Bacterial strain composition, radix paeoniae rubra fermentation liquor prepared from bacterial strain composition and application of bacterial strain composition in cosmetics

Abstract

The invention belongs to the technical field of preparation of microorganisms and cosmetics, and particularly relates to a strain composition, radix paeoniae rubra fermentation liquor prepared from the strain composition and application of the strain composition in cosmetics. Specifically, Pseudoalteromonas sp. SE9 and Schizoscharomyces pombe JM2-16 are utilized to perform fermentation treatment on radix paeoniae rubra root powder, and the finally obtained radix paeoniae rubra fermentation liquor has multiple effects of resisting oxidation, promoting proliferation and migration of keratinocytes, promoting skin injury repair, resisting allergy, resisting inflammation, relieving and the like, is high in safety, can be used as a cosmetic component, and can be used as a cosmetic preparation. Therefore, the method has good practical application value.

Description

Technical Field

[0001] The invention belongs to the technical field of microorganisms and cosmetic preparation, and particularly relates to a strain composition and a red peony root fermentation liquid prepared therefrom and application thereof in cosmetics. Background Art

[0002] Red Peony Root Paeonia veitchii Lynch ) is a traditional Chinese medicinal material with a bitter taste and the effects of clearing heat and cooling blood, dispersing blood stasis and relieving pain. Studies have found that red peony root has good antioxidant, soothing and skin barrier repair capabilities. However, when red peony root extract is directly applied, its bioavailability and stability of active ingredients are poor, which limits its wide application in the cosmetic field.

[0003] In order to solve this problem, fermentation technology has received widespread attention in recent years as an important means to improve the efficacy of plant active ingredients. Through microbial fermentation, the active ingredients in plants can be transformed by microbial metabolism, their bioavailability and stability can be improved, and their antioxidant properties can be enhanced. In particular, certain unique microbial strains, with their unique metabolic characteristics, can effectively improve the antioxidant effect of plant ingredients. However, there are few reports on the use of microorganisms to ferment red peony root to obtain corresponding products. Summary of the invention

[0004] Based on the above-mentioned deficiencies in the prior art, the present invention provides a strain composition and a red peony root fermented liquid prepared therefrom and its application in cosmetics. Specifically, the present invention utilizes Pseudoalteromonas and Schizosaccharomyces pombe to ferment red peony root, promotes the conversion and synergism of antioxidant components in red peony root through the fermentation process, significantly improves its biological activities such as antioxidant, and provides a new technical path for the development and application of red peony root and. Based on the above-mentioned research results, the present invention is completed.

[0005] In order to achieve the above technical objectives, the present invention relates to the following technical solutions: One aspect of the present invention provides a strain composition, the strain composition comprises Pseudoalteromonas ( Pseudoalteromonas sp.) SE9 and Schizosaccharomyces pombe ( Schizosaccharomyces pombe )JM2-16; Among them, Pseudoalteromonas Pseudoalteromonas sp.) SE9, which was deposited in the China Center for Type Culture Collection (address: Wuhan University, Luojia Mountain, Wuchang, Wuhan City, Hubei Province) on December 13, 2024, and its preservation number is CCTCC NO: M 20242809; specifically, the Pseudoalteromonas SE9 is actually Antarctic Pseudoalteromonas, a bacterium that can grow stably in extremely cold environments and has a strong ability to synthesize antioxidants.

[0006] Schizosaccharomyces pombe ( Schizosaccharomyces pombe) JM2-16, which was deposited in the China Center for Type Culture Collection (address: Wuhan University, Luojia Mountain, Wuchang, Wuhan City, Hubei Province) on December 13, 2024, and its deposit number is CCTCC NO: M 20242810. It can produce a variety of beneficial metabolites through the fermentation process, which helps to enhance the antioxidant properties of red peony root.

[0007] The second aspect of the present invention provides the use of the above-mentioned strain composition in red peony root fermentation.

[0008] The third aspect of the present invention provides a method for fermenting red peony root, which comprises: adding the above-mentioned strain composition to a liquid culture medium containing red peony root powder for fermentation culture, and separating and purifying to obtain red peony fermentation liquid.

