Skin metabolism regulation fermented vegetable oil rich in phosphatide glycolipid and preparation process of skin metabolism regulation fermented vegetable oil

Abstract

The invention discloses skin metabolism regulation fermented vegetable oil rich in phosphatide glycolipid and a preparation process thereof, and relates to the technical field of daily chemicals and medical skin care. The skin metabolism regulation and control fermented vegetable oil rich in phospholipid glycolipid comprises a core strain and a culture medium formula, and the culture medium formula comprises a plate screening culture medium (YM culture medium), a strain activation and primary seed culture medium, a secondary seed culture medium and a fermentation culture medium. The method comprises the following processes: S1, strain screening and purification, S2, fermentation production, S3, post-treatment purification and S4, quality detection, the vegetable oil autonomously generates multi-element active components, skin metabolism is accurately regulated and controlled, and a core strain drives synthesis of the active components, the formula design is simplified, the stimulation risk is reduced, and the interfacial activity is improved through strain metabolism.

Description

Technical Field

[0001] This invention relates to the fields of daily chemical and medical skincare technology, and in particular to a skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids and its preparation process. Background Technology

[0002] In the fields of daily chemical and medical skincare, natural plant oils have long been used as core moisturizing ingredients due to their good biocompatibility and the presence of essential nutrients such as unsaturated fatty acids. However, as consumers' demands for precise efficacy, skin feel compatibility, and ingredient safety have increased, traditional plant oils and existing fermentation oil technologies have gradually revealed multiple technical bottlenecks, making it difficult to meet the needs of high-end products. Specific issues are as follows: Traditional plant oils have limited effects and poor skin feel and stability. Traditional plant oils (such as olive oil and meadowfoam seed oil) primarily consist of high-molecular-weight triglycerides (accounting for over 90%), which only provide basic moisturizing and cannot specifically address skin metabolic imbalances. A reduction in the intercellular matrix (composed of 55% ceramides, 25% cholesterol, and 15% fatty acids) leads to disordered stratum corneum renewal and barrier damage, resulting in dryness and wrinkles. Furthermore, traditional plant oils cannot activate key genes regulating cell metabolism, such as PPARα / β / γ, making it difficult to repair the skin at the molecular level. Simultaneously, the high molecular weight triglycerides result in low skin absorption (only 10%-15%), leading to a heavy and greasy feel upon application. The high-temperature refining process also easily triggers oil oxidation, increasing the peroxide value (the peroxide value of conventional olive oil is approximately 0.6-0.8g / 100g), which not only shortens shelf life but may also produce oxidative products that irritate the skin. Summary of the Invention

[0003] The purpose of this invention is to at least solve one of the technical problems existing in the prior art, and to provide a skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids and its preparation process, which can solve the problems of traditional plant oils having single efficacy and poor skin feel and stability.

[0004] To achieve the above objectives, the present invention provides the following technical solution: a skin metabolism regulating fermented plant oil rich in phospholipids and glycolipids, comprising a core strain and a culture medium formulation, wherein the culture medium formulation comprises a plate screening medium (YM medium), a strain activation and primary seed medium, a secondary seed medium, and a fermentation medium; The core strain is Ustilago scitaminea MEL03, which was screened from oil-rich sunflower seeds. After being identified by the Institute of Microbiology, Chinese Academy of Sciences, it is now deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC34427. This strain has the ability to metabolize oils efficiently and can naturally produce phospholipids and glycolipids, which are beneficial to the skin, during the fermentation process. It is the key biological basis for the preparation of this fermented plant oil.

[0005] Preferably, the plate screening medium (YM medium) is used for the initial culture and screening of strains. Its components include 10 g / L glucose, 5 g / L tryptone, 3 g / L malt extract, and 3 g / L yeast extract, which can provide the necessary carbon source, nitrogen source and other nutrients for the initial growth of strains, and help the strains form observable colonies.

[0006] Preferably, the microbial activation and primary seed culture medium consists of 30 g / L glucose, 1 g / L NaNO3, 3 g / L yeast extract, 3 g / L KH2PO4, and 0.3 g / L MgSO4. Glucose provides the primary carbon source, NaNO3 and yeast extract provide the nitrogen source, and KH2PO4 and MgSO4 serve as inorganic salts, providing a suitable nutrient environment for the rapid proliferation of the activated microbial strain and ensuring sufficient activity and quantity in the primary seed culture.

[0007] Preferably, the secondary seed culture medium has the same composition as the primary seed culture medium. Continuing with the same nutrient formula ensures a stable growth environment for the microorganisms during proliferation, maintaining high activity levels and meeting the quantity and quality requirements of the secondary seed culture, thus laying a solid foundation for subsequent fermentation stages.

[0008] Preferably, the fermentation medium is a crucial environment for the fermentation of microorganisms to produce active ingredients. Its components include 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. This formula has a high oil content to meet the carbon source requirements of oleophilic bacteria, while adjusting the ratio of nitrogen source and inorganic salts to better suit the production of phospholipids, glycolipids, and other functional components. In the later stages of fermentation, natural vegetable oil (olive oil or meadowfoam seed oil) needs to be added, with the volume ratio of the added vegetable oil to the primary fermentation broth controlled at 0.5:1-2 to further enhance the fermentation effect and the content of active ingredients in the product.

[0009] Preferably, a process for preparing skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids includes the following steps: S1 strain screening and purification, S2 fermentation production, S3 post-treatment purification, and S4 quality detection. Among them, the S1 strain screening and purification includes S101 sample collection and pretreatment, S102 seed crushing and mixing, S103 culture and screening, and S104 strain identification. Among them, S2 fermentation production includes S201 primary seed liquid preparation, S202 secondary seed liquid preparation, S203 primary fermentation broth preparation and S204 secondary fermentation broth preparation; The S3 post-treatment purification includes S301 centrifugation, S302 flash dehydration and S303 membrane filtration purification. The S4 quality testing includes S401 specific gravity testing, S402 refractive index testing, S403 viscosity testing, S404 acid value testing, S405 peroxide value testing, S406 total triterpenoid content testing, S407 phospholipid content testing, S408 glycolipid content testing, S409 lipid composition analysis, and S4010 skin efficacy-related testing.

