Green biological surfactant with oil control and dandruff removal effects as well as preparation method and application of green biological surfactant

Abstract

The invention belongs to the technical field of daily chemicals, and discloses a green biological surfactant with oil control and dandruff removal effects and a preparation method and application thereof. The biological surfactant comprises the following components in parts by mass: 30-60 parts of sophorolipid, 10-40 parts of tea saponin and the balance of water, wherein the total mass part of the water is 100. The surface tension of a system is remarkably reduced through the composite effect between sophorolipid molecules and tea saponin molecules, so that the surfactant has good foam stability and good oil control and dandruff removal effects, is suitable for being applied to cosmetic emulsions and daily chemical systems, and can meet the green, safe and efficient market requirements.

Description

Technical Field

[0001] This invention belongs to the field of daily chemical products technology, and specifically relates to a green biosurfactant with oil-controlling and dandruff-removing effects, its preparation method, and its application. Background Technology

[0002] Surfactants are the cornerstone of daily chemical products (such as shampoos, shower gels, and facial cleansers), playing core roles in cleaning, foaming, and emulsification. Their development has evolved from soap-based surfactants to chemically synthesized surfactants, and now to the much-discussed biosurfactants. The driving force behind this evolution is the continuous pursuit of safety, environmental compatibility, and functional diversification.

[0003] For a long time, the personal care market has been dominated by chemically synthesized surfactants, mainly including sodium lauryl sulfate (SLS), sodium laureth sulfate (SLES), and cocamidopropyl betaine (CAB). These surfactants are widely used because of their readily available raw materials, low cost, strong cleaning power, and excellent foaming performance. However, as consumers' requirements for product experience and safety have increased, the inherent defects of chemically synthesized surfactants have become increasingly prominent: (1) Irritation problem: Anionic surfactants such as SLS / SLES have a strong degreasing ability and may excessively strip the natural sebum of the stratum corneum of the skin, damaging the skin barrier function, leading to dry and tight skin, and even causing sensitivity, stinging and other problems; (2) Environmental pressure: The production of chemically synthesized surfactants depends on petrochemical raw materials, and their biodegradability is relatively poor. Some metabolites may have a lasting impact on the environment, which is contrary to the concept of global sustainable development.

[0004] To address these challenges, the industry has turned its attention to biosurfactants. Biosurfactants are surface-active amphiphilic compounds produced by microorganisms or plants, such as sophorolipids, rhamnolipids, alkyl glycosides (APGs), and tea saponins. They are hailed as "green" surfactants due to their eco-friendly nature, biodegradability, environmentally friendly production processes, and extremely low toxicity and irritation, representing an important direction for future surfactant development. Despite the clear advantages of biosurfactants in safety and environmental friendliness, current industry and consumer perception of them remains primarily based on the labels of "mild" and "green." In commercial applications, the vast majority of biosurfactants have not demonstrated significant and stable specific effects beyond their basic cleaning function (such as strong oil control, dandruff removal, and acne treatment). This often results in these biosurfactants only being used as auxiliary ingredients or conceptual additives, making it difficult for them to become key functional ingredients driving the core efficacy of products.

[0005] The market urgently needs new solutions that combine the gentle and environmentally friendly properties of biosurfactants with precise and powerful efficacy. Currently, when used alone, biosurfactants often cannot match the efficacy of traditional chemical functional ingredients, thus rendering the addition of biosurfactants merely a conceptual addition that cannot truly play a role. Summary of the Invention

[0006] In order to overcome the shortcomings and deficiencies of the prior art, the primary objective of this invention is to provide a green biosurfactant with oil control and dandruff removal effects.

[0007] Another objective of this invention is to provide a method for preparing the aforementioned green biosurfactant with oil-controlling and dandruff-removing effects.

[0008] Another object of the present invention is to provide the application of the above-mentioned green biosurfactant with oil-controlling and dandruff-removing effects.

[0009] The objective of this invention is achieved through the following solution: A green biosurfactant with oil-controlling and dandruff-removing effects, comprising, by weight, 30-60 parts sophorolipid, 10-40 parts tea saponin, and water to a total weight of 100.

[0010] More preferably, the biosurfactant with oil-controlling and dandruff-removing effects comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0011] Optionally, in some embodiments of the present invention, the method for preparing the sophorolipid includes: B1. Inoculate the seed culture of *Candida bacillus* into a fermentation medium and ferment to obtain a crude fermentation broth; B2. The crude fermentation broth is sterilized, filtered, purified, and dried to obtain the sophorolipid.

[0012] Optionally, in some embodiments of the present invention, in step B1, the method for preparing the bumblebee Candida seed solution includes: inoculating bumblebee Candida colonies into YM medium, and obtaining the bumblebee Candida seed solution after expansion culture.

[0013] Optionally, in some embodiments of the present invention, the temperature of the expansion culture is 28-33°C, the time of the expansion culture is 36-60h, and the rotation speed of the fermentation shaker is 120-180rpm.

[0014] Optionally, in some embodiments of the present invention, in step B1, the fermentation medium includes oil, carbon source, nitrogen source, inorganic salt and water.

[0015] Optionally, in some embodiments of the present invention, the oil includes rapeseed oil and / or sunflower oil; the amount of oil added is 2-10% of the total mass of the fermentation medium.

[0016] Optionally, in some embodiments of the present invention, the carbon source includes at least one of glucose, fructose, and sucrose; the amount of carbon source added is 1-5% of the total mass of the fermentation medium.

