Preparation method of tremella aurantialba fermentation liquor and application of tremella aurantialba fermentation liquor in cosmetics

Abstract

The invention belongs to the technical field of cosmetics, and particularly discloses a preparation method of tremella aurantialba fermentation liquor and application of the tremella aurantialba fermentation liquor in cosmetics, and the preparation method comprises the following steps: (1) adding tremella aurantialba powder, cellulase and pectinase into water, performing enzymolysis, and filtering to obtain enzymatic hydrolysate; (2) uniformly mixing the enzymatic hydrolysate, a carbon source, a nitrogen source and a basal culture medium, adjusting the pH value to 5.5-6 by using inorganic salt, and sterilizing to obtain a fermentation substrate; (3) inoculating saccharomyces cerevisiae into the fermentation substrate, fermenting, sterilizing, centrifuging, filtering, performing ultrafiltration on supernate by using an ultrafiltration membrane with the molecular weight cutoff of 3000Da, and collecting filtrate to obtain a precursor; and (4) uniformly mixing the precursor, p-hydroxyacetophenone and 1, 2-hexanediol to obtain the tremella aurantialba fermentation liquor. The tremella aurantialba fermentation liquor disclosed by the invention has excellent effects of resisting oxidation, relieving and repairing.

Description

Technical Field

[0001] This invention relates to the field of cosmetic technology, specifically to a method for preparing a fungus fermentation liquid and its application in cosmetics. Background Art

[0002] Golden ear fungus is a traditional Chinese medicinal and edible fungus. Its fruiting body contains 37.8%-40.55% polysaccharides, which have significant antioxidant, immune-regulating and moisturizing effects.

[0003] Traditional extraction processes for *Auricularia auricula-judae* fruiting bodies rely on organic solvents (such as alcohol precipitation), leading to selective loss of active ingredients, complex processes, high safety risks due to solvent residues, and inability to fully extract water-soluble components, resulting in low resource utilization. Traditional *Auricularia auricula-judae* fermentation processes have the following drawbacks: 1. Low fermentation efficiency: Conventional liquid fermentation requires 7-10 days, and polysaccharide yield is unstable; 2. Limited active ingredients (i.e., limited efficacy); 3. Limited applications: Existing fermentation broths are mostly used in the food industry, and the antioxidant and anti-allergic properties of cosmetic-grade fermentation broths do not meet high industry standards.

[0004] Therefore, this application is submitted. Summary of the Invention

[0005] This invention provides a method for preparing a fungus fermentation liquid and its application in cosmetics. The fungus fermentation liquid described in this invention has excellent antioxidant, soothing and repairing effects.

[0006] The present invention solves its technical problem by adopting the following technical solution: A method for preparing a fungus fermentation broth includes the following steps: (1) Add the golden ear powder, cellulase and pectinase to water, hydrolyze, filter, and obtain the hydrolysate; (2) Mix the enzyme hydrolysate, carbon source, nitrogen source and basic culture medium evenly, adjust the pH to 5.5-6 with inorganic salts, sterilize, and obtain the fermentation substrate; (3) Inoculate the yeast into the fermentation substrate, ferment, sterilize, centrifuge, filter, ultrafilter the supernatant with an ultrafiltration membrane with a molecular weight cutoff of 3000 Da, collect the filtrate, and obtain the precursor. (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol evenly to obtain the fermentation broth of Auricularia auricula-judae.

[0007] This invention first enzymatically hydrolyzes the golden ear fungus powder, effectively promoting the dissolution rate of active ingredients and facilitating subsequent fermentation. The powder is then mixed with a carbon source, a nitrogen source, and a basal culture medium to obtain a fermentation substrate. Subsequently, fermentation is carried out using Saccharomyces cerevisiae to convert the large molecules of golden ear fungus (such as polysaccharides and proteins) into small molecule active substances (such as short-chain fatty acids and polypeptides). By utilizing gentle enzymatic hydrolysis combined with microbial transformation, the full range of components is efficiently released, while avoiding the use of solvents, significantly enhancing antioxidant, soothing, and repairing effects.

[0008] Compared to complex purification processes that rely on organic solvent precipitation (such as alcohol precipitation), the extraction method of this invention is simpler. It mainly includes mild enzymatic hydrolysis, microbial fermentation transformation, and necessary solid-liquid separation steps (centrifugation, filtration, ultrafiltration). It avoids the use of organic solvents, reduces purification costs and complexity, reduces industrial pollution, and has a high raw material utilization rate, which can effectively reduce production costs. It can replace high-priced active ingredients (such as nicotinamide and bosine).

