Preparation method of Ganoderma lucidum composite fermented scalp care liquid rich in gamma-aminobutyric acid

Abstract

The invention discloses a preparation method of Ganoderma lucidum composite fermented scalp care liquid rich in gamma-aminobutyric acid. Crushing ganoderma lucidum sporocarp which does not release spores into ganoderma lucidum powder, and then adding the ganoderma lucidum powder into a gamma-aminobutyric acid precursor substance solution for sterilization treatment to obtain a fermentation substrate solution; lactobacillus acidophilus, candida utilis and yarrowia lipolytica are selected and transferred to respective purification culture media to be cultured, three purified strains are obtained and then inoculated to respective seed culture media to be cultured, and seed solutions of the three strains are obtained; seed solutions of the three strains are mixed into a mixed seed solution, then the mixed seed solution is inoculated into a fermentation substrate solution for compound fermentation, and a compound fermentation solution is obtained; and centrifuging the composite fermentation liquid, adding citric acid, sorbitol and cholesterol, and mixing to obtain the Ganoderma lucidum composite fermentation scalp care liquid rich in gamma-aminobutyric acid. The invention provides the preparation method of the scalp care liquid which has the effects of improving the oxidation resistance of the scalp and improving the lipoid secretion of the skin surface layer, and is more stable and safer.

Description

1. Technical field:

[0001] The present invention belongs to the field of biological fermentation technology in cosmetic products, and specifically relates to a method for preparing a gamma-aminobutyric acid-rich ganoderma lucidum composite fermented scalp care solution. The method of the present invention aims to combine the natural nourishing ingredients of ganoderma lucidum with the physiological regulation function of gamma-aminobutyric acid to provide an efficient and safe care solution for scalp health. 2. Background technology:

[0002] Ganoderma lucidum has a history of more than 2,000 years in China. It is highly respected for its unique morphology and medicinal value and is regarded as a "magic grass". It is widely used in traditional medicine and is also a medicinal and edible fungus. It is rich in a variety of essential vitamins and amino acids, polysaccharides, triterpenes and other bioactive substances. It can not only directly remove free radicals, but also resist oxidative stress by enhancing the body's own antioxidant defense system. These characteristics make Ganoderma lucidum a potential source of natural antioxidants, showing broad application prospects in the development of cosmeceuticals and health care fields such as scalp health.

[0003] Gamma-aminobutyric acid (GABA), as an important neurotransmitter, can effectively relieve stress, promote blood circulation in the scalp, and help slow down scalp aging. However, some studies have shown that it plays a certain role in the anti-oxidation process. In 2015, my country has included GABA in the list of cosmetic raw materials. GABA can reduce oxidative stress by reducing the accumulation of reactive oxygen species (ROS), thereby protecting scalp epidermal cells from oxidative damage. As a GABA precursor: monosodium glutamate (monosodium glutamate content greater than 99%), it can not only provide the nitrogen source required for strain growth. In addition, monosodium glutamate also has a fermentation-promoting effect and improves yield and quality.

[0004] As people pay more attention to scalp health, the demand for natural and effective ingredients in the scalp care product market is growing. The scalp is the soil for hair growth, and its health directly affects the quality and growth environment of hair. At present, there are many scalp care products containing Ganoderma lucidum on the market, but their efficacy is general and single. The lack of a scalp care product that effectively combines Ganoderma lucidum with γ-aminobutyric acid provides an opportunity for innovative scalp care solutions. In addition, in terms of probiotic fermentation technology, how to optimize fermentation conditions to achieve the best scalp care effect is a key issue. Finally, when developing scalp care products, product safety and stability are crucial. It is necessary to ensure that the ingredients and fermentation process used will not cause skin irritation or allergic reactions. 3. Summary of the invention:

[0005] The technical problem to be solved by the present invention is: Based on the above problems, i.e., the deficiencies in the existing scalp care product preparation technology, the present invention provides a method for preparing a scalp care solution rich in γ-aminobutyric acid from a composite fermented Ganoderma lucidum. The technical solution of the present invention effectively combines Ganoderma lucidum with γ-aminobutyric acid through a specific microbial fermentation technology, providing a method for preparing a scalp care solution that has the functions of improving the scalp's antioxidant capacity, improving the secretion of lipids on the surface of the skin, and being more stable and safe.

[0006] In order to solve the above problems, the technical solution adopted by the present invention is:

[0007] The present invention provides a method for preparing a gamma-aminobutyric acid-rich Ganoderma lucidum composite fermented scalp care liquid, the preparation method comprising the following steps:

[0008] (1) drying and crushing the Ganoderma lucidum fruiting bodies that have not released spores in sequence, and sieving the crushed Ganoderma lucidum to obtain Ganoderma lucidum powder;

[0009] (2) adding the ganoderma powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 15 to 20 minutes, and cooling to room temperature to obtain a fermentation substrate solution;

[0010] (3) using an inoculation loop to select Lactobacillus acidophilus, Candida utilis, and Yarrowia lipolytica respectively, and then transferring them to their respective purification medium, culturing them at 30-35° C. for 48-72 hours, and obtaining the three purified strains;

[0011] (4) inoculating the three purified strains into their respective seed culture media at an inoculation rate of 1 to 2% (mass percentage of the seed culture medium) for cultivation, and obtaining seed solutions of the three strains after cultivation;

[0012] (5) detecting absorbance of the seed solutions of the three strains by a spectrophotometer, and after the detection, mixing the seed solutions of the three strains to obtain a mixed seed solution;

[0013] (6) inoculating the mixed seed solution into a fermentation matrix solution for composite fermentation to obtain a composite fermentation solution;

[0014] (7) The obtained composite fermentation liquid is centrifuged (5000-6000 rpm for 8-10 minutes) to remove the bacteria and precipitated impurities; then citric acid, sorbitol and cholesterol (to adjust the stability of the formula) are added and mixed to obtain a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid.

