Stowartia elegans and uses thereof

Abstract

The present application relates to the field of microbial technology, and particularly to Sparassis crispa Stowart and its application. The Sparassis crispa Stowart provided by the present application is preserved in China Center for Type Culture Collection, and the preservation number is CCTCC NO: M 20211503. The polysaccharide and glycoprotein content of the mycelium of the Sparassis crispa Stowart is significantly higher than that of the original strain, and the Sparassis crispa Stowart has high genetic stability. The extract prepared through fermentation and extraction has good effects of repairing internal and external structure damage of damaged hair and nourishing and caring hair and scalp, and can be applied to the field of cosmetics, and a hair cosmetic with hair nourishing and caring function is developed, and the effective utilization of Sparassis crispa resources is improved.

Description

Technical Field

[0001] This invention relates to the field of microbial technology, specifically to Stowartella asiatica and its applications. Background Technology

[0002] *Sparassis crispa*, a large edible and medicinal fungus belonging to the phylum Basidiomycota, class Agaricales, order Apocynomorales, family Sprastrophaceae, and genus *Sparassis*, is scarce in the wild. Although artificial cultivation has been achieved, the slow mycelial growth rate and low biomass result in a long preparation cycle for the mother culture. Increasing the content of active ingredients and enhancing the efficacy of *Sparassis crispa* is beneficial for solving the resource utilization problem and expanding its application areas. Summary of the Invention

[0003] The purpose of this invention is to provide *Stowart* fungus. This invention also provides applications of *Stowart* fungus and its products.

[0004] This invention uses the *Sparassis crispa* strain SC031 (accessed on May 3, 2015 at the China Center for Type Culture Collection, Wuhan University, Wuhan, China, and classified as *Sparassis crispa* SC031, accession number CCTCC NO:M 2015275; this strain has been disclosed in patent application CN104928189A) as the starting strain. Ultraviolet mutagenesis was performed on this strain, and after screening, a *Sparassis crispa* mutant strain was obtained and named *Sparassis crispa* Stowart. This strain exhibits significantly increased polysaccharide and glycoprotein content in its mycelium, resulting in significantly enhanced efficacy for hair and scalp, and also demonstrates good genetic stability.

[0005] Specifically, this invention provides *Sparassis crispa* Stowart, a strain that was deposited on November 29, 2021, at the China Center for Type Culture Collection (CCTCC, Wuhan University, Wuhan, China 430072, China), and classified as *Sparassis crispa* Stowart, with accession number CCTCC NO: M20211503.

[0006] The present invention provides a microbial agent containing the aforementioned Stowart fungus.

[0007] Preferably, the *Stowartia lycopersica* fungus in the above-mentioned fungal agent exists in the form of live bacteria. More preferably, it exists in the form of spores.

[0008] Preferably, the above-mentioned microbial agent also contains a freeze-drying protectant and / or a carrier.

[0009] The above-mentioned microbial agents can be liquid or solid (e.g., dry powder microbial agents).

[0010] The present invention provides a method for preparing the above-mentioned bacterial agent, the method comprising the step of culturing Stowartella asiatica to obtain a bacterial solution.

[0011] The present invention also provides a culture of *Stowart*, which is obtained by culturing the *Stowart* fungus.

[0012] The above-mentioned *Stowart* culture preferably contains one or more components selected from *Stowart* mycelium, *Stowart* metabolites, and culture medium components.

[0013] The Stowart mycelium described above is preferably inactivated mycelium.

[0014] The metabolites of Stowartia salina mentioned above may include extracellular metabolites and / or intracellular metabolites. The intracellular metabolites are obtained by processing the mycelium of Stowartia salina through methods such as cell wall disruption.

[0015] The present invention also provides a *Stowart* extract, which is prepared by a method comprising the following steps: culturing the *Stowart* fungus to obtain a culture, treating the culture with cellulase to obtain an enzymatic hydrolysate, subjecting the enzymatic hydrolysate to ultrasonic treatment, and then separating the solid and liquid phases to obtain the supernatant.

[0016] Preferably, the cellulase hydrolysis treatment is carried out at a temperature of 40–50°C, a pH of 4.5–5.5, and a hydrolysis time of 10–20 min.

