Moisturizing and repairing fermented oil with self-emulsifying delivery function as well as preparation method and application of moisturizing and repairing fermented oil

Abstract

The invention belongs to the technical field of microbial fermentation and daily chemicals, and particularly discloses moisturizing and repairing fermented oil with a self-emulsifying delivery function as well as a preparation method and application of the moisturizing and repairing fermented oil. According to the invention, the grease is used as a substrate, the fermented grease produced by fermenting the Pseudozyma aphis MEL02 strain has independent self-emulsifying capacity, and when the fermented grease is used for preparing cosmetics, grease and an emulsifier do not need to be additionally added, so that the use of the emulsifier can be reduced; a self-emulsifying system prepared from the fermented oil can promote delivery of active matters and improve the absorption and utilization degree of the active matters, and compared with a traditional penetration enhancer, the self-emulsifying system does not affect the completeness of a skin barrier; the barrier repairing and moisturizing capacities of the fermented oil are improved compared with those before fermentation, the absorption and utilization of grease can be improved, and the skin feeling of the grease is improved. The fermented oil provided by the invention can effectively solve the problems of natural oil, emulsifiers and traditional penetration enhancers in existing cosmetics.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fermentation and daily chemical products, and in particular to a moisturizing and repairing fermented oil with a self-emulsifying delivery function, and a preparation method and application thereof. Background Art

[0002] The skin is the largest organ in the human body, and its main function is to protect the body from exogenous substances and excessive water loss. The barrier function of the skin is mainly undertaken by the stratum corneum located in the outermost layer of the epidermis, which blocks allergens, irritants, microorganisms or toxic substances from entering through the skin surface. External environmental factors such as ultraviolet radiation and surfactants can easily cause damage to the skin barrier, resulting in a decrease in the skin's moisturizing function and damage to the skin tissue. Therefore, it is crucial to use cosmetics to moisturize and repair the skin and enhance the skin's barrier function to resist harmful external environmental factors.

[0003] Natural plant oils and fats are used as emollients in cosmetic systems. They can form a protective layer on the epidermis, help prevent skin moisture loss, and moisturize the skin. Natural plant oils and fats have good compatibility with human skin, but most natural plant oils and fats have long fatty acid chains, which are difficult to absorb through the skin and feel thick and sticky. In the process of preparing cosmetics, the emulsion needs to mix the oil phase and the water phase uniformly. Natural plant oils and fats are used as oil phases. Because they lack amphiphilicity, emulsifiers need to be added to disperse them to stabilize the water-oil interface and obtain a stable emulsion. However, the use of emulsifiers has the following disadvantages: in cosmetic formulas, emulsifiers, preservatives, and fragrances are the three main irritants that cause skin sensitivity and are easy to cause damage to sensitive skin. As a medium for the combination of oil and water, emulsifiers are difficult to replace in emulsions. Therefore, the use of natural raw materials with emulsifying properties can reduce the damage to the skin caused by the use of emulsifiers. The self-emulsifying formulas in the prior art all require the additional addition of emulsifiers (such as the self-emulsifying cream base disclosed in CN111821256B and the self-emulsifying seaweed cleansing oil disclosed in CN109730945A), and the emulsifiers are synthetic emulsifiers, not naturally derived amphiphilic substances. Therefore, the introduction of emulsifiers will still cause skin sensitivity problems.

[0004] In addition, traditional penetration enhancers in cosmetics also have drawbacks. Transdermal absorption in cosmetics refers to the process in which the functional ingredients in cosmetics penetrate the skin tissue barrier according to the different effects of the product, act on the targeted location, and accumulate and exert their effects at that location. There are some poorly soluble functional ingredients in cosmetic raw materials, which are difficult to dissolve by ordinary natural oils and are even more difficult to be absorbed and utilized by the skin. While promoting the deep absorption of functional ingredients, traditional penetration enhancers will cause damage to the skin barrier. For example, ethanol will cause a decrease in stratum corneum lipids and change the skin barrier properties; azone will dissolve lipids to form gaps or reduce the orderliness of lipid arrangement between cells in this layer, increase the fluidity of membrane lipids, cause the cell structure to become loose, and the outer keratin is easy to fall off; dimethyl sulfoxide (DMSO) will destroy the spatial arrangement structure of lipids in the skin barrier. Summary of the invention

[0005] The purpose of the present invention is to provide a moisturizing and repairing fermented oil with self-emulsifying delivery function and a preparation method and application thereof, so as to solve the above-mentioned problems existing in natural oils, emulsifiers and traditional penetration enhancers in existing cosmetics.

