Ganoderma lucidum AD256, ganoderma lucidum fermentation product prepared from ganoderma lucidum AD256 and application of ganoderma lucidum fermentation product

Abstract

The invention provides a ganoderma lucidum AD256 strain, a ganoderma lucidum fermentation product prepared from the ganoderma lucidum AD256 strain and application of the ganoderma lucidum fermentation product, and belongs to the technical field of microbial fermentation. The invention provides a strain of ganoderma lucidum AD256, the preservation number of which is CCTCC NO: M 20252522, the ganoderma lucidum AD256 is fermented in a ganoderma lucidum fermentation culture medium containing plant powder to prepare a ganoderma lucidum fermented product, efficient fermentation of ganoderma lucidum and increase of the content of active substances are synchronously realized through the promotion effect of the plant powder on ganoderma lucidum fermentation and the catalytic degradation effect of ganoderma lucidum on the plant powder, and the ganoderma lucidum has a good application prospect. The preparation method of the ganoderma lucidum fermentation product provided by the invention is green, environment-friendly, simple and feasible, and suitable for large-scale production. The prepared ganoderma lucidum fermentation product with high active substance content has good antioxidant activity and an anti-aging effect, and has a wide market application prospect in the field of anti-aging cosmetics.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, specifically relating to a Ganoderma lucidum strain AD256 and its prepared Ganoderma lucidum fermentation products and their applications. Background Technology

[0002] Reishi mushroom (Ganoderma lucidum), also known as the "immortal herb," ​​is described in the *Compendium of Materia Medica* as "tonifying the middle energizer, boosting qi, enhancing wisdom, improving complexion, and with prolonged consumption, promoting longevity and immortality." Reishi contains various bioactive substances such as polysaccharides and triterpenes, which exert antioxidant, anti-aging, soothing, anti-inflammatory, moisturizing, and repairing effects in cosmetics. During its growth, reishi also produces a large number of cellulases, hemicellulases, and glycosidases, which have excellent biocatalytic effects on active substances. Honeysuckle, the dried flower buds or unopened flowers of *Lonicera japonica* Thunb. (family Caprifoliaceae), contains polysaccharides, phenolic acids, flavonoids, and other bioactive substances, exerting antioxidant, anti-aging, soothing, anti-inflammatory, barrier repair, moisturizing, and antibacterial effects in cosmetics. Wild chrysanthemum is the capitulum of the perennial herbaceous plant *Chrysanthemum indicum*, belonging to the Asteraceae family. It contains various bioactive substances such as polysaccharides, flavonoids, volatile oils, and alkaloids, and plays a role in cosmetics with antibacterial, anti-inflammatory, whitening, moisturizing, soothing, and repairing effects. Sage is the dried whole herb of *Salvia japonica* Thunb., belonging to the Lamiaceae family. It contains various bioactive substances such as polysaccharides, phenolic acids, flavonoids, and terpenes, and plays a role in cosmetics with antioxidant, anti-aging, antibacterial, anti-inflammatory, moisturizing, and soothing effects. Centella asiatica is the dried whole herb of the perennial herb *Centella asiatica*, belonging to the Apiaceae family. It contains various bioactive substances such as triterpenes, phenolic acids, and flavonoids, and plays a role in cosmetics with barrier repair, anti-inflammatory and soothing, antioxidant, anti-aging, and whitening effects. Purslane is the dried aerial part of the Portulaca oleracea plant, which contains a variety of bioactive substances such as polysaccharides, flavonoids, polyphenols, and alkaloids. In cosmetics, it plays a role in anti-oxidation, anti-inflammatory, skin barrier repair, and moisturizing.

[0003] Two-way fermentation of medicinal fungi uses Chinese medicinal herbs with active ingredients as a substrate, and utilizes medicinal fungi for fermentation to transform the effective components of the substrate, producing new active ingredients and functions. Two-way fermentation not only increases the content and utilization rate of active ingredients in the fungal substrate, thus improving efficacy, but also transforms them into new active compounds with new efficacies. It can also reduce the toxic side effects of Chinese medicinal herbs and improve their taste. It has advantages such as low production cost, high production efficiency, controllable production process, and suitability for industrial production. While research on the two-way fermentation of Ganoderma lucidum (Reishi mushroom), a high-quality edible and medicinal fungus, is existing, research on the liquid fermentation of Ganoderma lucidum into plant powder is relatively limited.

[0004] Furthermore, solid-state fermentation in the current two-way fermentation process of Ganoderma lucidum has problems such as long production cycle, fermentation using water extracts of traditional Chinese medicine has problems such as complex extraction process, two-way fermentation combined with organic solvent extraction has problems such as loss of polysaccharides and other nutritional components and organic solvent residue, and adding traditional Chinese medicine powder or water extracts during the fermentation process has problems such as complex fermentation process and easy contamination.

[0005] Therefore, there is an urgent need to develop a Ganoderma lucidum strain that can efficiently ferment plant powders. Utilizing this strain to economically and efficiently ferment plant powders with good efficacy in cosmetics and prepare novel Ganoderma lucidum ferments for application in cosmetics has significant commercial prospects and application value. Summary of the Invention

[0006] To address the shortcomings of existing technologies, the purpose of this invention is to provide a Ganoderma lucidum plant AD256, its prepared Ganoderma lucidum ferment, and its applications. This invention employs a two-way liquid fermentation process involving Ganoderma lucidum and plant powder. Through the promoting effect of plant powder on the fermentation of Ganoderma lucidum and the catalytic degradation of plant powder by Ganoderma lucidum, a Ganoderma lucidum ferment with anti-aging effects is prepared economically and efficiently, providing a new approach in the field of cosmetics with antioxidant activity and anti-aging properties.

[0007] To achieve the above-mentioned objectives, the present invention employs the following technical solution:

[0008] This invention provides a Ganoderma lucidum AD256, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: M 20252522.

[0009] Furthermore, the colonies of Ganoderma lucidum AD256 are white, round, with dense hyphae, and the hyphae have clamp connections.

[0010] Furthermore, the Ganoderma lucidum AD256 has the ability to produce cellulase, and the cellulase activity is 0.8-1.5 U / mL, based on the volume of its bacterial culture and the carboxymethyl cellulase activity.

[0011] The present invention also provides Ganoderma lucidum fermentation product produced by fermentation of Ganoderma lucidum AD256, wherein the Ganoderma lucidum fermentation product is prepared by Ganoderma lucidum AD256 in a Ganoderma lucidum fermentation medium containing plant powder.