[0017] Optionally, in some embodiments of the present invention, the nitrogen source includes at least one of yeast extract, urea, soybean peptone, tryptone, and ammonium sulfate; the amount of nitrogen source added is 0.2-1% of the total mass of the fermentation medium.

[0018] Optionally, in some embodiments of the present invention, the inorganic salt includes at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate, wherein the inorganic salt adjusts the pH of the fermentation medium to 4.5-5.5.

[0019] Optionally, in some embodiments of the present invention, the amount of water added is to make up to 100% of the total mass of the fermentation medium.

[0020] Optionally, in some embodiments of the present invention, in step B1, the inoculation amount of the bumblebee Candida seed liquid is 2-6% (v / v).

[0021] Optionally, in some embodiments of the present invention, the fermentation conditions are as follows: fermentation temperature of 28-33℃, fermentation time of 3-7 days, stirring speed of 350-450 rpm, and sterile air ventilation rate of 0.3-0.6 vvm.

[0022] Optionally, in some embodiments of the present invention, in step B2, the sterilization conditions are: sterilization temperature of 85-95°C and sterilization time of 20-40 min.

[0023] Optionally, in some embodiments of the present invention, in step B2, the filtration is diatomaceous earth filter filtration.

[0024] Optionally, in some embodiments of the present invention, in step B2, the purification step includes: passing the filtrate obtained after filtration sequentially through a cation exchange resin, an anion exchange resin, and a macroporous adsorption resin, and collecting the purified solution.

[0025] Optionally, in some embodiments of the present invention, in step B2, the drying process is preferably spray drying, wherein the spray drying conditions are: inlet temperature of 110-130°C and outlet temperature of 70-80°C.

[0026] Optionally, in some embodiments of the present invention, the method for preparing the tea saponin includes: S1. Inoculate the Lactobacillus fermentation seed liquid into the fermentation substrate and culture to obtain a fermentation broth containing bacteria; S2. Add camellia oleifera residue powder to the fermentation broth containing bacteria, ferment and extract, and then heat treat to obtain crude fermentation broth containing tea saponins; S3. Add auxiliary materials to the crude fermentation liquid containing tea saponin, and obtain the tea saponin after subcritical cycle extraction and drying.

[0027] Optionally, in some embodiments of the present invention, in step S1, the method for preparing the Lactobacillus fermentum seed liquid includes: inoculating Lactobacillus fermentum colonies into MRS medium, and obtaining the Lactobacillus fermentum seed liquid after expansion culture.

[0028] Optionally, in some embodiments of the present invention, the temperature of the expansion culture is 33-40°C, the time of the expansion culture is 36-60 h, and the rotation speed of the fermentation shaker is 50-120 rpm.

[0029] Optionally, in some embodiments of the present invention, in step S1, the inoculation amount of the Lactobacillus fermentation seed liquid is 1-3% (v / v).

[0030] Optionally, in some embodiments of the present invention, in step S1, the fermentation substrate includes a nitrogen source, a carbon source, inorganic salts, and water.

[0031] Optionally, in some embodiments of the present invention, the nitrogen source includes at least one of soybean peptone, tryptone, peanut meal, and soybean meal; the amount of nitrogen source added is 1-3% of the total mass of the fermentation substrate.

[0032] Optionally, in some embodiments of the present invention, the carbon source includes at least one of glucose, maltose, and sucrose; the amount of carbon source added is 1-3% of the total mass of the fermentation substrate.

[0033] Optionally, in some embodiments of the present invention, the inorganic salt includes at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate, wherein the inorganic salt adjusts the pH of the fermentation substrate to 3.5-5.5.

[0034] Optionally, in some embodiments of the present invention, in step S1, the culture temperature is 33-40°C, the culture time is 18-30 h, and the sterile air ventilation rate is 0.05–0.2 vvm.

[0035] Optionally, in some embodiments of the present invention, in step S2, the mass ratio of the fermentation broth containing bacteria to the camellia oleifera residue powder is (1-3):1.

[0036] Optionally, in some embodiments of the present invention, in step S2, the fermentation extraction temperature is 33-40°C, the fermentation extraction time is 18-30 h, and the sterile air ventilation rate is 0.05-0.2 vvm.

[0037] Optionally, in some embodiments of the present invention, in step S2, the temperature of the heat treatment is 85-95°C, and the time of the heat treatment is 1-3 hours.

[0038] Optionally, in some embodiments of the present invention, in step S3, the auxiliary material includes silica granulation, and the mass ratio of the auxiliary material to the crude fermentation liquid containing tea saponin is (2-4):1.

[0039] Optionally, in some embodiments of the present invention, in step S3, the conditions for the subcritical cyclic extraction include: the extractant includes methanol and / or ethanol; the temperature is 40-50°C; and the pressure is 0.4-0.6 MPa.

[0040] Optionally, in some embodiments of the present invention, in step S3, the drying method is spray drying, and the conditions for spray drying include: an inlet temperature of 110-130°C and an outlet temperature of 70-80°C.

[0041] Another aspect of the present invention provides a method for preparing the above-mentioned green biosurfactant with oil-controlling and dandruff-removing effects, comprising: Sophorolipids and tea saponins are added to water and mixed evenly to obtain a green biosurfactant with oil-controlling and dandruff-removing effects.

