Eurotium cristatum and application thereof in skin care products

Abstract

The invention provides eurotium cristatum and an application mode thereof in skin care products, and belongs to the technical field of microorganisms, the name of the eurotium cristatum is eurotium cristatum K8824, the eurotium cristatum is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC M 2024643. The tea-hippophae rhamnoides leaf compound fermentation liquor of the strain has the functions of resisting oxidation, resisting aging and whitening, and can be used for preparing functional skin care products such as emulsion, skin care cream and facial masks with the characteristics. The eurotium cristatum fermentation process disclosed by the invention is simple and easy to operate and controllable in cost, and has certain technical advantages. Meanwhile, the growth speed of the eurotium cristatum in a tea-hippophae rhamnoides leaf fermentation system is remarkably increased, the fermentation liquor is good in oxidation resistance and skin aging resistance effect, typical rot-causing fungi in the skin care product are remarkably inhibited, the problem that the skin care product is prone to bacterial contamination can be effectively solved, and the skin care product has remarkable whitening and anti-aging effects after being tried on a human body.

Description

Technical Field

[0001] The invention belongs to the technical field of microbial fermentation, and particularly relates to Eurotium cristatum and application thereof in skin care products. Background Art

[0002] In the application and development of daily chemical products, natural plant raw materials have some defects, such as the content of active ingredients cannot meet the requirements of functional product development, the conventional extraction and enrichment technology is difficult, and there is even a risk of residual harmful substances in the extraction process, which restricts the application of plant raw materials in daily chemical products. By combining extraction and fermentation, the metabolism and transformation of microorganisms are used to ferment plant raw materials and regulate the process, providing a high-performance technical solution for the enrichment and preparation of natural active ingredients. The advantage of using microorganisms for plant fermentation is that, on the one hand, it can promote the release of active components in plant raw materials, the transformation and synthesis of new active derivatives, and the microorganisms themselves can produce a variety of functional ingredients, which ultimately increases the content of target active components. On the other hand, it can reduce the content of toxic and harmful substances, reduce the cost of detoxification of plant active substances, and improve the safety of plant fermentation active raw materials.

[0003] my country has abundant summer and autumn tea resources, but more than 30% of summer and autumn tea is not fully utilized every year, and the raw material value is low. Summer and autumn tea has a high content of tea polyphenols and rich carbohydrates, which can provide a good carbon source for microbial growth. The cellulose, pectin, tea polyphenols, protein, flavonoids and other substances in summer and autumn tea can produce a variety of active metabolites such as oligosaccharides, phenolic acids, flavonoid glycosides, amino acids, peptides, etc. through microbial fermentation metabolism, which enhances skin absorption and improves the antioxidant effect of the product. Seabuckthorn is recognized as the "king of vitamin C" among plants in the world. Seabuckthorn leaves were listed as new resource foods in 2013. They contain a variety of active compounds that are beneficial to the human body, such as vitamin C, proteins, alkaloids, polyphenols, flavonoids and mineral elements. Among them, flavonoids are one of its important functional active ingredients. Flavonoids can effectively scavenge free radicals and inhibit the growth of melanin, protect dermal cells, and prevent cell oxidation from producing wrinkles. Seabuckthorn leaves are rich in resources, but the current level of development and utilization is still relatively low.

[0004] Traditionally, commonly used production bacteria include lactic acid bacteria and yeast, which occupy a dominant position in the raw materials of daily chemical products. The probiotics of the genus Eurotium are the dominant bacteria that are naturally formed by the "flowering" process of Fuzhuan tea under specific temperature and humidity conditions. Scientific research has found that the extracellular enzymes such as amylase and oxidase secreted by Eurotium can catalyze the conversion of starch in tea into soluble sugars, catalyze the conversion of polyphenols, and produce small molecular active substances. Therefore, Fuzhuan tea has the effects of promoting digestion, reducing fat and weight, and regulating blood sugar metabolism. Summary of the invention