[0015] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the powder in step (1) is sieved through a 50-70 mesh screen.

[0016] According to the above-mentioned method for preparing the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the γ-aminobutyric acid precursor solution in step (2) is a monosodium glutamate solution (the content of sodium glutamate in monosodium glutamate is greater than 99%) prepared by diluting monosodium glutamate with distilled water to form a monosodium glutamate solution with a concentration of 0.2 to 0.3 g / L.

[0017] According to the method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid, the mass percentage of Ganoderma lucidum fruiting bodies in the fermentation matrix solution obtained in step (2) is 4-5%.

[0018] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the preparation method of the purified culture medium in step (3) is: 15-20 g of glucose, 4-5 g of peptone, 2.5-3 g of yeast extract powder and 18-20 g of agar powder, double distilled water is added to 1 L, sterilized at 121° C. for 15-20 min, and cooled to room temperature to obtain a solid purified culture medium;

[0019] The selected amounts of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica are all 0.01-0.07 g, and the used amount of the purification culture medium is all 40-60 mL.

[0020] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the preparation method of the seed culture medium in step (4) is: 15-20 g of glucose, 10-15 g of soy protein, 10-15 g of corn flour, 3-8 g of sodium chloride, 1-4 g of dipotassium hydrogen phosphate and 0.3-0.6 g of L-cysteine, double distilled water is added to 1 L, sterilized at 121° C. for 15-20 min, and cooled to room temperature to obtain a seed culture medium.

[0021] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the conditions for culturing in the seed culture medium in step (4) are: initial pH 5-6, fermentation temperature 30-33°C, rotation speed in the shaking incubator 100-120 rpm, and culturing time 48-54 h.

[0022] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the absorbance values ​​of the seed liquids of the three strains of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica detected in step (5) are all 2.0-2.2 (OD 600 ); the volume ratio of the three when mixed is 1~3:1~3:1~3.

[0023] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, when the mixed seed liquid is added to the fermentation matrix solution in step (6), the mass percentage of the mixed seed liquid in the fermentation matrix solution is 3-5%; the composite fermentation conditions are: the fermentation temperature is 28-33°C and the fermentation time is 5-7d.

[0024] According to the preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid, the amounts of citric acid, sorbitol and cholesterol added in step (7) are 0.5-1%, 5-8% and 1-1.2% by mass of the obtained Ganoderma lucidum composite fermented scalp care liquid, respectively.

[0025] In the technical solution of the present invention, the fermentation matrix solution is prepared by using the Ganoderma lucidum fruiting body without releasing spores and monosodium glutamate with a content of more than 99%. Compared with the Ganoderma lucidum fruiting body with released spores, the Ganoderma lucidum fruiting body without releasing spores has higher antioxidant active ingredients such as polysaccharides and triterpenes, and the monosodium glutamate with a content of more than 99% has a higher conversion rate of γ-aminobutyric acid than low-concentration monosodium glutamate, so that the antioxidant activity is better.

[0026] In the technical scheme of the present invention, three strains of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica are used for composite fermentation. First, Lactobacillus acidophilus has a strong ability to produce lactic acid, produce antioxidant peptides, improve the scalp microenvironment, and improve the antioxidant capacity of the scalp composite fermentation liquid; the fermentation amount of Candida utilis is high, and it produces proteins and vitamins, which can improve the antioxidant activity of Ganoderma lucidum polysaccharides and triterpenes; in addition, the bacteria itself can produce antioxidant peptides and glutathione, and has obvious antioxidant activity; Yarrowia lipolytica has a good lipolytic effect and can regulate the oil secretion of the scalp. Furthermore, compared with single fermentation, the composite fermentation of the three strains can significantly improve the ability of monosodium glutamate, a precursor of γ-aminobutyric acid, to convert into γ-aminobutyric acid. Finally, the composite fermentation liquid for maintaining the scalp through the composite fermentation of the three strains can not only improve its antioxidant activity, but also regulate the oil secretion of the scalp, improve the scalp environment and other functions, thereby maximizing the scalp maintenance effect.

[0027] The positive beneficial effects of the present invention are:

[0028] 1. Enhanced antioxidant capacity: The scalp care solution prepared by the present invention is rich in antioxidant components extracted from Ganoderma lucidum. Ganoderma lucidum's polysaccharides, triterpenes and other active ingredients and γ-aminobutyric acid itself have antioxidant properties, which can reduce the accumulation of reactive oxygen species (ROS) and protect scalp epidermal cells from oxidative damage. γ-aminobutyric acid, as a neurotransmitter, can effectively relieve stress, promote scalp blood circulation, stimulate fibroblasts to synthesize collagen and hyaluronic acid, and promote the repair of the skin barrier, thereby helping to slow down scalp aging.

[0029] 2. Regulating oil secretion: The care solution prepared by the present invention can regulate the oil secretion of the scalp and maintain the healthy state of the scalp. The oil secretion inhibition rate in Example 7-C reaches 28.02%, which can effectively prevent scalp problems caused by excessive oil secretion.

[0030] 3. Safety: The technical solution of the present invention uses natural green ingredients and probiotic fermentation, avoiding the side effects that may be caused by chemical synthetic ingredients. By combining the natural nourishing ingredients of Ganoderma lucidum and the physiological regulation function of γ-aminobutyric acid, it improves the scalp environment and provides an efficient and safe maintenance product for scalp health.

[0031] 4. Innovative fermentation technology: The technical solution of the present invention adopts the composite fermentation of three efficient microbial strains, namely Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica, to increase the yield of GABA, wherein the GABA concentration of Example 7-A reaches 6.75 g / L; at the same time, it increases the antioxidant active ingredients of Ganoderma lucidum, wherein the DPPH free radical scavenging rate of Example 7-B is 92.79%, and the ABTS free radical scavenging rate is 92.34%.