[0009] The liquid culture medium may be yeast extract peptone dextrose medium (YPD), which can be obtained commercially.

[0010] The specific method of the fermentation culture comprises: Schizosaccharomyces pombe ) JM2-16 was first added to the liquid medium containing red peony root powder for the first stage of fermentation culture, and then Pseudoalteromonas ( Pseudoalteromonas sp.)SE9 for the second stage of fermentation culture; Wherein, in the first stage of fermentation culture, the fermentation culture temperature is 25-35°C (preferably 30°C), the rotation speed is controlled to 150-200 rpm (preferably 180 rpm), the fermentation culture time is 10-60h (preferably 48h); the fermentation pH is 5.0-6.5; In the second stage of fermentation culture, the fermentation culture temperature is 10-20°C (preferably 15°C), the rotation speed is controlled at 150-200 rpm (preferably 180 rpm), and the fermentation culture time is 10-60h (preferably 48h).

[0011] The specific method for separating and purifying the red peony root fermentation liquid comprises: subjecting the fermentation liquid to centrifugation, high temperature sterilization, decolorization and filtering.

[0012] Wherein, the specific conditions of the centrifugation are: centrifugation at 5000-15000 r / min (preferably 11000 r / min) for 5-60 min (preferably 20 min); The specific conditions of high temperature sterilization are: sterilization at 121°C for 30 min; The decolorization can be carried out by using activated carbon.

[0013] The filtration may be performed using a 0.22 μm filter.

[0014] A fourth aspect of the present invention provides a red peony root fermentation liquid prepared by the above method.

[0015] A fifth aspect of the present invention provides the use of the red peony root fermented liquid in the preparation of cosmetics, wherein the cosmetics have any one or more of the following effects: (a) Antioxidant; (b) Promote the proliferation and migration of keratinocytes; (c) Promote skin damage repair; (d) Anti-allergic and soothing; (e) Anti-inflammatory and soothing.

[0016] The cosmetics may specifically be skin care products. The cosmetics may also contain any other raw material ingredients allowed to be added in the cosmetics field, and different cosmetic dosage forms may be prepared therefrom, which are not specifically limited here. At the same time, the cosmetics obviously do not contain live bacteria.

[0017] Beneficial technical effects of one or more of the above technical solutions: The above technical solution utilizes Pseudoalteromonas ( Pseudoalteromonas sp.) SE9 and Schizosaccharomyces pombe ( Schizosaccharomyces pombe ) JM2-16 ferments red peony root powder, and the final red peony fermentation liquid has multiple functions such as anti-oxidation, promoting the proliferation and migration of keratinocytes, promoting skin damage repair, anti-allergy, anti-inflammatory and soothing, etc. It is highly safe and can be used as a cosmetic ingredient, so it has good practical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The DPPH radical scavenging rate test results of the red peony root fermentation liquid in the embodiment of the present invention are shown.

[0019] Figure 2 The figure is the test result of hydroxyl radical scavenging rate of red peony root fermentation liquid in the embodiment of the present invention.

[0020] Figure 3 The results are shown in Table 1. These are the superoxide anion clearance test results of the red peony root fermentation liquid in the embodiments of the present invention.

[0021] Figure 4 It is the cell survival rate of different concentrations of A2~C2 in the embodiment of the present invention.

[0022] Figure 5 This is the effect of the red peony root fermentation liquid on the proliferation of HaCaT cells in the example of the present invention.

[0023] Figure 6 The figure shows the result of cell scratch test in the embodiment of the present invention.

[0024] Figure 7The results are the results of the permease inhibition test of the red peony root fermentation broth in the embodiments of the present invention.

[0025] Figure 8 The effect of the fermented broth of red peony root on the content of inflammatory factors in cells in the embodiment of the present invention is shown in Figure 1. A is tumor necrosis factor (TNF-α); B is interleukin (IL-6). DETAILED DESCRIPTION

[0026] It should be noted that the following detailed descriptions are exemplary and are intended to provide further explanation of the present invention. Unless otherwise specified, all technical and scientific terms used in the present invention have the same meanings as those commonly understood by those skilled in the art to which the present invention belongs.