[0012] Furthermore, the Ganoderma lucidum fermentation product includes at least one of the following: honeysuckle powder Ganoderma lucidum fermentation product, wild chrysanthemum powder Ganoderma lucidum fermentation product, sage powder Ganoderma lucidum fermentation product, centella asiatica powder Ganoderma lucidum fermentation product, and purslane powder Ganoderma lucidum fermentation product.

[0013] Furthermore, the polysaccharide content in the Ganoderma lucidum ferment is not less than 0.8 g / L, the total polyphenol content is not less than 80 mg / L, and the total flavonoid content is not less than 100 mg / L.

[0014] Furthermore, the preparation method of the Ganoderma lucidum ferment includes the following steps:

[0015] (1) The Ganoderma AD256 was inoculated into a liquid seed culture medium and fermented to obtain a seed liquid;

[0016] (2) The seed liquid is inoculated into a Ganoderma lucidum fermentation medium containing plant powder and fermented to obtain a fermentation liquid; the plant powder includes at least one of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder and purslane powder.

[0017] (3) Homogenize the fermentation liquid to obtain Ganoderma lucidum fermentation homogenate;

[0018] (4) Let the Ganoderma lucidum fermentation homogenate stand at room temperature, centrifuge and filter to obtain Ganoderma lucidum fermentation product.

[0019] Furthermore, based on the volume of the Ganoderma lucidum fermentation medium, the inoculation amount of the seed liquid in step (2) is 10%.

[0020] Furthermore, the pressure for homogenizing the fermentation broth is 80~120MPa.

[0021] Furthermore, the settling time is 6-8 hours.

[0022] Furthermore, the centrifugation operation steps include centrifugation at 8000~10000 rpm for 10~20 min.

[0023] Furthermore, the filtration is performed using a 0.45μm filter membrane.

[0024] Furthermore, based on the volume of the Ganoderma lucidum fermentation culture medium, the mass-volume concentration of the plant powder is 5~20 g / L.

[0025] Furthermore, the particle size of the plant powder is 40-200 mesh.

[0026] The present invention also provides the application of the aforementioned Ganoderma lucidum ferment in the preparation of cosmetics with anti-aging effects.

[0027] Furthermore, the Ganoderma lucidum ferment can promote the production of collagen and elastin by scavenging free radicals, thereby delaying aging.

[0028] Furthermore, the cosmetics include at least one of face masks, serums, lotions, creams, sprays, and liquids.

[0029] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0030] 1. This invention utilizes the Ganoderma lucidum strain AD256, which highly expresses cellulase, to liquid ferment a Ganoderma lucidum fermentation medium containing 40-200 mesh plant powders (honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder). During fermentation, the plant powders in the medium effectively disperse the Ganoderma lucidum mycelium and provide nutrients, promoting liquid fermentation and significantly increasing the biomass and polysaccharide yield of Ganoderma lucidum strain AD256. The strain AD256 can efficiently biodegrade the plant powders (honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder), promoting the release of active substances such as polysaccharides, polyphenols, and flavonoids. The prepared Ganoderma lucidum fermented product has a high content of active substances.

[0031] 2. This invention achieves efficient simultaneous fermentation of Ganoderma lucidum and green production of active substances through bidirectional liquid fermentation of Ganoderma lucidum and plant powder, which is economical and efficient. Moreover, the process does not involve exogenous enzymes and / or organic solvents, making it safe and environmentally friendly.

[0032] 3. The Ganoderma lucidum ferment prepared by this invention has a high content of active substances. The polysaccharides in the prepared Ganoderma lucidum ferment are not less than 0.8 g / L, the total polyphenols are not less than 80 mg / L, and the total flavonoids are not less than 100 mg / L. It has good DPPH free radical and hydroxyl free radical scavenging activity. At the cellular level, it can significantly increase the production of collagen (COL-I) and elastin (ELN) in human dermal fibroblasts, exerting antioxidant and anti-aging effects. It has obvious advantages and broad application prospects in the preparation of anti-aging cosmetics. Attached Figure Description

[0033] Figure 1 The DPPH free radical scavenging activity of the Ganoderma lucidum fermentation product is measured.

[0034] Figure 2 The hydroxyl radical scavenging activity of the Ganoderma lucidum fermentation product is described.

[0035] Figure 3 The cytotoxicity of the Ganoderma lucidum fermentation product is determined.

[0036] Figure 4 The effect of the Ganoderma lucidum fermentation product on COL-I production in HSF cells.

[0037] Figure 5 The effect of the Ganoderma lucidum fermentation product on ELN production in HSF cells. Detailed Implementation

[0038] The present invention will be further described below with reference to specific embodiments, but the scope of protection of the present invention is not limited to the following embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0039] 1. Culture medium

[0040] PDA medium: 200 g / L potato, 20 g / L glucose, 20 g / L agar, natural pH.

[0041] Carboxymethyl cellulose agar medium: sodium carboxymethyl cellulose (CMC-Na) 10 g / L, yeast extract 2 g / L, potassium dihydrogen phosphate 1.5 g / L, magnesium sulfate heptahydrate 0.5 g / L, sterilized at 121℃ for 20 minutes.

[0042] Liquid seed culture medium: glucose 20 g / L, yeast extract 5 g / L, potassium dihydrogen phosphate 1.5 g / L, magnesium sulfate heptahydrate 0.5 g / L, thiamine 5 mg / L, sterilized at 121℃ for 20 minutes.

[0043] Fermentation basal medium: glucose 30g / L, yeast powder 8g / L, potassium dihydrogen phosphate 2g / L, magnesium sulfate heptahydrate 1g / L, thiamine 8mg / L, sterilized at 121℃ for 20 minutes.

[0044] Ganoderma lucidum fermentation medium containing plant powder: Add plant powder to the basic fermentation medium and sterilize at 121℃ for 20 minutes.

[0045] 2. Cellulase Activity Assay

[0046] Cellulase activity was determined using a carboxymethyl cellulase assay. A 0.5% sodium carboxymethyl cellulose (CMC-Na) solution was prepared using 0.1 mol / L acetate buffer (pH 4.5). 1.5 mL of the CMC-Na solution was used as the substrate, and 0.5 mL of Ganoderma lucidum fermentation product was added. The mixture was incubated in a 50°C water bath for 30 min. The reaction was terminated by adding 2 mL of DNS. The mixture was then heated in a boiling water bath for 5 min to develop color. After cooling to room temperature, the volume was adjusted to 25 mL. A standard curve was plotted using glucose, and the absorbance at 540 nm was measured (OD). 540 An inactivated enzyme solution was used as a blank control. Enzyme activity was calculated by plotting a standard curve using glucose.