[0032] 5. Commercial application value: The technical solution of the present invention uses monosodium glutamate as a precursor of γ-aminobutyric acid, which not only provides the nitrogen source required for the growth of the strain, but also has a fermentation-promoting effect, which helps to reduce production costs. The product prepared by the present invention can be used as both a health product and a cosmetic, which broadens the scope of application, meets the needs of different consumers, and provides users with a variety of choices.

[0033] In summary, the technical solution of the present invention uses specific microorganisms: Candida utilis, Lactobacillus acidophilus, and Yersinia lipolytica to perform complex fermentation on Ganoderma lucidum fruiting bodies and γ-aminobutyric acid precursors, which not only reflects the value of the strain itself in improving the scalp microenvironment, but also improves the activity of the natural nourishing ingredients of Ganoderma lucidum and the physiological regulation ability of γ-aminobutyric acid, thereby maximizing the scalp care effect. In addition, as moisturizing and stabilizing supplementary materials: citric acid, sorbitol, and cholesterol are edible grade materials, thereby providing an efficient and safe scalp care product for the preparation of a scalp care composite fermentation liquid with functions such as high-efficiency anti-oxidation and regulation of scalp oil secretion. 4. Specific implementation methods:

[0034] In order to better explain and illustrate the technical solution of the present invention, the present invention is further described below in conjunction with embodiments and comparative examples, but the protection scope of the technical solution of the present invention is not limited.

[0035] The fermentation materials and strains used in the following embodiments of the present invention: Ganoderma lucidum fruiting bodies and monosodium glutamate were purchased from supermarkets or markets; reagents and consumables were purchased from professional reagent or consumable companies; strains were purchased from the Beina Biological Strain Collection Center.

[0036] Preparation method of purification medium: 20 g of glucose, 5 g of peptone, 2.5 g of yeast extract powder, 20 g of agar powder, add double distilled water to 1 L, sterilize at 121° C. for 20 min, cool to room temperature, and obtain purification medium.

[0037] Preparation method of seed culture medium: 20 g of glucose, 10 g of soy protein, 15 g of corn flour, 5 g of sodium chloride, 3 g of dipotassium hydrogen phosphate and 0.5 g of L-cysteine, add double distilled water to 1 L, sterilize at 121° C. for 20 min, cool to room temperature, and obtain seed culture medium.

[0038] The strain combinations and proportions used in the embodiments and comparative examples of the present invention are shown in Table 1.

[0039] Table 1 The strain combinations and proportions used in the embodiments and comparative examples of the present invention

[0040] Group Strain combination Example 1 Lactobacillus acidophilus Example 2 Candida utilis Example 3 Yarrowia lipolytica Example 4 Lactobacillus acidophilus + Candida utilis Example 5 Candida utilis + Yarrowia lipolytica Example 6 Lactobacillus acidophilus + Yarrowia lipolytica Example 7 Lactobacillus acidophilus + Candida utilis + Yarrowia lipolytica Example 7-A Lactobacillus acidophilus + Candida utilis + Yarrowia lipolytica (3:1:1) Example 7-B Lactobacillus acidophilus + Candida utilis + Yarrowia lipolytica (1:3:1) Example 7-C Lactobacillus acidophilus + Candida utilis + Yarrowia lipolytica (1:1:3) Comparative Example 1 No inoculum strain (distilled water instead) Comparative Example 2 Lactococcus lactis Comparative Example 3 Saccharomyces boulardii Comparative Example 4 Lactococcus lactis + Saccharomyces boulardii Comparative Example 5 Saccharomyces boulardii + Yarrowia lipolytica Comparative Example 6 Lactococcus lactis + Yarrowia lipolytica Comparative Example 7 Lactococcus lactis + Saccharomyces boulardii + Yarrowia lipolytica

[0041] Embodiment 1:

[0042] The preparation method of the ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention comprises the following detailed steps:

[0043] (1) selecting a Ganoderma lucidum fruiting body that has not released spores as a material, drying it, crushing it, and then passing it through a 60-mesh sieve to obtain Ganoderma lucidum powder;

[0044] (2) adding the ganoderma lucidum powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 20 min, and cooling to room temperature to obtain a fermentation substrate solution;

[0045] The γ-aminobutyric acid precursor solution is prepared by adding monosodium glutamate to dilute distilled water to form a monosodium glutamate concentration of 0.2 g / L (the content of sodium glutamate in monosodium glutamate is greater than 99%); the mass percentage of ganoderma lucidum powder in the fermentation matrix solution is 5%;

[0046] (3) using an inoculation loop to select 0.05 g of Lactobacillus acidophilus and transfer it to 50 mL of purified culture medium, and culturing at 33° C. for 60 h to obtain a purified Lactobacillus acidophilus strain;

[0047] (4) inoculating the purified Lactobacillus acidophilus strain into the seed culture medium at an inoculum amount of 1% (mass percentage of the seed culture medium), culturing for 50 h under the conditions of an initial pH of 5, a fermentation temperature of 30° C., and a rotation speed of 120 rpm in a shaking incubator, and obtaining a Lactobacillus acidophilus seed liquid after culturing;

[0048] (5) The obtained Lactobacillus acidophilus seed solution was measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 );

[0049] (6) inoculating the obtained Lactobacillus acidophilus seed solution into the fermentation matrix solution at an inoculum amount of 5% (the mass percentage of the fermentation matrix solution), and fermenting at 28-33° C. for 7 days to obtain a fermentation liquid;

[0050] (7) The obtained fermentation liquid was centrifuged at 5000 rpm for 10 min to remove the bacteria and precipitated impurities; then 0.7% citric acid, 6% sorbitol and 1% cholesterol (used to adjust the stability of the formula; the added amounts of citric acid, sorbitol and cholesterol are the mass percentages of the obtained Ganoderma lucidum composite fermented scalp care liquid) were added and mixed to obtain a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid.