[0017] Preferably, the enzymatic hydrolysis treatment includes: mixing the fermentation broth of *Stowartia lycopersica* with water, adding a pH adjuster to adjust the pH to 4.5-5.5, and adding cellulase for enzymatic hydrolysis. The amount of water added is 5-10 times the mass of the fermentation broth; the amount of pH adjuster added is 0.2-1% of the total mass of the enzymatic hydrolysis system; and the amount of cellulase added is 1-3% of the total mass of the enzymatic hydrolysis system.

[0018] Preferably, the ultrasonic treatment conditions are: ultrasonic treatment frequency of 15-25KHZ, power of 150-170W, temperature of 30-35℃, and time of 30-40min.

[0019] Preferably, the culture is sterilized before enzymatic hydrolysis.

[0020] The above-described method for preparing *Hydrangea macrophylla* extract employs cellulase enzymatic hydrolysis combined with ultrasonic treatment. The process conditions are controllable, resulting in a high extraction rate of active ingredients from *Hydrangea macrophylla*. The obtained *Hydrangea macrophylla* extract is rich in small molecule active ingredients that can be absorbed through the scalp, effectively nourishing the scalp and hair and repairing damage to the internal and external structures of damaged hair.

[0021] As a preferred embodiment of the present invention, the preparation method of the above-described *Hydrangea macrophylla* extract includes the following steps:

[0022] (1) Preparation of Stowart seed culture: Inoculate Stowart of the hydrangea into liquid seed culture medium and culture for 4-5 days at 23-29℃ and 120-160r / min on a shaker.

[0023] (2) Inoculate the seed liquid obtained in step (1) into the fermentation medium at an inoculation rate of 4-10%, and ferment it in a shaker at 23-29℃ and 120-160r / min for 4-5 days to obtain the fermentation liquid;

[0024] (3) After sterilizing the fermentation broth obtained in step (2), mix it with cellulase and water, add citric acid to adjust the pH to 4.5-5.5, and then carry out enzymatic hydrolysis to obtain the enzymatic hydrolysate. The amount of cellulase added is 1-3% of the total mass of the enzymatic hydrolysis system; the amount of water added is 5-10 times the mass of the fermentation broth; and the amount of citric acid added is 0.2-1% of the total mass of the enzymatic hydrolysis system.

[0025] The enzymatic hydrolysis treatment temperature is 40–50℃, and the time is 10–20 min;

[0026] (4) The enzymatic hydrolysate obtained in step (3) is subjected to ultrasonic treatment, and the supernatant is taken after solid-liquid separation to obtain the extract of Hydrangea macrophylla.

[0027] In step (3) above, citric acid is preferably used for pH adjustment.

[0028] The carbon source for the seed culture medium and fermentation culture medium described above is selected from one or more of sucrose, glucose, ribose, lactose, galactose, fructose, maltose, mannose, trehalose, mannitol, and sorbitol. Sucrose is preferred.

[0029] The nitrogen source for the seed culture medium and fermentation culture medium described above is selected from one or more of peptone, beef extract, and yeast extract. Peptone and yeast extract are preferred.

[0030] The seed culture medium and fermentation culture medium described above also contain one or more selected from inorganic salts and vitamins.

[0031] Preferably, the seed culture medium comprises the following components (g:ml): sucrose 0.5-5%, peptone 0.5-1.5%, yeast extract 0.2-1.5%, dipotassium hydrogen phosphate 0.01-0.05%, magnesium sulfate 0.01-0.05%, and V... B1 0.01% to 0.05%.

[0032] Preferably, the fermentation medium comprises the following components (g: ml): sucrose 0.5-5%, peptone 0.5-1.5%, yeast extract 0.2-1.5%, dipotassium hydrogen phosphate 0.01-0.05%, and magnesium sulfate 0.01-0.05%.

[0033] Stowartella asiatica exhibits better growth performance in the aforementioned culture medium.

[0034] Based on the functions of *Stowartia lycopersicum*, this invention provides the following uses for this strain:

[0035] The present invention provides the use of the *Stowartia lycopersica* fungal agent, the *Stowartia lycopersica* culture, or the *Stowartia lycopersica* extract in the preparation of food or medicine.