[0005] The purpose of the present invention is to provide a Eurotium cristatum, the strain name is Eurotium cristatum K8824, which has the characteristics of fast growth rate and high fermentation activity, the tea fermentation liquid of the strain has good anti-skin aging effect, and the tea-sea buckthorn leaf composite fermentation liquid has significant antioxidant, anti-aging and whitening effects. The strain has been preserved in the China Center for Type Culture Collection on April 7, 2024, and the preservation number of the strain is: CCTCC M 2024643, Latin name: Eurotiumcristatum, address: Wuhan University, Wuhan City, Hubei Province, Postal Code: 430072, Telephone: 027-68754052.

[0006] The inventor isolated and cultured the high antioxidant activity of Eurotium crown-like fungus isolated from Shaanxi Fuzhuan tea, screened and obtained the K8824 strain, and studied and expanded it. The isolated colony is milky white to light yellow, with a smooth and moist surface, slender hyphae, sparse branches, light yellow edges, and a convex center. The closer to the center of the colony, the darker the color becomes, yellow and dark yellow. The spores are oval, the cap is bowl-shaped, the inner wall is smooth, and the outer wall has a radial texture.

[0007] The tea fermentation liquid of Eurotium cristatum K8824 preserved in the present invention is used for preparing anti-aging skin care products, and the tea-sea buckthorn leaf composite fermentation liquid is used for preparing anti-oxidation, anti-aging and whitening skin care products.

[0008] The preparation method of tea fermentation liquid is as follows:

[0009] ① Grind the tea leaves into 40-100 mesh fine powder, mix with purified water in a ratio of 3:90-10:90 and stir evenly to obtain a fermentation medium, and sterilize at 121°C for 15 minutes;

[0010] ② The method for preparing the activated seed solution of Eurotium cristatum K8824 is as follows:

[0011] The preserved Eurotium cristatum K8824 was streaked on a PDA plate medium and activated at 28±1℃ for 48-72h. The spores on the plate were scraped and washed with sterile water. The spore liquid was fully shaken and mixed to form a spore suspension, and the concentration of the spore suspension was adjusted to 1.0×10 7 CFU / mL, and the activated seed solution of Eurotium cristatum K8824 was obtained.

[0012] ③ Inoculate the seed liquid of Eurotium cristatum into the fermentation medium for fermentation, with the inoculation amount being 3-8%, and culture in a shaking table at 28±2°C for 40-84h at a shaking speed of 120-200r / min.

[0013] The preparation method of tea-sea buckthorn leaf composite fermentation liquid is as follows:

[0014] ① Grind the tea and seabuckthorn leaves into 40-100 mesh fine powders respectively, and then mix the tea powder and seabuckthorn leaf powder in a mass ratio of 7:3-5:5 to obtain a tea-seabuckthorn leaf fermentation substrate;

[0015] ② Mix the tea-sea buckthorn leaf fermentation substrate with purified water in a ratio of 3:90 to 10:90 and stir evenly to obtain a fermentation medium, and sterilize at 121° C. for 15 min;

[0016] ③ The preparation method of the spore suspension of Eurotium cristatum is the same as that of the tea fermentation liquid. The seed liquid of Eurotium cristatum is inoculated into the fermentation medium for fermentation. The inoculation amount is 3-8%. The culture is shaken at 28±2°C for 40-84h at a shaking speed of 120-200r / min.