[0047] An enzyme activity unit (U) is defined as the amount of enzyme required to catalyze the production of 1 μmol of reducing sugar from a substrate per minute under experimental conditions. It is expressed in U / mL.

[0048] 3. Biomass determination

[0049] After fermentation, 50 mL of fermentation broth was centrifuged at 10,000 r / min for 10 min. The mycelium was washed twice with purified water to remove residual fermentation broth and fermentation residue. It was then dried at 80 ℃ for 48 h, weighed, and the biomass was calculated.

[0050] 4. Polysaccharide content determination

[0051] A certain volume of Ganoderma lucidum fermentation product was taken, and three times its volume of 95% ethanol was added. The mixture was thoroughly mixed, allowed to stand overnight at 4°C, and the precipitate was collected by centrifugation. The precipitate was washed twice with 95% ethanol. The precipitate was redissolved in purified water to a suitable concentration, and the polysaccharide content was determined using the benzoic acid-sulfuric acid method. A standard curve was plotted with glucose concentration as the abscissa (x) and absorbance at 490 nm as the ordinate (y). The polysaccharide concentration (calculated as glucose) was calculated. The standard curve was y = 10.731x - 0.027, R0. 2 =0.9997.

[0052] 5. Determination of total polyphenol content

[0053] A certain volume of Ganoderma lucidum fermentation product was taken, and the total polyphenol content was determined using the Folin-Ciocalteu method. A standard curve was plotted with gallic acid concentration as the abscissa (x) and absorbance at 765 nm as the ordinate (y). The total polyphenol concentration (calculated as gallic acid) was calculated. The standard curve was: y = 2.2464x + 0.0145, R0 2 =0.9997.

[0054] 6. Determination of total flavonoid content

[0055] A certain volume of Ganoderma lucidum ferment was taken and freeze-dried. Twice the volume of anhydrous ethanol was added to the ferment, and the mixture was refluxed at 70℃ for 2 hours. The supernatant was collected by centrifugation, and the extraction was repeated twice. The supernatants were combined. The total flavonoid content was determined using the sodium nitrite-aluminum nitrate-sodium hydroxide method. A standard curve was plotted with rutin concentration as the x-axis and absorbance at 510 nm as the y-axis. The total flavonoid concentration (calculated as rutin) was calculated. The standard curve was: y = 1.746x + 0.0004, R0 2 =0.9994.

[0056] 7. Sources of plant powder

[0057] The honeysuckle powder of this invention is obtained by drying and pulverizing the honeysuckle flower buds, separating them through sieves of 40 mesh and 200 mesh, and collecting honeysuckle powder with a particle size of 40-200 mesh; the wild chrysanthemum powder is obtained by drying and pulverizing the capitulum of the wild chrysanthemum plant, separating it through sieves of 40 mesh and 200 mesh, and collecting wild chrysanthemum powder with a particle size of 40-200 mesh; the sage powder is obtained by drying and pulverizing the whole sage plant, separating it through sieves of 40 mesh and 200 mesh. The sage powder with a particle size of 40-200 mesh is obtained by sieving and separating the whole plant of Centella asiatica through a 0-mesh sieve; the Centella asiatica powder is obtained by drying and pulverizing the whole plant of Centella asiatica, sieving and separating it through a 40-mesh and a 200-mesh sieve, and collecting the Centella asiatica powder with a particle size of 40-200 mesh; the purslane powder is obtained by drying and pulverizing the above-ground parts of the purslane plant, sieving and separating it through a 40-mesh and a 200-mesh sieve, and collecting the purslane powder with a particle size of 40-200 mesh.

[0058] In addition, collect plant powders of various mesh sizes, from <40 mesh to >200 mesh, for later use.

[0059] Honeysuckle buds, wild chrysanthemum, sage, centella asiatica, and purslane were all obtained through commercial channels.

[0060] Example 1: Isolation, directed evolution, identification and preservation of strain Ganoderma lucidum AD256

[0061] 1. Strain isolation

[0062] Wild Ganoderma lucidum was harvested from Changbai Mountain in Antu County, Jilin Province. After surface disinfection, the fruiting bodies were aseptically picked and inoculated onto 30 PDA plates. The plates were cultured at 25°C for 10 days. The strain G12, which had the fastest growth rate, the largest colony diameter, and clamp connections in the mycelium (methylene blue staining and microscopic observation), was selected for mutagenesis experiments.

[0063] 2. ARTP mutagenesis

[0064] Take 0.5 cm of strain G12 2 Ten mycelial fragments were aseptically inoculated into shake flasks containing liquid seed culture medium and incubated at 25°C and 150 rpm for 7 days. The mycelia were then collected by centrifugation at 4000 rpm for 5 minutes. The mycelia were washed twice with 0.01 mol / L PBS buffer (pH 7.4) and collected by centrifugation at 4000 rpm for 5 minutes. Ganoderma lucidum protoplasts were prepared using a filamentous fungus protoplast preparation kit (Bestbio) according to the kit instructions. The protoplasts were resuspended in sterile STC solution (Coolaber) to approximately 5 × 10⁻⁵. 7 Quantity / mL. Refrigerate at 2-8℃ for later use.

[0065] Inside the biosafety cabinet, 10 μL of protoplast suspension was evenly spread onto a sterile metal slide. Under aseptic conditions, the slide was placed in an ambient pressure, room temperature plasma mutagenesis breeding instrument (ARTP-M, Tianmu Biotechnology). The power was set to 120 W, the gas flow rate to 10 SLM, and the mutagenesis time to 80 s (lethality approximately 90%). After mutagenesis, the metal slide was placed in an EP tube containing 1 mL of 0.01 mol / L PBS buffer (pH 7.4), and shaken for 1 minute. The resuspended bacterial solution was diluted 100-fold, and 0.1 mL of the diluted solution was spread onto PDA plates (20 plates). The plates were incubated at 25°C for 7 days. Ten strains (G12A1~G12A10) with rapid growth, large colony diameter, and clamp connections in the mycelium (methylene blue staining, microscopic observation) were selected for further screening.

[0066] The mutant strains G12A1 to G12A10 were inoculated onto carboxymethyl cellulose agar medium and cultured at 25°C for 15 days. The strain G12A2 with a large clear zone / colony diameter was selected for further screening.

[0067] 3. Directed Evolution

[0068] Directed evolution of strains was carried out by continuously conducting plate rejuvenation-shake flask fermentation.

[0069] (1) Plate culture: Take 0.5 cm of plate culture of strain G12A2 2 Mycelia were inoculated onto PDA agar plates (20 plates) and cultured at 25°C for 7 days. Ten strains with larger colony diameters were selected.

