A dual-purpose medicinal and edible fungus co-culture fermentation liquor with whitening effect, a preparation method and application thereof

Abstract

This invention relates to the field of microbial fermentation technology, and particularly to a co-culture fermentation broth of medicinal and edible fungi with whitening effects, its preparation method, and its application. The preparation method of the co-culture fermentation broth of medicinal and edible fungi with whitening effects includes: inoculating the seed liquid of a first fungus and the seed liquid of a second fungus into a co-fermentation medium for fermentation and co-culture to obtain the co-culture fermentation broth of medicinal and edible fungi with whitening effects; the first fungus includes at least one of *Ganoderma lucidum* strain, *Trametes versicolor* strain, *Poria cocos* strain, *Auricularia auricula-judae* strain, *Phellinus linteus* strain, and *Aspergillus schavar*; the second fungus includes *Schizophyllum commune*. This invention co-cultures *Schizophyllum commune* with other specific strains and found that co-culturing the two strains can exert a synergistic effect, thereby significantly improving the tyrosinase inhibition effect of the fermentation broth. The optimal combination of fermentation broth can achieve a tyrosinase inhibition rate of over 90%, and even 1 wt% of the fermentation broth has a tyrosinase inhibition effect of approximately 90%.

Description

Technical Field

[0001] This invention relates to the field of microbial fermentation technology, and in particular to a co-culture fermentation broth of medicinal and edible fungi with whitening effects, its preparation method and application. Background Technology

[0002] Co-culture fermentation refers to the cultivation of two or more microorganisms (bacteria, fungi, yeasts, etc.) in the same fermentation system. Through metabolic synergy, niche complementarity, and signal molecule regulation among the microorganisms, functions that cannot be achieved by a single microbial species can be realized. Its core value lies in: overcoming the metabolic limitations of single microbial species, efficiently utilizing complex substrates (such as agricultural waste and industrial by-products); achieving the integration of "substrate degradation-product synthesis-metabolic regulation" to shorten the fermentation process; and synthesizing novel or high-value-added composite products (such as synergistic active ingredients and special enzyme preparations).

[0003] Co-culture fermentation is an important technique in the field of industrial biotechnology and is now widely used in food processing, preparation of functional products, and other fields. For example, yeast co-culture fermentation can be used to regulate the chemical composition of wine and improve its flavor quality; co-culture fermentation of filamentous fungi and lactic acid bacteria can be used to increase the types and contents of aroma compounds and improve the flavor quality of yogurt; probiotics can be used to ferment medicinal and edible materials such as Poria cocos, white hyacinth bean, coix seed, almond, and licorice to increase the content of active ingredients such as polysaccharides, proteins, polyphenols, and flavonoids in dehumidifying soups; and co-culture fermentation of Aspergillus, Trichoderma, and yeast can degrade straw and increase cellulase yield and substrate conversion rate.

[0004] Medicinal and edible fungi (such as Ganoderma lucidum, shiitake mushroom, Cordyceps militaris, Poria cocos, Hericium erinaceus, etc.) are special fungal resources that combine "food safety" and "medicinal efficacy". They are rich in active ingredients such as polysaccharides, triterpenes, polypeptides, and sterols, and have effects such as immune regulation, anti-tumor, antioxidant, and lipid-lowering. They are widely used in health food, functional food and medicine.

[0005] Currently, the industrial production of fungi that are both medicinal and edible mainly relies on the pure culture of single strains, such as liquid fermentation of Ganoderma lucidum and solid cultivation of shiitake mushrooms. Summary of the Invention

[0006] In view of this, the technical problem to be solved by the present invention is to provide a co-culture fermentation broth of medicinal and edible fungi with whitening effect, its preparation method and application, which can significantly improve the tyrosinase inhibition effect of the fermentation broth.

[0007] This invention provides a method for preparing a co-culture fermentation broth of medicinal and edible fungi with whitening effects, comprising the following steps:

[0008] The seed liquid of the first fungus and the seed liquid of the second fungus were inoculated into a co-fermentation medium for co-fermentation and co-culture to obtain a co-culture fermentation broth of medicinal and edible fungi with whitening effects.

[0009] The first fungus includes at least one of the following: Ganoderma lucidum strain, Trametes versicolor strain, Poria cocos strain, Auricularia auricula-judae strain, Phellinus linteus strain, and Aspergillus schavar;

[0010] The second fungus includes Schizophyllum commune.

[0011] Preferably, the co-fermentation medium includes a carbon source, malt extract, a nitrogen source, and water;

[0012] The co-fermentation medium contains 10-30 g / L of carbon source, 10-30 g / L of malt extract, and 1-3 g / L of nitrogen source.

[0013] Preferably, the carbon source includes at least one of glucose, sucrose, and starch;

[0014] In some embodiments of the present invention, the nitrogen source includes at least one of yeast extract, peptone, and ammonium sulfate.

[0015] Preferably, the method for preparing the co-fermentation medium includes the following steps:

[0016] The carbon source, malt extract powder, nitrogen source and water are mixed and sterilized to obtain the co-fermentation medium.

[0017] The sterilization temperature is 115~125℃, the pressure is 0.05~0.15 MPa, and the time is 10~30 min.

[0018] Preferably, the inoculum amount of the first fungus in the seed culture of the first fungus is 2.5 wt% to 7.5 wt%.

[0019] In the seed culture of the second fungus, the inoculum amount of the second fungus is 2.5 wt% to 7.5 wt%.

[0020] Preferably, the method for preparing the seed liquid of the first fungus includes the following steps:

[0021] The mycelium of the first fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the first fungus.

[0022] The method for preparing the seed solution of the second fungus includes the following steps:

[0023] The mycelium of the second fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the second fungus.

[0024] The seed culture medium includes a carbon source, malt extract powder, a nitrogen source, and water;

[0025] The seed culture medium contains 10-30 g / L of carbon source, 10-30 g / L of malt extract powder, and 1-3 g / L of nitrogen source.

[0026] Preferably, in the co-fermentation medium, the inoculation amount of the seed liquid of the first fungus is 2.5 wt% to 7.5 wt%, and the inoculation amount of the seed liquid of the second fungus is 2.5 wt% to 7.5 wt%.

[0027] Preferably, the fermentation co-culture temperature is 28~30℃, the stirring rate is 160~180 rpm, and the time is 1~10 days.

[0028] The present invention also provides a co-culture fermentation broth of medicinal and edible fungi with whitening effect prepared by the preparation method described above.

[0029] This invention also provides an application of the co-culture fermentation broth of the medicinal and edible fungi with whitening effects described above in the preparation of skin care products.

[0030] This invention co-cultures *Schizophyllum commune* with other specific strains and found that co-culturing the two strains can exert a synergistic effect, thereby significantly improving the tyrosinase inhibition effect of the fermentation broth. The optimal combination of fermentation broth can achieve a tyrosinase inhibition rate of over 90%, and 1 wt% of the fermentation broth also has a tyrosinase inhibition effect of about 90%. Detailed Implementation

[0031] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0032] This invention provides a method for preparing a co-culture fermentation broth of medicinal and edible fungi with whitening effects, comprising the following steps:

[0033] The seed liquid of the first fungus and the seed liquid of the second fungus were inoculated into a co-fermentation medium for co-fermentation and co-culture to obtain a co-culture fermentation broth of medicinal and edible fungi with whitening effects.