[0036] This invention provides the use of the *Stowartia lycopersica* fungal agent, the *Stowartia lycopersica* culture, or the *Stowartia lycopersica* extract in the preparation of cosmetics.

[0037] The present invention provides the use of the *Stowartia styracifolium*, the fungal agent, the *Stowartia styracifolium* culture, or the *Stowartia styracifolium* extract in the preparation of products with the function of repairing damaged hair and / or nourishing and moisturizing the scalp.

[0038] Preferably, the product is a cosmetic. More preferably, it is a hair cosmetic, including but not limited to shampoo, hair growth tonic, hair dye, conditioner, hair styling agent, hair lotion, and hair nourishing lotion.

[0039] The present invention provides a cosmetic containing the *Hydrangea macrophylla* extract.

[0040] Preferably, the *Hydrangea macrophylla* extract in the cosmetic has a mass percentage content of 0.1% to 15%.

[0041] In one embodiment of the present invention, the cosmetic is a shampoo product, which contains the *Hydrangea macrophylla* extract and also contains a shampoo base permitted in the art.

[0042] The beneficial effects of the present invention include at least the following:

[0043] 1. The polysaccharide and glycoprotein content of the mycelium of *Stowartia salina* was significantly higher than that of the original strain.

[0044] 2. Stowart fungus does not degenerate after more than 10 subcultures, demonstrating high genetic stability.

[0045] 3. The extract obtained from the fermentation and extraction of Stowartella asiatica has a good effect on repairing the internal and external structural damage of hair and nourishing and caring for hair and scalp. It can be applied in the cosmetic field to develop hair cosmetics with hair and scalp nourishing and caring functions. It has high application value in hair care products and is of great significance for solving scalp and hair health problems. At the same time, it improves the effective utilization of Stowartella asiatica resources. Attached Figure Description

[0046] Figure 1 The results of the growth performance test of Stowartella asiatica in Example 2 of this invention are shown.

[0047] Figure 2 This is the detection result of the hair contact angle in Embodiment 5 of the present invention.

[0048] Figure 3 The results are the test results of the shampoo's ability to repair the combability of dry hair in Example 5 of the present invention.

[0049] Figure 4 The results are the test results of the shampoo's ability to repair the combability of wet hair in Example 5 of the present invention. Detailed Implementation

[0050] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the invention.

[0051] Example 1: Obtaining Stowart fungus

[0052] Using *Stowart* strain SC031 (accession number CCTCC NO:M 2015275) as the starting strain, this strain was subjected to ultraviolet (UV) irradiation mutagenesis and screening. This invention optimized the UV irradiation mutagenesis conditions and conducted numerous mutagenesis and screening experiments, ultimately obtaining *Stowart*. The basic steps of mutagenesis and screening are as follows:

[0053] (1) After activating the *Hydrangea spp.* strain SC031, inoculate it onto a PDA slant and incubate it at 26°C for 6 days. Then, inject phosphate buffer (pH 6.0) into the slant culture medium, filter it through four layers of gauze, and dilute the spore concentration to 10. 6 -10 7 Individuals / mL, used for mutation breeding;

[0054] (2) Irradiate the spore suspension prepared in step (1) with 15W ultraviolet light for 2 minutes, dilute it, spread it on a plate, and incubate it in a constant temperature incubator at 26℃ for 2 days. Then, use sterile toothpicks to insert the spores point-to-point into a 96-well plate containing PDA solid medium. Inoculate the cultured mutant strains one by one into a 250mL Erlenmeyer flask and incubate them at 26℃ and 150rpm for 5 days to obtain *Hydrangea hydrangea* mycelium.

[0055] Through mutagenesis and screening, a high-performance strain of *Sparassiscrispa* was obtained and named *Sparassiscrispa* Stowart. *Sparassiscrispa* Stowart was deposited on November 29, 2021, at the China Center for Type Culture Collection (CCTCC, Wuhan University, Wuhan, China 430072, China), and classified as *Sparassiscrispa* Stowart, with accession number CCTCC NO: M 20211503.

[0056] Example 2: Performance testing of Stowart fungus.