[0070] (2) Place each of the above 10 strains at 0.5 cm depth. 2 Mycelia were inoculated onto carboxymethyl cellulose agar medium and cultured at 25°C for 15 days. Five strains with larger clear zones / colony diameters were selected.

[0071] (3) Shake flask culture: Take 0.5 cm of each of the above 5 strains. 2 Ten mycelial fragments were aseptically inoculated into shake flasks containing basal fermentation medium. The mixture was incubated at 25°C and 150 rpm for 7 days as one generation, and then subcultured for three consecutive generations. The biomass and carboxymethyl cellulase activity of the shake-flask fermentation were measured.

[0072] (4) Select strains with dense mycelia, high biomass and high carboxymethyl cellulase activity from the shake flask liquid fermentation, pick 20 colonies from the shake flask and inoculate them onto PDA plate medium, and repeat steps (1)-(4) for five consecutive cycles of directed evolution of the strains.

[0073] After five consecutive cycles of directed evolution, strain AD256 exhibited high mycelial biomass and carboxymethyl cellulase activity, at 14.5±0.4 g / L and 1.38±0.12 U / mL, respectively, representing 142% and 306% of the initial strain.

[0074] 4. Strain identification

[0075] Strains AD256 were cultured on PDA plates at 25°C for 7 days. The colonies were white, round, with dense hyphae that adhered to the surface of the medium. Methylene blue staining revealed clamp connections in the hyphae.

[0076] Using total DNA from the mycelium of strain AD256 as a template, and universal primers ITS1 and ITS4 as primers, PCR amplification of the ITS region sequence was performed.

[0077] The primer sequences are:

[0078] ITS1: 5'-TCCGTAGGTGAACCTGCGG-3'

[0079] ITS4: 5'-TCCTCCGCTTATTGATATGC-3'

[0080] PCR reaction conditions were as follows: 50 μL PCR reaction system containing 1 μL DNA template; F (10 μM), 1 μL;

[0081] R (10μM), 1μL; dNTP (2.5mM each), 4μL; Taq (2U / μL), 1μL; 10×Taq buffer, 5μL; ddH2O, add to 50μL.

[0082] The PCR amplification program was as follows: 94℃ pre-denaturation for 3 min, 94℃ denaturation for 30 s, 60℃ annealing for 30 s, 72℃ extension for 2 min, 35 cycles of amplification, and 72℃ final extension for 5 min.

[0083] The PCR amplification products were sequenced for the ITS region by Qingdao Ruiboxingke Biotechnology Co., Ltd., and their nucleotide sequences are shown in SEQ ID NO. 1. BLAST alignment was performed on the National Center for Biotechnology Information (NCBI) website, and the results showed a 99.36% similarity to *Ganoderma lucidum* sequences (including ID: KX589250.1, ID: GU213479.1, and ID: GU213483.1). A phylogenetic tree constructed using the NJ method confirmed that strain AD256 belongs to *Ganoderma lucidum*. Based on the morphological and molecular biological results, the strain was named *Ganoderma AD256*, and its taxonomic name is *Ganoderma lucidum*.

[0084] 5. Preservation of microbial strains

[0085] The identified Ganoderma lucidum AD256 strain was deposited at the China Center for Type Culture Collection (CCTCC); address: Wuhan University, Wuhan, China; deposit date: November 10, 2025; the accession number for Ganoderma lucidum AD256 is CCTCC NO: M 20252522.

[0086] Example 2: Preparation of Ganoderma lucidum fermented product using plant powder fermentation process

[0087] The preparation of Ganoderma lucidum ferment using plant powder fermentation technology includes the following steps:

[0088] 1. Cultivate Ganoderma lucidum AD256, and take 0.5cm of freshly cultivated material. 2 Ten mycelial fragments were inoculated into 100 mL of liquid seed culture medium and cultured at 25°C and 150 rpm for 7 days to obtain seed culture.

[0089] 2. Inoculate the seed liquid into the Ganoderma lucidum fermentation medium containing plant powder at an inoculation rate of 10% (v / v). The plant powder is honeysuckle powder with a particle size of 40-200 mesh. The mass-volume concentration of the plant powder is 10 g / L based on the volume of the Ganoderma lucidum fermentation medium. Ferment and culture at 25℃ and 150 rpm for 7 days to obtain the fermentation broth.

[0090] 3. Homogenize the fermentation broth at 8000 rpm for 3 minutes and then homogenize it under high pressure at 100 MPa to obtain Ganoderma lucidum fermentation homogenate.

[0091] 4. The Ganoderma lucidum fermentation homogenate was left to stand at room temperature for 7 hours, centrifuged at 8000 rpm for 10 min, the supernatant was collected, filtered through a 0.45 μm filter membrane, and the filtrate was collected to obtain the Ganoderma lucidum fermentation product.

[0092] The biomass of Ganoderma lucidum fermentation was 18.2 g / L, the polysaccharide content in the fermented product was 1.26 g / L, the total polyphenol content was 110.62 mg / L, and the total flavonoid content was 224.94 mg / L.

[0093] Example 3: The effect of plant powder type on Ganoderma lucidum fermentation product

[0094] Based on the preparation process in Example 2, the plant powder in step 2 was replaced with wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder, respectively. The fermentation basal culture medium without plant powder was used as a control, and the other process conditions remained unchanged. The biomass of Ganoderma lucidum fermentation product and the content of polysaccharides, total polyphenols, and total flavonoids in Ganoderma lucidum fermentation product were used as indicators to compare the effects of plant powder type on Ganoderma lucidum fermentation product.

[0095] Table 1. Effects of plant powder type on Ganoderma lucidum fermentation products

[0096]

[0097] The results are shown in Table 1: The Ganoderma lucidum biomass in the basal culture medium without added plant powder was low, and the content of polysaccharides, total polyphenols, and total flavonoids in the fermented product was also low. After fermentation in the culture medium supplemented with 10 g / L of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder, the Ganoderma lucidum fermentation biomass increased to 16.5–18.2 g / L, which was 1.17–1.29 times that of the control. The prepared Ganoderma lucidum fermented product showed significantly increased content of polysaccharides, total polyphenols, and total flavonoids. Specifically, the polysaccharide concentration increased to 0.82–1.26 g / L, which was 1.58–2.42 times that of the control; the total polyphenol concentration increased to 90.89–115.96 mg / L, which was 1.77–2.26 times that of the control; and the total flavonoid concentration increased to 157.52–394.98 mg / L, which was 2.36–10.09 times that of the control. The results showed that honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder could all effectively promote Ganoderma lucidum fermentation, increase Ganoderma lucidum biomass, and increase the content of polysaccharides, total polyphenols, and total flavonoids in Ganoderma lucidum fermentation products.