[0027] It should be noted that the terms used herein are only for describing specific embodiments and are not intended to limit the exemplary embodiments of the present invention. As used herein, unless the context clearly indicates otherwise, the singular form is also intended to include the plural form. In addition, it should be understood that when the terms "comprising" and / or "including" are used in this specification, it indicates the presence of features, steps, operations, devices, components and / or combinations thereof.

[0028] The present invention is further explained by the following examples, but they are not intended to limit the present invention. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0029] Example 1. Preparation of Fermentation Broth Activation of strains: Pseudoalteromonas SE9 and Schizosaccharomyces pombe JM2-16 were inoculated at 1% in sea salt medium and yeast extract peptone glucose medium, respectively. The activation temperatures were 15°C and 30°C, respectively, and the culture was carried out at 180 rpm for 12 h.

[0030] Expanded culture: According to the formula in Table 1, the fermentation medium was prepared. Sea salt medium (g / 100ml) was added to the A1~A2 conical flasks: 0.3g of yeast powder, 0.5g of peptone, and 15g of artificial seawater (3.5% sea salt solution); Yeast Extract Peptone Dextrose Medium was used in the B1~B2 and C1~C2 conical flasks. 5g of red peony root powder (Nanjing Tongrentang red peony powder, product number: 10130793948953) was added to A2, B2, and C2 respectively. At the same time, red peony root extract (liquid-to-solid ratio 1:20) was set as the control group D. The activated bacterial solution of Pseudoalteromonas SE9 and Schizosaccharomyces pombe JM2-16 was diluted to 1% ( v / v) were inoculated into the fermentation medium in sequence. The culture temperature of Pseudoalteromonas SE9 was 15℃, and the culture temperature of Schizosaccharomyces pombe JM2-16 was 30℃. The flasks were sealed with gauze, fermented at 180 rpm, and the pH was controlled at 6.0-6.5. C1~C2 was fermented for 48h, and then Pseudoalteromonas SE9 was inoculated and the culture medium was placed in a 15℃ incubator for further fermentation.

[0031] Table 1 Formula of each strain

[0032] Determination of fermentation endpoint: The fermentation endpoint is determined by detecting the changes in antioxidant activity (DPPH free radical scavenging rate) during the fermentation process. Fermentation is 96 hours.

[0033] Bacteria collection treatment: After fermentation, the product was centrifuged at 11000 r / min for 20 min to obtain the clarified red peony root fermentation product. After sterilization in a 121 ℃ high-temperature autoclave for 30 min, it was decolorized with activated carbon and filtered with a 0.22 μm needle filter. The liquid was used as the fermentation test liquid.

[0034] 2. Analysis of Antioxidant Activity of Fermentation Broth Three groups of microbial fermentation broth samples (A1-C1), three groups of red peony root fermentation broth samples (A2-C2), and one group of red peony root water extract samples (marked as D) were set up as the experimental sample groups of this study to compare and analyze the differences between red peony root fermentation broth and other types of samples (i.e., microbial fermentation broth and red peony root water extract).