[0034] The first fungus includes at least one of the following: Ganoderma lucidum strain, Trametes versicolor strain, Poria cocos strain, Auricularia auricula-judae strain, Phellinus linteus strain, and Aspergillus schavar;

[0035] The second fungus includes Schizophyllum commune.

[0036] Specifically, the first fungus is *Ganoderma sinensis*, *Trametes versicolor*, *Poria cocos*, *Auricularia auricula-judae*, *Sanghuang*, or *Aspergillus schavar*. The second fungus is *Schizophyllum commune*.

[0037] This invention involves inoculating seed culture into a co-fermentation medium for fermentation and co-cultivation to obtain a co-culture fermentation broth of medicinal and edible fungi with whitening effects. The seed culture consists solely of the seed culture of a first fungus and the seed culture of a second fungus.

[0038] Neither the seed culture of the first fungus nor the seed culture of the second fungus underwent inactivation treatment.

[0039] In some embodiments of the present invention, the inoculum amount of the first fungus in the seed solution of the first fungus is 2.5 wt% to 7.5 wt%, for example 2.5 wt%.

[0040] In some embodiments of the present invention, the inoculum amount of the second fungus in the seed solution of the second fungus is 2.5 wt% to 7.5 wt%, for example, 2.5 wt%.

[0041] In some embodiments of the present invention, the inoculum amount of the seed liquid of the first fungus in the co-fermentation medium is 2.5 wt% to 7.5 wt%, for example 2.5 wt%.

[0042] In some embodiments of the present invention, the inoculum amount of the seed liquid of the second fungus in the co-fermentation medium is 2.5 wt% to 7.5 wt%, for example 2.5 wt%.

[0043] In some embodiments of the present invention, the method for preparing the seed solution of the first fungus includes the following steps:

[0044] The mycelium of the first fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the first fungus.

[0045] In some embodiments of the present invention, the method for preparing the seed solution of the second fungus includes the following steps:

[0046] The mycelium of the second fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the second fungus.

[0047] The seed culture medium comprises a carbon source, malt extract powder, a nitrogen source, and water. Specifically, the seed culture medium is composed of a carbon source, malt extract powder, a nitrogen source, and water; the carbon source content in the seed culture medium is 10-30 g / L, the malt extract powder content is 10-30 g / L, and the nitrogen source content is 1-3 g / L. The water is deionized water. Specifically, the carbon source content in the seed culture medium can be 10 g / L, 15 g / L, 20 g / L, 25 g / L, or 30 g / L; the malt extract powder content can be 10 g / L, 15 g / L, 20 g / L, 25 g / L, or 30 g / L; and the nitrogen source content can be 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, or 3 g / L. More specifically, the seed culture medium contains 20 g / L of carbon source, 20 g / L of malt extract powder, and 2 g / L of nitrogen source.

[0048] In some embodiments of the present invention, the carbon source is at least one of glucose, sucrose, and starch.

[0049] In some embodiments of the present invention, the nitrogen source is at least one selected from yeast extract, peptone, and ammonium sulfate. Specifically, the nitrogen source is yeast extract and peptone; the mass ratio is 0.5~1.5:0.5~1.5, for example, 1:1.

[0050] In some embodiments of the present invention, the method for preparing the seed culture medium includes the following steps:

[0051] The carbon source, malt extract powder, nitrogen source and water are mixed and sterilized to obtain the seed culture medium.

[0052] The mixing process involves stirring and mixing.

[0053] The sterilization temperature is 115~125℃, such as 115℃, 117℃, 121℃, 123℃, 121℃; the pressure is 0.05~0.15 MPa, such as 0.05 MPa, 0.09 MPa, 0.11 MPa, 0.13 MPa, 0.15 MPa; and the time is 10~30 min, such as 10 min, 20 min, 30 min.

[0054] The culture rotation speed is 160~180 rpm, for example, 160 rpm, 170 rpm, 180 rpm. The time is 2~10 days, for example, 2 days, 4 days, 6 days, 8 days, 10 days.

[0055] In some embodiments of the present invention, the co-fermentation medium comprises a carbon source, malt extract powder, a nitrogen source, and water. Specifically, the co-fermentation medium contains 10-30 g / L of carbon source, 10-30 g / L of malt extract powder, and 1-3 g / L of nitrogen source. The water is deionized water. More specifically, the co-fermentation medium may contain 10 g / L, 15 g / L, 20 g / L, 25 g / L, or 30 g / L of carbon source; 10 g / L, 15 g / L, 20 g / L, 25 g / L, or 30 g / L of malt extract powder; and 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, or 3 g / L of nitrogen source. For example, the co-fermentation medium may contain 20 g / L of carbon source, 20 g / L of malt extract powder, and 2 g / L of nitrogen source.

[0056] In some embodiments of the present invention, the carbon source is at least one of glucose, sucrose, and starch.

[0057] In some embodiments of the present invention, the nitrogen source is at least one selected from yeast extract, peptone, and ammonium sulfate. Specifically, the nitrogen source is yeast extract and peptone; the mass ratio is 0.5~1.5:0.5~1.5, for example, 1:1.

[0058] In some embodiments of the present invention, the method for preparing the co-fermentation culture medium includes the following steps:

[0059] The carbon source, malt extract powder, nitrogen source and water are mixed and sterilized to obtain the co-fermentation medium.

[0060] The mixing process involves stirring and mixing.

[0061] The sterilization temperature is 115~125℃, such as 115℃, 117℃, 121℃, 123℃, 121℃; the pressure is 0.05~0.15 MPa, such as 0.05 MPa, 0.09 MPa, 0.11 MPa, 0.13 MPa, 0.15 MPa; and the time is 10~30 min, such as 10 min, 20 min, 30 min.

[0062] The culture rotation speed is 160~180 rpm, for example, 160 rpm, 170 rpm, 180 rpm. The time is 2~10 days, for example, 2 days, 4 days, 6 days, 8 days, 10 days.

[0063] In some embodiments of the present invention, the temperature of the fermentation co-culture is 28~30℃, such as 28℃, 29℃, or 30℃; the stirring rate is 160~180 rpm, such as 160 rpm, 165 rpm, 170 rpm, 175 rpm, or 180 rpm; and the time is 1~10 days, such as 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, or 10 days.

[0064] In some embodiments of the present invention, after the co-fermentation, the process further includes centrifugation and filtration. The centrifugation speed is 8000~9000 rpm, for example, 8000 rpm, 8500 rpm, or 9000 rpm; the temperature is 15~25℃, for example, 15℃, 18℃, 20℃, 22℃, or 25℃. The supernatant after centrifugation is filtered.

[0065] The present invention also provides a co-culture fermentation broth of medicinal and edible fungi with whitening effect prepared by the preparation method described above.