[0010] Preferably, the S101 sample collection and pretreatment involves: firstly collecting fresh sunflower seeds rich in oil, as these seeds are more susceptible to the growth of oil-loving bacteria in their environment. After collection, the seeds undergo strict pretreatment: firstly, rinsing them 3-5 times with sterile water to remove surface dust and impurities; then rinsing them with 75% alcohol for 3-5 minutes for preliminary disinfection; followed by rinsing them 3 times with sterile water to remove residual alcohol; finally, drying them on a clean bench and sterilizing them with ultraviolet light for 30 minutes to ensure that subsequent operations are performed in a sterile environment and to avoid contamination by other microorganisms. S102 Seed Crushing and Mixing: Under sterile conditions in a clean bench, carefully remove the seed coat to obtain the embryo (kernel). Then, use a sterile scalpel to cut the kernel into small pieces roughly the size of sesame seeds. This process increases the contact area between the kernel and the subsequent solution, which is conducive to the release of microorganisms. After cutting, add a small amount of sterile water to the pieces and place them on a shaker to mix thoroughly, so that the microorganisms are evenly dispersed in the aqueous solution. S103 Cultivation and Screening: Take the supernatant of the above mixed aqueous solution and spread it evenly on the pre-prepared YM plate medium. Place the plate in a cultivation environment of 32-37℃ and incubate for 3-5 days to allow the microorganisms to grow fully and form colonies. Then, pick 50 single colonies with different morphologies from the plate and inoculate them into a liquid medium containing oil (the composition is the same as the culture medium for strain activation and primary seed culture, i.e., glucose 30g / L, NaNO3 1g / L, yeast extract 3g / L, KH2PO4 3g / L, MgSO4 0.3g / L). Incubate at 32-37℃ for 48-72 hours to allow the strains in the colonies to further proliferate in the liquid environment. Strain S104 identification: After fermentation culture, the fermentation broth of each strain was allowed to stand for a period of time until the oil phase and water phase separated. Then, a drop of the upper oil phase of each fermentation broth was taken and dropped onto a smooth surface, followed by a drop of red water. The state of the water droplets was observed to determine the oil-water miscibility. The strain with the best oil-water miscibility was selected, which is the target strain Ustilago scitaminea MEL03. It has the ability to efficiently metabolize oils and promote oil-water fusion, and is the core strain for subsequent preparation of fermented vegetable oil.

[0011] Preferably, the preparation of the S201 primary seed culture involves: activating the screened Ustilago scitaminea MEL03 strain, inoculating the activated strain into the primary seed culture medium, and placing the inoculated medium at 32-37°C for 24-36 hours. During this process, the strain will rapidly proliferate, ultimately forming a primary seed culture with high activity and sufficient quantity, providing the initial strain for the subsequent preparation of the secondary seed culture. S202 Secondary Seed Culture Preparation: According to the inoculation ratio of 10%, the prepared primary seed culture is inoculated into the secondary seed tank containing the secondary seed culture medium. The culture temperature is maintained at 32-37℃ and the culture is continued for 36-48 hours. Through the expansion culture, the number of strains is further increased and the activity is kept stable, and finally the secondary seed culture that meets the fermentation requirements is obtained. Preparation of S203 primary fermentation broth: Inoculate the secondary seed liquid into the fermentation equipment containing fermentation medium at an inoculation ratio of 10%, control the culture temperature at 32-37℃, and the culture time is 48-72 hours. During this stage, the strain will multiply in large quantities in a suitable nutrient environment and begin to metabolize lipids, producing preliminary active ingredients, and finally forming the primary fermentation broth. S204 Secondary Fermentation Broth Preparation: Add natural vegetable oil (olive oil or meadowfoam seed oil) to the primary fermentation broth, strictly control the volume ratio of vegetable oil to primary fermentation broth to be 0.5:1-2, continue to maintain the cultivation temperature of 32-37℃, and cultivate for 48-96 hours. During this process, the strain will further metabolize the added vegetable oil, producing more phospholipids and glycolipids, ultimately obtaining a secondary fermentation broth with richer components and stronger activity.

[0012] Preferably, the S301 centrifugal separation involves centrifuging the secondary fermentation broth to separate the microbial cells, fermentation waste liquid, and oil phase using centrifugal force. After centrifugation, the microbial cells precipitated at the bottom and the fermentation waste liquid at the top are discarded, leaving only the middle oil phase, which is the crude oil containing the target active ingredient. S302 Flash Dehydration: The oil phase obtained by centrifugation is sent to a flash evaporation device for flash dehydration. Flash dehydration utilizes the principle of rapid pressure reduction to quickly vaporize and remove the residual water in the oil phase, effectively avoiding the damage to the quality and active ingredients of the oil phase caused by prolonged heating. After processing, crude fermented oil is obtained. S303 Membrane Filtration Refining: Membrane filtration technology is used to filter crude fermented oil. By selecting a membrane with a suitable pore size, residual inorganic salts and protein impurities in the crude product can be effectively removed without causing loss of the target active ingredients such as phospholipids and glycolipids. After membrane filtration, a high-purity and stable-quality fermented vegetable oil product is finally obtained.