[0017] Beneficial Effects

[0018] First, the K8824 strain of the present invention can better adapt to the mixed fermentation system of tea and seabuckthorn leaves, and the fermentation speed is significantly higher than that of the crown-like spores strain K8821. When the fermentation substrate is tea, the elastase inhibition rate of its fermentation liquid is 74.19%, which has a good anti-aging effect. At the same time, the DPPH free radical scavenging rate of the tea-seabuckthorn composite fermentation liquid obtained by the present invention reaches 87.15%, the hydroxyl free radical scavenging rate reaches 85.22%, the FRAP antioxidant capacity reaches 3.6mmol / L, the ABTS free radical scavenging rate reaches 0.89mmol / L, the tyrosinase inhibition rate reaches 85.29%, and the elastase inhibition rate reaches 83.58%, which has good antioxidant, whitening and anti-aging effects, and is very suitable for preparing antioxidant, anti-aging and whitening skin care products. According to the results of preliminary experiments, if the Cremaster tremella K8824 of the present invention is cultured in PDB medium, the fermentation broth after culture does not have the anti-aging, antioxidant and whitening effects. The speculated mechanism may be that the Cremaster tremella K8824 preserved in the present invention and the phenolic, flavonoid and protein components in tea and sea buckthorn leaves are transformed under the action of biological enzymes during the fermentation process, thereby increasing the content of antioxidant, whitening and anti-aging substances such as flavonoid glycosides, organic acids, polypeptides, amino acids and other secondary metabolites, making the fermentation broth have stronger biological activity, thereby playing a role in whitening, antioxidant and anti-aging.

[0019] Secondly, the present invention uses the fermentation liquid obtained from the cristatum cristatum to prepare skin care products. Experimental results show that after being prepared into skin care products, the skin care products have sustained and significant anti-aging, antioxidant and whitening effects, and the processing and preparation process of the skin care products have no significant effect on the anti-aging, antioxidant and whitening effects of the fermentation liquid. At the same time, the skin care products prepared after adding the fermentation liquid also have good sunscreen (anti-ultraviolet effect) and anti-aging effects, and all experimenters responded that they have good skin feel and lubricating and moisturizing effects, and have strong market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 This is a colony morphology diagram of the Eurotium cristatum K8824 preserved in the present invention on PDA culture medium.

[0021] Figure 2 This is the growth curve of the Eurotium cristatum K8824 preserved in the present invention in tea-sea buckthorn leaves.

[0022] Figure 3 This is the growth curve of Eurotium cristatum K8821 isolated by the applicant in tea-sea buckthorn leaves.

[0023] Figure 4 The R2 value of skin elasticity of the anti-aging skin care product prepared by fermenting the Eurotium cristatum K8824 preserved in the present invention is tested. DETAILED DESCRIPTION

[0024] The present invention is described in detail below with reference to examples. All methods and techniques, unless otherwise specified, are conventional methods and techniques.

[0025] The preparation method of tea fermentation liquid is as follows:

[0026] ① Grind the tea leaves into 40-100 mesh fine powder, mix with purified water in a ratio of 3:90-10:90 and stir evenly to obtain a fermentation medium, and sterilize at 121°C for 15 minutes;

[0027] ② Inoculate the seed liquid of Eurotium cristatum into the fermentation medium for fermentation, with the inoculation amount being 3-8%, and culture in a shaking table at 28±2°C for 40-84h at a shaking speed of 120-200r / min.

[0028] The preparation method of seabuckthorn leaf fermentation liquid is as follows:

[0029] ① Grind the seabuckthorn leaves into 40-100 mesh fine powder, mix with purified water in a ratio of 3:90-10:90 and stir evenly to obtain a fermentation medium, and sterilize at 121°C for 15 minutes;

[0030] ② Inoculate the seed liquid of Eurotium cristatum into the fermentation medium for fermentation, with the inoculation amount being 3-8%, and culture in a shaking table at 28±2°C for 40-84h at a shaking speed of 120-200r / min.