[0006] In order to achieve the above object, the specific technical solution adopted by the present invention is as follows:

[0007] In the first aspect, the present invention provides a Pseudozyma aphidis MEL02 strain for preparing a moisturizing and repairing fermented oil with a self-emulsifying delivery function, which is classified and named Pseudozyma aphidis. The strain is deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, with a deposit number of CGMCC No. 31644 and a deposit date of August 8, 2024.

[0008] In a second aspect, the present invention provides a method for preparing a moisturizing and repairing fermented oil having a self-emulsifying delivery function using the above-mentioned strain, comprising the following steps:

[0009] S1. Inoculate the Pseudozyma aphidis MEL02 strain into a seed culture medium and culture to obtain a primary seed solution;

[0010] S2. The primary seed solution is connected to a seed tank containing a seed culture medium to obtain a secondary seed solution;

[0011] S3. The secondary seed solution is connected to the fermentation medium for fermentation culture to obtain a primary fermentation liquid;

[0012] S4. Continue to add oil to the primary fermentation broth and ferment it to obtain a secondary fermentation broth;

[0013] S5. The secondary fermentation broth is centrifuged, the cells and the fermentation waste liquid are discarded, and the fermentation oil is initially obtained by flash dehydration;

[0014] S6. Remove inorganic salts and proteins through membrane filtration to obtain fermented oil.

[0015] Preferably, in steps S1 and S2, the composition of the seed culture medium is: 30 g / L glucose, 31 g / L NaNO, 3 g / L yeast extract, 3 g / L KH2PO4, and 0.3 g / L MgSO4.

[0016] Preferably, in step S1, the culturing time is 20-28 hours.

[0017] Preferably, in step S2, the inoculation amount of the primary seed solution is 4-6vt%, and the culturing time is 20-28h.

[0018] Preferably, in step S3, the composition of the fermentation medium is: 20 g / L glucose, 100 g / L oil, 3 g / L NaNO3, 1 g / L yeast extract, 0.3 g / L KH2PO4, and 0.3 g / L MgSO4; the oil is a natural oil or a synthetic oil.

[0019] Furthermore, the natural oils include, but are not limited to, one or more of olive oil, camellia seed oil, sunflower seed oil, grape seed oil, sweet almond oil, macadamia seed oil, soybean oil, almond oil, linseed oil, rice bran oil, sesame seed oil, evening primrose oil, oat kernel oil, castor seed oil, wheat germ oil, corn germ oil, rice germ oil, and sea buckthorn oil. The synthetic oils include, but are not limited to, one or more of coconut oil caprylate and isopropyl palmitate.

[0020] Preferably, in step S3, the inoculation amount of the secondary seed solution is 5-10 vt%, and the culture time of the primary fermentation solution is 48-144 h.

[0021] Preferably, in step S4, the amount of oil added is 500-1000 g / L, and the secondary fermentation liquid is cultured for 48-96 h.

[0022] In a third aspect, the present invention provides a moisturizing and repairing fermented oil with a self-emulsifying delivery function obtained by the above preparation method.

[0023] In a fourth aspect, the present invention provides the use of the moisturizing and repairing fermented oil with self-emulsifying delivery function prepared by the above preparation method in the preparation of cosmetics. The fermented oil can be used to prepare cosmetics such as emulsions and creams. The present invention has proved through experiments that the fermented oil can achieve self-emulsification in the emulsion and cream system, and also provides an application example of using the fermented oil to prepare emulsions and creams.

[0024] The present invention has the following beneficial effects:

[0025] 1. The present invention provides a fermented oil with independent self-emulsification ability, which can achieve self-emulsification by adding an aqueous phase without the need to add additional oils and emulsifiers, thereby effectively reducing the use of emulsifiers.

[0026] 2. The self-emulsifying system prepared from the fermented oil of the present invention can enhance the delivery of effective active ingredients, solve the problem of difficult transdermal absorption of poorly soluble active ingredients, improve the absorption and utilization of active ingredients, and compared with traditional penetration enhancers, does not affect the integrity of the skin barrier.