[0013] Preferably, the specific gravity detection in S401 is performed by using an Anton Paar DMA35 handheld densitometer to measure the specific gravity of the fermented vegetable oil. This instrument is highly accurate and can quickly and accurately obtain the specific gravity data of the sample. Specific gravity is one of the important indicators reflecting the purity and composition of oil. S402 Refractive Index Test: The refractive index of fermented vegetable oil was tested using an ATAGO portable refractometer. The refractive index is closely related to the composition and purity of the oil. By testing the refractive index, the quality of the oil and the presence of impurities can be preliminarily determined. S403 Viscosity Testing: A BROOKFIELD digital touchscreen rotational viscometer was used to measure the viscosity of fermented vegetable oil at 25°C using a #2 rotor at 200 rpm. Viscosity affects the feel of the oil on the skin and the stability of the formula, and is an important parameter in the product application process. S404 Acid Value Test: The acid value of fermented vegetable oil is tested according to the national standard "GB 5009.229-2016 Determination of Acid Value in Food". The acid value reflects the content of free fatty acids in the oil. Excessively high acid value will affect the stability and safety of the oil. S405 Peroxide Value Test: The peroxide value of fermented vegetable oils is tested according to the standard "GB 5009.227-2023 Determination of Peroxide Value in Food". Peroxide value is a key indicator for measuring the degree of oxidation of oils. Excessive peroxide value indicates that the oil has undergone oxidative deterioration, which will affect the quality and efficacy of the product. S406 Total Triterpenoid Content Detection: The total triterpenoid content in fermented vegetable oil was detected according to "NYT3676-2020 Determination of Total Triterpenoid Content in Ganoderma lucidum by Spectrophotometry". Total triterpenoids are one of the important active ingredients in this fermented vegetable oil, and their content directly affects the efficacy of the product. S407 Phospholipid Content Detection: Phospholipid content is detected according to the molybdenum blue colorimetric method in "GB / T 5537-2008 Grain and Oil Inspection - Determination of Phospholipid Content". Phospholipids are key functional ingredients in products, and accurate detection of their content can ensure that product quality meets standards. S408 Glycolipid Content Detection: The glycolipid content is detected by high performance liquid chromatography (HPLC). HPLC has the characteristics of high separation efficiency and high detection accuracy, which can accurately determine the glycolipid content and ensure that the glycolipid components in the product meet the requirements. S409 Lipid Composition Analysis: TOF / MS (Time-of-Flight Mass Spectrometry) was used to analyze lipids in the raw oil and the fermented oil. This analytical method can clearly show the changes in various lipid components in the oil before and after fermentation, clarify the impact of the fermentation process on lipid components, and further verify the effectiveness of the fermentation process. S4010 Skin Efficacy Related Tests: Skin moisture content was measured to evaluate the moisturizing and barrier repair effects of fermented plant oils; its influence on the expression levels of PPARα, PPARβ, and PPARγ genes was also tested to verify its function in regulating skin cell metabolism; in addition, the liquid crystal phenomenon after the fermented oil was formulated into an emulsion was observed. The liquid crystal structure helps to improve the stability and skin permeability of the emulsion, further ensuring the application effect of the product.

[0014] Compared with the prior art, the beneficial effects of the present invention are: 1. This skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids and its preparation process: This plant oil autonomously generates multiple active ingredients, precisely regulates skin metabolism, and the core strain drives the synthesis of active ingredients: The selected exclusive strain Ustilago scitaminea MEL03 (accession number CGMCC34427) has a highly efficient lipid metabolism capacity. During the fermentation process, it can naturally synthesize functional components such as phospholipids, glycolipids, and total triterpenes without the need for exogenous addition. Among them, phospholipids can enhance the skin barrier's water-locking ability, glycolipids can improve the permeability of skin cells, and total triterpenes have anti-inflammatory and antioxidant effects; Activating key genes in skin metabolism: Skin efficacy testing has verified that this fermented plant oil can significantly upregulate the relative expression levels of PPARα, PPARβ, and PPARγ genes (as shown in Figure 6-11). Among them, PPARα regulates stratum corneum renewal, PPARβ promotes skin cell proliferation, and PPARγ repairs damaged barriers. It improves skin metabolic imbalance at the molecular level, overcoming the limitation of traditional plant oils that "only moisturize and do not regulate," and improving the repair effect on dry and sensitive skin by more than 30%.

[0015] 2. This skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids and its preparation process. This process improves oil-water miscibility, simplifies formulation design and reduces irritation risk. Microbial metabolism improves interfacial activity: Ustilago scitaminea MEL03 produces natural interfacial active substances (such as glycosylglycerol, ester peptides, etc., as shown in Figure 3) when metabolizing oils, which significantly improves the oil-water miscibility of the fermented plant oil. Experiments show that when this fermented plant oil is mixed with the aqueous phase, the separation time is extended to more than 5 times that of traditional plant oils, and stable fusion can be achieved without the addition of additional chemical emulsifiers. Reduced irritation and enhanced naturalness: Because it does not rely on exogenous emulsifiers, the product reduces skin irritation by 60% while retaining the natural properties of plant oils, which aligns with the trend of "gentle skincare" and is suitable for the needs of special groups such as those with sensitive skin and pregnant women.

[0016] 3. This skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids and its preparation process features an optimized purification process to ensure product quality and stability. Multi-step purification removes impurities and reduces safety risks: the post-treatment process (S3 step) of "centrifugal separation - flash dehydration - membrane filtration purification" effectively removes bacteria, waste liquid, inorganic salts, and residual proteins from the fermentation broth. Centrifugal separation first separates the crude oil phase; flash dehydration utilizes rapid pressure reduction to avoid high-temperature damage to active ingredients; and membrane filtration precisely traps impurities (pore size adaptability design). The final product's acid value meets GB 5009.229-2016 standard (≤1.0mg KOH / g), and its peroxide value meets GB 5009.227-2023 standard (≤0.15g / 100g), far exceeding the quality indicators of traditional plant oils. Extended shelf life: The low impurity content and stable active ingredient structure extend the product's shelf life to 24 months, solving the problem of easy oxidation and deterioration of traditional vegetable oils and reducing the company's storage and loss costs. Attached Figure Description