[0035] DPPH free radical scavenging rate detection: prepare 0.1 mM DPPH solution, take 0.002 g DPPH each time and dissolve it in 50 mL ethanol to prepare 0.1 mM DPPH solution, and store it in the dark. The experiment can be carried out in a 96-well plate, with three replicates for each group to reduce experimental errors, and anhydrous ethanol is used to adjust the zero. Take 180 μL of the sample group and add 90 μL of DPPH solution; the control group takes 90 μL of DPPH solution and adds 180 μL of distilled water; blank group: add 90 μL of anhydrous ethanol to 180 μL of sample solution. Shake in the dark for 10 min and test the absorbance. The DPPH free radical scavenging rate calculation formula is as follows: DPPH clearance rate = [A 对照组 -(A 样品组 -A 空白组 )] / A 对照组 ×100% Salicylic acid method was used to determine the scavenging ability of red peony root fermentation liquid: Hydroxyl radicals are generated by Fenton reaction, and ·OH oxidizes salicylic acid to produce 2,3-dihydroxybenzoic acid with characteristic absorption at 510 nm. The scavenging rate of ·OH was determined by measuring the product obtained by salicylic acid capturing ·OH. 0.1 mL 9 mM FeSO4, 0.1 mL 9 mM ethanol-salicylic acid, an appropriate amount of plasma water, and finally 0.1 mL 8.8 mM hydrogen peroxide and red peony root fermentation liquid were added to the colorimetric tube. The test object was shaken, and the sample was bathed in 37°C water for 15 min, and its absorbance was measured. The sample absorbance was measured as A. X , blank control is A0, sample background is A X0 , the scavenging rate of hydroxyl free radicals is calculated as follows: Hydroxyl radical scavenging rate = [A0-(A X -A X0 )] / A0×100% Determination of the ability to scavenge superoxide anions by the self-oxidation method of pyrogallol: This reaction can be carried out in a cuvette and is prepared by solution A and solution B. Solution A: First add 0.6 mL of 0.1 mol / l Tris-HCl (pH=8.2) hydrochloride buffer and 0.58 mL of distilled water, mix well and place in a water bath at 25℃ for 20 min, add red peony fermentation broth to wait for the reactant to be measured, and the final concentration is 20 μM. Solution B: Preheated 0.02 mL of 5 mM pyrogallol solution. After the preparation of solution A and solution B, quickly mix and shake well, and measure the absorbance of the solution at 325 nm as the first absorbance value. Measure once every 1 min, and measure continuously for 4 min to calculate the increase in absorbance per minute within the linear range. The absorbance is zeroed and 10 mM Tris-HCl is used to replace the Tris-HCl solution of pyrogallol. △A0 is the self-oxidation rate of pyrogallol; △A is the self-oxidation rate of pyrogallol after adding the sample solution. The superoxide anion clearance rate was calculated as follows: Superoxide anion clearance rate = [(△A0-△A) / △A0] × 100% Experimental results: The free radical scavenging ability test results of red peony root fermentation liquid are shown in ( Figures 1 to 3 ), compared with red peony root water extract and the same type of microbial fermentation liquid, the ability of red peony root fermentation liquid to scavenge free radicals is stronger, among which the red peony root fermentation liquid (A2, C2) fermented by Pseudoalteromonas and Schizosaccharomyces pombe has a particularly obvious DPPH free radical scavenging effect, which is 92.68% and 93.76% respectively. The hydroxyl free radical scavenging effect of red peony root fermentation liquid (B2) fermented by Schizosaccharomyces pombe is as high as 91.38%. The ability of red peony root fermentation liquid to scavenge superoxide anion free radicals is stronger, and there are significant differences ( p<0.01), and the superoxide anion scavenging rates of red peony root fermentation broth (A2, B2, C2) were 65.50%, 74.59%, and 65.28%, respectively. Among them, the red peony root fermentation broth (B2) fermented by Schizosaccharomyces pombe had the most obvious effect. In summary, compared with the red peony root water extract, the red peony root fermentation broth had a stronger free radical scavenging ability. The free radical scavenging rate of red peony root could be improved by fermentation with Pseudoalteromonas and Schizosaccharomyces pombe, and its antioxidant capacity was enhanced, which was not affected by the microbial fermentation broth itself.

[0036] 3. Toxicity test of red peony root fermentation broth on HaCaT cells HaCaT cell model establishment: Keratinocytes (HaCaT) were cultured in DMEM medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS). The cells were incubated at 37°C in a constant temperature incubator containing 5% CO2.

[0037] Cell seeding: 6×10 3 Keratinocytes (HaCaT) were seeded at a seeding density of 10 cells / well in a 96-well plate and incubated overnight in an incubator (37°C, 5% CO2).

[0038] Experimental grouping: set up a blank group, a control group and a sample group. In the sample group, set up seven concentration gradients, and set up three replicate wells under each concentration gradient.

[0039] Solution preparation: According to the set test concentration, use samples A2, B2, and C2 to prepare working solutions of the test substance with different concentrations. Dosing: Dosing was performed when the cell plating rate in the 96-well plate reached 40%-60%. The blank group had no cell inoculation and only 200 μL of cell culture medium was added; 200 μL of culture medium was added to each well of the control group; 200 μL of culture medium containing the corresponding concentration of sample was added to each well of the sample group. After the administration was completed, the 96-well plate was placed in an incubator (37°C, 5% CO2) for incubation.