[0051] Embodiment 2:

[0052] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 1, except that:

[0053] In step (3): using an inoculation loop, select 0.05 g of Candida utilis and transfer to 50 mL of purification medium;

[0054] In step (4): the purified Candida utilis strain is inoculated into the seed culture medium at an inoculation amount of 1% (mass percentage of the seed culture medium) for cultivation;

[0055] In step (5), the obtained Candida utilis seed solution was measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 );

[0056] In step (6): the obtained Candida utilis seed solution is inoculated into the fermentation matrix solution at an inoculation amount of 5% (mass percentage of the fermentation matrix solution) for composite fermentation.

[0057] Embodiment 3:

[0058] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 1, except that:

[0059] In step (3): using an inoculation loop, 0.05 g of Yarrowia lipolytica was selected and transferred to 50 mL of purification medium;

[0060] In step (4): the purified Yarrowia lipolytica strain is inoculated into the seed culture medium at an inoculation rate of 1% (mass percentage of the seed culture medium) for cultivation;

[0061] In step (5): the obtained Yarrowia lipolytica seed solution was measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 );

[0062] In step (6): the obtained Yarrowia lipolytica seed solution is inoculated into the fermentation substrate solution at an inoculation amount of 5% (mass percentage of the fermentation substrate solution) for composite fermentation.

[0063] Embodiment 4:

[0064] The preparation method of the ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention comprises the following detailed steps:

[0065] (1) selecting a Ganoderma lucidum fruiting body that has not released spores as a material, drying it, crushing it, and then passing it through a 60-mesh sieve to obtain Ganoderma lucidum powder;

[0066] (2) adding the ganoderma lucidum powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 20 min, and cooling to room temperature to obtain a fermentation substrate solution;

[0067] The γ-aminobutyric acid precursor solution is prepared by adding monosodium glutamate to dilute distilled water to form a monosodium glutamate concentration of 0.2 g / L (the content of sodium glutamate in monosodium glutamate is greater than 99%); the mass percentage of ganoderma lucidum powder in the fermentation matrix solution is 5%;

[0068] (3) using an inoculation loop to select 0.05 g of Lactobacillus acidophilus and 0.05 g of Candida utilis and transfer them to 50 mL of purification medium, respectively, and culture them at 33° C. for 60 h to obtain purified Lactobacillus acidophilus strains and Candida utilis strains;

[0069] (4) inoculating the purified Lactobacillus acidophilus strain and Candida utilis strain into their respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media), culturing for 50 h under the conditions of an initial pH of 5, a fermentation temperature of 30° C., and a rotation speed of 120 rpm in a shaking incubator, and obtaining Lactobacillus acidophilus seed liquid and Candida utilis seed liquid after culturing;

[0070] (5) The obtained Lactobacillus acidophilus seed solution and Candida utilis seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were both 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution;

[0071] (6) inoculating the obtained mixed seed solution into the fermentation matrix solution at an inoculation amount of 5% (mass percentage of the fermentation matrix solution), and fermenting at 28-33° C. for 7 days to obtain a composite fermentation solution;

[0072] (7) The obtained composite fermentation liquid was centrifuged at 5000 rpm for 10 min to remove the bacteria and precipitated impurities; then 0.7% citric acid, 6% sorbitol and 1% cholesterol (used to adjust the stability of the formula; the added amounts of citric acid, sorbitol and cholesterol are the mass percentages of the obtained Ganoderma lucidum composite fermentation scalp care liquid) were added and mixed to obtain a Ganoderma lucidum composite fermentation scalp care liquid rich in γ-aminobutyric acid.

[0073] Embodiment 5:

[0074] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 4, except that:

[0075] In step (3): using an inoculation loop, 0.05 g of Yarrowia lipolytica and 0.05 g of Candida utilis were selected and transferred to 50 mL of purification medium respectively;

[0076] In step (4): the purified Yarrowia lipolytica and Candida utilis strains are inoculated into respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0077] In step (5): the obtained Yarrowia lipolytica seed solution and Candida utilis seed solution were measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution.

[0078] Embodiment 6:

[0079] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 4, except that:

[0080] In step (3): using an inoculation loop, 0.05 g of Yarrowia lipolytica and 0.05 g of Lactobacillus acidophilus were selected and transferred to 50 mL of purification medium respectively;

[0081] In step (4): the purified Yarrowia lipolytica and Lactobacillus acidophilus strains are inoculated into their respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0082] In step (5): the obtained Yarrowia lipolytica seed solution and Lactobacillus acidophilus seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were both 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution.

[0083] Embodiment 7:

[0084] The preparation method of the ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention comprises the following detailed steps:

[0085] (1) selecting a Ganoderma lucidum fruiting body that has not released spores as a material, drying it, crushing it, and then passing it through a 60-mesh sieve to obtain Ganoderma lucidum powder;

[0086] (2) adding the ganoderma lucidum powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 20 min, and cooling to room temperature to obtain a fermentation substrate solution;

[0087] The γ-aminobutyric acid precursor solution is prepared by adding monosodium glutamate to dilute distilled water to form a monosodium glutamate concentration of 0.2 g / L (the content of sodium glutamate in monosodium glutamate is greater than 99%); the mass percentage of ganoderma lucidum powder in the fermentation matrix solution is 5%;

[0088] (3) using an inoculation loop to select 0.05 g of Lactobacillus acidophilus, 0.05 g of Candida utilis, and 0.05 g of Yarrowia lipolytica, and transfer them to 50 mL of purification medium, respectively, and culture them at 33° C. for 60 h to obtain purified Lactobacillus acidophilus strains, Candida utilis strains, and Yarrowia lipolytica strains;