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments: Figure 1 This is a process flow diagram of the present invention; Figure 2 This is the high-performance liquid chromatogram of the present invention; Figure 3 This is the negative TOF / MS TIC spectrum of this invention; Figure 4 This is a comparison chart of skin moisture content according to the present invention; Figure 5 This is a comparison chart of transdermal water loss (TWEL) according to the present invention; Figure 6 This is a schematic diagram showing the relative expression levels of the PPARα gene in olive oil (model control group) and fermented olive oil (sample group) of the present invention; Figure 7 This is a schematic diagram showing the relative expression levels of the PPARβ gene in olive oil (model control group) and fermented olive oil (sample group) of the present invention; Figure 8 This is a schematic diagram showing the relative expression levels of the PPARγ gene in olive oil (model control group) and fermented olive oil (sample group) of the present invention; Figure 9 This is a schematic diagram showing the relative expression levels of the PPARα gene in meadowfoam seed oil (model control group) and fermented meadowfoam seed oil (sample group) of the present invention. Figure 10 This is a schematic diagram showing the relative expression levels of the PPARβ gene in meadowfoam seed oil (model control group) and fermented meadowfoam seed oil (sample group) of the present invention; Figure 11 This is a schematic diagram showing the relative expression levels of the PPARγ gene in meadowfoam seed oil (model control group) and fermented meadowfoam seed oil (sample group) of the present invention. Figure 12 This is a liquid crystal phenomenon diagram of the fermented olive oil emulsion preparation according to the present invention; Figure 13 This is a diagram illustrating the liquid crystal phenomenon in the preparation of an emulsion from fermented meadowfoam seed oil according to the present invention. Detailed Implementation

[0018] This section will describe in detail specific embodiments of the present invention. Preferred embodiments of the present invention are shown in the accompanying drawings. The purpose of the drawings is to supplement the textual description with graphics, so that people can intuitively and vividly understand each technical feature and overall technical solution of the present invention, but they should not be construed as limiting the scope of protection of the present invention.

[0019] In the description of this invention, it should be understood that the orientation descriptions, such as up, down, front, back, left, right, etc., are based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limiting this invention.

[0020] In the description of this invention, terms such as greater than, less than, and exceeding are understood to exclude the stated number, while terms such as above, below, and within are understood to include the stated number. The use of "first" and "second" in the description is merely for distinguishing technical features and should not be construed as indicating or implying relative importance, or implicitly indicating the number of indicated technical features, or implicitly indicating the order of the indicated technical features.

[0021] In the description of this invention, unless otherwise explicitly defined, terms such as "set up," "install," and "connect" should be interpreted broadly, and those skilled in the art can reasonably determine the specific meaning of the above terms in this invention in conjunction with the specific content of the technical solution.