[0027] 3. The fermented oil of the present invention can enhance the efficacy of the oil itself, that is, the barrier repair and moisturizing abilities are enhanced compared to before fermentation.

[0028] 4. The fermented oil of the present invention can enhance the absorptive and spreadable properties of natural oils and fats, and improve the heavy and greasy skin feel of the natural oils and fats themselves.

[0029] 5. The fermented oil of the present invention can inhibit the secretion of inflammatory factors and has a soothing effect.

[0030] 6. The present invention provides the use of the independently screened Pseudozyma aphidis MEL02 strain for the production of fermented oil. With oil as a substrate, the strain is used to ferment and produce a moisturizing and repairing fermented oil with a self-emulsifying delivery function. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1 : The initial appearance of the emulsions prepared according to emulsion recipes 1-5 in Example 3; in the figure, from left to right corresponds to the emulsions of recipes 1-5.

[0032] Figure 2 : The view of the emulsion prepared according to emulsion recipes 1-5 in Example 3 under an optical microscope.

[0033] Figure 3 : Appearance of the emulsions prepared according to emulsion recipes 1-5 in Example 3 after standing for 10 days; the emulsions of recipes 1-5 correspond from left to right in the figure.

[0034] Figure 4 : The view of the fermented oil-containing cream prepared in Example 4 under an optical microscope.

[0035] Figure 5 : The expression levels of TGM1 gene and CLDN1 gene after keratinocytes were treated with fermented oil and olive oil in Example 5; in the figure, BC is a blank control and PC is a positive control.

[0036] Figure 6 : LOR immunofluorescence (left) and LOR content (right) test results in Example 5.

[0037] Figure 7: TGM1 immunofluorescence (left) and TGM1 content (right) results in Example 6.

[0038] Figure 8 : The results of Caspase-14 immunofluorescence (left) and Caspase-14 content (right) in Example 6.

[0039] Fig. 9 : Claudin-1 immunofluorescence (left) and Claudin-1 content (right) results in Example 6.

[0040] Fig.10 : Barrier lipid immunofluorescence (left) and barrier lipid content (right) results in Example 6.

[0041] Fig.11 : Test results of the effect of fermented oil on IL-1α content in Example 7.

[0042] Fig.12 : Comparative test results of spreadability of fermented oil and olive oil in Example 9; in the figure, A and B are respectively a top view and a side view, and in Figures A and B, the left is the fermented oil and the right is the olive oil.

[0043] Fig.13 : Comparative test results of the absorbency of fermented oil and olive oil in Example 9; the left side of the figure is olive oil and the right side is fermented oil. DETAILED DESCRIPTION

[0044] The present invention is further described below in conjunction with the accompanying drawings and specific embodiments.

[0045] Example 1: Strain screening

[0046] The screening of the original strain includes the following steps:

[0047] 1. Collection and processing of oil-producing plant leaf samples

[0048] Collect comfrey leaves, camphor leaves, olive leaves, and perilla leaves, screen the leaves, remove diseased and worm-eyed samples, and obtain fresh and healthy sample leaves.

[0049] The leaves obtained by screening were rinsed with running water for 5-10 minutes to remove dust and other foreign matter on the surface. Then the leaves were cleaned with distilled water for 1-2 minutes. Finally, they were rinsed with sterile water for 3 times in a sterile operating table to obtain leaf samples with clean surfaces.

[0050] 2. Cultivation of sample leaves to obtain oleophilic bacteria

[0051] Mix 2-5g of sample and cut the leaves in a sterile operating table to a size of about 0.2-0.3cm in length and width. Then transfer the cut leaves into a conical bottle filled with sterile water and culture them in a shaker at 25±2℃ and 80-100rpm for 30min.

[0052] 1 mL of the liquid was inoculated into an oil-producing medium (olive oil 20 g / L, urea 0.2 g / L, NaNO3 1 g / L, soy peptone 5 g / L, KH2PO4 0.3 g / L, MgSO4 0.3 g / L) and cultured for 24-48 h;

[0053] 100uL of bacterial solution was spread on a YM (10g / L glucose, 5g / L tryptone, 3g / L malt extract powder and 3g / L yeast extract powder) solid plate and cultured for 48h; 200 single colonies with regular and uniform colony morphology were selected.