[0040] Detection: After the cells have been incubated for 24 hours, 10 μL CCK-8 is added and incubated at 37°C in the dark for 2-4 hours. After the incubation, the OD value is read at 490 nm. The cell viability calculation formula is as follows: Cell viability = (OD 样品组 -OD 空白组 / OD 对照组 -OD 空白组 )×100% Experimental results: According to the results of CCK-8 cytotoxicity test, A2 showed no cytotoxicity to keratinocytes within a volume fraction of 0.5%; B2 and C2 had no cytotoxicity within a mass concentration of 0.25% ( Figure 4). This result shows that low concentrations of red peony root fermentation liquid have almost no toxic effects on cells, will not cause irritation or damage, and has good safety.

[0041] 4. EdU Detection Cell seeding: 5×10 HaCaT cells were plated per well. 4 The cells were seeded in a 24-well plate and cultured in a cell culture incubator at 37 °C and 5% CO2 for 24 h.

[0042] Experimental grouping: set up a control group (Control) and three sample groups A2~C2.

[0043] Solution preparation: According to the CCK-8 test results, the concentration at the inflection point of cell viability was selected as the detection concentration of A2~C2 cell proliferation ability. A DMEM solution with a volume fraction of 0.5% for A2 and 0.25% for B2 and C2 was used.

[0044] Drug administration: When the cell plating rate of the 24-well plate reaches 40%-60%, drug administration is performed. 500 μL of cell culture medium containing samples of corresponding concentrations is added to the sample group, and an equal volume of cell culture medium is added to the control group, and the cells are incubated in an incubator (37°C, 5% CO2) for 24 h.

[0045] EdU labeling, fixation, washing and permeabilization of cultured cells: 20 μmol·L -1 The cells were incubated with fresh medium containing EdU for 2 h, washed three times with PBS containing 3% BSA, and fixed with 4% paraformaldehyde for 15 min. After washing with the washing solution, they were incubated with an appropriate amount of permeabilization solution (PBS containing 0.3% Triton X-100) at room temperature for 10-15 min, and incubated with Click staining reaction solution containing Azide 488 at room temperature in the dark for 30 min.

[0046] Nuclear staining: Wash three times with PBS containing 3% BSA, each wash for 3-5 min, use Hoechst3342 for nuclear staining, incubate in the dark for 10 min, observe and collect images under a fluorescence microscope.

[0047] Experimental results: EdU staining (see Figure 5 ) showed that compared with the control group, the proliferation of HaCaT cells in group C2 was significantly enhanced (p<0.01), indicating that red peony root fermentation broth (A2, B2, C2) all promoted the proliferation of HaCaT cells, and the two-step fermentation broth C2 had a significant promoting effect on the proliferation and migration of HaCaT cells.

[0048] 5. HaCaT cell scratch assay Cell inoculation: HaCaT cells in logarithmic phase were taken at a rate of 0.8×10 4 The cells were inoculated into 6-well plates at a density of 100 cells / well and incubated overnight in an incubator (37°C, 5% CO2). The experiment could be started when the cells grew into a monolayer of adherent cells.

[0049] Experimental groups: set up a control group (Control) and experimental groups A2~C2, and set up 3 replicate wells in each group.

[0050] Solution preparation: According to the CCK-8 test results, the concentration at the inflection point of cell viability was selected as the scratch healing ability test concentration of A2~C2. A DMEM solution with a volume fraction of 0.5% for A2 and 0.25% for B2 and C2 was used.

[0051] Scratch and drug administration: draw a "+" line in the center of the plate, wash away the floating cells, observe and take pictures under a microscope, record it as 0 h, and continue to culture for 24 h after drug administration, observe and take pictures under a microscope. Calculation of healing rate: ImageJ was used to calculate the gray value of the scratch and the healing rate of the scratch.

[0052] Experimental results: like Figure 6 As shown in the results, compared with the control group, the fermented broth of red peony root had no significant inhibitory effect on the cell healing ability, and the cell scratch healing rates of the three experimental groups were all improved. Among them, the two-step fermented broth of red peony root C2 group had a significant promoting healing and repair function on HaCaT cells ( p <0.001), further confirming its excellent potential as a natural skin care raw material in the field of cell proliferation and skin damage repair.