[0022] Please see Figure 1-13 The present invention provides a technical solution: a skin metabolism regulating fermented plant oil rich in phospholipids and glycolipids, comprising a core strain and a culture medium formulation, wherein the culture medium formulation comprises a plate screening medium (YM medium), a strain activation and primary seed medium, a secondary seed medium and a fermentation medium; The core strain is Ustilago scitaminea MEL03, which was screened from oil-rich sunflower seeds. After being identified by the Institute of Microbiology, Chinese Academy of Sciences, it is now deposited at the China General Microbiological Culture Collection Center, with accession number CGMCC34427. This strain has the ability to metabolize oil efficiently and can naturally produce phospholipids and glycolipids, which are beneficial to the skin, during the fermentation process. It is the key biological basis for the preparation of this fermented plant oil. Plate screening medium (YM medium): This medium is used for the initial culture and screening of strains. Its components include 10 g / L glucose, 5 g / L tryptone, 3 g / L malt extract, and 3 g / L yeast extract. It can provide the necessary carbon source, nitrogen source and other nutrients for the initial growth of strains, and help the strains form observable colonies. Strain activation and primary seed culture medium: This culture medium is used for strain activation and primary seed culture. The composition consists of glucose 30 g / L, NaNO3 1 g / L, yeast extract 3 g / L, KH2PO4 3 g / L, and MgSO4 0.3 g / L. Glucose provides the main carbon source, NaNO3 and yeast extract provide the nitrogen source, and KH2PO4 and MgSO4 serve as inorganic salts. This provides a suitable nutrient environment for the rapid proliferation of the strain after activation, ensuring that the primary seed culture has sufficient activity and quantity. Secondary seed culture medium: The composition of the secondary seed culture medium is exactly the same as that of the primary seed culture medium, and the same nutrient formula is used to ensure that the growth environment of the strain is stable during the proliferation process and to maintain a high activity level, so as to meet the quantity and quality requirements of the secondary seed liquid and lay a good foundation for the subsequent fermentation process. Fermentation medium: The fermentation medium is the key environment for the fermentation of microorganisms to produce active ingredients. Its components include glucose 20g / L, oil 100g / L, NaNO3 3g / L, yeast extract 1g / L, KH2PO4 0.3g / L, and MgSO4 0.3g / L. This formula has a high oil content to meet the carbon source requirements of oleophilic bacteria. At the same time, the ratio of nitrogen source and inorganic salts has been adjusted to be more suitable for the microorganisms to produce phospholipids, glycolipids and other functional components. In the later stage of the fermentation process, natural vegetable oil (olive oil or meadowfoam seed oil) needs to be added. The volume ratio of the added vegetable oil to the primary fermentation liquid is controlled at 0.5:1-2 to further improve the fermentation effect and the content of active ingredients in the product. A process for preparing skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids includes the following steps: S1 strain screening and purification: S101 Sample Collection and Pretreatment: First, collect fresh sunflower seeds rich in oil, as these seeds are more susceptible to the growth of oil-loving bacteria in their environment. After collection, the seeds undergo strict pretreatment: first, rinse with sterile water 3-5 times to remove surface dust and impurities; then, rinse with 75% alcohol for 3-5 minutes for preliminary disinfection; then rinse with sterile water 3 times to remove residual alcohol; finally, drain the water on a clean bench and sterilize with ultraviolet light for 30 minutes to ensure that subsequent operations are carried out in a sterile environment and avoid contamination by other microorganisms. S102 Seed Crushing and Mixing: Under sterile conditions in a clean bench, carefully remove the seed coat to obtain the embryo (kernel). Then, use a sterile scalpel to cut the kernel into small pieces roughly the size of sesame seeds. This process increases the contact area between the kernel and the subsequent solution, which is conducive to the release of microorganisms. After cutting, add a small amount of sterile water to the pieces and place them on a shaker to mix thoroughly, so that the microorganisms are evenly dispersed in the aqueous solution. S103 Cultivation and Screening: Take the supernatant of the above mixed aqueous solution and spread it evenly on the pre-prepared YM plate medium. Place the plate in a cultivation environment of 32-37℃ and incubate for 3-5 days to allow the microorganisms to grow fully and form colonies. Then, pick 50 single colonies with different morphologies from the plate and inoculate them into a liquid medium containing oil (the composition is the same as the culture medium for strain activation and primary seed culture, i.e., glucose 30g / L, NaNO3 1g / L, yeast extract 3g / L, KH2PO4 3g / L, MgSO4 0.3g / L). Incubate at 32-37℃ for 48-72 hours to allow the strains in the colonies to further proliferate in the liquid environment. Strain S104 identification: After fermentation culture, the fermentation broth of each strain was allowed to stand for a period of time until the oil phase and water phase separated. Then, a drop of the upper oil phase of each fermentation broth was taken and dropped onto a smooth surface. A drop of red water was then added to each surface. The state of the water droplets was observed to determine the oil-water miscibility. The strain with the best oil-water miscibility was selected. This strain is the target strain Ustilago scitaminea MEL03. It has the ability to efficiently metabolize oils and promote oil-water fusion, and is the core strain for subsequent preparation of fermented vegetable oil. S2 fermentation production ( Figure 1 (Process flow diagram) S201 Primary Seed Culture Preparation: The screened Ustilago scitaminea MEL03 strain was activated and then inoculated into the primary seed culture medium. The inoculated medium was placed in a culture environment of 32-37℃ for 24-36 hours. During this process, the strain will rapidly proliferate and eventually form a primary seed culture with high activity and sufficient quantity, providing the initial strain for the subsequent preparation of secondary seed culture. S202 Secondary Seed Culture Preparation: According to the inoculation ratio of 10%, the prepared primary seed culture is inoculated into the secondary seed tank containing the secondary seed culture medium. The culture temperature is maintained at 32-37℃ and the culture is continued for 36-48 hours. Through the expansion culture, the number of strains is further increased and the activity is kept stable, and finally the secondary seed culture that meets the fermentation requirements is obtained. Preparation of S203 primary fermentation broth: Inoculate the secondary seed liquid into the fermentation equipment containing fermentation medium at an inoculation ratio of 10%, control the culture temperature at 32-37℃, and the culture time is 48-72 hours. During this stage, the strain will multiply in large quantities in a suitable nutrient environment and begin to metabolize lipids, producing preliminary active ingredients, and finally forming the primary fermentation broth. S204 secondary fermentation broth preparation: Add natural vegetable oil (olive oil or meadowfoam seed oil) to the primary fermentation broth, strictly control the volume ratio of vegetable oil to primary fermentation broth to be 0.5:1-2, continue to maintain the cultivation temperature of 32-37℃, and cultivate for 48-96 hours. During this process, the strain will further metabolize the added vegetable oil, producing more phospholipids and glycolipids, and finally obtain a secondary fermentation broth with richer components and stronger activity. S3 post-processing purification: S301 Centrifugal Separation: The secondary fermentation broth is centrifuged to separate the bacterial cells, fermentation waste liquid and oil phase in the fermentation broth using centrifugal force. After centrifugation, the bacterial cells at the bottom and the fermentation waste liquid at the top are discarded, and only the middle oil phase is retained. This oil phase is the crude oil containing the target active ingredient. S302 Flash Dehydration: The oil phase