[0031] The preparation method of tea-sea buckthorn leaf composite fermentation liquid is as follows:

[0032] ① After grinding tea and seabuckthorn leaves into 40 mesh fine powder respectively, the tea and seabuckthorn leaves are mixed evenly in a mass ratio of 7:3 to 5:5 to obtain a tea-seabuckthorn leaf fermentation substrate;

[0033] ② Mix the tea-sea buckthorn leaf fermentation substrate with purified water in a ratio of 3:90 and stir evenly to obtain a fermentation medium, and sterilize at 121°C for 15 minutes;

[0034] ③ Inoculate the seed liquid of Eurotium cristatum into the fermentation medium for fermentation, with the inoculation amount being 3-8%, and culture in a shaking table at 28±2°C for 40-84h at a shaking speed of 120-200r / min.

[0035] Test Example 1: DPPH-free radical scavenging experiment

[0036] Prepare 0.5mmol / L DPPH-ethanol solution, dilute the fermentation sample solution 10 times, take 0.1mL sample and add 0.9mL DPPH solution to mix, the control group is 0.1mL distilled water and add 0.9mL DPPH solution to mix, react in the dark for 30min, measure its absorbance at a wavelength of 517nm, and the DPPH free radical scavenging rate is calculated by the formula:

[0037] DPPH free radical scavenging rate (%) = (1-A1 / A0) × 100%, A1 is the sample absorbance; A0 is the blank control absorbance. The higher the scavenging rate, the better the antioxidant effect of the fermentation broth.

[0038] Test Example 2: Hydroxyl free radical scavenging ability determination

[0039] Experimental method: A colorimetric kit (Nanjing Jiancheng Bioengineering Institute) was used to detect the hydroxyl radical scavenging ability. According to the instructions of the hydroxyl radical scavenging ability assay kit (A018-1-1), 0.2 mL of a 10-fold diluted fermentation sample solution was added to the Fenton system to generate hydroxyl radicals. The absorbance of each sample was measured at 550 nm using the Griess reagent.

[0040] Test Example 3: Determination of ABTS free radical scavenging ability

[0041] Experimental method: A colorimetric kit (Nanjing Jiancheng Bioengineering Institute) was used to detect the antioxidant capacity of ABTS free radicals. According to the kit instructions (A015-2), the concentration of the sample was diluted 10 times, and the antioxidant capacity of ABTS free radicals was determined. The absorbance was detected at 734 nm, and the value was expressed as the value equivalent to 1 mmol / L Trolox.

[0042] Test Example 4: FRAP Total Antioxidant Capacity Assay

[0043] Experimental method: A colorimetric kit (Nanjing Jiancheng Bioengineering Institute) was used to detect the FRAP free radical antioxidant capacity. According to the kit instructions (A015-3-1), a 10-fold diluted sample solution (5 μL) was added to 2,4,6-tripyridine-s-triazine (TPTZ) in an acidic medium to detect FRAP. The absorbance was measured at 593 nm, and the FRAP value was expressed as the equivalent of 1 mmol / L FeSO4-7H2O. The FRAP total antioxidant capacity results are shown in Table 1.

[0044] Test Example 5: Tyrosinase Inhibition Ability Determination

[0045] Experimental method: Refer to "T / GDCA006-2021 Daily Chemical Raw Materials Tyrosinase Activity Inhibition Test Method (In Vitro Method)" to conduct tyrosinase activity inhibition test, and the fermentation broth sample was diluted 10 times for measurement.

[0046] Test Example 6: Determination of Elastase Activity Inhibition Rate

[0047] Experimental method: 85μL 50mmol / L Tris-HCL buffer (pH 8.0) was mixed with 15μL of 100-fold diluted sample solution, and then 25μL of elastase solution (60mU / mL) was added. The mixture was incubated at 25℃ for 15min, and then 25uL 1.015mmol / L N-succinyl-alanine-alanine-alanine-p-nitroaniline (AAAPAN) solution was added. After 15min, the absorbance was measured at 410nm. Both the elastase solution and AAAPAN solution were prepared with 50mmol / LTris-HCl buffer (pH8.0), and the inhibition rate of the tested sample on the elastase activity was calculated.