[0057] 1. Growth performance testing

[0058] *Stowart* and *SC031* were inoculated into liquid seed culture medium and cultured at 26℃ and 150 rpm for 1, 3, and 5 days, respectively. Mycelial yield was measured at the end of the culture, and growth curves of *Stowart* and *SC031* were plotted. The seed culture medium composition was: 3% sucrose, 1% yeast extract, 1% peptone, 0.02% KH₂PO₄, 0.01% MgSO₄, V₂O₃ ... B1 0.01%.

[0059] The results are as follows Figure 1 As shown, the growth rate of Stowart fungus was significantly higher than that of SC031, and the mycelial yield was also significantly higher than that of the starting strain Stowart fungus SC031.

[0060] 2. Detection of polysaccharide and glycoprotein content in mycelium

[0061] The mycelial polysaccharides and glycoproteins of *Stowartia stowartica* were extracted and their contents were determined. The specific methods are as follows:

[0062] The *Stowart* fermentation broth was centrifuged at 5000 rpm to collect mycelia. The mycelia were washed twice with water, freeze-dried, pulverized, and passed through an 80-mesh sieve. Three 0.1 g portions of *Stowart* mycelial powder were weighed and each portion was added to 5 mL of distilled water. After extraction in an 80℃ water bath for 3 hours, the mixture was centrifuged at 5000 g / min for 6 minutes. The supernatant was collected to determine the polysaccharide and glycoprotein content of the mycelia. The results are shown in Table 1. *Stowart* strains 1, 2, 3, and 4 were other relatively good-performing strains obtained during the mutagenesis screening process in Example 1. The starting strain and all other mutagenic strains were cultured under the same fermentation conditions.

[0063] The fermentation medium used for fermentation culture includes the following components (g:ml): 3% sucrose, 1% yeast extract, 1% peptone, 0.02% KH2PO4, and 0.01% MgSO4.

[0064] Fermentation conditions were: 26℃, 150 r / min for 5 consecutive days. Results showed that the polysaccharide and glycoprotein contents of *Stowart* mycelium were significantly higher than those of the starting strain and other screened strains.

[0065] Table 1. Polysaccharide and glycoprotein content

[0066]

[0067]

[0068] 3. Genetic stability analysis

[0069] Stowart was passaged, and the polysaccharide and glycoprotein content of the mycelium was measured every other generation. After 10 generations, the genetic stability of Stowart was investigated. Some of the test results are shown in Table 2. The results show that Stowart has high genetic stability.

[0070] Table 2. Investigation of genetic stability of the mutagenic strains

[0071] generation polysaccharide g / g glycoprotein mg / g Generation 1 0.46 122 3rd generation 0.43 123 6th generation 0.46 120 10th generation 0.47 121

[0072] Example 3: Fermentation culture of Stowartella asiatica and preparation of its extract

[0073] The fermentation culture of *Stowartia lycopersicum* and the preparation of *Stowartia lycopersicum* extract are carried out using the following method:

[0074] (1) Preparation of Hydrangea spp. seed liquid

[0075] Stowartella asiatica was inoculated into liquid seed culture medium and cultured for 5 days at 23-29℃ and 120-160 rpm on a shaker. The seed culture medium consisted of 3% sucrose, 1% yeast extract, 1% peptone, 0.02% KH₂PO₄, 0.01% MgSO₄, and V. B1 0.01%;

[0076] (2) Fermentation

[0077] The seed culture of *Hydrangea macrophylla* was inoculated into the fermentation medium at an inoculation rate of 5% and cultured at 26℃ and 150 rpm for 5 days to obtain the fermentation broth. The fermentation medium consisted of 3% sucrose, 1% yeast extract, 1% peptone, 0.02% KH2PO4, and 0.01% MgSO4.

[0078] (3) Extraction of fermentation broth

[0079] The fermentation broth obtained in step (2) (mycelium content approximately 1.68 g / L) was sterilized at 121°C for 20 min. Pure water was added, and citric acid was added to adjust the pH to 5.0. Cellulase was then added for enzymatic hydrolysis to obtain the hydrolysate. The amount of cellulase added was 2% of the total mass of the hydrolysis system; the amount of water added was 8 times the mass of the fermentation broth; the amount of citric acid added was 0.6% of the total mass of the hydrolysis system; the hydrolysis temperature was 43°C, and the time was 15 min. The hydrolysate was then ultrasonically treated at a frequency of 20 kHz, a power of 160 W, a temperature of 35°C, and a time of 35 min. After cooling to room temperature, the *Hydrangea macrophylla* extract was obtained.