[0009] As a preferred embodiment of the present invention, the ratio of the auricularia auricula-judae powder, cellulase, pectinase and water is 1g:(0.004~0.008)g:(0.002~0.006)g:(4~10)mL.

[0010] In a preferred embodiment of the present invention, the enzymatic hydrolysis temperature is 45~55℃ and the enzymatic hydrolysis time is 2~6h.

[0011] As a preferred embodiment of the present invention, the mass ratio of the enzymatic hydrolysate, carbon source, nitrogen source and basal culture medium is (2~5):(0.1~2):(0.4~1):(92~97.5).

[0012] In a preferred embodiment of the present invention, the carbon source includes at least one of glucose and sucrose; The nitrogen source includes at least one of peptone and yeast extract; The basal culture medium includes YPD culture medium; The inorganic salt includes at least one of sodium chloride, magnesium sulfate, and potassium dihydrogen phosphate.

[0013] In a preferred embodiment of the present invention, the amount of brewer's yeast inoculated is 2-5% of the total mass of the fermentation substrate.

[0014] In a preferred embodiment of the present invention, the fermentation temperature is 26~30℃ and the time is 72~96h.

[0015] As a preferred embodiment of the present invention, the sterilization temperature in step (2) is 120~125℃ and the sterilization time is 20~40min; Step (3) The sterilization temperature is 65~70℃ and the time is 8~20min.

[0016] In a preferred embodiment of the present invention, the mass ratio of the precursor, p-hydroxyacetophenone, and 1,2-hexanediol is 1:(0.005~0.01):(0.005~0.02).

[0017] In a preferred embodiment of the present invention, the particle size of the gold ear powder is 80-100 mesh.

[0018] The present invention also provides an application of the *Auricularia auricula-judae* fermentation liquid prepared by the above preparation method in the preparation of cosmetics.

[0019] In a preferred embodiment of the present invention, the mass percentage of the *Auricularia auricula-judae* fermentation liquid in cosmetics is 0.1-10%.

[0020] The beneficial effects of this invention are as follows: This invention first enzymatically hydrolyzes the golden ear fungus powder, effectively promoting the dissolution rate of active ingredients and facilitating subsequent fermentation. The powder is then mixed with carbon sources, nitrogen sources, and a basic culture medium to obtain a fermentation substrate. Subsequently, fermentation is carried out using Saccharomyces cerevisiae to convert the large molecules of golden ear fungus (such as polysaccharides and proteins) into small molecule active substances (such as short-chain fatty acids and polypeptides). By utilizing gentle enzymatic hydrolysis combined with microbial transformation, the full components are efficiently released, while avoiding the use of solvents, significantly enhancing the antioxidant, soothing, and repairing effects. Detailed Implementation

[0021] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0022] In this invention, the technical features described in an open-ended manner include both closed-ended technical solutions composed of the listed features and open-ended technical solutions that include the listed features.

[0023] In this invention, numerical ranges are involved. Unless otherwise specified, the numerical ranges are considered continuous and include the minimum and maximum values ​​of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Additionally, when multiple ranges are provided to describe features or characteristics, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are included.

[0024] In this invention, there are no particular limitations on the specific dispersion and stirring methods.

[0025] Unless otherwise specified, all reagents or instruments used in this invention are commercially available conventional products. Unless otherwise specified, the raw materials used in each comparative example and the parallel experiments of each embodiment are the same commercially available products.

[0026] Example 1 A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.006 g: 0.004 g: 5 mL.

[0027] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 4:0.8:0.5:94.7.

[0028] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 4% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0029] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0030] Example 2 A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.008 g: 0.002 g: 5 mL.

[0031] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 2:0.1:0.4:97.5.

[0032] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 5% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0033] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.01:0.01 to obtain the fermentation broth of Auricularia auricula-judae.

[0034] Example 3 A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.004 g: 0.006 g: 5 mL.

[0035] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 5:2:1:92.

[0036] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 2% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0037] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.02 to obtain the fermentation broth of Auricularia auricula-judae.

[0038] Comparative Example 1

[0039] A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; (2) Mix the auricularia auricula powder, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of auricularia auricula powder, glucose, peptone and YPD medium is 4:0.8:0.5:94.7.