[0054] 3. Screening of strains for oleophilicity

[0055] 200 single colonies were inoculated into oil production screening medium (olive oil 40 g / L, urea 0.2 g / L, NaNO3 1 g / L, soy peptone 5 g / L, KH2PO4 0.3 g / L, MgSO4 0.3 g / L) and cultured for 168 h;

[0056] Observe the state of the culture medium with naked eyes and select 50 shake flasks with the best bacterial concentration. Take the culture medium and test the OD value of the fermentation liquid respectively, and select the strain with the highest bacterial concentration, which is recorded as MEL02. The strain has been deposited in CGMCC, with the deposit number CGMCC No. 31644.

[0057] Example 2: Preparation of fermented oil

[0058] A method for preparing fermented oil comprises the following steps:

[0059] S1. Inoculate Pseudozyma aphidis MEL02 strain into 50 mL seed culture medium (glucose 30 g / L, NaNO3 1 g / L, yeast extract 3 g / L, KH2PO4 3 g / L, MgSO4 0.3 g / L), and culture for 24 h to obtain primary seed solution;

[0060] S2. 100 mL of the first-level seed solution was inoculated into a seed tank containing 2 L of seed culture medium, and the second-level seed solution was obtained after culturing for 24 h;

[0061] S3. 2L of the secondary seed solution was inoculated into 30L of fermentation medium (glucose 20g / L, olive oil 100g / L, NaNO33g / L, yeast extract 1g / L, KH2PO4 0.3g / L, MgSO4 0.3g / L), without adding defoaming agent to avoid its residue affecting product quality, and fermented for 96h to obtain the primary fermentation liquid;

[0062] S4. Add 800 g / L of olive oil to the primary fermentation broth and continue fermentation for 48 h to obtain a secondary fermentation broth; the culture / fermentation temperature in steps S1-S4 was 25 ° C;

[0063] S5. The secondary fermentation broth is centrifuged, the cells and the fermentation waste liquid are discarded, and the fermentation oil is initially obtained by flash dehydration;

[0064] S6. Remove inorganic salts and proteins through membrane filtration (membrane pore size 50 nm) to obtain the fermented oil.

[0065] Example 3: Low-energy emulsification ability test of fermented oil

[0066] The fermented oil prepared in Example 2 was prepared into oil phases of different proportions according to Table 1 (the component contents in the table are all mass fractions), and the water phases were prepared accordingly. The oil phases and the water phases were mixed respectively at room temperature and shaken for 6 seconds to form emulsions of formulas 1-5.

[0067] Table 1

[0068]

[0069] Take photos to observe the emulsification conditions of each emulsion, such as Figure 1 As shown, with the increase of fermented oil content, the color of the emulsion becomes purer and whiter, and the emulsification effect becomes better. Among them, the emulsification effect of formula 5 is the best after shaking the oil phase and the water phase, indicating that the self-emulsification effect of 10% fermented oil is the most excellent.

[0070] Observe the particle size of the emulsion under an optical microscope and take photos. Figure 2 As shown in the figure, with the increase of fermentation oil content, the particle size of the emulsion gradually becomes finer and the emulsification effect becomes better. The emulsion particle size of formula 5 is the smallest, that is, 10% fermentation oil is self-emulsified, and the average particle size is about 20μm, indicating that the self-emulsification effect of 10% fermentation oil is the best.

[0071] After the obtained emulsion was placed at room temperature for 10 days, photographs were taken to observe the emulsification conditions. Figure 3 As shown, the color difference of 10% fermented oil after emulsification is the smallest after 10 days, and the emulsification effect is the most stable.

[0072] Example 4: Test on the high energy homogenization self-emulsification effect of fermented oil

[0073] The fermented oil prepared in Example 2 was used to prepare a cream according to the formula and test process described in Table 2.

[0074] Table 2

[0075]

[0076] Observe the particle size of the cream under an optical microscope and take photos. Figure 4 As shown, 18% fermented oil in a cream system can form a self-emulsifying body without adding additional emulsifiers, and the particle size of the emulsion can reach 5-10μm.