[0080] Example 4: Shampoo containing Hydrangea macrophylla extract

[0081] This embodiment provides a shampoo with the following composition (by weight): 1% of the *Hydrangea macrophylla* extract from Example 3, and 99% shampoo base.

[0082] The composition of the shampoo base is as follows (by weight percentage):

[0083] Ammonium lauryl ether sulfate 15%, sodium methyl myristoyl taurate 3%, lauramide propyl betaine 7%, cocamide MEA 0.5%, polyquaternium-47 2%, cocoyl hydrolyzed soy protein 0.1%, dilinoleamide propyl PG-dimethylammonium chloride sodium phosphate 0.2%, guar hydroxypropyltrimethylammonium chloride 0.2%, flavor 0.1%, iodopropynyl butylcarbamate 0.1%, deionized water 71.8%.

[0084] Example 5: Functional Test of Hydrangea macrophylla Extract in Repairing Damaged Hair

[0085] 1. The effect of *Hydrangea macrophylla* extract in repairing damage to the outer cuticles of damaged hair.

[0086] Healthy hair samples from volunteers were washed with sodium lauryl sulfate solution, rinsed thoroughly, and air-dried. Four samples of the original hair were soaked in 0.5 g / L NaOH solution at 30°C for 1 hour, then removed, rinsed until neutral, and air-dried to prepare damaged hair samples.

[0087] Damaged hair was treated with the shampoo of Example 4. After washing, the hair was rinsed with clean water and air-dried. Untreated (blank sample) and treated hair strands were soaked in 0.2 mol / L copper sulfate solution for 15 min, and then the amount of copper ions adsorbed by the hair was tested.

[0088] A shampoo containing an extract of the starting strain *Hydrangea spp.* SC031 was used as a control. The preparation method of the extract of *Hydrangea spp.* SC031 was the same as that of the extract of *Hydrangea stowart* in Example 3, except that *Hydrangea stowart* was replaced with *Hydrangea spp.* SC031. The formula of the shampoo containing the extract of the starting strain *Hydrangea spp.* SC031 was different from that of the shampoo formula in Example 4, except that the extract of *Hydrangea stowart* was replaced with the extract of *Hydrangea spp.* SC031. A shampoo base (formulation as described in Example 4) was used as a control.

[0089] Table 3 Copper ion adsorption capacity

[0090]

[0091] Damaged hair becomes negatively charged and adsorbs copper ions. Measuring the amount of copper ions adsorbed by hair can characterize the degree of hair damage. The greater the degree of hair damage, the more copper ions are adsorbed. The results in Table 3 show that, compared with the blank sample and the shampoo matrix, the amount of copper ions adsorbed by the hair treated with the shampoo of Example 4 was significantly reduced, indicating that the extract of *Stowartia lycopersica* has a significant repairing effect on damage to the outer cuticle of hair. Moreover, the amount of copper ions adsorbed by the hair treated with the shampoo of Example 4 was significantly lower than that of the shampoo containing the extract of the starting strain *Stowartia lycopersica* SC031, indicating that the repair effect of the extract of *Stowartia lycopersica* on damage to the outer cuticle of hair is significantly better than that of its starting strain SC031.

[0092] 2. Repairs damaged hair with both hydrophilic and lipophilic properties.

[0093] Thirty volunteers who had undergone hair dyeing and perming within the past three months were selected, with 10 volunteers in each group. Hair was collected from each volunteer and recorded as a blank group. The damaged hair was treated with the shampoo described in Example 4, three times a week, and hair was collected from the volunteers once every weekend.

[0094] A shampoo containing an extract of the starting strain *Hydrangea spp.* SC031 was used as a control. The preparation method of the extract of *Hydrangea spp.* SC031 was the same as that of the extract of *Hydrangea stowart* in Example 3, except that *Hydrangea stowart* was replaced with *Hydrangea spp.* SC031. The formula of the shampoo containing the extract of the starting strain *Hydrangea spp.* SC031 was different from that of the shampoo formula in Example 4, except that the extract of *Hydrangea stowart* was replaced with the extract of *Hydrangea spp.* SC031. A shampoo base (formulation as described in Example 4) was used as a control.