[0048] The inhibition rate of elastase activity was calculated by the following formula:

[0049] Elastase activity inhibition rate (%) = [1-(CD) / AB] x 100%,

[0050] Wherein, A is the absorbance of the reaction solution without sample; B is the absorbance of the reaction solution without sample and enzyme;

[0051] C is the absorbance of the reaction solution containing the sample and the enzyme; D is the absorbance of the reaction solution without the enzyme.

[0052] The higher the elastase inhibition rate, the better the anti-aging effect of the fermentation liquid.

[0053] Test Example 7: Determination of antibacterial ability

[0054] Under sterile conditions, pour beef extract peptone agar medium into a sterilized test tube to prepare a slant medium. Inoculate the test bacteria (Escherichia coli) on the medium by streaking, and culture at 37°C for 24 hours. Dilute the activated bacteria with high-temperature sterilized distilled water, and adjust the bacterial suspension to 1.0×10 8 CFU / mL is ready for use and is prepared before use. Soak blank drug-sensitive paper sheets (6.0mm in diameter and 1.0mm in thickness) in different fermentation broth samples respectively. After fully soaking for 1 hour, take out the filter paper sheets and wait for the solvent to evaporate. Take 0.1mL of the bacterial suspension and add it to a sterile plate. Pour 15-20mL of beef extract peptone agar medium to mix the bacterial suspension with the medium. After the medium is condensed, use sterile tweezers to clamp the dried filter paper sheets and stick them on the surface of the bacterial culture dish, 3 sheets per dish. Put the culture dish in a 37℃ constant temperature incubator and invert it for 24 hours, then take it out and measure the diameter of the antibacterial ring (including the patch) with a vernier caliper and record it.

[0055] Example 1

[0056] A kind of Eurotium cristatum K8824, which is isolated from Fuzhuan tea produced in Shaanxi, grows fast in a mixture of tea and sea buckthorn leaves, and the fermented liquid has antioxidant, anti-aging and whitening effects, and can be used to prepare skin care products. The strain was deposited in the China Center for Type Culture Collection on April 7, 2024. The preservation number of K8824 is: CCTCC M 2024643, Latin name: Eurotium cristatum. Address: Wuhan University, Wuhan City, Hubei Province, Postal Code: 430072, Telephone: 027-68754052.

[0057] The Eurotium cristatum K8824 preserved in the present invention is isolated from Fuzhuan tea produced in Shaanxi Province, and its strain morphological characteristics are shown in Figure 1 The inventors also isolated another strain of Eurotium cristatum in their laboratory, named Eurotium cristatum K8821. The fermentation rates of the strains in the mixture of green tea and seabuckthorn leaves were measured respectively. The results are shown in Figure 2 , Figure 3 shown.

[0058] The morphological characteristics of the preserved Eurotium cristatum K8824 of the present invention are as follows: the isolated colonies are milky white to light yellow, the surface is smooth and moist, the hyphae are slender, the branches are sparse, the edges of the colonies are light yellow, the center is convex, and the color becomes darker to yellow and dark yellow as it approaches the center of the colony. The spores are oval, the cap is bowl-shaped, the inner wall is smooth, and the outer wall has radial textures.

[0059] In Example 1, tea leaves and sea buckthorn leaves were crushed to 40 mesh, the mixing ratio of tea powder or sea buckthorn leaf powder or the mixture to pure water was 3:90, the mixing ratio of tea powder to sea buckthorn leaf powder was 6:4, the inoculation amount was 5%, and the culture was shaken at 28° C. for 48 hours at a shaking speed of 150 r / min.

[0060] According to the results of the preliminary experiment, the fermentation time of the fermentation broth is 72h.

[0061] Tea-sea buckthorn leaf fermentation substrate, pure green tea fermentation substrate and pure sea buckthorn leaf fermentation substrate were used as culture medium, and strains K8821 and K8824 were used for fermentation. After 72 hours of culture, the fermentation liquid was taken out and the data were measured as shown in Table 1.