[0077] Example 5: Gene expression test related to the moisturizing and repairing effect of fermented oil

[0078] The fermented oil and unfermented olive oil (100% olive oil) prepared in Example 2 were respectively used to treat keratinocytes at a concentration of 0.01%. The blank control BC was not treated. The positive control (PC) WY14643 was used to treat keratinocytes at a concentration of 50 μM. The control group and the experimental sample were incubated for 24 hours. After the incubation, the old liquid was discarded, PBS was washed twice, 1 mL AG RNAex Pro Reagent was added to each well, and the cells were lysed by blowing and collecting the samples. RNA was extracted, reverse transcribed to cDNA, and then fluorescent quantitative PCR was performed. The expression amount was calculated using the 2-△△CT method, and the gene expression upregulation rate was calculated according to the following formula:

[0079]

[0080] The results showed that the fermented oil prepared in Example 2 could significantly increase the expression of TGM1 and CLDN1 genes in keratinocytes, while the unfermented olive oil had no significant difference. Figure 5 As shown, compared with the BC group, the fermented oil described in Example 2 can up-regulate the expression of TGM1 gene by 62% and can up-regulate the expression of CLDN1 gene by 12%.

[0081] The up-regulation of TGM1 gene expression indicates that the fermented oil in Example 2 can promote the formation of cornified capsule and enhance the skin barrier function; the up-regulation of CLDN1 gene expression indicates that the fermented oil in Example 2 can enhance tight junctions and strengthen the skin barrier. Enhanced barrier function can play a role in skin repair and moisturizing.

[0082] Example 6: 3D epidermal skin model with moisturizing and repairing effects test

[0083] 1. Test Grouping

[0084]

[0085] 2. Model Drug Administration

[0086] According to the test grouping, the model was transferred to a 6-well plate (0.9 mL EpiGrowth culture medium was added in advance). 25 μL of 0.1% SLS solution was added to the surface of the NC group, PC group and sample group and incubated for 30 minutes. After the incubation, the working solution of the corresponding concentration was added to the PC group, and the sample group evenly distributed the sample working solution on the surface of the model and placed in a CO2 incubator (37°C, 5% CO2) for 24 hours. After the incubation, the residual test substance was washed with sterile PBS solution and the residual liquid was wiped off with a sterile cotton swab.

[0087] 3. Immunofluorescence test

[0088] The model to be tested was fixed with 4% paraformaldehyde for 24 hours, and then immunofluorescence detection was performed. The model was observed and photographed under a microscope, and the images were collected and analyzed.

[0089] 4. Nile red staining

[0090] The paraffin sections were baked, dewaxed and hydrated, and washed with PBS for later use; the Nile red staining solution was prepared by diluting the diluted Nile red stock solution 600 times with 75% glycerol; 50 μL of Nile red staining solution was added to the tissue sections, and after staining for 20 minutes at room temperature in the dark, the sections were rinsed and wiped, and the sections were placed in glycerol gelatin and observed under a fluorescence microscope.

[0091] 5. Calculation of promotion rate

[0092]

[0093] 6. Experimental Results

[0094] like Figure 6-Figure 9 As shown, the fluorescence microscope (Olympus, BX43) was used to shoot, and the shooting magnification was 20×. The blue fluorescence was the cell nucleus; the green fluorescence was the target protein. The stronger the green fluorescence intensity, the higher the target protein content. Example 2 The fermented oil obtained at a concentration of 3%, relative to the NC negative control group, loricrin (LOR), glutaminase 1 (TGM1), cysteine ​​aspartate specific protease 14 (Caspase-14), tight junction protein 1 (Claudin-1) content were significantly increased, the increase rate was 222.22%, 315.79%, 192.59%, 238.10%, respectively, indicating that it has a moisturizing and repairing effect on the skin.

[0095] like Fig.10As shown, the images were taken using a fluorescence microscope (Olympus, BX43) at a magnification of 40×. The blue fluorescence represents the cell nucleus; the red fluorescence represents the barrier lipids. The stronger the red fluorescence, the higher the barrier lipid content. The fermented oil obtained in Example 2 has a significant increase in barrier lipid content at a concentration of 3% relative to the NC negative control group, with an increase rate of 80.95%, indicating that it has a repairing effect.