[0042] In another aspect, the present invention provides the application of the above-mentioned green biosurfactant with oil-controlling and dandruff-removing effects in the preparation of daily chemical products.

[0043] Optionally, in some embodiments of the present invention, the amount of the green biosurfactant with oil-controlling and dandruff-removing effects added to the daily chemical product is 0.1-20 wt%.

[0044] Compared with the prior art, the present invention has the following advantages and beneficial effects: Currently, the extraction of tea saponins mainly relies on solvent systems such as water or ethanol. However, traditional methods generally suffer from low extraction rates, high energy consumption, and insufficient purity of the obtained products, making it difficult to meet the demands of green and efficient production. This invention applies a beneficial bacteria fermentation system and low-temperature phase change extraction technology as an innovative process to replace traditional solvent extraction. This method uses Lactobacillus fermentation to initially and effectively reduce impurities such as polysaccharides and fiber in tea cake residue, further improving the purity of tea saponins. On the other hand, low-boiling-point liquid methanol is used as the extractant, and subcritical cyclic extraction is carried out in a closed extractor under mild conditions of approximately 40-50 °C and 0.5 MPa. The liquefied gas remains liquid under pressure, which can efficiently dissolve tea saponins, greatly increasing the extraction rate and fully extracting tea saponins from the tea cake residue. During decompression, it rapidly vaporizes, achieving instantaneous and complete separation of the solvent and the target product, avoiding the risk of residual organic solvents.

[0045] This invention proposes a novel strategy for constructing a natural surfactant by combining sophorolipids and tea saponins, resulting in a green biosurfactant. The green biosurfactant of this invention uses water as a solvent and sophorolipids and tea saponins as surfactants. Sophorolipid molecules have high hydrophilicity and dissociability, while tea saponins molecules are based on a triterpenoid steroidal skeleton, possessing both hydrophobic steroidal rings and multiple hydrophilic hydroxyl groups. At gas-liquid and liquid-liquid interfaces, the two can form complementary adsorption layers through hydrophobic interactions, hydrogen bonding, and electrostatic interactions, enhancing the close arrangement of molecules at the interface. This composite effect significantly reduces the surface tension of the system, thus enabling the green biosurfactant to exhibit excellent foam stability and superior surfactant performance without the need for high concentrations of irritating surfactants.

[0046] Tests have shown that the green biosurfactant of this invention has excellent oil control and dandruff removal effects, making it suitable for use in cosmetic emulsions and daily chemical systems, and meeting the market demand for green, safe and efficient products. Detailed Implementation

[0047] The present invention will be further described in detail below with reference to embodiments, but the implementation of the present invention is not limited thereto. Unless otherwise specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments used, unless otherwise specified, are all commercially available products.

[0048] The bumblebee Candida yeast used in this invention is numbered ATCC 22214 and comes from Wuhan Gray Algae Biotechnology Co., Ltd.

[0049] The Lactobacillus fermentum used in this invention is numbered ATCC 11739 and originated from Wuhan Gray Algae Biotechnology Co., Ltd.

[0050] The *Lactobacillus plantarum* used in this invention is designated GIM1.648 and is sourced from Ningbo Mingzhou Biotechnology Co., Ltd.

[0051] The Lactobacillus casei used in this invention is designated GIM1.204 and is sourced from Ningbo Mingzhou Biotechnology Co., Ltd.

[0052] The rhamnolipin used in this invention is sourced from Shanghai Aladdin Biochemical Technology Co., Ltd., and its model number is R488080.

[0053] The camellia oleifera residue powder used in this invention is obtained by grinding waste camellia oleifera residue.

[0054] The YM liquid culture medium used in this invention is a commonly used culture medium in the prior art, and its formula is as follows: glucose 10g / L, peptone 5g / L, yeast extract 3g / L, malt extract 3g / L, and water as the solvent. If necessary, the pH value can be adjusted to 6.2±0.2 with 1mol / L NaOH solution or 1mol / L HCl.

[0055] The MRS liquid culture medium used in this invention is a commonly used culture medium in the prior art, and its formula is as follows: peptone 10 g / L, beef extract 10 g / L, yeast extract 5 g / L, glucose 20 g / L, dipotassium hydrogen phosphate 2 g / L, sodium acetate 5 g / L, triammonium citrate 2 g / L, magnesium sulfate 0.1 g / L, manganese sulfate 0.05 g / L, Tween 80 1 g / L, and water as the solvent. If necessary, the pH value can be adjusted to 6.4 ± 0.2 with 1 mol / L NaOH solution or 1 mol / L HCl.

[0056] The cation exchange resin used in this invention is D202 anion exchange resin.

[0057] The anion exchange resin used in this invention is HZD-2 cation exchange resin.

[0058] The macroporous adsorption resin used in this invention is LSA-10 macroporous adsorption resin.

[0059] The commercially available sophorolipid used in this invention is sourced from Shanghai Boku Biotechnology Co., Ltd., and its model number is EcolifeSL-02.

[0060] The commercially available tea saponin used in this invention is sourced from Xi'an Aosai Biotechnology Co., Ltd., and its trade name is Tea Tree Seed Plant Extract (containing 60% tea saponin). Unless otherwise specified, all water used in this invention is deionized water.

[0061] Unless otherwise specified, all percentages used in the description of this invention are mass percentages.

[0062] Example 1: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0063] The preparation method includes adding sophorolipids and tea saponins to water and mixing them evenly to obtain a green biosurfactant.