[0053] Determination of Hyaluronidase Activity Inhibition Experiment in Red Peony Fermentation Broth Hyaluronidase is a specific hydrolase of hyaluronic acid, which is related to the permeability and inflammation of blood vessels. When the multivalent allergen enters the body for the second time and specifically binds to IgE, FcεR1 cross-linking occurs, which leads to mast cell degranulation, releases mediators such as histamine in the granules, and triggers allergic inflammatory reactions. Therefore, the hyaluronidase in vitro activity inhibition experiment can be used to screen the anti-allergic soothing performance of red peony root fermented liquid. The present embodiment adopts in vitro inhibition experiment Elson-Morgan method to measure the inhibition rate of prepared red peony root fermented liquid to hyaluronidase, and the hyaluronidase inhibition rate is calculated according to the following formula.

[0054] Inhibition rate = [(A1-A2)-(B1-B2)] / (A1-A2)×100% Among them, A1 is the absorbance value of the reference solution; A2 is the absorbance value of the reference blank solution, in which the hyaluronidase and sodium hyaluronate solution are replaced by acetate buffer solution; B1 is the absorbance value of the sample solution; B2 is the absorbance value of the sample blank solution, in which the hyaluronidase and sodium hyaluronate solution are replaced by acetate buffer solution.

[0055] Experimental results: Hyaluronidase is a key enzyme that regulates vascular permeability and triggers inflammation. In allergic reactions, it promotes mast cells to release inflammatory mediators. Therefore, the experiment of inhibiting hyaluronidase activity is an important means to evaluate whether a substance has anti-allergic and soothing effects. The results of the hyaluronidase activity inhibition experiment of red peony root fermentation liquid are shown in Figure 2-4 Compared with red peony root water extract, red peony root fermented liquid has a stronger inhibitory effect on hyaluronidase activity, among which the red peony root fermented liquid (B2) fermented by Schizosaccharomyces pombe has a significantly stronger inhibitory effect on hyaluronidase activity ( p <0.001), and its hyaluronidase inhibition rate was 89.24%. It can be seen that red peony root fermented liquid has better anti-allergic and soothing effects.

[0056] 7. TNF-α and IL-6 inflammatory factor release inhibition experiment Lipopolysaccharide (LPS) induced RAW264.7 cell inflammation model: RAW264.7 cells were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS). The cells were incubated at 37°C in an incubator containing 5% CO2.

[0057] Cell seeding: 1.6x10 4 RAW264.7 cells were seeded into 12-well plates at a seeding density of 10 cells / well and incubated in an incubator (37°C, 5% CO2) for 24 h.

[0058] Experimental groups: set up a blank control group (Control), a positive control group (LPS), and experimental groups A2~C2, and set up 3 replicates in each group.

[0059] Liquid preparation: Use DMEM solution with a volume fraction of 0.5% for A2 and 0.25% for B2 and C2.

[0060] Drug administration: Add 2.5 μL of LPS, stimulate for 2 h, and then divide the drug administration into groups, 1 ml per well, and continue culturing for 24 h.

[0061] Detection: The cell culture supernatant was added to the ELISA plate to measure the release of inflammatory factors IL-6 and TNF-α.

[0062] Experimental results: In this experiment, lipopolysaccharide (LPS) was used to induce an inflammatory cell model and the changes in the levels of proinflammatory cytokines tumor necrosis factor TNF-α and interleukin IL-6 were measured. Figure 8 As shown in A to B, red peony root fermented liquid has a regulatory effect on both cytokines and can effectively reduce the content of the two pro-inflammatory factors. The content of TNF-α in the blank control group was 110.80 pg / ml, and the content of TNF-α in the LPS group was 140.36 pg / ml; under the action of red peony root fermentation broth (A2, B2, C2), the content of proinflammatory factor TNF-α was 117.92 pg / ml, 117.26 pg / ml, and 120.10 pg / ml, respectively, and its content decreased by 15.99%, 16.46%, and 14.43%, respectively, with good significance (P<0.001); at the same time, the content of proinflammatory factor IL-6 was analyzed and it was found that the content of IL-6 in the blank control group was 106.80 pg / ml, and the content of IL-6 in the LPS group was 147.03 pg / ml; under the action of red peony root fermentation broth (A2, B2, C2), the content of proinflammatory factor IL-6 was 126.99 pg / ml, 123.63 pg / ml, 120.61 pg / ml, and their contents decreased by 13.63%, 15.92%, and 17.97%, respectively. In summary, red peony root fermented liquid can inhibit the release of TNF-α and IL-6 in HaCaT cells to a certain extent, which can be judged that red peony root fermented liquid has certain anti-inflammatory and soothing effects.