[0089] (4) The purified Lactobacillus acidophilus strain, Candida utilis strain and Yarrowia lipolytica strain were inoculated into their respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture medium), and cultured for 50 hours under the conditions of an initial pH of 5, a fermentation temperature of 30° C., and a rotation speed of 120 rpm in a shaker, to obtain Lactobacillus acidophilus seed liquid, Candida utilis seed liquid and Yarrowia lipolytica seed liquid;

[0090] (5) The obtained Lactobacillus acidophilus seed solution, Candida utilis seed solution and Yarrowia lipolytica seed solution were measured by spectrophotometer to determine the absorbance values ​​of the strains to be 2.0 (OD 600 ); then the seed solutions of the three strains were mixed in a volume ratio of 1:1:1 to obtain a mixed seed solution;

[0091] (6) inoculating the obtained mixed seed solution into the fermentation matrix solution at an inoculation amount of 5% (mass percentage of the fermentation matrix solution), and fermenting at 28-33° C. for 7 days to obtain a composite fermentation solution;

[0092] (7) The obtained composite fermentation liquid was centrifuged at 5000 rpm for 10 min to remove the bacteria and precipitated impurities; then 0.7% citric acid, 6% sorbitol and 1% cholesterol (used to adjust the stability of the formula; the added amounts of citric acid, sorbitol and cholesterol are the mass percentages of the obtained Ganoderma lucidum composite fermentation scalp care liquid) were added and mixed to obtain a Ganoderma lucidum composite fermentation scalp care liquid rich in γ-aminobutyric acid.

[0093] Example 7-A:

[0094] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 7, except that:

[0095] In step (5), the Lactobacillus acidophilus seed solution, the Candida utilis seed solution and the Yarrowia lipolytica seed solution are mixed in a volume ratio of 3:1:1.

[0096] Example 7-B:

[0097] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 7, except that:

[0098] In step (5), the Lactobacillus acidophilus seed solution, the Candida utilis seed solution and the Yarrowia lipolytica seed solution are mixed in a volume ratio of 1:3:1.

[0099] Example 7-C:

[0100] The preparation method of the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid of the present invention is basically the same as that of Example 7, except that:

[0101] In step (5), the Lactobacillus acidophilus seed solution, the Candida utilis seed solution and the Yarrowia lipolytica seed solution are mixed in a volume ratio of 1:1:3.

[0102] Comparative Example 1:

[0103] A method for preparing a gamma-aminobutyric acid-rich Ganoderma lucidum composite fermented scalp care liquid, the detailed steps of which are as follows:

[0104] (1) selecting a Ganoderma lucidum fruiting body that has not released spores as a material, drying it, crushing it, and then passing it through a 60-mesh sieve to obtain Ganoderma lucidum powder;

[0105] (2) adding the ganoderma lucidum powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 20 min, and cooling to room temperature to obtain a fermentation substrate solution;

[0106] The γ-aminobutyric acid precursor solution is prepared by adding monosodium glutamate to dilute distilled water to form a monosodium glutamate concentration of 0.2 g / L (the content of sodium glutamate in monosodium glutamate is greater than 99%); the mass percentage of ganoderma lucidum powder in the fermentation matrix solution is 5%;

[0107] (3) Sterilize distilled water at 121°C for 20 min and cool to room temperature;

[0108] (4) adding sterilized distilled water to the fermentation substrate solution at an inoculation rate of 5% (mass percentage of the fermentation substrate solution), and standing at 28-33° C. for 7 days to obtain a composite fermentation liquid;

[0109] (5) The obtained composite fermentation liquid was centrifuged at 5000 rpm for 10 min to remove the bacteria and precipitated impurities; then 0.7% citric acid, 6% sorbitol and 1% cholesterol (used to adjust the stability of the formula; the added amounts of citric acid, sorbitol and cholesterol are the mass percentages of the obtained Ganoderma lucidum composite fermentation scalp care liquid) were added and mixed to obtain a Ganoderma lucidum composite fermentation scalp care liquid rich in γ-aminobutyric acid.

[0110] Comparative Example 2:

[0111] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 1, except that:

[0112] In step (3): using an inoculation loop, select 0.05 g of Lactococcus lactis and transfer to 50 mL of purified culture medium;

[0113] In step (4): the purified Lactococcus lactis strain is inoculated into the seed culture medium at an inoculation amount of 1% (mass percentage of the seed culture medium) for cultivation;

[0114] In step (5): the obtained Lactococcus lactis seed solution is measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 );

[0115] In step (6): the obtained Lactococcus lactis seed solution is inoculated into the fermentation matrix solution at an inoculation amount of 5% (mass percentage of the fermentation matrix solution) for composite fermentation.

[0116] Comparative Example 3:

[0117] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 1, except that:

[0118] In step (3): using an inoculation loop, 0.05 g of Saccharomyces boulardii was selected and transferred to 50 mL of purification medium;

[0119] In step (4): the purified Saccharomyces boulardii strain is inoculated into the seed culture medium at an inoculation rate of 1% (mass percentage of the seed culture medium) for cultivation;

[0120] In step (5), the obtained Saccharomyces boulardii seed solution was measured by spectrophotometer to obtain a strain absorbance value of 2.0 (OD 600 );

[0121] In step (6): the obtained Saccharomyces boulardii seed solution is inoculated into the fermentation matrix solution at an inoculation amount of 5% (mass percentage of the fermentation matrix solution) for composite fermentation.