[0098] Example 4: Effect of plant powder particle size on Ganoderma lucidum fermentation product

[0099] The dried and pulverized honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder were separated by sieving through 40-mesh and 200-mesh sieves, collecting plant powders with particle sizes >40 mesh, 40-200 mesh, and <200 mesh. Based on the preparation process in Example 2, the plant powders and their particle sizes from step 2 were used to prepare Ganoderma lucidum fermentation culture media with honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder with particle sizes >40 mesh, 40-200 mesh, and <200 mesh, respectively. The mass-volume concentration of different plant powders was 10 g / L, and other process conditions remained unchanged. The fermentation basal culture medium without plant powder was used as a control. The effects of different plant powder particle sizes on Ganoderma lucidum fermentation products were studied using Ganoderma lucidum fermentation biomass and the content of polysaccharides, total polyphenols, and total flavonoids in the fermented product as indicators.

[0100] Table 2. Effect of plant powder particle size on Ganoderma lucidum fermentation products

[0101]

[0102] As shown in Table 2, when the plant powder particle size is <40 mesh, the particle size is relatively large, and the promoting effect of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder on Ganoderma lucidum fermentation is weak, resulting in lower biomass and lower polysaccharide, total polyphenol, and total flavonoid content in the fermented product. When the plant powder particle size is >200 mesh, the particle size is relatively small, and the promoting effect of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder on Ganoderma lucidum fermentation is strong, resulting in vigorous mycelial growth, high biomass, and strong biocatalytic effect of Ganoderma lucidum on plant powder. The total polyphenol and total flavonoid content in the fermented product is relatively high. However, due to the vigorous mycelial growth, a large amount of nutrients are consumed, which limits the biosynthesis of Ganoderma lucidum polysaccharides, resulting in a decrease in polysaccharide content in the fermented product. When the particle size of the plant powder is between 40 and 200 mesh, the particle size is suitable, and the biomass of Ganoderma lucidum fermentation and the content of polysaccharides, total polyphenols, and total flavonoids in the fermented product are all at a high level. Specifically, the biomass of Ganoderma lucidum fermentation increased to 17.2-18.1 g / L, which is 1.20-1.27 times that of the control; the polysaccharide concentration in the fermented product increased to 0.83-1.17 g / L, which is 1.48-2.09 times that of the control; the total polyphenol concentration increased to 90.94-124.89 mg / L, which is 1.60-2.20 times that of the control; and the total flavonoid concentration increased to 169.82-554.51 mg / L, which is 2.45-7.99 times that of the control. The results showed that honeysuckle powder (40-200 mesh), wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder could all effectively increase the biomass of Ganoderma lucidum fermentation and the content of polysaccharides, total polyphenols, and total flavonoids in Ganoderma lucidum fermentation products.

[0103] Example 5: Effect of plant powder concentration on Ganoderma lucidum fermentation products

[0104] Based on the preparation process in Example 2, the particle sizes of the plant powders in step 2 were respectively prepared using honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder with particle sizes of 40-200 mesh to prepare Ganoderma lucidum fermentation medium. The mass-volume concentrations of each plant powder were 5 g / L, 10 g / L, and 20 g / L, respectively, based on the volume of the Ganoderma lucidum fermentation medium. Other process conditions remained unchanged. The fermentation basal medium without plant powder was used as a control. The effects of plant powder concentration on Ganoderma lucidum fermentation product were studied using Ganoderma lucidum fermentation biomass and the contents of polysaccharides, total polyphenols, and total flavonoids in the Ganoderma lucidum fermentation product as indicators.

[0105] Table 3 Effect of plant powder concentration on Ganoderma lucidum fermentation products

[0106]

[0107] The results are shown in Table 3: Ganoderma lucidum fermentation culture medium was prepared using honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder at concentrations ranging from 5 g / L to 20 g / L. The biomass of Ganoderma lucidum fermentation and the contents of polysaccharides, total polyphenols, and total flavonoids in the fermented product increased with the increase of plant powder concentration. Among them, the biomass of fermented honeysuckle powder, sage powder, and centella asiatica powder was significantly increased, the polysaccharide content of fermented honeysuckle powder and wild chrysanthemum powder was significantly increased, the total polyphenol content of fermented sage powder and centella asiatica powder was significantly increased, and the total flavonoid content of fermented sage powder and purslane powder was significantly increased. The biomass of Ganoderma lucidum fermentation increased to 15.7–20.2 g / L, which is 1.13–1.45 times that of the control; the polysaccharide concentration in the fermented Ganoderma lucidum increased to 0.82–2.13 g / L, which is 1.44–3.74 times that of the control; the total polyphenol concentration increased to 82.78–192.09 mg / L, which is 1.54–3.57 times that of the control; and the total flavonoid concentration increased to 102.96–1637.91 mg / L, which is 1.58–25.06 times that of the control. This indicates that a plant powder concentration of 5–20 g / L has a good promoting effect on Ganoderma lucidum fermentation and can effectively increase the biomass of Ganoderma lucidum fermentation and the content of polysaccharides, total polyphenols, and total flavonoids in the fermented Ganoderma lucidum.

[0108] To investigate the functional characteristics of Ganoderma lucidum fermented products, this invention provides Ganoderma lucidum fermented products obtained by fermentation with different concentrations and types of plant powders for experimental development. The Ganoderma lucidum fermented product prepared by fermentation with the basal fermentation medium without added plant powder as described in Example 5 serves as the control group. See the following examples for details.

[0109] Example 6: Preparation of honeysuckle powder and Ganoderma lucidum fermentation product

[0110] The preparation of honeysuckle powder and Ganoderma lucidum fermentation product includes the following steps:

[0111] 1. Ganoderma lucidum AD256 was cultured to obtain 0.5cm... 2 Ten mycelial fragments were inoculated into 100 mL of liquid seed culture medium and cultured at 25°C and 150 rpm for 7 days to obtain seed liquid.

[0112] 2. Inoculate the seed liquid into the Ganoderma lucidum fermentation medium containing plant powder at an inoculation rate of 10% (v / v). The plant powder is honeysuckle powder with a particle size of 40-200 mesh. The mass-volume concentration of the plant powder is 5 g / L, 7.5 g / L, 10 g / L, 15 g / L, and 20 g / L, respectively, based on the volume of the Ganoderma lucidum fermentation medium. Ferment and culture at 25℃ and 150 rpm for 7 days to obtain the fermentation broth.

[0113] 3. Homogenize the fermentation broth at 8000 rpm for 3 minutes and then homogenize it under high pressure at 100 MPa to obtain Ganoderma lucidum fermentation homogenate.