[0062] Comparative Example:

[0063] The tea leaves were crushed to 40 meshes, the tea powder and purified water were mixed in a ratio of 3:90, and soaked in hot water at 80°C for 2 hours to obtain unfermented tea.

[0064] The sea buckthorn leaves were crushed into 40 meshes, the sea buckthorn leaf powder was mixed with purified water in a ratio of 3:90, and the mixture was soaked in hot water at 80° C. for 2 hours to obtain unfermented sea buckthorn leaves.

[0065] The tea leaves and sea buckthorn leaves are both crushed to 40 meshes, the mixing ratio of tea powder or sea buckthorn leaf powder or the mixture with purified water is 3:90, and the mixing ratio of tea powder to sea buckthorn leaf powder is 6:4. The mixture is soaked in hot water at 80°C for 2 hours to obtain unfermented tea-sea buckthorn leaves.

[0066] Table 1 The results of the determination of various data of different strains and different culture media

[0067]

[0068] The data in Table 1 show that the antioxidant, whitening and anti-aging properties of K8824 strain in tea-sea buckthorn leaves are relatively balanced, significantly higher than those of K8821 strain, and the antioxidant capacity, tyrosinase inhibition rate and elastase inhibition rate of K8824 strain after tea-sea buckthorn leaf fermentation are significantly higher than those of unfermented tea-sea buckthorn leaves, indicating that K8824 strain produces antioxidant, anti-aging and whitening substances during the fermentation process. The most notable feature of K8824 is that it can produce anti-aging substances during the fermentation process of tea-sea buckthorn leaves, which is not obvious in the environment of pure sea buckthorn leaves as fermentation liquid, while K8821 has weak anti-aging, antioxidant and whitening effects in the environment of tea-sea buckthorn leaves. It may be that during the fermentation process, K8824 strain converts active substances such as phenolic substances and flavonoids in tea leaves and sea buckthorn leaves into more effective anti-aging substances.

[0069] From the data in Table 1, K8824 has a significantly stronger ability to transform antioxidant, whitening and anti-aging substances than K8821, and its fermentation speed is fast. According to the results of mycelium dry weight determination ( Figure 2 , Figure 3 ), the growth rate of K8824 was significantly higher than that of K8821, which may be because the strain K8824 is more adaptable to the fermentation liquid environment of seabuckthorn leaves and green tea, and it also has a strong antibacterial effect.

[0070] Table 2 Inhibition zones of Escherichia coli in different groups

[0071] Strains and fermentation broth Escherichia coli inhibition zone (mm) K8824 Tea-Seabuckthorn Leaf 10.72 K8824 Tea 10 K8824 Sea buckthorn leaves 9.33 K8821 Tea-Seabuckthorn Leaf 8.21 K8821 Tea 7.44 K8821 Sea buckthorn leaves 7.20

[0072] According to the test results in Table 2, the tea-sea buckthorn leaf composite fermentation liquid obtained by composite fermentation of Eurotium cristatum K8824 and tea-sea buckthorn leaf has a good inhibitory effect on Escherichia coli. The inhibitory effect of K8821 tea-sea buckthorn leaf composite fermentation liquid on Escherichia coli is significantly lower than that of K8824. When tea or sea buckthorn leaf is fermented alone, the inhibitory effect on Escherichia coli is weakened. Using tea and sea buckthorn leaf as composite raw materials, the tea-sea buckthorn leaf composite fermentation liquid obtained by Eurotium cristatum fermentation has a better inhibitory effect on Escherichia coli.