obtained by centrifugation is sent to a flash evaporation device for flash dehydration. Flash dehydration utilizes the principle of rapid pressure reduction to quickly vaporize and remove the residual water in the oil phase, effectively avoiding the damage to the quality and active ingredients of the oil phase caused by prolonged heating. After processing, crude fermented oil is obtained. S303 membrane filtration refining: Membrane filtration technology is used to filter crude fermented oil. By selecting a membrane with a suitable pore size, residual inorganic salts and protein impurities in the crude product can be effectively removed without causing loss of the target active ingredients such as phospholipids and glycolipids. After membrane filtration, a high-purity and stable-quality fermented vegetable oil product is finally obtained. S4 Quality Inspection: S401 Specific Gravity Test: The specific gravity of fermented vegetable oil was measured using an Anton Paar DMA35 handheld densitometer. This instrument is highly accurate and can quickly and accurately obtain the specific gravity data of the sample. Specific gravity is one of the important indicators reflecting the purity and composition of oil. S402 Refractive Index Test: The refractive index of fermented vegetable oil was tested using an ATAGO portable refractometer. The refractive index is closely related to the composition and purity of the oil. By testing the refractive index, the quality of the oil and the presence of impurities can be preliminarily determined. S403 Viscosity Testing: A BROOKFIELD digital touchscreen rotational viscometer was used to measure the viscosity of fermented vegetable oil at 25°C using a #2 rotor at 200 rpm. Viscosity affects the feel of the oil on the skin and the stability of the formula, and is an important parameter in the product application process. S404 Acid Value Test: The acid value of fermented vegetable oil is tested according to the national standard "GB 5009.229-2016 Determination of Acid Value in Food". The acid value reflects the content of free fatty acids in the oil. Excessively high acid value will affect the stability and safety of the oil. S405 Peroxide Value Test: The peroxide value of fermented vegetable oils is tested according to the standard "GB 5009.227-2023 Determination of Peroxide Value in Food". Peroxide value is a key indicator for measuring the degree of oxidation of oils. Excessive peroxide value indicates that the oil has undergone oxidative deterioration, which will affect the quality and efficacy of the product. S406 Total Triterpenoid Content Detection: The total triterpenoid content in fermented vegetable oil was detected according to "NYT3676-2020 Determination of Total Triterpenoid Content in Ganoderma lucidum by Spectrophotometry". Total triterpenoids are one of the important active ingredients in this fermented vegetable oil, and their content directly affects the efficacy of the product. S407 Phospholipid Content Detection: Phospholipid content is detected according to the molybdenum blue colorimetric method in "GB / T 5537-2008 Grain and Oil Inspection - Determination of Phospholipid Content". Phospholipids are key functional ingredients in products, and accurate detection of their content can ensure that product quality meets standards. S408 Glycolipid Content Detection: Glycolipid content was detected by high performance liquid chromatography (HPLC). Figure 2 High-performance liquid chromatography (HPLC) is characterized by high separation efficiency and high detection accuracy, which can accurately determine the content of glycolipids and ensure that the glycolipid components in the product meet the requirements. S409 Lipid Composition Analysis: Lipid analysis of raw oil and post-fermentation oil was performed using TOF / MS (Time-of-Flight Mass Spectrometry). Figure 3 (TOF / MS TIC negative spectrum). This analytical method can clearly show the changes in various lipid components in the oil before and after fermentation, clarify the impact of the fermentation process on lipid components, and further verify the effectiveness of the fermentation process. S4010 Skin Efficacy Related Testing: Testing Skin Moisture Content ( Figure 4 (Comparison chart of skin moisture content) and transepidermal water loss (TEWL) value ( Figure 5 Transdermal water loss (TWEL) comparison chart was used to evaluate the moisturizing and barrier repair effects of fermented plant oils; simultaneously, its effect on the expression levels of PPARα, PPARβ, and PPARγ genes was detected (Figure 6-11: Comparison chart of relative PPAR gene expression levels of different oils), verifying its function in regulating skin cell metabolism; in addition, the liquid crystal phenomenon after the fermented oil was formulated into an emulsion was observed (…). Figure 12 : Liquid crystal phenomenon diagram of emulsion preparation with fermented olive oil; Figure 13 (Illustration of liquid crystal structure in emulsion formulated with fermented meadowfoam seed oil) The liquid crystal structure helps to improve the stability and skin permeability of the emulsion, further ensuring the application effect of the product; Furthermore, this plant oil autonomously generates multiple active ingredients, precisely regulating skin metabolism. The core microbial strain drives the synthesis of active ingredients: the selected exclusive microbial strain Ustilago scitaminea MEL03 (accession number CGMCC34427) has a highly efficient lipid metabolism capacity. During fermentation, it can naturally synthesize functional components such as phospholipids, glycolipids, and total triterpenes without the need for exogenous addition. Among them, phospholipids can enhance the skin barrier's water-locking ability, glycolipids can improve the permeability of skin cells, and total triterpenes have anti-inflammatory and antioxidant effects. Activating key genes in skin metabolism: Skin efficacy testing has verified that this fermented plant oil can significantly upregulate the relative expression levels of PPARα, PPARβ, and PPARγ genes (as shown in Figure 6-11). Among them, PPARα regulates stratum corneum renewal, PPARβ promotes skin cell proliferation, and PPARγ repairs damaged barriers. It improves skin metabolic imbalance at the molecular level, overcoming the limitation of traditional plant oils that "only moisturize and do not regulate," and improving the repair effect on dry and sensitive skin by more than 30%. Furthermore, this process improves oil-water miscibility, simplifies formulation design, and reduces the risk of irritation. Microbial metabolism improves interfacial activity: Ustilago scitaminea MEL03 produces natural interfacial active substances (such as glycosylglycerol, ester peptides, etc., as shown in Figure 3) when metabolizing oils, which significantly improves the oil-water miscibility of fermented vegetable oils. Experiments show that when this fermented vegetable oil is mixed with the aqueous phase, the separation time is extended to more than 5 times that of traditional vegetable oils, and stable fusion can be achieved without the addition of additional chemical emulsifiers. Reduced irritation and enhanced naturalness: Because it does not rely on exogenous emulsifiers, the product reduces skin irritation by 60% while retaining the natural properties of plant oils, which aligns with the trend of "gentle skincare" and is suitable for the needs of special groups such as those with sensitive skin and pregnant women. Furthermore, this process optimizes the purification process to ensure product quality and stability. Multi-step purification removes impurities and reduces safety risks: the post-treatment process (S3 step) of "centrifugal separation - flash dehydration - membrane filtration purification" effectively removes bacteria, waste liquid, inorganic salts, and residual proteins from the fermentation broth. Centrifugal separation first separates the crude oil phase; flash dehydration utilizes rapid pressure reduction to avoid high-temperature damage to active ingredients; and membrane filtration precisely traps impurities (pore size adaptability design). The final product's acid value meets GB 5009.229-2016 standard (≤1.0mg KOH / g), and its peroxide value meets GB 5009.227-2023 standard (≤0.15g / 100g), far exceeding the quality indicators of traditional vegetable oils. Extended shelf life: The low impurity content and stable active ingredient structure extend the product's shelf life to 24 months, solving the problem of easy oxidation and deterioration of traditional vegetable oils and reducing the company's storage and loss costs.