[0114] 4. The Ganoderma lucidum fermentation homogenate was left to stand at room temperature for 7 hours, centrifuged at 8000 rpm for 10 min, the supernatant was collected, filtered through a 0.45 μm filter membrane, and the filtrate was collected to obtain the Ganoderma lucidum fermentation product.

[0115] Using the obtained honeysuckle powder and Ganoderma lucidum fermentation biomass, as well as the contents of polysaccharides, total polyphenols, and total flavonoids in the honeysuckle powder and Ganoderma lucidum fermentation products, as indicators, the corresponding Ganoderma lucidum fermentation products were measured, as follows:

[0116] Table 4. Biochemical indicators of Ganoderma lucidum fermentation products prepared with different concentrations of honeysuckle powder

[0117]

[0118] Example 7: Preparation of Wild Chrysanthemum Pollen and Ganoderma Fermentation Product

[0119] The preparation method of the wild chrysanthemum powder Ganoderma lucidum fermentation product in this embodiment differs from that in embodiment 6 in that the plant powder in step 2 is wild chrysanthemum powder, and the mass-volume concentration of the plant powder is 5 g / L, 7.5 g / L, 10 g / L, 15 g / L, and 20 g / L, respectively; the pressure of high-pressure homogenization in step 3 is 120 MPa; and the Ganoderma lucidum fermentation homogenate in step (4) is left to stand at room temperature for 6 hours; the remaining operation methods are the same as in embodiment 6.

[0120] Using the biomass of wild chrysanthemum pollen fermented with Ganoderma lucidum and the contents of polysaccharides, total polyphenols, and total flavonoids in the fermented product as indicators, the corresponding Ganoderma lucidum fermented products were measured, as follows:

[0121] Table 5. Biochemical indicators of Ganoderma lucidum fermentation products prepared with different concentrations of wild chrysanthemum pollen.

[0122]

[0123] Example 8: Preparation of Sage Powder and Ganoderma Fermentation Product

[0124] The preparation method of the sage powder Ganoderma lucidum fermentation product in this embodiment differs from that in Example 6 in that the plant powder in step 2 is sage powder, and the mass-volume concentration of the plant powder is 5 g / L, 7.5 g / L, 10 g / L, 15 g / L, and 20 g / L, respectively; the pressure of high-pressure homogenization in step 3 is 110 MPa; and the Ganoderma lucidum fermentation homogenate in step 4 is allowed to stand at room temperature for 6.5 hours; the remaining operation methods are the same as in Example 6.

[0125] Using the biomass of Ganoderma lucidum fermented with sage powder and the contents of polysaccharides, total polyphenols, and total flavonoids in the fermented product of Ganoderma lucidum fermented with sage powder as indicators, the corresponding Ganoderma lucidum fermented products were measured, as follows:

[0126] Table 6. Biochemical indicators of Ganoderma lucidum fermentation products prepared with different concentrations of Sage powder

[0127]

[0128] Example 9: Preparation of Centella Asiatica Powder and Ganoderma Lucidum Fermentation Product

[0129] The preparation method of the Centella asiatica powder and Ganoderma lucidum fermentation product in this embodiment differs from that in Example 6 in that the plant powder in step 2 is Centella asiatica powder, and the mass-volume concentration of the Centella asiatica powder is 5 g / L, 7.5 g / L, 10 g / L, 15 g / L, and 20 g / L, respectively; the pressure of high-pressure homogenization in step 3 is 90 MPa; and the Ganoderma lucidum fermentation homogenate is allowed to stand at room temperature for 7.5 hours in step 4; the remaining operation methods are the same as in Example 6.

[0130] Using the biomass of Ganoderma lucidum fermented with Centella asiatica powder and the contents of polysaccharides, total polyphenols, and total flavonoids in the fermented product as indicators, the corresponding Ganoderma lucidum fermented products were measured, as follows:

[0131] Table 7. Biochemical indicators of Ganoderma lucidum fermentation products prepared from different concentrations of Centella asiatica powder

[0132]

[0133] Example 10: Preparation of purslane powder and Ganoderma lucidum fermentation product

[0134] The preparation method of the purslane powder and Ganoderma lucidum fermentation product in this embodiment differs from that in Example 6 in that the plant powder in step 2 is purslane powder, and the mass-volume concentrations of the plant powder are 5 g / L, 7.5 g / L, 10 g / L, 15 g / L, and 20 g / L, respectively; the pressure of high-pressure homogenization in step 3 is 80 MPa; and the Ganoderma lucidum fermentation homogenate is left to stand at room temperature for 8 hours in step 4; the remaining operation methods are the same as in Example 6.

[0135] Using the biomass of purslane powder and Ganoderma lucidum fermentation, and the contents of polysaccharides, total polyphenols, and total flavonoids in the purslane powder and Ganoderma lucidum fermentation products as indicators, the corresponding Ganoderma lucidum fermentation products were measured, as follows:

[0136] Table 8. Biochemical indicators of Ganoderma lucidum fermentation products prepared from different concentrations of purslane powder

[0137]

[0138] Example 11: Antioxidant activity of Ganoderma lucidum fermentation products

[0139] Take 100 mL of each of the Ganoderma lucidum fermentation products prepared by fermentation in different concentrations of plant powder (5 g / L, 7.5 g / L, 10 g / L, 15 g / L, 20 g / L) in Examples 6-10, mix them evenly according to the same type, and obtain samples of Ganoderma lucidum fermentation products of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder.

[0140] The Ganoderma lucidum fermentation product prepared by fermentation in the fermentation medium without the addition of plant powder in Example 5 was used as a control sample.

[0141] 1. DPPH free radical scavenging activity

[0142] The five Ganoderma lucidum fermentation samples and the control sample were diluted with purified water to volume fractions of 0.6%, 1.2%, 2.4%, 4.8%, and 9.6%, respectively. The DPPH free radical scavenging rate of the samples at different concentrations was measured. 2 mL of the same concentration of sample solution was added to both the sample tube (T) and the background sample tube (T0), and 2 mL of water was used instead of water in the DPPH tube (C). 2 × 10⁻⁶ ppm of water was added to both the sample tube (T) and the DPPH tube (C). -4 Add 2 mL of 1 mol / L DPPH methanol solution, replacing the sample background (T0) with 2 mL of methanol solution. Vortex to mix, react at room temperature in the dark for 30 min, then zero the sample with purified water as a blank and measure the absorbance at 517 nm.

[0143] DPPH radical scavenging rate (%) = [1 - (T - T0) / C] × 100%;

[0144] T - Absorbance of the sample tube;

[0145] T0 - Absorbance of the methanol solution with methanol instead of DPPH;

[0146] C - The absorbance of the sample solution is replaced by purified water.