[0064] The preparation methods of sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 4% (v / v) into the fermentation medium (the fermentation medium consists of 5% sunflower oil, 3% glucose, 0.6% soybean peptone, and the remainder water; the pH of the fermentation medium is adjusted to 5.0 with sodium dihydrogen phosphate). Ferment for 5 days at 30°C, with a stirring speed of 400 rpm and a sterile air flow rate of 0.45 vvm to obtain the crude fermentation broth.

[0065] B2. The crude fermentation broth was sterilized by incubating at 90℃ for 30 min, and then filtered through a diatomaceous earth filter. The filtrate was then passed through a cation exchange resin, anion exchange resin and macroporous adsorption resin in sequence. The purified solution was collected and spray-dried (inlet air temperature 120℃, outlet air temperature 75℃). The resulting solid was sophorolipid.

[0066] The preparation methods for tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 2% (v / v) into the fermentation substrate (the fermentation substrate consists of 2% peanut cake powder, 2% maltose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate), and incubate at 37°C with sterile air at a ventilation rate of 0.12 vvm for 24 h to obtain the fermentation broth containing bacteria.

[0067] S2. Sterilized camellia oleifera residue powder (mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 2:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 37℃ and sterile air flow rate of 0.12 vvm for 24 hours, the broth is heat-treated at 90℃ for 2 hours to obtain crude fermentation broth containing tea saponins.

[0068] S3. Add three times the mass of silica to the crude fermentation broth containing tea saponin to granulate, and then obtain tea saponin by subcritical cycle extraction (extractant is methanol, temperature is 45℃, pressure is 0.5 MPa) and spray drying (inlet air temperature is 120℃, outlet air temperature is 75℃).

[0069] The preparation method of the bumblebee Candida seed liquid in this embodiment includes: inoculating bumblebee Candida colonies into YM medium, and expanding the culture at 30°C and a shaking speed of 150 rpm for 48 h to obtain bumblebee Candida seed liquid.

[0070] The method for preparing Lactobacillus fermentation seed liquid in this embodiment includes: inoculating Lactobacillus fermentation colonies into MRS medium, and culturing them at 37°C and a shaking speed of 80 rpm for 48 hours to obtain Lactobacillus fermentation seed liquid.

[0071] Examples 2-4: A green biosurfactant with oil-controlling and dandruff-removing effects The preparation methods of sophorolipids and tea saponins in the green biosurfactants of Examples 2-4 are the same as those in Example 1, the only difference being the mass ratio of sophorolipids, tea saponins and water. The proportions of each component in the green biosurfactants with oil-controlling and dandruff-removing effects of Examples 1-4 are shown in Table 1 below.

[0072] Table 1

[0073] Example 5: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0074] The preparation methods for green biosurfactants and tea saponins are the same as in Example 1.

[0075] The preparation methods of sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 4% (v / v) into the fermentation medium (the fermentation medium consists of 5% rapeseed oil, 3% glucose, 0.6% soybean peptone, and the remainder water; the pH of the fermentation medium is adjusted to 5.0 with sodium dihydrogen phosphate). Ferment for 5 days at 30°C, with a stirring speed of 400 rpm and a sterile air flow rate of 0.45 vvm to obtain the crude fermentation broth.

[0076] B2. The crude fermentation broth was sterilized by incubating at 90℃ for 30 min, and then filtered through a diatomaceous earth filter. The filtrate was then passed through a cation exchange resin, anion exchange resin and macroporous adsorption resin in sequence. The purified solution was collected and spray-dried (inlet air temperature 120℃, outlet air temperature 75℃). The resulting solid was sophorolipid.

[0077] The preparation method of the bumblebee Candida seed liquid is the same as in Example 1.

[0078] Example 6: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0079] The preparation methods for green biosurfactants and tea saponins are the same as in Example 1.

[0080] The preparation methods of sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 4% (v / v) into the fermentation medium (the fermentation medium consists of 5% sunflower seed oil, 3% glucose, 0.6% yeast extract, and the remainder water; the pH of the fermentation medium is adjusted to 5.0 with sodium dihydrogen phosphate). Ferment for 5 days at 30°C, with a stirring speed of 400 rpm and a sterile air flow rate of 0.45 vvm to obtain the crude fermentation broth.

[0081] B2. The crude fermentation broth was sterilized by incubating at 90℃ for 30 min, and then filtered through a diatomaceous earth filter. The filtrate was then passed through a cation exchange resin, anion exchange resin and macroporous adsorption resin in sequence. The purified solution was collected and spray-dried (inlet air temperature 120℃, outlet air temperature 75℃). The resulting solid was sophorolipid.

[0082] The preparation method of the bumblebee Candida seed liquid is the same as in Example 1.

[0083] Example 7: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0084] The preparation methods for green biosurfactants and sophorolipids are the same as in Example 1.

[0085] The preparation methods for tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture into the fermentation substrate (the fermentation substrate consists of 2% soybean peptone, 2% maltose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate) at an inoculation rate of 2% (v / v) and culture at 37°C with sterile air at a ventilation rate of 0.12 vvm for 24 h to obtain the fermentation broth containing bacteria.

[0086] S2. Sterilized camellia oleifera residue powder (mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 2:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 37℃ and sterile air flow rate of 0.12 vvm for 24 hours, the broth is heat-treated at 90℃ for 2 hours to obtain crude fermentation broth containing tea saponins.