[0096] Example 7: 3D epidermal skin model with anti-inflammatory and soothing effects test

[0097] 1. Test Grouping

[0098]

[0099] 2. Model Drug Administration

[0100] According to the test grouping, the model was transferred to a 6-well plate (0.9 mL EpiGrowth culture medium was added in advance). 25 μL of 0.1% SLS solution was added to the surface of the NC group, PC group and sample group and incubated for 30 minutes. After the incubation, the working solution of the corresponding concentration was added to the PC group, and the sample group evenly distributed the sample working solution on the surface of the model and placed in a CO2 incubator (37°C, 5% CO2) for 24 hours. After the incubation, the residual test substance was washed with sterile PBS solution and the residual liquid was wiped off with a sterile cotton swab.

[0101] 3.ELISA test

[0102] After the incubation, the culture medium of the 3D epidermal skin model was collected in a centrifuge tube. After collection, the samples for ELISA detection were placed in a -80°C refrigerator for freezing and stored, and the detection and analysis were performed according to the operating instructions of the ELISA kit.

[0103] 4. Calculation of inhibition rate

[0104]

[0105] 5. Experimental Results

[0106] like Fig.11 As shown, the fermented oil obtained in Example 2 can significantly reduce the content of inflammatory factors (IL-1α) at ​​a concentration of 3% relative to the NC negative control group, with an inhibition rate of 21.46%, indicating that the sample can inhibit the secretion of inflammatory factors (IL-1α) at ​​this concentration and has a soothing effect.

[0107] Example 8: Transdermal delivery test of fermented oil self-emulsifying delivery system

[0108] 1. Test samples

[0109] 1) The fermented oil obtained in Example 2 was formulated into a self-emulsifying delivery system emulsion containing 1% pterostilbene according to Table 3. 2) Olive oil containing 1% pterostilbene.

[0110] Table 3

[0111]

[0112] 2. Test Method

[0113] The back skin of a one-month-old suckling pig stored at -20°C was thawed at room temperature with deionized water and repeatedly rinsed with PBS buffer. The suckling pig skin was fixed between the supply chamber and the receiving chamber of the Franz diffusion cell, with the skin stratum corneum facing the supply chamber and the dermis facing the receiving chamber. 7.0 mL of receiving solution was added to the receiving chamber. After the suckling pig skin was tightened and fixed, 1.0 mL of receiving solution (PBS) was added to the receiving chamber through a sampler, and the air was exhausted to make the skin dermis layer in close contact with the receiving solution. The sample was added to the skin surface in the supply chamber, and the effective penetration area S was about 3.14 cm 2 . Add the sample to the surface of the pig skin, spread the sample evenly from the center of the skin to the edge, and repeat 3 times for each sample in parallel. After the application, put it into the constant temperature water bath set in the (32±1)℃ receiving pool for infiltration, turn on the electromagnetic stirrer and stir at a speed of 300rpm to ensure that there are no bubbles in the water bath interlayer. After 8h of infiltration, HPLC detection of the content of the substance to be tested on the skin, in the skin, and under the skin of different experimental groups was performed.

[0114] 3. Statistics and computing

[0115] 1) Cumulative penetration

[0116] Q=Cn×V+∑Ci×V0(i=1···n-1)

[0117] Where: Q: cumulative permeation amount, V: volume of receiving solution in the receiving chamber, V0: volume of each sampling, Ci: drug concentration in the receiving solution from the first sampling to the last sampling, Cn: sample concentration measured at the nth sampling point.

[0118] 2) Diffusion percentage

[0119] P=Q / Po×100%

[0120] Where: P: diffusion percentage, Q: cumulative permeation amount of the sample in the receiving chamber at each time point, Po: the content of the sample in the release pool.

[0121] 3) Permeation rate

[0122] The cumulative permeation amount (Y axis) is used to make a regression equation against the permeation time (X axis). The slope of the equation is the permeation rate K. P .

[0123] 4. Experimental Results

[0124] The cumulative penetration amount of pterostilbene in olive oil was 1.64 μg, the diffusion percentage was 0.60%, and the penetration rate K P The cumulative permeation amount of pterostilbene in the fermented oil self-emulsification emulsion was 4.98 μg, the diffusion percentage was 1.79%, and the permeation rate K P The value was 0.7362 μg / h, indicating that the self-emulsifying delivery system using the fermented oil obtained in Example 2 had a better transdermal delivery effect on pterostilbene than olive oil.