[0066] The present invention also provides an application of the co-culture fermentation broth of the medicinal and edible fungi with whitening effect described above in the preparation of skin care products, wherein the skin care products have whitening effect.

[0067] This invention co-cultures *Schizophyllum commune* with other specific strains and found that co-culturing the two strains can exert a synergistic effect, thereby significantly improving the tyrosinase inhibition effect of the fermentation broth. The optimal combination of fermentation broth can achieve a tyrosinase inhibition rate of over 90%, and 1 wt% of the fermentation broth also has a tyrosinase inhibition effect of about 90%.

[0068] To further illustrate the present invention, the following detailed description, in conjunction with embodiments, describes a co-culture fermentation broth of medicinal and edible fungi with whitening effects, its preparation method, and its application, but this should not be construed as limiting the scope of protection of the present invention.

[0069] In the examples and comparative examples, the seed culture medium was prepared according to the following method:

[0070] Glucose, malt extract, yeast powder, peptone, and deionized water were mixed evenly and sterilized at 121℃ and 0.11 MPa for 20 min to obtain seed culture medium. The seed culture medium contained 20 g / L carbon source, 20 g / L malt extract, 1 g / L yeast powder, and 1 g / L peptone.

[0071] Example 1

[0072] A co-culture fermentation broth of Schizophyllum commune and Ganoderma sinensis is obtained by inoculating the seed liquid of Ganoderma sinensis and the seed liquid of Schizophyllum commune into a co-fermentation medium for fermentation and co-culture.

[0073] 1) Preparation of seed culture of Schizophyllum commune: Mycelium of Schizophyllum commune was inoculated into sterilized seed culture medium and cultured at 30℃ and 180rpm for 4 days to obtain seed culture of Schizophyllum commune; The inoculation amount of Schizophyllum commune in the seed culture of Schizophyllum commune was 2.5wt%.

[0074] 2) Preparation of seed culture of Ganoderma sinensis: The mycelium of Ganoderma sinensis was inoculated into a sterilized seed culture medium and cultured at 30℃ and 180 rpm for 4 days to obtain the seed culture of Ganoderma sinensis; the inoculation amount of Ganoderma sinensis in the seed culture of Ganoderma sinensis was 2.5 wt.

[0075] 3) Preparation of co-fermentation medium: Glucose, malt extract powder, yeast powder, peptone and deionized water are stirred evenly and sterilized at 121℃ and 0.11 MPa for 20 min to obtain co-fermentation medium; the co-fermentation medium contains 20 g / L carbon source, 20 g / L malt extract powder, 1 g / L yeast powder and 1 g / L peptone.

[0076] 4) The seed liquid of Ganoderma sinensis and the seed liquid of Schizophyllum commune were inoculated into a co-fermentation medium (the inoculation amount of the seed liquid of Ganoderma sinensis in the co-fermentation medium was 2.5 wt%, and the inoculation amount of the seed liquid of Schizophyllum commune was 2.5 wt%). The co-fermentation was carried out at 30℃ for 3 days. After co-fermentation, the mixture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-culture fermentation broth of Schizophyllum commune and Ganoderma sinensis.

[0077] Example 2

[0078] A co-culture fermentation broth of Schizophyllum commune and Coriolus versicolor is obtained by inoculating the seed liquid of Coriolus versicolor and the seed liquid of Schizophyllum commune into a co-fermentation medium for fermentation and co-culture.

[0079] The difference from Example 1 is that Ganoderma lucidum is replaced with Ganoderma yunnanensis;

[0080] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Trametes versicolor is obtained.

[0081] Example 3

[0082] A co-culture fermentation broth of Schizophyllum commune and Auricularia aurea is obtained by inoculating the seed liquid of Auricularia aurea and the seed liquid of Schizophyllum commune into a co-fermentation medium for fermentation and co-culture.

[0083] The difference from Example 1 is that Ganoderma sinensis is replaced with Auricularia aurea; and the seed culture time for Auricularia aurea is 10 days.

[0084] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Auricularia aurea is obtained.

[0085] Example 4

[0086] A co-culture fermentation broth of Schizophyllum commune and Poria cocos is obtained by inoculating the seed liquid of Poria cocos and the seed liquid of Schizophyllum commune into a co-fermentation medium for fermentation and co-culture.

[0087] The difference from Example 1 is that Ganoderma lucidum is replaced with Poria cocos; and the culture time for the seed liquid of Poria cocos is 10 days.

[0088] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Poria cocos is obtained.

[0089] Example 5

[0090] A co-culture fermentation broth of Schizophyllum commune and Phellinus linteus is obtained by inoculating the seed liquid of Phellinus linteus and the seed liquid of Schizophyllum commune into a co-fermentation medium for fermentation and co-culture.

[0091] The difference from Example 1 is that Ganoderma sinensis is replaced with Phellinus linteus; the seed culture time of Phellinus linteus is 10 days in the preparation of the seed culture.

[0092] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Phellinus linteus is obtained.

[0093] Example 6

[0094] A co-culture fermentation broth of Schizophyllum commune and Aspergillus serrata is obtained by inoculating Aspergillus serrata seed liquid and Schizophyllum commune seed liquid into a co-fermentation medium for fermentation and co-culture.

[0095] The difference from Example 1 is that *Ganoderma sinensis* is replaced with *Aspergillus serrata*; in the preparation of the seed culture of *Aspergillus serrata*, the culture time is 7 days.

[0096] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Aspergillus schwannoides is obtained.

[0097] Comparative Example 1

[0098] A fermentation broth of a Schizophyllum commune strain is obtained by inoculating a seed culture of Schizophyllum commune into a fermentation medium and then fermenting it.

[0099] 1) Preparation of seed culture of Schizophyllum commune: Mycelium of Schizophyllum commune was inoculated into sterilized seed culture medium and cultured at 30℃ and 180rpm for 4 days to obtain seed culture of Schizophyllum commune; The inoculation amount of Schizophyllum commune in the seed culture of Schizophyllum commune was 2.5wt%.

[0100] 2) Preparation of fermentation medium: Glucose, malt extract powder, yeast powder, peptone and deionized water are stirred evenly and sterilized at 121℃ and 0.11 MPa for 20 min to obtain fermentation medium; the fermentation medium contains 20 g / L carbon source, 20 g / L malt extract powder, 1 g / L yeast powder and 1 g / L peptone.

[0101] 3) The seed culture of Schizophyllum commune was inoculated into the fermentation medium (the inoculation amount of the seed culture of Schizophyllum commune in the fermentation medium was 5 wt%), and fermented and co-cultured at 30℃ for 3 days. After fermentation, the culture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the fermentation broth of Schizophyllum commune strain.

[0102] Comparative Example 2

[0103] A fermentation broth of a strain of Ganoderma sinensis is obtained by inoculating the seed liquid of Ganoderma sinensis into a fermentation medium for fermentation culture;

[0104] The difference from Comparative Example 1 is that *Schizophyllum commune* was replaced with *Ganoderma sinense*.

[0105] The remaining steps are the same as in Example 1, and the fermentation broth of Ganoderma sinense strain is obtained.