[0073] Example 2

[0074] Prepare materials by mass percentage: 5.0% lanolin, 8.0% tea-sea buckthorn leaf composite fermentation liquid, 2.5% white vaseline, 4.0% stearic acid, 0.05% propyl parahydroxybenzoate, 0.1% methyl parahydroxybenzoate, 0.05% EDTA-2Na, 5.0% 1,2-propylene glycol, 1.0% triethanolamine, 74.3% deionized water. Heat lanolin, tea-sea buckthorn leaf composite fermentation liquid, white vaseline, stearic acid, propyl parahydroxybenzoate at 72-82°C, stir and mix to obtain a first mixture; heat methyl parahydroxybenzoate, EDTA-2Na, 1,2-propylene glycol, triethanolamine, deionized water at 72-82°C, stir and mix to obtain a second mixture; add the first mixture to the second mixture under stirring, keep warm and emulsify for 20 minutes, let stand, cool to room temperature 15-30°C, and fill to obtain an anti-ultraviolet, anti-oxidant, whitening skin care product.

[0075] Comparative Example 1

[0076] Preparation of blank anti-ultraviolet antioxidant whitening skin care product: Same as Example 2, except that the tea-sea buckthorn leaf composite fermentation liquid is not contained, the amount of water added is 74.8%, and the others are the same.

[0077] Comparative Example 2

[0078] Preparation of control anti-ultraviolet antioxidant whitening skin care product: same as Example 2, except that the tea-sea buckthorn leaf composite fermentation liquid was not contained, 8.0% trimethylol salicylate was added, and the rest were the same.

[0079] Anti-ultraviolet whitening test

[0080] The anti-ultraviolet antioxidant whitening skin care product obtained in Example 2 was tested on volunteers. Test population: 30 volunteers, all female, aged 18 to 55, healthy, for 30 days. The test subjects applied the sunscreen skin care product to the exposed skin 15 minutes before going out, and reapplied it if they were outdoors for more than 3 hours. The trial results are shown in Table 3.

[0081] Table 3 Trial results of anti-ultraviolet and anti-oxidant whitening skin cream

[0082]

[0083] As can be seen from Table 3, according to the corresponding test standards, after the 30-day trial, all the test subjects of the anti-ultraviolet antioxidant whitening skin care products did not find redness, itching and irritation reactions, and they reported that they felt smooth and comfortable, without greasy feeling, and were particularly suitable for use in summer. From the above examples, it can be seen that the tea-sea buckthorn leaf composite fermentation liquid can well inhibit the activity of tyrosinase, and the skin care products added with the tea-sea buckthorn leaf composite fermentation liquid can control the formation of melanin, and the anti-ultraviolet whitening effect is better than the same proportion of trimethylol salicylate, and has a good whitening effect.

[0084] Example 3

[0085] The materials were prepared by mass percentage: 3.0% butylene glycol, 4.0% glycerol, 2.0% trehalose, 0.1% xanthan gum, 5% β-glucan, 2.0% glyceryl stearate, 3.0% capric triglyceride, 4.0% cyclohexylsiloxane, 1.5% shea butter, 66.8% deionized water, 8.0% tea-sea buckthorn leaf composite fermentation liquid, 0.1% essence and 0.5% phenoxyethanol.

[0086] Glyceryl stearate, capric triglyceride, cyclohexylsiloxane and shea butter are mixed and heated to melt to obtain a first mixture. Butylene glycol, glycerol, xanthan gum and deionized water are heated and stirred to mix, β-glucan and trehalose are added, and stirred evenly to obtain a second mixture; the second mixture is poured into the first mixture, stirred and homogenized and emulsified. When the temperature drops below 50°C, the tea-sea buckthorn leaf composite fermentation liquid and essence prepared in Example 1 are added in sequence and allowed to stand, cooled, and filled to obtain an anti-aging skin care product.

[0087] Comparative Example 3

[0088] Preparation of control anti-aging skin care product: Same as Example 3, except that the tea-sea buckthorn leaf composite fermentation liquid was not contained, 8.0% commercially available tea fermentation extract was added, and the rest were the same.

