight of Phellinus linteus mycelia increased with the increase of fermentation time; (2) DPPH of Phellinus igniarius mycelium at different stages The DPPH scavenging ability of Phellinus igniarius mycelia at both concentrations first increased and then decreased with the extension of fermentation time; (3) Reducing power of Phellinus igniarius mycelium at different stages The reducing power of the Phellinus igniarius solution at both concentrations showed a trend of first increasing and then decreasing.

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