[0106] Comparative Example 3

[0107] A fermentation broth of a strain of *Trametes versicolor* is obtained by inoculating the seed liquid of *Trametes versicolor* into a fermentation medium and then fermenting it.

[0108] The difference from Comparative Example 1 is that Schizophyllum commune is replaced with Trametes versicolor;

[0109] The remaining steps are the same as in Example 1, and the fermentation broth of the *Trametes versicolor* strain is obtained.

[0110] Comparative Example 4

[0111] A fermentation broth of a strain of Auricularia auricula-judae is obtained by inoculating the seed liquid of Auricularia auricula-judae into a fermentation medium and then fermenting it.

[0112] The difference from Comparative Example 1 is that Schizophyllum commune is replaced with Auricularia aurea.

[0113] The remaining steps are the same as in Example 1, and the fermentation broth of Auricularia auricula-judae strain is obtained.

[0114] Comparative Example 5

[0115] A fermentation broth of a strain of Poria cocos is obtained by inoculating the seed liquid of Poria cocos into a fermentation medium and then fermenting it.

[0116] The difference from Comparative Example 1 is that Schizophyllum commune is replaced with Poria cocos.

[0117] The remaining steps are the same as in Example 1, and the fermentation broth of the Poria cocos strain is obtained.

[0118] Comparative Example 6

[0119] A fermentation broth of a strain of Phellinus linteus is obtained by inoculating the seed liquid of Phellinus linteus into a fermentation medium and then fermenting it.

[0120] The difference from Comparative Example 1 is that Schizophyllum commune is replaced with Phellinus linteus;

[0121] The remaining steps are the same as in Example 1, and the fermentation broth of Phellinus linteus strain is obtained.

[0122] Comparative Example 7

[0123] A fermentation broth of a strain of Aspergillus schwannii is obtained by inoculating the seed liquid of Aspergillus schwannii into a fermentation medium and then fermenting it.

[0124] The difference from Comparative Example 1 is that *Schizophyllum commune* was replaced with *Aspergillus chevaleri*.

[0125] The remaining steps are the same as in Example 1, and the fermentation broth of Aspergillus chevallis strain is obtained.

[0126] Comparative Example 8

[0127] A co-culture fermentation broth of Ganoderma lucidum and Trametes versicolor is obtained by inoculating the seed liquid of Ganoderma lucidum and Trametes versicolor into a co-fermentation medium for fermentation and co-culture.

[0128] 1) Preparation of seed culture of Ganoderma sinensis: The mycelium of Ganoderma sinensis was inoculated into a sterilized seed culture medium and cultured at 30℃ and 180 rpm for 4 days to obtain the seed culture of Ganoderma sinensis; the inoculation amount of Ganoderma sinensis in the seed culture of Ganoderma sinensis was 2.5 wt.

[0129] 2) Preparation of seed liquid of Ganoderma lucidum: The difference from step 1) is that Ganoderma sinensis is replaced with Ganoderma lucidum.

[0130] 3) Preparation of co-fermentation medium: Same as in Example 1.

[0131] 4) The seed liquid of Ganoderma lucidum and the seed liquid of Trametes versicolor were inoculated into a co-fermentation medium (the inoculation amount of seed liquid of Ganoderma lucidum was 2.5 wt%, and the inoculation amount of seed liquid of Trametes versicolor was 2.5 wt%). The co-fermentation was carried out at 30℃ for 3 days. After co-fermentation, the mixture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-culture fermentation broth of Ganoderma lucidum and Trametes versicolor.

[0132] Comparative Example 9

[0133] A co-culture fermentation broth of Ganoderma lucidum and Poria cocos is obtained by inoculating the seed liquid of Ganoderma lucidum and the seed liquid of Poria cocos into a co-fermentation medium for fermentation and co-culture.

[0134] The difference from Comparative Example 8 is that *Trametes versicolor* was replaced with *Poria cocos*; the culture time for the seed culture of *Poria cocos* was 10 days.

[0135] The remaining steps are the same as in Comparative Example 8, and the co-culture fermentation broth of Ganoderma lucidum and Poria cocos is obtained.

[0136] Comparative Example 10

[0137] A co-culture fermentation broth of Ganoderma lucidum and Auricularia auricula-judae is obtained by inoculating the seed liquid of Ganoderma lucidum and the seed liquid of Auricularia auricula-judae into a co-fermentation medium for fermentation and co-culture.

[0138] The difference from Comparative Example 8 is that *Trametes versicolor* was replaced with *Auricularia aurea*; the seed culture time for *Auricularia aurea* was 10 days.

[0139] The remaining steps are the same as those in Comparative Example 8, and the co-culture fermentation broth of Ganoderma lucidum and Auricularia auricula-judae is obtained.

[0140] Comparative Example 11

[0141] A co-culture fermentation broth of Ganoderma lucidum and Phellinus linteus is obtained by inoculating the seed liquid of Ganoderma lucidum and Phellinus linteus into a co-fermentation medium for fermentation and co-culture.

[0142] The difference from Comparative Example 8 is that *Trametes versicolor* was replaced with *Sanghuang*; the seed culture time for *Sanghuang* was 10 days.

[0143] The remaining steps are the same as those in Comparative Example 8, and the co-culture fermentation broth of Ganoderma lucidum and Phellinus linteus is obtained.

[0144] Comparative Example 12

[0145] A co-culture fermentation broth of Ganoderma sinensis and Aspergillus serrata is obtained by inoculating the seed liquid of Ganoderma sinensis and the seed liquid of Aspergillus serrata into a co-fermentation medium for fermentation and co-culture.

[0146] The difference from Comparative Example 8 is that *Trametes versicolor* was replaced with *Aspergillus serrata*; in the preparation of the seed culture of *Aspergillus serrata*, the culture time was 7 days.

[0147] The remaining steps are the same as those in Comparative Example 8, and the co-culture fermentation broth of Ganoderma lucidum and Aspergillus sieboldii is obtained.

[0148] Comparative Example 13

[0149] A co-culture fermentation broth of *Trametes versicolor* and *Poria cocos* is obtained by inoculating the seed liquid of *Trametes versicolor* and the seed liquid of *Poria cocos* into a co-fermentation medium for fermentation and co-culture.

[0150] 1) Preparation of seed culture of Trametes versicolor: Mycelium of Trametes versicolor was inoculated into sterilized seed culture medium and cultured at 30℃ and 180 rpm for 4 days to obtain seed culture of Trametes versicolor; The inoculation amount of Trametes versicolor in the seed culture of Trametes versicolor is 2.5 wt%.

[0151] 2) Preparation of seed culture of Poria cocos: The difference from step 1) is that Poria cocos is replaced with Trametes versicolor and the culture time is 10 days.

[0152] 3) Preparation of co-fermentation medium: Same as in Example 1.

[0153] 4) The seed liquid of *Trametes versicolor* and the seed liquid of *Poria cocos* were inoculated into a co-fermentation medium (the inoculation amount of the seed liquid of *Trametes versicolor* was 2.5 wt%, and the inoculation amount of the seed liquid of *Poria cocos* was 2.5 wt%). The co-fermentation was carried out at 30℃ for 3 days. After co-fermentation, the mixture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-culture fermentation broth of *Trametes versicolor* and *Poria cocos*.