[0089] Anti-aging test

[0090] The anti-aging test selected people with wrinkles on the outer corners of the eyes (wrinkle levels on both sides were between 1 and 7, and the wrinkle levels on the left and right sides were the same. 20 people aged 35 to 50 years old, with a male and female ratio of 50%. No products (skin care products, external medicines, internal health care products) should be used on the test area 15 days before. Before the test, the volunteers were required to wash their faces and enter the climate-controlled room (22±1℃, relative humidity 50%) 2 hours after washing to sit for more than 20 minutes and remain relaxed. In the test, half of the face was tested according to the random table, one side was the sample and the other side was the control. , the left and right corners of the volunteers were selected for skin elasticity test using ElastiMeter skin elasticity measuring instrument (Delfin, Finland), and the average value of the three tests was taken as the skin elasticity data of the left and right corners of the eyes. After the test, the volunteers were required to use it as required, applying the sample (anti-aging skin care product in Example 3) on one corner of the eye and the control product (control anti-aging skin care product) on the other corner of the eye. The test was used continuously for 63 days and the subjects were required to return for a visit on the 7th, 14th, 21st, 28th, 35th, 42nd, 49th, 56th and 63rd days of the test. The results of measuring the skin elasticity R2 value are shown in Figure 4 .

[0091] from Figure 4 It can be clearly seen that the anti-aging skin care product prepared by the present invention has a very significant effect on the skin at the corner of the eye. As the use time increases, the skin elasticity R2 value shows a significant upward trend, while the control has no significant effect on the skin elasticity R2 value. This shows that the anti-aging skin care product prepared by the present invention can significantly improve skin elasticity and has anti-aging effects.

Claims

1. A cristatum cristatum, characterized in that: The name of the described Eurotium cristatum is Eurotium cristatum K8824, and its Latin name is: Eurotium cristatum , deposited in China Center for Type Culture Collection with the accession number: CCTCC M 2024643.

2. A use of the Eurotium cristatum according to claim 1, characterized in that: The tea fermentation liquid of Eurotium cristatum K8824 is suitable for producing anti-aging skin care products.

3. A use of the Eurotium cristatum according to claim 1, characterized in that: The tea-sea buckthorn leaf composite fermentation liquid of Eurotium cristatum K8824 is suitable for producing skin care products with antioxidant, anti-aging and whitening effects.

4. The use of Eurotium cristatum according to claim 3, characterized in that: The preparation method of the tea-sea buckthorn leaf composite fermentation liquid is as follows: ① Grind the tea and seabuckthorn leaves separately and sieve them, then mix the tea powder and seabuckthorn leaf powder evenly to obtain a tea-seabuckthorn leaf fermentation substrate; ② Mix the tea-sea buckthorn leaf fermentation substrate with purified water and stir evenly to obtain a fermentation medium, and sterilize it; ③Inoculate the seed liquid of Eurotium cristatum into the fermentation medium and ferment for 40 to 84 hours.

5. The use of Eurotium cristatum according to claim 2, characterized in that: The preparation method of the tea fermentation liquid is as follows: Grinding the tea leaves into powder and mixing with purified water and stirring evenly to obtain a tea fermentation medium, and sterilizing the medium; The seed liquid of Eurotium cristatum was inoculated into the tea fermentation medium and fermented for 40 to 84 hours.

6. The use of Eurotium cristatum according to claim 4 or 5, characterized in that: The inoculation amount of the Eurotium cristatum is 3-8%, and the culture condition is shaking culture.

7. The use of Eurotium cristatum according to claim 4 or 5, characterized in that: The shaking culture is specifically carried out on a shaking table at 28±2°C, with a shaking speed of 120-200 r / min.

8. The use of Eurotium cristatum according to claim 4, characterized in that: The mass ratio of the tea powder to the seabuckthorn leaf powder is 7:3-5:

5.

9. The use of Eurotium cristatum according to claim 4 or 5, characterized in that: The mixing ratio of the tea-sea buckthorn leaf fermentation substrate or tea powder to the purified water is 3:90-10:90.

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