[0095] After the 3-week experiment, the Wilhelmy equilibrium method was used. Hair from each group was immersed in water in both dry and wet states. The contact angle of the water on the hair surface was calculated based on the forces exerted. The results are as follows: Figure 2 As shown.

[0096] The degree of hair damage is directly related to the hydrophilicity and hydrophobicity of the hair surface; the greater the damage, the stronger the hydrophilicity. When a liquid spreads on a solid surface, the angle between the solid / liquid interface and the gas / liquid interface is called the contact angle. The smaller the contact angle of water on the hair surface, the easier it spreads, and the stronger its hydrophilicity. Figure 2 As shown, compared with dyed and permed hair (blank group), the hair treated with the shampoo of Example 4 showed a significant increase in contact angle, indicating that the extract of *Stowartia lycopersica* can significantly repair hair damage and restore the hair's hydrophilic and lipophilic properties. Furthermore, the contact angle of hair treated with the shampoo of Example 4 was significantly higher than that of shampoo containing the extract of the original strain *Stowartia lycopersica* SC031, indicating that the extract of *Stowartia lycopersica* has a significantly better effect on repairing the hydrophilic and lipophilic properties of damaged hair than its original strain SC031.

[0097] 3. Improves hair's combability

[0098] The effect of the shampoo of Example 4 on hair combability repair was analyzed. A shampoo containing the extract of the starting strain *Hydrangea hydrangea* SC031 was used as a control. The preparation method of the extract of *Hydrangea hydrangea* SC031 was the same as that of the extract of *Hydrangea hydrangea* Stowart in Example 3, except that *Hydrangea hydrangea* Stowart was replaced with *Hydrangea hydrangea* SC031. The formula of the shampoo containing the extract of the starting strain *Hydrangea hydrangea* SC031 was different from that of the shampoo formula of Example 4, except that the extract of *Hydrangea hydrangea* Stowart was replaced with the extract of *Hydrangea hydrangea* SC031. The shampoo base (formulation as described in Example 4) was used as a control.

[0099] The specific method is as follows:

[0100] Thirty volunteers who had dyed or permed their hair within the past three months were selected, with 10 volunteers in each group. Hair was collected from each volunteer and washed with the shampoos used in Example 4 and the control group, respectively. Hair was collected from the volunteers again after one week of use.

[0101] Rinse your hair with tap water, towel dry until the hair is no longer dripping, comb it through, and test the combability of wet hair with a tensile strength tester.

[0102] Then, use a hairdryer to blow-dry the hair from a distance of 20cm until it is half-dry. Comb it smooth and let it air dry naturally. Use a multi-purpose tensile strength tester to test the combability of the dry hair. The test results are as follows: Figure 3 and Figure 4 As shown.

[0103] The results showed that the shampoo sample of Example 4 had a significantly better effect on hair combability (wet hair combability and dry hair combability) than the control groups.

[0104] 4. Repairs damaged hair from dyeing and perming

[0105] To verify the effect of the shampoo in Example 4 on repairing dyed and permed hair, a shampoo containing an extract of the starting strain *Hydrangea hydrangea* SC031 was used as a control. The preparation method of the *Hydrangea hydrangea* SC031 extract was the same as that of the *Hydrangea hydrangea* Stowart extract in Example 3, except that *Hydrangea hydrangea* Stowart was replaced with *Hydrangea hydrangea* SC031. The formula of the shampoo containing the extract of the starting strain *Hydrangea hydrangea* SC031 was different from that of the shampoo in Example 4, except that the extract of *Hydrangea hydrangea* Stowart was replaced with the extract of *Hydrangea hydrangea* SC031. At the same time, the shampoo base (formulation as described in Example 4) was used as a control.