[0154] Comparative Example 14

[0155] A co-culture fermentation broth of *Trametes versicolor* and *Auricularia auricula-judae* is obtained by inoculating the seed liquid of *Trametes versicolor* and the seed liquid of *Auricularia auricula-judae* into a co-fermentation medium for fermentation and co-culture.

[0156] The difference from Comparative Example 13 is that Poria cocos was replaced with Auricularia aurea.

[0157] The remaining steps are the same as in Comparative Example 13, to obtain the co-culture fermentation broth of *Trametes versicolor* and *Auricularia auricula-judae*.

[0158] Comparative Example 15

[0159] A co-culture fermentation broth of *Trametes versicolor* and *Aspergillus serrata* is obtained by inoculating the seed liquid of *Trametes versicolor* and the seed liquid of *Aspergillus serrata* into a co-fermentation medium for fermentation and co-culture.

[0160] The difference from Comparative Example 13 is that Poria cocos was replaced with Aspergillus chevaleri.

[0161] The remaining steps were the same as in Comparative Example 13, and the co-culture fermentation broth of *Trametes versicolor* and *Aspergillus serrata* was obtained.

[0162] Comparative Example 16

[0163] A co-culture fermentation broth of *Trametes versicolor* and *Sanghuang* is obtained by inoculating the seed liquid of *Trametes versicolor* and the seed liquid of *Sanghuang* into a co-fermentation medium for fermentation and co-culture.

[0164] The difference from Comparative Example 13 is that Poria cocos was replaced with Phellinus linteus.

[0165] The remaining steps are the same as in Comparative Example 13, to obtain the co-culture fermentation broth of *Trametes versicolor* and *Sanghuang*.

[0166] Comparative Example 17

[0167] A co-culture fermentation broth of Poria cocos and Auricularia aurea is obtained by inoculating the seed liquid of Poria cocos and the seed liquid of Auricularia aurea into a co-fermentation medium for fermentation and co-culture.

[0168] 1) Preparation of seed culture of Poria cocos: Mycelium of Poria cocos was inoculated into sterilized seed culture medium and cultured at 30℃ and 180rpm for 10 days to obtain seed culture of Poria cocos; The inoculation amount of Poria cocos in the seed culture of Poria cocos was 2.5 wt.

[0169] 2) Preparation of seed culture of Auricularia aurea: The difference from step 1) is that Auricularia aurea is replaced with Poria cocos.

[0170] 3) Preparation of co-fermentation medium: Same as in Example 1.

[0171] 4) The seed culture of Poria cocos and the seed culture of Auricularia auricula-judae were inoculated into a co-fermentation medium (the inoculation amount of the seed culture of Poria cocos was 2.5 wt%, and the inoculation amount of the seed culture of Auricularia auricula-judae was 2.5 wt%). The co-fermentation was carried out at 30℃ for 3 days. After the co-fermentation, the mixture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-fermentation broth of Poria cocos and Auricularia auricula-judae.

[0172] Comparative Example 18

[0173] A co-culture fermentation broth of Poria cocos and Aspergillus serrata is obtained by inoculating the seed liquid of Poria cocos and the seed liquid of Aspergillus serrata into a co-fermentation medium for fermentation and co-culture.

[0174] The difference from Comparative Example 17 is that *Auricularia aurea* was replaced with *Aspergillus chevaleri*. In the preparation of the seed culture of *Aspergillus chevaleri*, the culture time was 7 days.

[0175] The remaining steps are the same as in Comparative Example 17, to obtain the co-culture fermentation broth of Poria cocos and Aspergillus serrata.

[0176] Comparative Example 19

[0177] A co-culture fermentation broth of Poria cocos and Phellinus linteus is obtained by inoculating the seed liquid of Poria cocos and the seed liquid of Phellinus linteus into a co-fermentation medium for fermentation and co-culture.

[0178] The difference from Comparative Example 17 is that Auricularia auricula-judae was replaced with Phellinus linteus.

[0179] The remaining steps are the same as in Comparative Example 17, and the co-culture fermentation broth of Poria cocos and Phellinus linteus is obtained.

[0180] Comparative Example 20

[0181] A co-culture fermentation broth of Auricularia aurea and Aspergillus serrata is obtained by inoculating the seed liquid of Auricularia aurea and the seed liquid of Aspergillus serrata into a co-fermentation medium for fermentation and co-culture.

[0182] 1) Preparation of seed culture of Auricularia aurea: Mycelium of Auricularia aurea was inoculated into sterilized seed culture medium and cultured at 30℃ and 180 rpm for 10 days to obtain seed culture of Auricularia aurea; The inoculation amount of Auricularia aurea in the seed culture of Auricularia aurea was 2.5 wt.

[0183] 2) Preparation of seed culture of *Aspergillus serrata*: The difference from step 1) is that *Auricularia aurea* is replaced with *Aspergillus serrata*. The incubation time for the seed culture of *Aspergillus serrata* is 7 days.

[0184] 3) Preparation of co-fermentation medium: Same as in Example 1.

[0185] 4) The seed culture of Auricularia aurea and the seed culture of Aspergillus serrata were inoculated into a co-fermentation medium (the inoculation amount of seed culture of Auricularia aurea was 2.5 wt%, and the inoculation amount of seed culture of Aspergillus serrata was 2.5 wt%). The co-fermentation was carried out at 30°C for 3 days. After co-fermentation, the mixture was centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-culture fermentation broth of Auricularia aurea and Aspergillus serrata.

[0186] Comparative Example 21

[0187] A co-culture fermentation broth of Auricularia auricula-judae and Phellinus linteus is obtained by inoculating the seed liquid of Auricularia auricula-judae and the seed liquid of Phellinus linteus into a co-fermentation medium for fermentation and co-culture.

[0188] The difference from Comparative Example 20 is that *Aspergillus chevaleri* was replaced with *Phellinus linteus*. In the preparation of the *Phellinus linteus* seed culture, the culture time was 10 days.

[0189] The remaining steps were the same as in Comparative Example 20, and the co-culture fermentation broth of Auricularia auricula-Sanghuang was obtained.

[0190] Comparative Example 22

[0191] A co-culture fermentation broth of Aspergillus serrata and Phellinus linteus is obtained by inoculating the seed liquid of Aspergillus serrata and the seed liquid of Phellinus linteus into a co-fermentation medium for fermentation and co-culture.

[0192] 1) Preparation of seed culture of Aspergillus serrata: The mycelium of Aspergillus serrata was inoculated into sterilized seed culture medium and cultured at 30℃ and 180 rpm for 7 days to obtain the seed culture of Aspergillus serrata; the inoculation amount of Aspergillus serrata in the seed culture of Aspergillus serrata was 5 wt.

[0193] 2) Preparation of *Sanghuang* seed culture: The difference from step 1) is that *Aspergillus chevaleri* is replaced with *Sanghuang*. The incubation time for the *Sanghuang* seed culture is 10 days.