[0122] Comparative Example 4:

[0123] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 4, except that:

[0124] In step (3): using an inoculation loop, 0.05 g of Lactococcus lactis and 0.05 g of Saccharomyces boulardii were selected and transferred to 50 mL of purification medium respectively;

[0125] In step (4): the purified Lactococcus lactis strain and Saccharomyces boulardii strain are inoculated into their respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0126] In step (5): the obtained Lactococcus lactis seed solution and Saccharomyces boulardii seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were both 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution.

[0127] Comparative Example 5:

[0128] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 4, except that:

[0129] In step (3): using an inoculation loop, 0.05 g of Saccharomyces boulardii and 0.05 g of Yarrowia lipolytica were selected and transferred to 50 mL of purification medium respectively;

[0130] In step (4): the purified Saccharomyces boulardii strain and Yarrowia lipolytica strain are inoculated into their respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0131] In step (5), the obtained Saccharomyces boulardii seed solution and the Yarrowia lipolytica seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were both 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution.

[0132] Comparative Example 6:

[0133] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 4, except that:

[0134] In step (3): using an inoculation loop, 0.05 g of Lactococcus lactis and 0.05 g of Yarrowia lipolytica were selected and transferred to 50 mL of purification medium respectively;

[0135] In step (4): the purified Lactococcus lactis strain and Yarrowia lipolytica strain are inoculated into respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0136] In step (5): the obtained Lactococcus lactis seed solution and Yarrowia lipolytica seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were both 2.0 (OD 600 ); then the seed solutions of the two strains were mixed in a volume ratio of 1:1 to obtain a mixed seed solution.

[0137] Comparative Example 7:

[0138] A method for preparing a Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid is basically the same as that in Example 7, except that:

[0139] In step (3): using an inoculation loop, 0.05 g of Lactococcus lactis, 0.05 g of Saccharomyces boulardii, and 0.05 g of Yarrowia lipolytica were selected and transferred to 50 mL of purification medium respectively;

[0140] In step (4): the purified Lactococcus lactis strain, Saccharomyces boulardii strain and Yarrowia lipolytica strain are inoculated into respective seed culture media at an inoculation rate of 1% (mass percentage of the seed culture media);

[0141] In step (5), the obtained Lactococcus lactis seed solution, Saccharomyces boulardii seed solution and Yarrowia lipolytica seed solution were measured by spectrophotometer to determine that the absorbance values ​​of the strains were all 2.0 (OD 600 ); then the seed solutions of the three strains were mixed in a volume ratio of 1:1:1 to obtain a mixed seed solution.

[0142] In order to further illustrate that the technical solution of the present invention can achieve good technical effects, the following tests were conducted on the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid prepared in each embodiment and comparative example:

[0143] 1. In vitro antioxidant activity test:

[0144] The example groups involved in the test are: Examples 1 to 7, Examples 7-A to 7-C and Comparative Examples 1 to 7.

[0145] (1) DPPH free radical scavenging rate detection (determined by DPPH free radical scavenging ability detection kit):

[0146] Test method: Pipette 100 μL sample and add 900 μL extract, vortex to mix, centrifuge at 10000 rpm for 10 min at room temperature, take the supernatant and place on ice for testing. 空白 ): Extraction solution + DPPH working solution, determination tube (A 测定 ): supernatant + DPPH working solution, control tube (A 对照 ): supernatant + anhydrous ethanol, positive control tube (A 阳性对照 ): Vitamin C + DPPH working solution. Vortex to mix, stand at room temperature away from light for 30 minutes, preheat the spectrophotometer for more than 30 minutes, and adjust the wavelength to OD 515 , anhydrous ethanol was used to adjust the zero value, and all experiments were repeated three times.

[0147] The DPPH free radical scavenging rate was calculated by the following formula:

[0148] Sample: DPPH free radical scavenging rate (%) = [[A 空白 -(A 测定 -A 对照 )]÷A 空白 ]×100%.

[0149] Positive control: DPPH free radical scavenging rate (%) = [(A 空白 -A 阳性对照 )÷A 空白 ]×100%.

[0150] (2) ABTS free radical scavenging rate test (determined by ABTS free radical scavenging ability test kit):

[0151] Test method: Pipette 100 μL sample and add 900 μL extract, vortex to mix, centrifuge at 10000 rpm for 10 min at room temperature, take the supernatant and place on ice for testing. 空白 ): distilled water + assay working solution + ABTS working solution, assay tube (A 测定 ): supernatant + assay working solution + ABTS working solution, control tube (A 对照 ): supernatant + anhydrous ethanol, positive control tube (A 阳性对照 ): Vitamin C + ABTS working solution. Vortex to mix, stand at room temperature away from light for 30 minutes, preheat the spectrophotometer for more than 30 minutes, and adjust the wavelength to OD 405 , anhydrous ethanol was used to adjust the zero value, and all experiments were repeated three times.

[0152] The ABTS free radical scavenging rate was calculated by the following formula:

[0153] Sample: ABTS free radical scavenging rate (%) = [[A 空白 -(A测定 -A 对照 )]÷A 空白 ]×100%.

[0154] Positive control: ABTS free radical scavenging rate (%) = [(A 空白 -A 阳性对照 )÷A 空白 ]×100%.

[0155] The in vitro antioxidant activity test results of the products obtained from the above embodiments and comparative examples are shown in Table 2.

[0156] Table 2 In vitro antioxidant activity test results of the products obtained in each embodiment and comparative example

[0157] Group DPPH free radical scavenging rate (%) ABTS free radical scavenging rate (%) Positive Control 95.85 96.91 Example 1 69.48 68.34 Example 2 71.45 70.65 Example 3 65.58 65.93 Example 4 88.61 85.52 Example 5 82.29 80.38 Example 6 81.76 79.11 Example 7 91.33 87.57 Example 7-A 91.78 87.16 Example 7-B 92.79* 89.34 Example 7-C 91.13 86.87 Comparative Example 1 55.25 60.39 Comparative Example 2 65.35 66.27 Comparative Example 3 63.61 63.67 Comparative Example 4 78.43 74.28 Comparative Example 5 73.58 71.94 Comparative Example 6 76.21 72.81 Comparative Example 7 86.57 84.17

[0158] (*: p<0.05) The result is significant.