[0147] The measurement results are shown below. Figure 1 The results showed that the DPPH radical scavenging rate gradually increased with the increase of the volume fraction of different plant powder Ganoderma lucidum fermentation samples, and this increase was concentration-dependent. The IC50 values ​​for the DPPH radical scavenging rates of different plant powder Ganoderma lucidum fermentation samples were calculated. 50 The results are shown in Table 9. As can be seen from the table, compared with the control group without added plant powder, the IC50 values ​​of the DPPH free radical scavenging rates of different plant powder Ganoderma lucidum ferments were significantly different. 50 All decreased, IC 50 The concentrations decreased by 29.45% to 59.09% respectively, while the dilution factors increased by 8-26 times. The decreases were particularly noticeable in the fermented products containing honeysuckle powder, centella asiatica powder, and purslane powder. This indicates that the addition of plant powders significantly improved the DPPH free radical scavenging rate of the fermented products.

[0148] Table 9 DPPH free radical scavenging activity (IC50) 50 )

[0149]

[0150] 2. Hydroxyl radical scavenging activity

[0151] The five Ganoderma lucidum fermentation samples and the control sample were diluted with purified water to volume fractions of 0.8%, 1.6%, 3.2%, 6.4%, and 12.8%, respectively. The hydroxyl radical scavenging rate of the samples at different concentrations was measured. The reaction system consisted of 1 mL of 9 mmol / L FeSO4·7H2O solution, 1 mL of 9 mmol / L salicylic acid-ethanol solution, and 1 mL of sample solutions of different concentrations. Then, 1 mL of 8 mmol / L H2O2 solution was added to initiate the reaction. Considering the absorbance of the samples themselves, the background absorbance (T0) of the system was measured using purified water instead of H2O2. After incubation at 37 ℃ for 0.5 h, the absorbance (T) at 510 nm was measured using purified water as the zeroing point.

[0152] Hydroxyl radical scavenging rate (%) = [1 - (T - T0) / C] × 100%;

[0153] T - Absorbance of the sample tube;

[0154] T0 - Absorbance value when purified water is used instead of H2O2;

[0155] C - The absorbance of the sample solution is replaced by purified water.

[0156] The measurement results are shown below. Figure 2 The results showed that the hydroxyl radical scavenging rate gradually increased with the increase of the volume fraction of different plant powder Ganoderma lucidum fermentation samples, and this increase was concentration-dependent. The IC50 values ​​for the hydroxyl radical scavenging rates of different plant powder Ganoderma lucidum fermentation samples were calculated. 50 The results are shown in Table 10. As can be seen from the table, compared with the control group without added plant powder, the IC50 values ​​of the hydroxyl radical scavenging rates of different plant powder Ganoderma lucidum ferments were significantly higher. 50 All decreased, IC 50 The concentrations decreased by 29.45% to 41.27% respectively, while the dilution factor increased by 8-13 times. The decreases were particularly noticeable in honeysuckle powder and Ganoderma lucidum fermentation products, as well as Centella asiatica and Ganoderma lucidum fermentation products. This indicates that the addition of plant powder significantly improved the hydroxyl radical scavenging rate of Ganoderma lucidum fermentation products, demonstrating good antioxidant activity.

[0157] Table 10 Hydroxyl radical scavenging activity (IC50) 50 )

[0158]

[0159] 3. Summary

[0160] The above experimental results show that the fermented products of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder exhibit good DPPH and hydroxyl radical scavenging activities and have good antioxidant activities.

[0161] Example 12: Safety of Ganoderma lucidum fermentation products

[0162] Take 100 mL of each of the Ganoderma lucidum fermentation products prepared by fermentation in different concentrations of plant powder (5 g / L, 7.5 g / L, 10 g / L, 15 g / L, 20 g / L) in Examples 6-10, mix them evenly according to the same type, and obtain samples of Ganoderma lucidum fermentation products of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder.

[0163] The Ganoderma lucidum fermentation product prepared by fermentation in the fermentation medium without the addition of plant powder in Example 5 was used as a control sample.

[0164] The cytotoxicity of Ganoderma lucidum ferment broth to human skin fibroblasts (HSF) was determined using a CCK-8 assay kit (CCK-8, Dojindo, Kumamoto, Japan). Generally, a cell viability of over 80% was considered to indicate no significant toxicity to the cells. Logarithmic growth phase cells were collected, counted, diluted with complete culture medium, and the cell pellet was resuspended. 10 μL of the solution was added to each well of a 96-well plate. 4 HSF cells were cultured for 24 hours in a 5% CO2, 37°C incubator. Different plant-based Ganoderma lucidum ferments were concentrated 10-fold, filtered through a 0.22 μm sterile membrane, and serially diluted twice with complete culture medium to achieve initial sample volume fractions of 2.5%, 5%, 10%, 20%, 40%, and 80%. Culture medium containing different volume fractions of Ganoderma lucidum ferments was added to each well, with three replicates for each concentration. The control group received complete culture medium. Cells were cultured for 24 hours in a 5% CO2, 37°C incubator. CCK-8 diluent (10-fold dilution of the stock solution with complete culture medium) was added to each well, and the cells were cultured for another 2-4 hours in a 5% CO2, 37°C incubator. The absorbance at 450 nm was then measured. Cell viability % = (OD value of the treated group / OD value of the control group) * 100%.

[0165] Cytotoxicity results are shown in Figure 3 As shown in the figure, the cell survival rate of different plant powder Ganoderma lucidum fermentation products was higher than 80% when the volume fraction was 0-80%, with no obvious cytotoxicity and high safety.

[0166] Example 13: Effects of Ganoderma lucidum fermentation products on collagen in HSF cells under oxidative stress

[0167] Take 100 mL of each of the Ganoderma lucidum fermentation products prepared by fermentation in different concentrations of plant powder (5 g / L, 7.5 g / L, 10 g / L, 15 g / L, 20 g / L) in Examples 6-10, mix them evenly according to the same type, and obtain samples of Ganoderma lucidum fermentation products of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder, and purslane powder.

[0168] The Ganoderma lucidum fermentation product prepared by fermentation in the fermentation medium without the addition of plant powder in Example 5 was used as a control sample.

[0169] A decrease in type I collagen (COL-I) and elastin (ELN) levels leads to sagging skin and wrinkles, which are important indicators of skin aging. This invention utilizes H2O2 to induce a damaged cell model of human dermal fibroblasts (HSF) and uses collagen and elastin as indicators to study the anti-aging effects of Ganoderma lucidum fermentation products.