[0194] 3) Preparation of co-fermentation medium: Same as in Example 1.

[0195] 4) The seed culture of Aspergillus schavar and the seed culture of Phellinus linteus were inoculated into a co-fermentation medium (the inoculation amount of the seed culture of Aspergillus schavar is 2.5 wt% and the inoculation amount of the seed culture of Phellinus linteus is 2.5 wt%). The co-fermentation was carried out at 30℃ for 3 days. After co-fermentation, the mixture was centrifuged at 20℃ and 8500 rpm. The supernatant after centrifugation was filtered to obtain the co-culture fermentation broth of Aspergillus schavar and Phellinus linteus.

[0196] 1. Determination of tyrosinase inhibition rate of the fermentation broth described in this invention

[0197] 1.1 Reagent Preparation

[0198] (1) Prepare a phosphate buffer solution with a pH of 6.8: Weigh 6.8g of potassium dihydrogen phosphate and 0.94g of sodium hydroxide, add 800mL of distilled water to dissolve them, adjust the pH to 6.8, and make up to 1000mL in a volumetric flask.

[0199] (2) Prepare L-tyrosine solution (prepare immediately): accurately weigh 0.0272 g L-tyrosine, add it to a phosphate buffer solution with a pH of 6.8 to dissolve and dilute to 100 mL in a volumetric flask.

[0200] (3) Prepare a 200U enzyme activity tyrosinase solution (prepare immediately): Dissolve 25KU of Solarbio tyrosinase in 12.5mL of phosphate buffer solution with a pH of 6.8 to obtain a 2KU tyrosinase solution. Store at -20℃. When using, take an appropriate amount of the 2KU tyrosinase solution and dilute it 10 times with phosphate buffer solution with a pH of 6.8.

[0201] 1.2 Experimental Procedure

[0202] (1) Using a pipette, add solutions A, B, and C to the 96-well plate according to the dosage in Table 1. Finally, add solution D, mix thoroughly, incubate at 37°C for 10 min, measure the absorbance A of the reaction solution at 475 nm, and calculate its inhibition rate on tyrosinase.

[0203] The calculation formula is shown in equation (1):

[0204] Inhibition rate = [1-(A)] 样 -A 空白 Equation (1): ) / (A0-A1)]×100%

[0205] Among them, A 样 : Sample tube absorbance, i.e., the absorbance of the solution after the sample reacts with tyrosinase;

[0206] A 空白 : Sample background absorbance;

[0207] A0: Absorbance of the reaction between tyrosinase and tyrosine without sample added;

[0208] A1: Solvent background absorbance.

[0209] Table 1 Composition of the reaction solution

[0210]

[0211] The tyrosinase inhibition rate of the fermentation broths in Examples 1-6 and Comparative Examples 1-22 was detected, and the results are shown in Table 2.

[0212] Table 2. Tyrosinase inhibition rates of fermentation broths in Examples 1-6 and Comparative Examples 1-22

[0213]

[0214] The results of the tyrosinase inhibition rate of the fermentation broth above show that mixed fermentation of Schizophyllum commune with any other strain can enhance the tyrosinase inhibition effect of the fermentation broth, while the combination of other strains in pairs has no such effect.

[0215] To investigate the effect of fermentation time on the efficacy of co-culture fermentation broth, fermentation studies were conducted on the six combinations with superior whitening effects at different times.

[0216] Example 7

[0217] The difference from Example 1 is that in step 4), the fermentation co-culture time is 1 day.

[0218] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Ganoderma sinense is obtained.

[0219] Example 8

[0220] The difference from Example 1 is that in step 4), the fermentation co-culture time is 2 days.

[0221] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Ganoderma sinense is obtained.

[0222] Example 9

[0223] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 1.

[0224] Example 10

[0225] The difference from Example 1 is that in step 4), the fermentation co-culture time is 4 days.

[0226] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum commune and Ganoderma sinense is obtained.

[0227] Example 11

[0228] The difference from Example 2 is that in step 4), the fermentation co-culture time is 1 day.

[0229] The remaining steps are the same as in Example 2, and the co-culture fermentation broth of Schizophyllum commune and Trametes versicolor is obtained.

[0230] Example 12

[0231] The difference from Example 2 is that in step 4), the fermentation co-culture time is 2 days.

[0232] The remaining steps are the same as in Example 2, and the co-culture fermentation broth of Schizophyllum commune and Trametes versicolor is obtained.

[0233] Example 13

[0234] The raw materials and preparation process are the same as those used in Example 2 for preparing the fermentation broth.

[0235] Example 14

[0236] The difference from Example 2 is that in step 4), the fermentation co-culture time is 4 days.

[0237] The remaining steps are the same as in Example 2, and the co-culture fermentation broth of Schizophyllum commune and Trametes versicolor is obtained.

[0238] Example 15

[0239] The difference from Example 3 is that in step 4), the fermentation co-culture time is 1 day.

[0240] The remaining steps are the same as in Example 3, and the co-culture fermentation broth of Schizophyllum commune and Auricularia aurea is obtained.

[0241] Example 16

[0242] The difference from Example 3 is that in step 4), the fermentation co-culture time is 2 days.

[0243] The remaining steps are the same as in Example 3, and the co-culture fermentation broth of Schizophyllum commune and Auricularia aurea is obtained.

[0244] Example 17

[0245] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 3.

[0246] Example 18

[0247] The difference from Example 3 is that in step 4), the fermentation co-culture time is 4 days.

[0248] The remaining steps are the same as in Example 3, and the co-culture fermentation broth of Schizophyllum commune and Auricularia aurea is obtained.

[0249] Example 19

[0250] The difference from Example 4 is that in step 4), the fermentation co-culture time is 1 day.

[0251] The remaining steps are the same as in Example 4, and the co-culture fermentation broth of Schizophyllum commune and Poria cocos is obtained.

[0252] Example 20

[0253] The difference from Example 4 is that in step 4), the fermentation co-culture time is 2 days.

[0254] The remaining steps are the same as in Example 4, and the co-culture fermentation broth of Schizophyllum commune and Poria cocos is obtained.

[0255] Example 21

[0256] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 4.

[0257] Example 22

[0258] The difference from Example 4 is that in step 4), the fermentation co-culture time is 4 days.

[0259] The remaining steps are the same as in Example 4, and the co-culture fermentation broth of Schizophyllum commune and Poria cocos is obtained.

[0260] Example 23

[0261] The difference from Example 5 is that in step 4), the fermentation co-culture time is 1 day.

[0262] The remaining steps are the same as in Example 5, and the co-culture fermentation broth of Schizophyllum commune and Phellinus linteus is obtained.

[0263] Example 24

[0264] The difference from Example 5 is that in step 4), the fermentation co-culture time is 2 days.

[0265] The remaining steps are the same as in Example 5, and the co-culture fermentation broth of Schizophyllum commune and Phellinus linteus is obtained.

[0266] Example 25

[0267] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 5.

[0268] Example 26

[0269] The difference from Example 5 is that in step 4), the fermentation co-culture time is 4 days.