[0106] Thirty volunteers aged 25-60 who had dyed or permed their hair within the past year were selected. Their hair was damaged during the dyeing and perming process. They were divided into three groups of 10 each, and were provided with the shampoo described in Example 4 and the control groups, respectively. The shampoo was used for two months, every two days, to examine its effect on repairing the internal structure of the hair and strengthening it. Before the experiment, the maximum breaking force and breaking energy of the hair were measured. After two months of shampoo use, the maximum breaking force and breaking energy were measured again to compare changes in hair strength. A Diastron tensiometer was used to test hair strength, and the results are shown in Table 4.

[0107] Table 4. Changes in hair breaking strength

[0108]

[0109]

[0110] As shown in Table 4, compared with the control group and before use, the maximum breaking force and breaking strength of hair increased significantly after using the shampoo of Example 4, indicating that the extract of Stowartella asiatica can significantly repair the internal structure of hair and improve hair strength.

[0111] Example 6: Analysis of the scalp nourishing and moisturizing effect of *Hydrangea macrophylla* extract.

[0112] The scalp nourishing and moisturizing effect of the *Hydrangea stowart* extract in Example 3 was analyzed. The extract of the starting strain *Hydrangea stowart* SC031 was used as a control. The preparation method of the extract of *Hydrangea stowart* SC031 was the same as that of the extract of *Hydrangea stowart* in Example 3, except that *Hydrangea stowart* was replaced by *Hydrangea stowart* SC031.

[0113] The specific method is as follows:

[0114] Twenty volunteers were selected and divided into two groups of ten. A 5*5cm section of hair was shaved to expose the scalp for the experiment. A Corneometer CM825 (German CK) skin moisture analyzer was used to apply *Hydrangea macrophylla* extract to the exposed scalp. Before application, the scalp moisture content of each subject was measured using the skin moisture analyzer under constant temperature and humidity (21℃ and 40% humidity). The initial value (T0) and changes in scalp moisture content at 30 minutes (T1) and 8 hours (T2) after application were measured.

[0115] Table 5. Scalp Moisture Content Detection

[0116]

[0117] The test results are shown in Table 5. The results show that the skin water content after 30 minutes and 8 hours of use with the Stowart extract of Example 3 was significantly higher than that of the Stowart SC031 extract. This indicates that the Stowart extract can nourish and moisturize the scalp, provide sufficient moisture to the scalp, and delay scalp aging.

[0118] Although the present invention has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, all such modifications or improvements made without departing from the spirit of the present invention fall within the scope of protection claimed by the present invention.

Claims

1. *Sparassis crispa* Stowart, characterized by, It is deposited at the China Center for Type Culture Collection, with accession number CCTCC NO: M 20211503.

2. A microbial agent, characterized in that, It contains the *Sparassis crispa* Stowart as described in claim 1.

3. The method for preparing the microbial agent according to claim 2, characterized in that, The method includes the step of culturing the *Sparassiscrispa* Stowart to obtain a bacterial solution.

4. A culture of *Hydrangea macrophylla*, characterized in that, It is obtained by culturing *Sparassis crispa* Stowart as described in claim 1.

5. A *Hydrangea macrophylla* extract, characterized in that, The Sprassis crispa extract is prepared by a method comprising the following steps: culturing the Sprassis crispa Stowart as described in claim 1 to obtain a culture, subjecting the culture to enzymatic hydrolysis with cellulase to obtain an enzymatic hydrolysate, subjecting the enzymatic hydrolysate to ultrasonic treatment, and then separating the solid and liquid phases to obtain the supernatant.

6. The *Hydrangea macrophylla* extract according to claim 5, characterized in that, The cellulase hydrolysis treatment is carried out at a temperature of 40-50℃, a pH of 4.5-5.5, and a hydrolysis time of 10-20 min. And / or, the ultrasonic treatment is performed at a frequency of 15-25 kHz, a power of 150-170 W, a temperature of 30-35 °C, and a time of 30-40 min.

7. The use of the *Hydrangea macrophylla* extract according to claim 5 or 6 in the preparation of cosmetics.

8. The application according to claim 7, characterized in that, The cosmetic product has the function of repairing damaged hair and / or nourishing and moisturizing the scalp.

9. A cosmetic product, characterized in that, The cosmetic contains the *Hydrangea macrophylla* extract as described in claim 5 or 6.

Images