[0125] Example 9: Fermented oil performance test

[0126] 1. Spreadability

[0127] One drop of the fermented oil obtained in Example 2 and one drop of olive oil were respectively added to the plane of polymethyl methacrylate (acrylic plate), and then two drops of distilled water containing red pigment were respectively added, and the spreading of the oil droplets in water was observed.

[0128] like Fig.12 As shown, the oil droplet on the left is fermented oil, and the oil droplet on the right is olive oil. The fermented oil has a larger spreading area after meeting water, a smaller contact angle when viewed from the side, and has better spreading properties.

[0129] 2. Absorbency

[0130] Add 1 drop of the fermented oil obtained in Example 2 and 1 drop of olive oil to the forearm respectively, and apply it in a circular motion for 3 minutes until the skin completely absorbs it. Apply black oil-absorbing paper to the two areas respectively, press with a heavy object for the same time, and observe the absorption of residual oil by the oil-absorbing paper.

[0131] like Fig.13 As shown, the left side is olive oil and the right side is fermented oil. The amount of residual oil on the fermented oil side is obviously less, indicating that it has better absorbency.

[0132] Example 10: Application of fermented oil in the preparation of cosmetics

[0133] This embodiment provides examples of preparing emulsions and creams using the fermented oil of the present invention, specifically a self-emulsifying system cream containing fermented oil as shown in Table 4 and a self-emulsifying system emulsion containing fermented oil as shown in Table 5.

[0134] Table 4

[0135]

[0136] Table 5

[0137]

[0138]

[0139] This specific implementation is only an explanation of the present invention, not a limitation of the present invention. Any changes made by those skilled in the art after reading the specification of the present invention will be protected by the patent law as long as they are within the scope of the claims of the present invention.

Claims

1. Application of Pseudozyma aphidis MEL02 strain in the preparation of moisturizing and repairing fermented oil with self-emulsifying delivery function, characterized in that: The Pseudozyma aphidis MEL02 strain is deposited in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit number CGMCC No.31644 and the deposit date August 8, 2024.

2. A method for preparing a moisturizing and repairing fermented oil with self-emulsifying delivery function, characterized in that: The following steps are involved: S1. Inoculate the Pseudozyma aphidis MEL02 strain into a seed culture medium and culture to obtain a primary seed solution; S2. The primary seed solution is connected to a seed tank containing a seed culture medium to obtain a secondary seed solution; S3. The secondary seed solution is connected to the fermentation medium for fermentation culture to obtain a primary fermentation liquid; S4. Continue to add oil to the primary fermentation broth and ferment it to obtain a secondary fermentation broth; S5. The secondary fermentation broth is centrifuged, the cells and the fermentation waste liquid are discarded, and the fermentation oil is initially obtained by flash dehydration; S6. Remove inorganic salts and proteins through membrane filtration to obtain fermented oil.

3. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In steps S1 and S2, the composition of the seed culture medium is: 30 g / L glucose, NaNO3 1g / L, yeast extract 3g / L, KH2PO4 3g / L, MgSO4 0.3g / L.

4. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In step S1, the culturing time is 20-28 hours.

5. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In step S2, the inoculation amount of the primary seed solution is 4-6vt%, and the culturing time is 20-28h.

6. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In step S3, the composition of the fermentation medium is: glucose 20 g / L, oil 100 g / L, NaNO3 3 g / L, yeast extract 1 g / L, KH2PO4 0.3 g / L, MgSO4 0.3 g / L; The oil is natural oil or synthetic oil.

7. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In step S3, the inoculation amount of the secondary seed solution is 5-10 vt%, and the culture time of the primary fermentation solution is 48-144 h.

8. The method for preparing the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 2, characterized in that: In step S4, the amount of oil added is 500-1000 g / L, and the secondary fermentation liquid is cultured for 48-96 hours.

9. The moisturizing and repairing fermented oil with self-emulsifying delivery function obtained by the preparation method according to any one of claims 2 to 8.

10. Use of the moisturizing and repairing fermented oil with self-emulsifying delivery function according to claim 9 in the preparation of cosmetics.

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