[0159] Result analysis: By comparing the test data between Examples 1 to 7, Examples 7-A to 7-C and Comparative Examples 1 to 7, it can be seen that the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid prepared in the Examples of the present invention has good in vitro antioxidant activity. Among them, the antioxidant activity of the Examples is better than that of the Comparative Examples. Under the composite fermentation of the three strains, the antioxidant activity is significantly improved, among which Example 7-B has the best antioxidant activity, with a DPPH free radical scavenging rate of 92.79% and an ABTS free radical scavenging rate of 89.34%.

[0160] 2. GABA concentration test:

[0161] The example groups involved in the test are: Examples 1 to 7, Examples 7-A to 7-C and Comparative Examples 2 to 7 (since Comparative Example 1 is a sterile fermentation and cannot convert the GABA precursor sodium glutamate into GABA, no GABA concentration detection test was performed).

[0162] Experimental method: First, prepare GABA standard solutions with concentration gradients of 0.5, 1.5, 3, 4, 5, 6, 8, 9, and 10 g / L, and perform quantitative analysis by paper chromatography pre-staining method, with GABA mass concentration as the horizontal axis and OD as the horizontal axis. 512 . Draw a standard curve with y as the vertical axis. Then take 1 mL of sample, centrifuge at 5000 rpm for 10 min, draw 0.5 μL of supernatant and spot on filter paper, directly develop the chromatography paper with a developing agent containing 1.2% ninhydrin (n-butanol, glacial acetic acid, water volume ratio 5:3:2), color at 80°C for 15 min, cut out the GABA spot and elute it in an eluent (75% ethanol, 0.6% CuS04·5H20 volume ratio 38:2) at 40°C and 50 rpm for 4 h, and measure the OD of the eluent 512. Quantitative analysis of GABA.

[0163] The GABA concentration test results of the products obtained in the above embodiments and comparative examples are shown in Table 3.

[0164] Table 3 GABA concentration test results of the products obtained in each embodiment and comparative example GABA concentration test results

[0165]

[0166] (*: p<0.05) The result is significant.

[0167] Result analysis: By comparing the test data between Examples 1 to 7, Examples 7-A to 7-C and Comparative Examples 2 to 7, it can be seen that the GABA content under the composite fermentation of three strains is higher than that of single strain fermentation and double strain fermentation, among which the GABA concentration in the case of Lactobacillus acidophilus is obviously advantageous, among which the GABA concentration of Example 7-A is the highest, reaching 6.75 g / L.

[0168] 3. Oil secretion inhibition rate test (determined by 5α-reductase kit):

[0169] The example groups involved in the test: Examples 1 to 7, Examples 7-A to 7-C (referring to the in vitro antioxidant test and the GABA concentration detection test, it can be seen that the antioxidant activity of the comparative example is low and the GABA concentration is not high, so the examples are selected for further testing).

[0170] This test determines the 5α-reductase inhibition rate of the sample according to the laboratory method QTTZ-XZ-39 "Cosmetic Oil Control Efficacy Verification Test Cells" and determines whether the sample has oil control effect by comparing it with the negative control group.

[0171] Test method: Accurately weigh the sample and dilute it into a 1% aqueous solution; react the pretreated sample with the 5α-reductase solution, and set up a negative control and a blank control; operate according to the test kit and measure the OD 340 ; The content of 5α-reductase was obtained based on the standard curve.

[0172] The calculation formula of 5α-reductase inhibition rate is: inhibition rate (%) = (1-T / C) × 100%;

[0173] T: 5α-reductase content of sample, C-5α-reductase content of negative control.

[0174] The test results of the oil secretion inhibition rate of the products prepared in the above embodiments are shown in Table 4.

[0175] Table 4 Test results of oil secretion inhibition rate of the products prepared in each embodiment

[0176]

[0177] (*: p<0.05) The result was significant. When calculating the 5α-reductase inhibition rate, the average value of 5α-reductase content was taken for calculation.

[0178] Result analysis: The 5α-reductase inhibition rate of Example 7-C is 28.02%, indicating that the enzyme content of the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid prepared in Example 7-C is significantly lower than that of the negative control, indicating that the Ganoderma lucidum fermented scalp care solution rich in γ-aminobutyric acid prepared in Example 7-C has the most ideal oil control effect.

[0179] 4. Human skin safety test:

[0180] The example groups involved in the test are: Examples 1 to 7, Examples 7-A to 7-C.

[0181] This trial complies with the human skin patch test method in the "Technical Specifications for Safety of Cosmetics" revised in 2023, and 35 subjects are selected according to the subject inclusion criteria.

[0182] (1) Skin occlusive patch test:

[0183] Test method: The selected area shall not exceed 50mm 2 , a qualified spot tester with a depth of about 1mm. Place the sample in the small chamber of the spot tester, with a dosage of about 0.02mL. Apply the spot tester with scalp care solution to the curved side of the subject's forearm with hypoallergenic tape, and gently press it with the palm of your hand to evenly apply it to the skin for 24 hours. Observe the skin reaction according to the skin reaction evaluation criteria 30 minutes (after the indentation disappears), 24 hours and 48 hours after removing the test object spot tester, and record the observation results.

[0184] (2) Repeated open smear test:

[0185] Test method: The flexed side of the forearm is used as the test site, with an area of ​​3×3cm 2 The test area should be kept dry and avoid contact with other topical preparations. Take about 0.05 mL of the sample and apply it evenly to the test area twice a day for 7 consecutive days. Observe according to the skin reaction evaluation criteria and record the results.