[0087] S3. Add three times the mass of silica to the crude fermentation broth containing tea saponin to granulate, and then obtain tea saponin by subcritical cycle extraction (extractant is methanol, temperature is 45℃, pressure is 0.5 MPa) and spray drying (inlet air temperature is 120℃, outlet air temperature is 75℃).

[0088] The preparation method of fermented Lactobacillus seed liquid is the same as in Example 1.

[0089] Example 8: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

[0090] The preparation methods for green biosurfactants and sophorolipids are the same as in Example 1.

[0091] The preparation methods for tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 2% (v / v) into the fermentation substrate (the fermentation substrate consists of 2% peanut cake powder, 2% glucose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate), and incubate at 37°C with sterile air at a ventilation rate of 0.12 vvm for 24 h to obtain the fermentation broth containing bacteria.

[0092] S2. Sterilized camellia oleifera residue powder (mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 2:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 37℃ and sterile air flow rate of 0.12 vvm for 24 hours, the broth is heat-treated at 90℃ for 2 hours to obtain crude fermentation broth containing tea saponins.

[0093] S3. Add three times the mass of silica to the crude fermentation broth containing tea saponin to granulate, and then obtain tea saponin by subcritical cycle extraction (extractant is methanol, temperature is 45℃, pressure is 0.5 MPa) and spray drying (inlet air temperature is 120℃, outlet air temperature is 75℃).

[0094] The preparation method of fermented Lactobacillus seed liquid is the same as in Example 1.

[0095] Example 9: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 40 parts sophorolipid, 10 parts tea saponin and 50 parts water.

[0096] The preparation method includes adding sophorolipids and tea saponins to water and mixing them evenly to obtain a green biosurfactant.

[0097] The preparation methods of sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 2% (v / v) into the fermentation medium (the fermentation medium consists of 5% sunflower oil, 5% rapeseed oil, 5% fructose, 0.2% urea, and the remainder water; adjust the pH of the fermentation medium to 5.5 with dipotassium hydrogen phosphate). Ferment for 3 days at 28°C, with a stirring speed of 450 rpm and a sterile air flow rate of 0.6 vvm to obtain the crude fermentation broth.

[0098] B2. The crude fermentation broth was sterilized by incubating at 85℃ for 40 min, and then filtered through a diatomaceous earth filter. The filtrate was then passed through a cation exchange resin, anion exchange resin, and macroporous adsorption resin in sequence. The purified solution was collected and spray-dried (inlet air temperature 130℃, outlet air temperature 80℃). The resulting solid was sophorolipid.

[0099] The preparation methods for tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 1% (v / v) into the fermentation substrate (the fermentation substrate consists of 1% soybean meal, 3% sucrose and the remainder water, and the pH of the fermentation substrate is adjusted to 5.5 with potassium dihydrogen phosphate). Incubate at 40°C with sterile air at an aeration rate of 0.05 vvm for 18 h to obtain the fermentation broth containing bacteria.

[0100] S2. Sterilized camellia oleifera residue powder (the mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 1:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 40℃ and sterile air ventilation rate of 0.05 vvm for 30 h, the broth is heat-treated at 85℃ for 3 h to obtain crude fermentation broth containing tea saponins.

[0101] S3. Add twice the mass of silica to the crude fermentation broth containing tea saponin to granulate, and then obtain tea saponin by subcritical cycle extraction (extractant is ethanol, temperature is 40℃, pressure is 0.6 MPa) and spray drying (inlet air temperature is 130℃, outlet air temperature is 80℃).

[0102] The preparation method of the bumblebee Candida seed liquid in this embodiment includes: inoculating bumblebee Candida colonies into YM medium, and expanding the culture at 28°C and a shaking speed of 120 rpm for 36 h to obtain bumblebee Candida seed liquid.

[0103] The method for preparing Lactobacillus fermentation seed liquid in this embodiment includes: inoculating Lactobacillus fermentation colonies into MRS medium, and culturing them at 40°C and a shaking speed of 50 rpm for 60 h to obtain Lactobacillus fermentation seed liquid.

[0104] Example 10: A green biosurfactant with oil-controlling and dandruff-removing effects and its preparation method The green biosurfactant in this embodiment comprises, by weight, 20 parts sophorolipid, 40 parts tea saponin and 40 parts water.

[0105] The preparation method includes adding sophorolipids and tea saponins to water and mixing them evenly to obtain a green biosurfactant.

[0106] The preparation methods of sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 6% (v / v) into the fermentation medium (the fermentation medium consists of 1% sunflower oil, 1% rapeseed oil, 1% sucrose, 1% ammonium sulfate, and the remainder is water; the pH of the fermentation medium is adjusted to 4.5 with potassium dihydrogen phosphate). Ferment for 7 days at 33°C, with a stirring speed of 350 rpm and a sterile air flow rate of 0.3 vvm to obtain the crude fermentation broth.

[0107] B2. The crude fermentation broth was sterilized by incubating at 95℃ for 20 min, and then filtered through a diatomaceous earth filter. The filtrate was then passed through a cation exchange resin, anion exchange resin and macroporous adsorption resin in sequence. The purified broth was collected and spray-dried (inlet air temperature 110℃, outlet air temperature 70℃). The resulting solid was sophorolipid.

[0108] The preparation methods for tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 3% (v / v) into the fermentation substrate (the fermentation substrate consists of 3% soybean peptone, 1% glucose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with disodium hydrogen phosphate), and incubate at 33°C with sterile air at a ventilation rate of 0.2 vvm for 30 h to obtain the fermentation broth containing bacteria.