[0023] The embodiments of the present invention have been described in detail above with reference to the accompanying drawings. However, the present invention is not limited to the above embodiments. Within the scope of knowledge possessed by those skilled in the art, various changes can be made without departing from the spirit of the present invention.

Claims

1. A skin metabolism regulating fermentation plant oil enriched with phosphoglycolipid, comprising a core bacterial strain and a culture medium formulation, characterized in that: The culture medium formula includes plate screening medium (YM medium), strain activation and primary seed culture medium, secondary seed culture medium and fermentation medium; ​ The core strain is Ustilago scitaminea MEL03, which is screened from oil-rich sunflower seeds, identified by the Chinese Academy of Sciences Microbiology Institute, and preserved in the China General Microbiological Culture Collection Center with the preservation number CGMCC34427. The strain has the ability to efficiently metabolize oil and naturally produce phospholipids and glycolipids and other skin-beneficial ingredients during fermentation, and is the key biological basis for preparing the fermented plant oil.

2. The skin metabolism regulating fermented plant oil enriched with glycolipids according to claim 1, characterized by: The plate screening medium (YM medium) is used for preliminary culture and screening of the strain, and its components include glucose 10 g / L, tryptone 5 g / L, malt extract powder 3 g / L and yeast extract powder 3 g / L, which can provide the necessary carbon source, nitrogen source and other nutrients for the initial growth of the strain, and help the strain to form observable colonies.

3. The skin metabolism regulating fermented plant oil enriched with glycolipids according to claim 1, characterized by: The strain activation and primary seed culture medium is used for strain activation and primary seed liquid culture, and its components include glucose 30 g / L, NaNO3 1 g / L, yeast extract 3 g / L, KH2PO4 3 g / L and MgSO4 0.3 g / L. Glucose provides the main carbon source, NaNO3 and yeast extract provide the nitrogen source, and KH2PO4 and MgSO4 serve as inorganic salts to provide a suitable nutrient environment for the rapid proliferation of the activated strain, ensuring that the primary seed liquid has sufficient activity and quantity.

4. The skin metabolism regulating fermented plant oil enriched with glycolipids according to claim 1, characterized by: The secondary seed culture medium has the same components as the primary seed culture medium. The same nutrient formula is continued to ensure that the strain grows stably during the propagation process and maintains a high activity, thereby meeting the quantity and quality requirements of the secondary seed liquid and laying a good foundation for the subsequent fermentation process.

5. The skin metabolism regulating fermented plant oil enriched with glycolipids according to claim 1, characterized by: The fermentation medium is the key environment for the strain to produce active ingredients during fermentation, and its components include glucose 20 g / L, oil 100 g / L, NaNO3 3 g / L, yeast extract 1 g / L, KH2PO4 0.3 g / L and MgSO4 0.3 g / L. The oil content in this formula is high to meet the carbon source demand of oil-loving bacteria, and the ratio of nitrogen source and inorganic salt is adjusted to be more suitable for the strain to produce phospholipids, glycolipids and other functional ingredients. In the later stage of the fermentation process, natural plant oil (olive oil or white pool flower seed oil) needs to be added, and the volume ratio of the added plant oil to the primary fermentation liquid is controlled at 0.5:1-2 to further improve the fermentation effect and the content of active ingredients in the product.

6. A process for the preparation of skin metabolism regulating fermented plant oil enriched with phosphoglycolipids, characterized by: The process includes S1 strain screening and purification, S2 fermentation production, S3 post-treatment purification and S4 quality detection. S1 strain screening and purification includes S101 sample collection and pretreatment, S102 seed crushing and mixing, S103 culture and screening and S104 strain determination. S2 fermentation production includes S201 first seed liquid preparation, S202 second seed liquid preparation, S203 first fermentation liquid preparation and S204 second fermentation liquid preparation; S3 post-processing purification includes S301 centrifugal separation, S302 flash dehydration and S303 membrane filtration refining; S4 quality detection includes S401 specific gravity detection, S402 refractive index detection, S403 viscosity detection, S404 acid value detection, S405 peroxide value detection, S406 total triterpenoid content detection, S407 phospholipid content detection, S408 glycolipid content detection, S409 lipid component analysis and S4010 skin efficacy related detection.

7. The preparation process of a skin metabolism-regulating fermented plant oil rich in phospholipids and glycolipids according to claim 6, characterized in that: The S101 sample collection and pretreatment: first, collect fresh sunflower seeds rich in oil, because the environment of such seeds is more prone to oil-loving bacteria, after collection, the seeds are subjected to strict pretreatment, first washed with sterile water for 3-5 times to remove the surface dust and impurities; then rinsed with 75% disinfectant alcohol for 3-5 minutes for preliminary disinfection; then washed with sterile water for 3 times to remove the residual alcohol; finally, control the water content on the clean bench and perform ultraviolet irradiation sterilization for 30 minutes to ensure that the subsequent operation is carried out in a sterile environment to avoid bacterial contamination; S102 seed crushing and mixing: under the sterile state of the clean bench, carefully remove the seed coat to obtain the embryo (kernel), then cut the kernel into pieces with a volume roughly equivalent to that of a sesame seed using a sterile scalpel, which can increase the contact area of the kernel with the subsequent solution, facilitating the release of microorganisms, after cutting, add a small amount of sterile water to the pieces and place them in a shaker for thorough mixing to uniformly disperse the microorganisms into the aqueous solution; S103 culture and screening: take the supernatant of the mixed aqueous solution, evenly spread it on the pre-prepared YM plate culture medium, place the plate in a culture environment of 32-37°C, culture for 3-5 days to allow the microorganisms to grow and form colonies, then pick 50 single colonies with different morphologies from the plate and inoculate them into a liquid culture medium containing oil (the same composition as the strain activation and primary seed culture medium, i.e. glucose 30g / L, NaNO31g / L, yeast extract 3g / L, KH2PO43g / L, MgSO40.3g / L), culture at 32-37°C for 48-72 hours to allow the strains in the colonies to further proliferate in the liquid environment; S104 strain determination: after fermentation culture, let the fermentation broth of each strain stand for a period of time until the oil and water phases separate, then take one drop of the oil phase from the upper layer of each fermentation broth and add it to a smooth surface, then add one drop of red water drop to each, observe the state of the water drop to determine the oil-water miscibility, select the strain with the best oil-water miscibility, which is the target strain Ustilago scitaminea MEL03, which has the ability to efficiently metabolize oil and promote oil-water fusion, and is the core strain for subsequent preparation of fermented vegetable oil.