[0270] The remaining steps are the same as in Example 5, and the co-culture fermentation broth of Schizophyllum commune and Phellinus linteus is obtained.

[0271] Example 27

[0272] The difference from Example 6 is that in step 4), the fermentation co-culture time is 1 day.

[0273] The remaining steps are the same as in Example 6, and the co-culture fermentation broth of Schizophyllum commune and Aspergillus schwannoides is obtained.

[0274] Example 28

[0275] The difference from Example 6 is that in step 4), the fermentation co-culture time is 2 days.

[0276] The remaining steps are the same as in Example 6, and the co-culture fermentation broth of Schizophyllum commune and Aspergillus schwannoides is obtained.

[0277] Example 29

[0278] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 6.

[0279] Example 30

[0280] The difference from Example 6 is that in step 4), the fermentation co-culture time is 4 days.

[0281] The remaining steps are the same as in Example 6, and the co-culture fermentation broth of Schizophyllum commune and Aspergillus schwannoides is obtained.

[0282] The tyrosinase inhibition rate of the fermentation broth (original broth) of Examples 7-30 was detected, and the results are shown in Table 3.

[0283] Table 3. Tyrosinase inhibition rates of fermentation broths (original solutions) from Examples 7 to 30

[0284]

[0285] The tyrosinase inhibition rate of the fermentation broths (diluted 10 times: 10% fermentation broth) of Examples 7-30 was detected, and the results are shown in Table 4.

[0286] Table 4. Tyrosinase inhibition rate of fermentation broth diluted 10 times in Examples 7-30

[0287]

[0288] The tyrosinase inhibition rate of the fermentation broths (diluted 100 times: 1% fermentation broth) of Examples 7-30 was detected, and the results are shown in Table 5.

[0289] Table 5. Tyrosinase inhibition rate of fermentation broths (100-fold dilution: 1% fermentation broth) from Examples 7-30

[0290]

[0291] Based on the above tyrosinase inhibition rate results, it can be seen that during the fermentation process from day 1 to day 4, the mixed fermentation broth of Auricularia auricula-judae, Ganoderma lucidum, Trametes versicolor and Schizophyllum commune showed the best tyrosinase inhibition rate on day 3; the mixed fermentation broth of Aspergillus schwannii and Schizophyllum commune showed the best tyrosinase inhibition rate on day 2; the mixed fermentation broth of Poria cocos and Schizophyllum commune showed the best tyrosinase inhibition rate on day 1; and the mixed fermentation broth of Phellinus linteus and Schizophyllum commune showed the best tyrosinase inhibition rate on day 4.

[0292] To investigate whether inactivated strains can also play the same role, the following experiment was designed. Based on the experimental results, the subsequent fermentation time was set at 3 days.

[0293] Example 31

[0294] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 1.

[0295] Example 32

[0296] The raw materials and preparation process are the same as those used in Example 2 for preparing the fermentation broth.

[0297] Example 33

[0298] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 3.

[0299] Example 34

[0300] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 4.

[0301] Example 35

[0302] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 5.

[0303] Example 36

[0304] The raw materials and preparation process are the same as those used in the fermentation broth described in Example 6.

[0305] Comparative Example 23

[0306] The difference from Example 1 is as follows:

[0307] Step 4) involves inoculating the seed culture of Ganoderma sinensis and the inactivated seed culture of Schizophyllum commune into a co-fermentation medium (the inoculation amount of the seed culture of Ganoderma sinensis is 2.5 wt%, and the inoculation amount of the seed culture of Schizophyllum commune is 2.5 wt%). The mixture is then fermented and co-cultured at 30°C for 3 days. After fermentation and co-culture, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the co-culture fermentation broth of Schizophyllum commune and Ganoderma sinensis.

[0308] Comparative Example 24

[0309] The difference from Example 2 is as follows:

[0310] Step 4) involves inoculating the seed culture of *Trametes versicolor* and the inactivated seed culture of *Schizophyllum commune* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Trametes versicolor* seed culture is 2.5 wt%)). The mixture is then fermented at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant is filtered to obtain the *Schizophyllum commune*-*Trametes versicolor* co-culture fermentation broth.

[0311] Comparative Example 25

[0312] The difference from Example 3 is as follows:

[0313] Step 4) involves inoculating the seed culture of *Auricularia aurea* and the inactivated seed culture of *Schizophyllum commune* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Auricularia aurea* seed culture is 2.5 wt%)). The mixture is then fermented at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the *Schizophyllum commune*-*Auricularia aurea* co-culture fermentation broth.

[0314] Comparative Example 26

[0315] The difference from Example 4 is as follows:

[0316] Step 4) involves inoculating the seed culture of *Poria cocos* and the inactivated seed culture of *Schizophyllum commune* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Poria cocos* seed culture is 2.5 wt%), and co-culturing at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the co-culture fermentation broth of *Schizophyllum commune* and *Poria cocos*.

[0317] Comparative Example 27

[0318] The difference from Example 5 is as follows:

[0319] Step 4) involves inoculating the seed culture of *Sanghuang* fungus and the inactivated seed culture of *Schizophyllum commune* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Sanghuang* seed culture is 2.5 wt%)). The mixture is then fermented at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the *Schizophyllum commune*-*Sanghuang* co-culture fermentation broth.

[0320] Comparative Example 28

[0321] The difference from Example 6 is as follows:

[0322] Step 4) involves inoculating the seed culture of *Aspergillus schaferi* and the inactivated seed culture of *Schizophyllum commune* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Aspergillus schaferi* seed culture is 2.5 wt%). The mixture is then fermented at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant is filtered to obtain the *Schizophyllum commune*-*Aspergillus schaferi* co-culture fermentation broth.

[0323] Comparative Example 29

[0324] The difference from Example 1 is as follows:

[0325] Step 4) involves inoculating the seed culture of *Schizophyllum commune* and the inactivated seed culture of *Ganoderma sinense* into a co-fermentation medium (the inoculation amount of *Ganoderma sinense* seed culture is 2.5 wt%, and the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%), and co-fermenting at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the *Schizophyllum commune*-*Ganoderma sinense* co-culture fermentation broth.

[0326] Comparative Example 30

[0327] The difference from Example 2 is as follows:

[0328] Step 4) involves inoculating the seed culture of Schizophyllum commune and the inactivated seed culture of Trametes versicolor into a co-fermentation medium (the inoculation amount of the seed culture of Schizophyllum commune is 2.5 wt%, and the inoculation amount of the seed culture of Trametes versicolor is 2.5 wt%), and co-fermenting at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the Schizophyllum commune-Trametes versicolor co-culture fermentation broth.

[0329] Comparative Example 31

[0330] The difference from Example 3 is as follows:

[0331] Step 4) involves inoculating the seed culture of *Schizophyllum commune* and the inactivated seed culture of *Auricularia aurea* into a co-fermentation medium (the inoculation amount of the seed culture of *Schizophyllum commune* is 2.5 wt%, and the inoculation amount of the seed culture of *Auricularia aurea* is 2.5 wt%)). The co-fermentation is carried out at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the co-culture fermentation broth of *Schizophyllum commune* and *Auricularia aurea*.