[0109] S2. Sterilized camellia oleifera residue powder (the mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 3:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 33℃ and sterile air flow rate of 0.2 vvm for 18 hours, the broth is heat-treated at 95℃ for 1 hour to obtain crude fermentation broth containing tea saponins.

[0110] S3. Add 4 times the mass of silica to the crude fermentation broth containing tea saponin for granulation. After subcritical cycle extraction (extractant is methanol, temperature is 50℃, pressure is 0.4 MPa) and spray drying (inlet air temperature is 110℃, outlet air temperature is 70℃), tea saponin is obtained.

[0111] The preparation method of the bumblebee Candida seed liquid in this embodiment includes: inoculating bumblebee Candida colonies into YM medium, and expanding the culture at 33°C and a shaking speed of 180 rpm for 60 h to obtain bumblebee Candida seed liquid.

[0112] The method for preparing Lactobacillus fermentation seed liquid in this embodiment includes: inoculating Lactobacillus fermentation colonies into MRS medium, and culturing them at 33°C and a shaking speed of 120 rpm for 36 h to obtain Lactobacillus fermentation seed liquid.

[0113] Comparative Example 1 The green biosurfactant in this comparative example comprises, by weight, 65 parts sophorolipid and 35 parts water.

[0114] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0115] Comparative Example 2 The green biosurfactant in this comparative example comprises, by weight, 65 parts tea saponin and 35 parts water.

[0116] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0117] Comparative Example 3 The green biosurfactant in this comparative example comprises, by weight, 40 parts rhamnolipin, 25 parts tea saponin and 35 parts water.

[0118] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0119] Comparative Example 4 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0120] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0121] The preparation method of sophorolipid is similar to that in Example 1, except that sunflower seed oil is replaced with corn oil in the same mass ratio.

[0122] Comparative Example 5 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0123] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0124] The preparation method of sophorolipid is similar to that in Example 1, except that sunflower seed oil is replaced with palm oil in the same mass ratio.

[0125] Comparative Example 6 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0126] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0127] Methods for preparing sophorolipids include: B1. Inoculate the *Candida baccata* seed culture at a rate of 4% (v / v) into the fermentation medium (the fermentation medium consists of 5% sunflower oil, 3% glucose, 0.6% soybean peptone, and the remainder water; the pH of the fermentation medium is adjusted to 5.0 with sodium dihydrogen phosphate). Ferment for 5 days at 30°C, with a stirring speed of 400 rpm and a sterile air flow rate of 0.45 vvm to obtain the crude fermentation broth.

[0128] B2. Sterilize the crude fermentation broth by incubating at 90℃ for 30 min. Add ethyl acetate twice its volume to the sterilized fermentation broth for extraction. Remove the upper organic phase by rotary evaporation to remove the ethyl acetate. Wash the rotary evaporated liquid twice with n-hexane. Spray dry the obtained rotary evaporated liquid (inlet air temperature 120℃, outlet air temperature 75℃). The obtained solid is sophorolipid.

[0129] Comparative Example 7 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0130] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0131] The preparation method of tea saponin is similar to that of Example 1, except that the fermented Lactobacillus seed liquid used for fermentation is replaced with 1% (v / v) Lactobacillus plantarum seed liquid and 1% (v / v) Lactobacillus casei seed liquid. The remaining steps are the same as the preparation method of tea saponin in Example 1.

[0132] The preparation method of Lactobacillus plantarum seed liquid includes: inoculating Lactobacillus plantarum colonies into MRS medium, and expanding the culture at 37°C and a shaking speed of 80 rpm for 48 h to obtain Lactobacillus plantarum seed liquid.

[0133] The method for preparing Lactobacillus casei seed culture includes: inoculating Lactobacillus casei colonies into MRS medium and culturing them at 37°C and 80 rpm in a fermentation shaker for 48 hours to obtain Lactobacillus casei seed culture.

[0134] Comparative Example 8 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0135] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0136] The preparation method of tea saponin is similar to that of Example 1, except that the fermented Lactobacillus seed liquid used for fermentation is replaced with the same amount of plant Lactobacillus seed liquid. The remaining steps are the same as the preparation method of tea saponin in Example 1.

[0137] The preparation method of Lactobacillus plantarum seed liquid is the same as that of Comparative Example 7.

[0138] Comparative Example 9 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0139] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0140] The preparation method of tea saponin is similar to that of Example 1, except that the Lactobacillus fermentation seed liquid used for the fermentation agent is replaced with an equal amount of Lactobacillus casei seed liquid. The remaining steps are the same as the preparation method of tea saponin in Example 1.

[0141] The preparation method of Lactobacillus casei seed culture is the same as that of Comparative Example 7.

[0142] Comparative Example 10 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0143] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0144] Methods for preparing tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 2% (v / v) into the fermentation substrate (the fermentation substrate consists of 2% peanut cake powder, 2% maltose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate), and incubate at 37°C with sterile air at a ventilation rate of 0.12 vvm for 24 h to obtain the fermentation broth containing bacteria.

[0145] S2. Sterilized camellia oleifera residue powder (mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 2:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 37℃ and aeration rate of 0.12 vvm for 24 h, the broth is heat-treated at 90℃ for 2 h to obtain crude fermentation broth containing tea saponins.