[0063] In summary, the red peony root fermented liquid has multiple functions such as anti-oxidation, promotion of keratinocyte proliferation and migration, promotion of skin damage repair, anti-allergy, anti-inflammatory and soothing, and has high biological safety, and can be used as an important active ingredient in cosmetics.

[0064] The above embodiments are only for illustrating the technical concept and features of the present invention, and their purpose is to enable people familiar with the technology to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the protection scope of the present invention. Any equivalent changes or modifications made according to the spirit of the present invention should be included in the protection scope of the present invention.

Claims

1. A strain composition, characterized in that: The strain composition comprises Pseudoalteromonas ( Pseudoalteromonas sp.) SE9 and Schizosaccharomyces pombe ( Schizosaccharomyces pombe )JM2-16; Wherein, the Pseudoalteromonas ( Pseudoalteromonas sp.) SE9, which was deposited in the China Center for Type Culture Collection on December 13, 2024, and its deposit number is CCTCC NO: M 20242809; The Schizosaccharomyces pombe ( Schizosaccharomyces pombe ) JM2-16, the strain was deposited in the China Center for Type Culture Collection on December 13, 2024, and its deposit number is CCTCC NO: M 20242810.

2. Use of the strain composition as claimed in claim 1 in red peony root fermentation.

3. A method for fermenting red peony root, characterized in that: The method comprises: adding the strain composition according to claim 1 to a liquid culture medium containing red peony root powder for fermentation culture, and separating and purifying to obtain red peony fermentation liquid.

4. The method for fermenting red peony root as claimed in claim 3, characterized in that, The specific method of the fermentation culture comprises: Schizosaccharomyces pombe ) JM2-16 was first added to the liquid medium containing red peony root powder for the first stage of fermentation culture, and then Pseudoalteromonas ( Pseudoalteromonas sp.)SE9 for the second stage of fermentation.

5. The method for fermenting red peony root as claimed in claim 4, characterized in that, In the first stage of fermentation, the fermentation temperature is 25-35°C, the rotation speed is controlled at 150-200 rpm, the fermentation time is 10-60h, and the fermentation pH is 5.0-6.5; In the second stage of fermentation, the fermentation temperature is 10-20°C, the rotation speed is controlled at 150-200 rpm, and the fermentation time is 10-60h.

6. The method for fermenting red peony root as claimed in claim 4, characterized in that: The specific method for separating and purifying the red peony root fermentation liquid comprises: subjecting the fermentation liquid to centrifugation, high temperature sterilization, decolorization and filtering.

7. The method for fermenting red peony root as claimed in claim 6, characterized in that, The specific centrifugal conditions are: centrifugal treatment at 5000-15000 r / min for 5-60 min; The specific conditions of high temperature sterilization are: sterilization at 121°C for 30 min; The decolorization is carried out using activated carbon; The filtration is performed using a 0.22 μm filter.

8. A red peony root fermented liquid, characterized in that: The red peony root fermented liquid is prepared by the red peony root fermentation method according to any one of claims 3 to 7.

9. The use of the red peony root fermented liquid in the preparation of cosmetics as claimed in claim 8, characterized in that: The cosmetic has any one or more of the following effects: (a) Antioxidant; (b) Promote the proliferation and migration of keratinocytes; (c) Promote skin damage repair; (d) Anti-allergic and soothing; (e) Anti-inflammatory and soothing.

10. The use according to claim 9, characterized in that: The cosmetics are skin care products.

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