[0332] Comparative Example 32

[0333] The difference from Example 4 is as follows:

[0334] Step 4) involves inoculating the seed culture of Schizophyllum commune and the inactivated seed culture of Poria cocos into a co-fermentation medium (the inoculation amount of the seed culture of Schizophyllum commune is 2.5 wt%, and the inoculation amount of the seed culture of Poria cocos is 2.5 wt%), and co-fermenting at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the co-culture fermentation broth of Schizophyllum commune and Poria cocos.

[0335] Comparative Example 33

[0336] The difference from Example 5 is as follows:

[0337] Step 4) involves inoculating the seed culture of *Schizophyllum commune* and the inactivated seed culture of *Phellinus linteus* into a co-fermentation medium (the inoculation amount of *Schizophyllum commune* seed culture is 2.5 wt%, and the inoculation amount of *Phellinus linteus* seed culture is 2.5 wt%), and co-fermenting at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the *Schizophyllum commune*-*Phellinus linteus* co-culture fermentation broth.

[0338] Comparative Example 34

[0339] The difference from Example 6 is as follows:

[0340] Step 4) involves inoculating the seed culture of *Schizophyllum commune* and the seed culture of *Aspergillus serrata* (which has been inactivated) into a co-fermentation medium (the inoculation amount of the seed culture of *Schizophyllum commune* is 2.5 wt%, and the inoculation amount of the seed culture of *Aspergillus serrata* is 2.5 wt%)). The co-fermentation is carried out at 30°C for 3 days. After co-fermentation, the mixture is centrifuged at 20°C and 8500 rpm. The supernatant after centrifugation is filtered to obtain the co-culture fermentation broth of *Schizophyllum commune* and *Aspergillus serrata*.

[0341] The tyrosinase inhibition rate of the fermentation broths in Examples 31-36 and Comparative Examples 23-34 was detected, and the results are shown in Table 6.

[0342] Table 6. Tyrosinase inhibition rates of fermentation broths in Examples 31-36 and Comparative Examples 23-34

[0343]

[0344] Based on the above results of tyrosinase inhibition rate in the fermentation broth, it was found that only mixed fermentation of non-inactivated Schizophyllum commune and other non-inactivated strains could improve the tyrosinase inhibition effect of the fermentation broth.

[0345] It is possible that other fungi stimulated certain silent genes during co-fermentation with Schizophyllum commune, causing it to produce substances with whitening effects that would not normally be produced. Furthermore, different strains have different stimulation effects. For example, strains with similar culture cycles to Schizophyllum commune, such as Ganoderma lucidum and Trametes versicolor, can produce better effects, while strains with slightly longer culture cycles, such as Phellinus linteus, have relatively less prominent effects.

[0346] Comparative Example 35

[0347] A co-culture fermentation broth of Schizophyllum commune and Ganoderma lucidum is obtained by inoculating Ganoderma lucidum seed liquid and Schizophyllum commune seed liquid into a co-fermentation medium for fermentation and co-culture.

[0348] The difference from Example 1 is that Ganoderma lucidum is replaced with Ganoderma lucidum;

[0349] The remaining steps are the same as in Example 1, and the Schizophyllum co-culture fermentation broth is obtained.

[0350] Comparative Example 36

[0351] A co-culture fermentation broth of Schizophyllum commune and Pleurotus ostreatus is obtained by inoculating Pleurotus ostreatus seed liquid and Schizophyllum commune seed liquid into a co-fermentation medium for fermentation and co-culture.

[0352] The difference from Example 1 is that Ganoderma sinensis is replaced with Pleurotus ostreatus;

[0353] The remaining steps are the same as in Example 1, and the co-culture fermentation broth of Schizophyllum coulombia and Pleurotus ostreatus is obtained.

[0354] The tyrosinase inhibition rate of the fermentation broths prepared in comparative examples 35-36 was detected, and the results are shown in Table 7.

[0355] Table 7. Tyrosinase inhibition rate of fermentation broths prepared in Comparative Examples 35-36

[0356]

[0357] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for preparing a co-culture fermentation broth of medicinal and edible fungi with whitening effects, characterized in that, Includes the following steps: The seed liquid of the first fungus and the seed liquid of the second fungus were inoculated into a co-fermentation medium for co-fermentation and co-culture to obtain a co-culture fermentation broth of medicinal and edible fungi with whitening effects. The first fungus includes at least one of the following: Ganoderma lucidum strain, Trametes versicolor strain, Poria cocos strain, Auricularia auricula-judae strain, Phellinus linteus strain, and Aspergillus schavar; The second fungus includes Schizophyllum commune.

2. The preparation method according to claim 1, characterized in that, The co-fermentation medium includes a carbon source, malt extract, a nitrogen source, and water; The co-fermentation medium contains 10-30 g / L of carbon source, 10-30 g / L of malt extract, and 1-3 g / L of nitrogen source.

3. The preparation method according to claim 2, characterized in that, The carbon source includes at least one of glucose, sucrose, and starch; The nitrogen source includes at least one of yeast extract, peptone, and ammonium sulfate.

4. The preparation method according to claim 2, characterized in that, The preparation method of the co-fermentation medium includes the following steps: The carbon source, malt extract powder, nitrogen source and water are mixed and sterilized to obtain the co-fermentation medium. The sterilization temperature is 115~125℃, the pressure is 0.05~0.15 MPa, and the time is 10~30 min.

5. The preparation method according to claim 1, characterized in that, In the seed culture of the first fungus, the inoculum amount of the first fungus is 2.5 wt%~7.5 wt%; In the seed culture of the second fungus, the inoculum amount of the second fungus is 2.5 wt% to 7.5 wt%.

6. The preparation method according to claim 1, characterized in that, The method for preparing the seed solution of the first fungus includes the following steps: The mycelium of the first fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the first fungus. The method for preparing the seed solution of the second fungus includes the following steps: The mycelium of the second fungus was inoculated into a sterilized seed culture medium and cultured at 28-30℃ to obtain the seed liquid of the second fungus. The seed culture medium includes a carbon source, malt extract powder, a nitrogen source, and water; The seed culture medium contains 10-30 g / L of carbon source, 10-30 g / L of malt extract powder, and 1-3 g / L of nitrogen source.

7. The preparation method according to claim 1, characterized in that, In the co-fermentation medium, the inoculation amount of the seed liquid of the first fungus is 2.5 wt% to 7.5 wt%, and the inoculation amount of the seed liquid of the second fungus is 2.5 wt% to 7.5 wt%.

8. The preparation method according to claim 1, characterized in that, The fermentation co-culture temperature is 28~30℃, the stirring rate is 160~180 rpm, and the time is 1~10 days.

9. The co-culture fermentation broth of medicinal and edible fungi with whitening effect prepared by the preparation method according to any one of claims 1 to 8.

10. The application of the co-culture fermentation broth of medicinal and edible fungi with whitening effect as described in claim 9 in the preparation of skin care products.

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