[0146] S3. After mixing the crude fermentation broth containing tea saponin with 2 times the volume of n-butanol, let it stand in a pear-shaped separatory funnel for 8 hours, shaking it vigorously every 2 hours. After the extraction is completed, the extractant is vacuum rotary evaporated at 60°C and 100 rpm for 15 minutes. The evaporated liquid is then dried under vacuum at 70°C for 8 hours to obtain tea saponin.

[0147] Comparative Example 11 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0148] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0149] Methods for preparing tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 2% (v / v) into the fermentation substrate (the fermentation substrate consists of 2% peanut cake powder, 2% maltose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate). Then add camellia oleifera residue powder (the mass ratio of fermentation substrate to camellia oleifera residue powder is 2:1). Incubate at 37℃ and an aeration rate of 0.12 vvm for 48 h to obtain a crude fermentation broth containing tea saponins.

[0150] S2. Add three times the mass of silica to the crude fermentation broth containing tea saponin for granulation. After subcritical cycle extraction (extractant is methanol, temperature is 45℃, pressure is 0.5 MPa) and spray drying (inlet air temperature is 120℃, outlet air temperature is 75℃), tea saponin is obtained.

[0151] Comparative Example 12 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0152] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0153] Methods for preparing tea saponins include: S1. Inoculate the Lactobacillus fermentation seed culture at an inoculation rate of 2% (v / v) into the fermentation substrate (the fermentation substrate consists of 2% peanut cake powder, 2% maltose and the remainder water, and the pH of the fermentation substrate is adjusted to 4.5 with sodium dihydrogen phosphate), and incubate at 37°C with sterile air at a ventilation rate of 0.12 vvm for 24 h to obtain the fermentation broth containing bacteria.

[0154] S2. Sterilized camellia oleifera residue powder (mass ratio of fermentation broth containing bacteria to camellia oleifera residue powder is 2:1) is added to the fermentation broth containing bacteria. After fermentation and extraction at 37℃ and sterile air flow rate of 0.12 vvm for 24 hours, the broth is heat-treated at 90℃ for 2 hours to obtain crude fermentation broth containing tea saponins.

[0155] S3. Add three times the mass of silica to the crude fermentation broth containing tea saponin to granulate, and then obtain tea saponin by subcritical cycle extraction (extractant is tetrafluoroethane, temperature is 45℃, pressure is 0.5 MPa) and spray drying (inlet air temperature is 120℃, outlet air temperature is 75℃).

[0156] Comparative Example 13 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0157] The preparation methods for green biosurfactants and tea saponins are the same as those in Example 1.

[0158] The sophorolipid used in this comparative example is a commercially available sophorolipid.

[0159] Comparative Example 14 The green biosurfactant in this comparative example, by weight, comprises: 40 parts sophorolipid, 25 parts tea saponin, and 35 parts water.

[0160] The preparation methods for green biosurfactants and sophorolipids are the same as those in Example 1.

[0161] The tea saponin used in this comparative example is commercially available tea saponin.

[0162] Test Example 1: Foam Performance Test Foaming performance is related to the type, purity, and concentration of surfactants, and is also affected by water quality. High hardness (high calcium and magnesium ion content) hinders foam formation. This test simulates the foaming performance of various samples in tap water. Test samples: Green biosurfactants from Examples 1-10 and Comparative Examples 1-14, prepared into 0.05% solutions with deionized water. Simulated tap water formulation: 0.1 g / L CaCl2, 0.1 g / L MgCl2·6H2O.

[0163] Test procedure: The static foam of different test samples at 0 min and 5 min is tested using a Roche foam analyzer. The height difference of the static foam at 0 min and 5 min is calculated to determine the foam stability of the test samples.

[0164] The test results are shown in Table 2 below.

[0165] Table 2

[0166] Test results show that the green biosurfactant of this invention exhibits lower foam collapse after 5 minutes, indicating higher foam stability. This is because the green biosurfactant in this embodiment combines sophorolipid and tea saponin. Sophorolipid molecules have high hydrophilicity and dissociability, while tea saponin molecules are primarily composed of a triterpenoid steroidal skeleton, possessing both hydrophobic steroidal rings and multiple hydrophilic hydroxyl groups. At the gas-liquid and liquid-liquid interfaces, these two molecules can form complementary adsorption layers through hydrophobic interactions, hydrogen bonds, and electrostatic interactions, enhancing the close arrangement of molecules at the interface. This composite effect significantly reduces the surface tension of the system, allowing the green biosurfactant to maintain excellent surfactant performance in a simulated tap water environment, thereby improving foam stability.