8. The process for preparing a phosphoglycolipid-rich skin metabolism regulating fermented vegetable oil according to claim 6, characterized by: The S201 primary seed liquid preparation: the screened Ustilago scitaminea MEL03 strain is activated, the activated strain is inoculated into a primary seed culture medium, the inoculated culture medium is placed in a culture condition of 32-37 DEG C, and is cultured for 24-36 hours; in the process, the strain proliferates rapidly, and finally forms a primary seed liquid with high activity and sufficient quantity, providing an initial strain for subsequent preparation of a secondary seed liquid; The S202 secondary seed liquid preparation: according to a 10% inoculation ratio, the prepared primary seed liquid is inoculated into a secondary seed tank containing a secondary seed culture medium, a culture temperature of 32-37 DEG C is maintained, and the culture is continued for 36-48 hours; through scale-up culture, the strain quantity is further increased, and the activity is kept stable, and finally the secondary seed liquid meeting the fermentation requirement is obtained; The S203 primary fermentation liquid preparation: according to a 10% inoculation ratio, the secondary seed liquid is inoculated into a fermentation device containing a fermentation culture medium, the culture temperature is controlled at 32-37 DEG C, and the culture time is 48-72 hours; in this stage, the strain proliferates in a suitable nutrient environment, starts to metabolize oil and fat, and produces preliminary active ingredients, and finally forms a primary fermentation liquid; The S204 secondary fermentation liquid preparation: natural plant oil (olive oil or white pool flower seed oil) is added to the primary fermentation liquid, the volume ratio of the plant oil to the primary fermentation liquid is strictly controlled to be 0.5:1-2, the culture temperature of 32-37 DEG C is maintained, and the culture is continued for 48-96 hours; in the process, the strain further metabolizes the added plant oil, produces more phospholipid and glycolipid active ingredients, and finally obtains a secondary fermentation liquid with more components and higher activity.

9. The process for preparing a skin metabolism regulating fermented vegetable oil enriched with glycolipids according to claim 6, characterized in that: The S301 centrifugal separation: The secondary fermentation liquid is subjected to centrifugal treatment, the centrifugal force is used to separate the strain, fermentation waste liquid and oil phase in the fermentation liquid, after the centrifugal treatment is completed, the strain in the bottom sediment and the fermentation waste liquid in the upper layer are discarded, and only the middle oil phase part is reserved, and the oil phase part is the crude oil containing the target active ingredients; The S302 flash dehydration: the oil phase obtained through the centrifugal separation is sent into a flash device for flash dehydration treatment; the flash dehydration uses the principle of rapid pressure reduction to make the residual water in the oil phase vaporize rapidly and be removed, effectively avoiding the damage of long-time heating to the oil phase quality and active ingredients; after the treatment, the fermentation oil crude product is obtained; The S303 membrane filtration refining: the membrane filtration technology is used to filter the fermentation oil crude product; through selecting a membrane with a suitable pore size, the residual inorganic salt and protein impurities in the crude product can be effectively removed, and the phospholipid and glycolipid target active ingredients are not lost; after the membrane filtration, the fermentation plant oil finished product with high purity and stable quality is finally obtained.

10. The process for preparing a phosphoglycolipid-rich skin metabolism regulating fermented vegetable oil according to claim 6, characterized by: The S401 specific gravity detection: the specific gravity of the fermentation plant oil is measured by using an Anton Paar handheld density meter DMA35; the instrument has high precision, and can quickly and accurately obtain the specific gravity data of the sample; the specific gravity is one of important indexes reflecting the purity and composition of oil. S402 Refractive index detection: Use ATAGO portable refractometer to detect the refractive index of fermented vegetable oil, which is closely related to the composition and purity of oil. The quality of oil and the presence of impurities can be preliminarily judged by detecting the refractive index; S403 Viscosity detection: Use BROOKFIELD digital touch screen rotary viscometer to measure the viscosity of fermented vegetable oil at 25℃ temperature condition, using 2# rotor at 200rpm speed. Viscosity will affect the skin feel and formula stability of oil, which is an important parameter in product application process; S404 Acid value detection: According to the national standard "GB 5009.229-2016 Determination of acid value in food", the acid value of fermented vegetable oil is detected. Acid value reflects the content of free fatty acid in oil. High acid value will affect the stability and safety of oil; S405 Peroxide value detection: According to the standard "GB 5009.227-2023 Determination of peroxide value in food", the peroxide value of fermented vegetable oil is detected. Peroxide value is a key indicator to measure the degree of oil oxidation. High peroxide value indicates that the oil has been oxidized and deteriorated, which will affect the quality and efficacy of the product; S406 Total triterpenoid content detection: According to "NYT3676-2020 Determination of total triterpenoids in Ganoderma lucidum by spectrophotometry", the content of total triterpenoids in fermented vegetable oil is detected. Total triterpenoids are one of the important active ingredients in the fermented vegetable oil, and their content directly affects the strength of the product's efficacy; S407 Phospholipid content detection: According to the molybdenum blue colorimetric method in "GB / T 5537-2008 Grain and oil testing Determination of phospholipid content", the phospholipid content is detected. Phospholipid is a key active ingredient in the product, and accurate detection of its content can ensure that the product quality meets the standards; S408 Glycolipid content detection: High performance liquid chromatography is used to detect the content of glycolipid. High performance liquid chromatography has the characteristics of high separation efficiency and high detection precision, which can accurately determine the content of glycolipid and ensure that the glycolipid content in the product meets the requirements; S409 Lipid composition analysis: TOF / MS (Time of Flight Mass Spectrometry) is used to analyze the lipid composition of raw oil and fermented oil. Through this analysis method, the changes of various lipid components in oil before and after fermentation can be clearly understood, the influence of fermentation process on lipid composition can be determined, and the effectiveness of fermentation process can be further verified; S4010 Skin efficacy related detection: Detect the skin moisture content to evaluate the moisturizing and barrier repair effect of fermented vegetable oil; At the same time, detect the influence on the expression amount of PPARα, PPARβ and PPARγ genes to verify its function of regulating skin cell metabolism; In addition, observe the liquid crystal phenomenon of emulsion prepared by fermented oil. Liquid crystal structure helps to improve the stability and skin permeability of emulsion, further ensuring the application effect of the product.

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