[0040] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 4% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0041] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0042] Comparative Example 2

[0043] A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.001 g: 0.009 g: 5 mL.

[0044] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 4:0.8:0.5:94.7.

[0045] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 4% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0046] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0047] Comparative Example 3

[0048] A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.009 g: 0.001 g: 5 mL.

[0049] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 4:0.8:0.5:94.7.

[0050] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 4% of the total mass of the fermentation substrate. The brewer's yeast strain was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 2.1195.

[0051] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0052] Comparative Example 4

[0053] A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.006 g: 0.004 g: 5 mL.

[0054] (2) Mix the enzyme hydrolysate and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate to YPD medium is 4:96.

[0055] (3) Inoculate the brewing yeast into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of the brewing yeast is 4% of the total mass of the fermentation substrate. The brewer's yeast was purchased from Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC NO 2.1195.

[0056] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0057] Comparative Example 5

[0058] A method for preparing a fungus fermentation broth includes the following steps: (1) Grind the dried golden ear fungus into 100 mesh to obtain golden ear fungus powder; Golden ear fungus powder, cellulase, and pectinase were added to water and enzymatically hydrolyzed at 48°C for 3.5 hours. The mixture was then filtered to obtain the enzymatic hydrolysate. The ratio of the golden ear fungus powder, cellulase, pectinase, and water was 1 g: 0.006 g: 0.004 g: 5 mL.

[0059] (2) Mix the enzyme hydrolysate, glucose, peptone and YPD medium evenly, adjust the pH to 5.8 with sodium chloride, sterilize at 121℃ for 30 min to obtain the fermentation substrate; the mass ratio of the enzyme hydrolysate, glucose, peptone and YPD medium is 4:0.8:0.5:94.7.

[0060] (3) Inoculate Lactobacillus plantarum into the fermentation substrate, ferment on a shaker at 28°C for 96 h, sterilize at 68°C for 10 min, centrifuge, filter, and ultrafilter the supernatant using an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. Collect the filtrate to obtain the precursor; the inoculation amount of Saccharomyces cerevisiae is 4% of the total mass of the fermentation substrate. The *Lactobacillus plantarum* was purchased from the Guangdong Provincial Microbial Culture Collection Center, with the number GDMCC 1.1516.

[0061] (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol in a mass ratio of 1:0.005:0.005 to obtain the fermentation broth of Auricularia auricula-judae.

[0062] 1. DPPH free radical scavenging rate: Tested according to T / SHRH 006-2018 "Cosmetics - Experimental Method for Free Radical (DPPH) Scavenging".

[0063] 2. Determination of the inhibition rate of macrophage inflammatory factors (TNF-α / IL-6) TNF-α and IL-6, inflammatory factors released by macrophages, are core signaling molecules in skin sensitivity responses. These factors directly participate in skin redness, stinging, and barrier damage processes, and are widely used as key indicators in evaluating the soothing efficacy of cosmetics. Measuring the inhibition rate of TNF-α / IL-6 in a sample can objectively assess its soothing effect.

[0064] Experimental methods Sample processing: The fermentation broth samples obtained in Examples 1-3 and Comparative Examples 1-5 were prepared into experimental test solutions with a volume fraction of 1% using sterile PBS buffer (sterilized by passing through a 0.22 μm filter membrane).

[0065] Cell stimulation and treatment: Logarithmic growth phase mouse macrophages RAW264.7 were used, and 1×10⁻⁶ cells were used for stimulation and treatment. 5 Cells / mL were seeded at a density of 100 μL / well in 96-well plates and cultured for 24 hours until adherence was achieved. Control group: 100 μL of fresh culture medium was added. Model group: 100 μL of culture medium containing 1 μg / mL LPS was added.

[0066] Experimental group: Add 100 μL of culture medium containing 1 μg / mL LPS + 1% sample (6 replicates per group, incubate at 37°C for 24 hours).

[0067] Factor detection: Collect the supernatant and detect the contents of TNF-α and IL-6 separately according to the ELISA kit instructions (purchased from Shanghai Enzyme-Link Biotechnology Co., Ltd.). Measure the absorbance (OD value) at 450 nm using an enzyme-linked immunosorbent assay (ELISA) reader.

[0068] Results calculation: TNF-α inhibition rate (%) = 1 − (concentration of experimental group − mean of blank group) / (concentration of model group − mean of blank group) × 100%.

[0069] IL-6 inhibition rate (%) = calculated as above.