[0170] HSF cells were divided into 10 5 Cells were seeded per well in 24-well plates and cultured in complete culture medium for 12-24 hours to allow cell adhesion. All cells were then divided into four groups according to Table 11.

[0171] Table 11 Experimental Groups

[0172]

[0173] The above experimental groups were incubated for another 24 hours. The sample groups consisted of 10% (v / v) fermented products of Ganoderma lucidum (honeysuckle powder), Ganoderma lucidum (wild chrysanthemum powder), Ganoderma lucidum (sage powder), Ganoderma lucidum (centella asiatica powder), and Ganoderma lucidum (purslane powder) diluted with culture medium (samples), and 10% fermented product of Ganoderma lucidum without plant powder (control). After the culture and stimulation process was completed, the culture supernatant was carefully collected into centrifuge tubes and centrifuged at 3000 rpm for 10 min to remove cell debris and other impurities. The supernatant was then used to determine the COL-I and ELN contents using a human type I collagen ELISA kit (Jianglai Biotechnology, JL19111) and a human elastin ELISA kit (Jianglai Biotechnology, JL12704).

[0174] The effects of different plant powders and Ganoderma lucidum fermentation products on COL-I production in H2O2-treated HSF cells are shown in the figure. Figure 4As shown in the figure, H2O2 treatment of HSF cells significantly reduced COL-I production compared to the negative control group (p < 0.01), indicating successful model establishment. Treatment with different Ganoderma lucidum ferments significantly increased COL-I production in HSF cells compared to the model group (p < 0.05), with the most significant increases observed in fermented Ganoderma lucidum containing honeysuckle powder, sage powder, and purslane powder (p < 0.01). Compared to the control group without plant powder, the addition of different plant powder Ganoderma lucidum ferments increased COL-I production in HSF cells, with the most significant increases observed in fermented Ganoderma lucidum containing honeysuckle powder and sage powder (p < 0.01). This indicates that plant powder Ganoderma lucidum ferments can more effectively inhibit H2O2-induced COL-I loss in HSF cells.

[0175] The effects of different plant powders and Ganoderma lucidum fermentation products on ELN production in H2O2-treated HSF cells are shown in the figure. Figure 5 As shown in the figure, H2O2 treatment of HSF cells led to a decrease in HSF cell ELN production, which was 22.05% lower than the negative control group (p < 0.01), indicating successful model establishment. Treatment with different Ganoderma lucidum ferments increased HSF cell ELN production compared to the model group, with honeysuckle powder and sage powder Ganoderma lucidum ferments showing significantly higher ELN production (p < 0.01). Compared to the control group without plant powder, HSF cells with different plant powder Ganoderma lucidum ferments showed increased ELN production, with honeysuckle powder and sage powder Ganoderma lucidum ferments showing significantly higher ELN production (p < 0.05). This indicates that plant powder Ganoderma lucidum ferments can more effectively inhibit H2O2-induced HSF cell ELN loss.

[0176] Experiments have shown that different Ganoderma lucidum ferments can significantly promote the production of collagen and elastin under oxidative stress, thus playing a role in delaying aging.

[0177] In summary, the Ganoderma lucidum ferment prepared by this invention not only has a high content of active polysaccharides, but is also rich in active substances such as polyphenols and flavonoids. It has good DPPH free radical and hydroxyl free radical scavenging activity, and can significantly increase the production of collagen and elastin in human dermal fibroblasts at the cellular level. It has good antioxidant and anti-aging effects, and has obvious advantages and broad application prospects in the preparation of anti-aging cosmetics (such as masks, serums, lotions, creams, sprays or liquids).

[0178] The above examples only represent the technical solutions of this invention and are not intended to limit the experiments. Although we have improved the experimental scheme, researchers in the same field can still further improve the experimental scheme described above or make scientifically equivalent substitutions for the experimental steps. These changes do not cause the substance of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions claimed in this invention.

Claims

1. A Ganoderma lucidum plant, AD256, characterized in that, The Ganoderma lucidum AD256 is deposited at the China Center for Type Culture Collection (CCTCC), with accession number CCTCC NO: M 20252522.

2. The Ganoderma lucidum AD256 according to claim 1, characterized in that, The colonies of Ganoderma lucidum AD256 are white, round, with dense hyphae, and the hyphae have clamp connections.

3. The Ganoderma lucidum AD256 according to claim 1, characterized in that, The Ganoderma lucidum AD256 has the ability to produce cellulase.

4. The Ganoderma lucidum ferment produced by fermenting Ganoderma lucidum AD256 according to claim 1, characterized in that, The Ganoderma lucidum ferment was prepared by fermenting Ganoderma lucidum AD256 in a Ganoderma lucidum fermentation medium containing plant powder.

5. The Ganoderma lucidum fermented product according to claim 4, characterized in that, The Ganoderma lucidum fermented products include at least one of the following: honeysuckle powder Ganoderma lucidum fermented products, wild chrysanthemum powder Ganoderma lucidum fermented products, sage powder Ganoderma lucidum fermented products, centella asiatica powder Ganoderma lucidum fermented products, and purslane powder Ganoderma lucidum fermented products.

6. The Ganoderma lucidum fermented product according to claim 4, characterized in that, The preparation method of the Ganoderma lucidum ferment includes the following steps: (1) The Ganoderma AD256 was inoculated into a liquid seed culture medium and fermented to obtain a seed liquid; (2) The seed liquid is inoculated into a Ganoderma lucidum fermentation medium containing plant powder and fermented to obtain a fermentation liquid; the plant powder includes at least one of honeysuckle powder, wild chrysanthemum powder, sage powder, centella asiatica powder and purslane powder. (3) Homogenize the fermentation liquid to obtain Ganoderma lucidum fermentation homogenate; (4) Let the Ganoderma lucidum fermentation homogenate stand at room temperature, centrifuge and filter to obtain Ganoderma lucidum fermentation product.

7. The Ganoderma lucidum fermented product according to claim 6, characterized in that, The particle size of the plant powder is 40-200 mesh; the mass-volume concentration of the plant powder is 5-20 g / L, based on the volume of the Ganoderma lucidum fermentation culture medium.

8. The Ganoderma lucidum fermented product according to claim 7, characterized in that, The pressure for homogenizing the fermentation broth is 80~120 MPa.

9. The use of Ganoderma lucidum AD256 as described in claim 1 or Ganoderma lucidum fermentation product as described in claim 4 in the preparation of cosmetics with anti-aging effects.

10. The application according to claim 9, characterized in that, The cosmetics include at least one of the following: face masks, serums, lotions, creams, sprays, and liquids.

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