[0186] The human skin safety test results of the products prepared in the above examples are shown in Table 5.

[0187] Table 5 Human skin safety test results of the products prepared in each embodiment

[0188] Test items 24h 48h 7d Example 1 Level 0 Level 0 Level 0 Example 2 Level 0 Level 0 Level 0 Example 3 Level 0 Level 0 Level 0 Example 4 Level 0 Level 0 Level 0 Example 5 Level 0 Level 0 Level 0 Example 6 Level 0 Level 0 Level 0 Example 7 Level 0 Level 0 Level 0 Example 7-A Level 0 Level 0 Level 0 Example 7-B Level 0 Level 0 Level 0 Example 7-C Level 0 Level 0 Level 0

[0189] Table 6 Skin reaction evaluation criteria

[0190]

[0191] Result analysis: The data obtained through the human skin safety test show that the Ganoderma lucidum fermented scalp care solution rich in γ-aminobutyric acid prepared by the present invention will not affect the safety of human skin under various implementation methods, and is a safe care product.

[0192] In summary, through the comprehensive analysis of the results of the in vitro antioxidant test, the GABA content detection test, the oil secretion inhibition rate test and the human skin safety test, it can be seen that the scalp care solution fermented by the three strains of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica has the best data. In terms of the in vitro antioxidant activity, GABA content and oil secretion inhibition rate, the solution of Example 7-B is the best. Therefore, the gamma-aminobutyric acid-rich Ganoderma lucidum fermented scalp care solution prepared by the present invention has good antioxidant activity, high GABA content, and good inhibition of oil secretion, and is a green, safe and efficient cosmetic product.

Claims

1. A method for preparing a scalp care liquid containing Ganoderma lucidum compound fermentation and rich in γ-aminobutyric acid, characterized in that: The preparation method comprises the following steps: (1) drying and crushing the Ganoderma lucidum fruiting bodies that have not released spores in sequence, and sieving the crushed Ganoderma lucidum to obtain Ganoderma lucidum powder; (2) adding the ganoderma powder to the γ-aminobutyric acid precursor solution, sterilizing at 121° C. for 15 to 20 minutes, and cooling to room temperature to obtain a fermentation substrate solution; (3) using an inoculation loop to select Lactobacillus acidophilus, Candida utilis, and Yarrowia lipolytica respectively, and then transferring them to their respective purification medium, culturing them at 30-35° C. for 48-72 hours, and obtaining the three purified strains; (4) inoculating the three purified strains into their respective seed culture media at an inoculum amount of 1 to 2% for cultivation, and obtaining seed solutions of the three strains after cultivation; (5) detecting absorbance of the seed solutions of the three strains by a spectrophotometer, and after the detection, mixing the seed solutions of the three strains to obtain a mixed seed solution; (6) inoculating the mixed seed solution into a fermentation matrix solution for composite fermentation to obtain a composite fermentation solution; (7) The obtained composite fermentation liquid is centrifuged to remove the bacterial cells and precipitated impurities; then citric acid, sorbitol and cholesterol are added and mixed to obtain a Ganoderma lucidum composite fermentation scalp care liquid rich in γ-aminobutyric acid.

2. The method for preparing the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid according to claim 1, characterized in that: After the pulverization in step (1), the product is sieved through a 50-70 mesh sieve.

3. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: The γ-aminobutyric acid precursor solution in step (2) is prepared by diluting monosodium glutamate with distilled water to form a monosodium glutamate solution with a concentration of 0.2 to 0.3 g / L.

4. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: The mass percentage of the ganoderma lucidum fruiting bodies in the fermentation matrix solution obtained in step (2) is 4-5%.

5. The method for preparing the Ganoderma lucidum composite fermented scalp care liquid rich in γ-aminobutyric acid according to claim 1, characterized in that: The preparation method of the purification medium in step (3) is: 15-20 g of glucose, 4-5 g of peptone, 2.5-3 g of yeast extract powder and 18-20 g of agar powder, add double distilled water to 1 L, sterilize at 121° C. for 15-20 min, and cool to room temperature to obtain a solid purification medium; The selected amounts of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica are all 0.01-0.07 g, and the used amount of the purification culture medium is all 40-60 mL.

6. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: The preparation method of the seed culture medium in step (4) is: 15-20 g of glucose, 10-15 g of soy protein, 10-15 g of corn flour, 3-8 g of sodium chloride, 1-4 g of dipotassium hydrogen phosphate and 0.3-0.6 g of L-cysteine, double distilled water is added to 1 L, sterilized at 121° C. for 15-20 min, and cooled to room temperature to obtain a seed culture medium.

7. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: The conditions for culturing in the seed culture medium in step (4) are: initial pH of 5-6, fermentation temperature of 30-33° C., rotation speed of 100-120 rpm in the shaking incubator, and culturing time of 48-54 h.

8. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: The absorbance values ​​of the seed solutions of the three strains of Lactobacillus acidophilus, Candida utilis and Yarrowia lipolytica obtained in step (5) were all 2.0 to 2.2 (OD 600 ); the volume ratio of the three when mixed is 1~3:1~3:1~3.

9. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: When the mixed seed liquid in step (6) is added to the fermentation matrix solution, the mass percentage of the mixed seed liquid to the fermentation matrix solution is 3-5%; the composite fermentation conditions are: the fermentation temperature is 28-33° C., and the fermentation time is 5-7 days.

10. The method for preparing the Ganoderma lucidum composite fermented scalp care solution rich in γ-aminobutyric acid according to claim 1, characterized in that: In step (7), the amounts of citric acid, sorbitol and cholesterol added are 0.5-1%, 5-8% and 1-1.2% by mass of the obtained Ganoderma lucidum composite fermented scalp care liquid respectively.

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