[0070] 3. HaCaT cell scratch migration rate assay The migration capacity of HaCaT cells (human immortalized keratinocytes) is a core indicator for assessing skin barrier repair function. When the skin barrier is damaged, keratinocytes migrate to cover the wound, accelerating wound healing. Measuring the rate at which a sample promotes scratch closure can objectively assess its repair efficacy.

[0071] Sample processing: The fermentation broth samples obtained in Examples 1-3 and Comparative Examples 1-5 were prepared into experimental test solutions with a volume fraction of 1% using serum-free DMEM medium (sterilized by passing through a 0.22 μm filter membrane).

[0072] Establishment of the scratch model: HaCaT cells in the logarithmic growth phase were harvested and subjected to a 5×10⁻⁶ sclerotherapy. 5 Cells were seeded at a density of 500 μL / well in 24-well plates and cultured for 24 hours until fully adherent. Using a 200 μL sterile pipette tip, a uniform vertical scratch was made in the center of each well. Cells were gently washed three times with PBS to remove detached cells. Grouping was performed as follows: Control group: serum-free DMEM medium containing 1% FBS; Experimental group: serum-free DMEM medium containing 1% sample + 1% FBS (3 replicates per group, incubated at 37°C for 24 hours).

[0073] Image acquisition and analysis: Scratch images at fixed locations were captured under an inverted microscope (40×) at 0h and 24h, and the scratch area was measured using ImageJ software. The scratch closure rate (%) was calculated as: 1 − (scratch area at 24h / scratch area at 0h) × 100%.

[0074] Migration facilitation rate (%) = (Closed-off rate of experimental group - Closed-off rate of blank group) / Closed-off rate of blank group × 100%.

[0075] Table 1

[0076] As can be seen from Table 1, the auricularia auricula-judae fermentation liquid of the present invention has excellent antioxidant, soothing and repairing effects.

[0077] Finally, it should be noted that the above embodiments are used to illustrate the technical solutions of the present invention and not to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention without departing from the essence and scope of the technical solutions of the present invention.

Claims

1. A method for preparing a fungus fermentation broth, characterized in that, The following steps are involved: (1) Add the golden ear powder, cellulase and pectinase to water, hydrolyze, filter, and obtain the hydrolysate; (2) Mix the enzyme hydrolysate, carbon source, nitrogen source and basic culture medium evenly, adjust the pH to 5.5-6 with inorganic salts, sterilize, and obtain the fermentation substrate; (3) Inoculate the yeast into the fermentation substrate, ferment, sterilize, centrifuge, filter, ultrafilter the supernatant with an ultrafiltration membrane with a molecular weight cutoff of 3000 Da, collect the filtrate, and obtain the precursor. (4) Mix the precursor, p-hydroxyacetophenone and 1,2-hexanediol evenly to obtain the fermentation broth of Auricularia auricula-judae.

2. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The ratio of the auricularia auricula-judae powder, cellulase, pectinase, and water is 1g:(0.004~0.008)g:(0.002~0.006)g:(4~10)mL.

3. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The enzymatic hydrolysis temperature is 45~55℃, and the enzymatic hydrolysis time is 2~6h.

4. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The mass ratio of the enzymatic hydrolysate, carbon source, nitrogen source, and basal culture medium is (2~5):(0.1~2):(0.4~1):(92~97.5).

5. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The carbon source includes at least one of glucose and sucrose; The nitrogen source includes at least one of peptone and yeast extract; The basal culture medium includes YPD culture medium; The inorganic salt includes at least one of sodium chloride, magnesium sulfate, and potassium dihydrogen phosphate.

6. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The amount of Saccharomyces cerevisiae inoculated is 2-5% of the total mass of the fermentation substrate.

7. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The fermentation temperature is 26~30℃ and the time is 72~96h.

8. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, Step (2) The sterilization temperature is 120~125℃ and the sterilization time is 20~40min; Step (3) The sterilization temperature is 65~70℃ and the time is 8~20min.

9. The method for preparing the *Auricularia auricula-judae* fermentation broth according to claim 1, characterized in that, The mass ratio of the precursor, p-hydroxyacetophenone, and 1,2-hexanediol is 1:(0.005~0.01):(0.005~0.02).

10. The application of the *Auricularia auricula-judae* fermentation liquid prepared by any one of the preparation methods according to claims 1 to 9 in the preparation of cosmetics.

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