[0167] Among them, the foam stability of the green biosurfactants in Examples 1-10 was higher than that in Comparative Examples 1-3, indicating that the critical micelle concentration of the green biosurfactant formed by the combination of sophorolipid and tea saponin was lower. Sophorolipid molecules have high hydrophilicity and dissociability, while tea saponin molecules are mainly composed of triterpenoid steroidal skeletons, with both hydrophobic steroidal rings and multiple hydrophilic hydroxyl groups. At the gas-liquid and liquid-liquid interfaces, the two can form complementary adsorption layers through hydrophobic interactions, hydrogen bonds, and electrostatic interactions, enhancing the close arrangement of molecules at the interface. This composite effect significantly reduces the surface tension of the system. The green surfactant cannot achieve the same effect as the combination of sophorolipid and tea saponin by using only sophorolipid or tea saponin, or by replacing sophorolipid with a similar rhamnolipid. The foam stability of Examples 1-10 was higher than that in Comparative Examples 4-6, indicating that the preparation process of sophorolipid has a great influence on the surface tension of the final green biosurfactant. Comparative Examples 4 and 5 replaced the fermentation oil of *Candida bacillus*, and Comparative Example 6 changed the post-purification method of sophorolipid. These steps all alter the proportions of various sophorolipids in sophorolipids, thus changing the synergistic effect between sophorolipids and tea saponins, affecting the surface tension and foam stability of the final product. In this invention, the carbon source used for fermenting sophorolipids is preferably rapeseed oil and / or sunflower seed oil, and the purification step is preferably a sequential purification process using anion exchange resin, cation exchange resin, and macroporous adsorption resin. The foam stability of Examples 1-10 is higher than that of Comparative Examples 7-9, indicating that the bacterial strain used for extracting tea saponins has a significant impact on the type of tea saponins in the final green biosurfactant. Different bacterial strains result in different metabolic products, leading to differences in the structure of the purified tea saponins, which weakens the synergistic effect between sophorolipids and tea saponins, thereby affecting the foam stability of the final product. This invention preferably uses Lactobacillus fermentation to extract tea saponins through fermentation. The foam stability of Examples 1-10 was superior to that of Comparative Examples 10-12, indicating that the fermentation method, the purification steps of tea saponins, and the solvent used in subcritical circulating extraction affect the types of tea saponins extracted, ultimately impacting the foam stability of the green biosurfactant. Comparative Example 10 used n-butanol to extract tea saponins from the crude fermentation broth; Comparative Example 11 used a method of directly fermenting camellia oleifera residue powder with Lactobacillus seed culture; Comparative Example 12 modified the subcritical circulating extraction method. These steps resulted in differences in the structure and glycosides of the finally extracted tea saponins, leading to a decreased synergistic effect between sophorolipids and tea saponins. In this invention, tea saponins should be extracted by adding camellia oleifera residue powder after further fermentation of the Lactobacillus seed culture, and the crude fermentation broth containing tea saponins is preferably extracted using subcritical fluid extraction, with methanol and / or ethanol being the preferred extractant.Comparative Examples 13 and 14 used commercially available sophorolipids or tea saponins to form green biosurfactants. The preparation methods and final products of the commercially available products are very different from the sophorolipids or tea saponins of this invention, which reduces the synergistic effect between sophorolipids and tea saponins, resulting in poor foam stability of the final product.

[0168] Test Example 2: Oil Control Effect Test Test Principle: The secretory activity of sebaceous glands is mainly regulated by androgens, and dihydrotestosterone (DHT) is the most potent androgen. It strongly stimulates sebaceous gland cells to synthesize lipids and increases sebum secretion. DHT is produced from testosterone under the catalysis of 5α-reductase. The activity of 5α-reductase directly determines the local concentration of DHT, thus determining the degree of sebum secretion. Therefore, the oil-controlling potential of a test sample can be evaluated by directly detecting its inhibitory effect on 5α-reductase activity.

[0169] Test samples: Green biosurfactants from Examples 1-10 and Comparative Examples 1-14 were prepared into 1% solutions using 0.1M PBS buffer.

[0170] Test Procedure: Prepare NADPH working solution using 0.1M PBS buffer. Dilute the NADPH working solution to a series of concentration gradients and measure their OD values ​​at 340 nm to plot a standard curve. In a 96-well plate, add 0.1M PBS buffer sequentially to the blank control group, control group, sample group, and positive control group. Then add 5α-reductase working solution (2 mg / mL male SD rat liver microsomes) to the negative control group, sample group, and positive control group. Add an equal volume of 0.1M PBS buffer to the blank control group. After preheating at 37℃, add NADPH working solution, testosterone working solution, and an equal volume of solvent as the sample group to initiate the reaction in the control group. Add NADPH working solution, testosterone working solution, and sample solution to the sample group to initiate the reaction. Add NADPH working solution, testosterone working solution, and finasteride solution (10 μM) to the positive control group to initiate the reaction. After mixing, measure the OD value at 340 nm for each group. Incubate the 96-well plate at 37℃ in the dark for 1.5 hours, then measure the OD value at 340 nm again and record the relevant data. The change in OD value (ΔOD) for each group was calculated, and the ΔOD was converted into the change in NADPH content (ΔNADPH) using the NADPH standard curve, thereby calculating the inhibition rate of 5α-reductase. Each sample was tested three times, and the average value of the results was taken.

[0171] 5α-Reductase inhibition rate (%) = [1 - (ΔNADPH sample group / Positive control group - ΔNADPH control group) / (ΔNADPH blank control group – ΔNADPH control group)] × 100% The test results are shown in Table 3 below: Table 3

[0172] Test results show that the green biosurfactant of the present invention can effectively inhibit the activity of 5α-reductase, thereby weakening the secretion of dihydrotestosterone (DHT) and achieving the effect of controlling sebum secretion. Specifically, the oil-controlling effect of Examples 1-10 is better than that of Comparative Examples 1-3, indicating that there is a synergistic effect between sophorolipids and tea saponins in the present invention. The absence of either component, or the use of rhamnolipin (which has a similar effect to sophorolipids) as a component, results in a poorer oil-controlling effect of the green biosurfactant. The oil-controlling effect of Examples 1-10 is better than that of Comparative Examples 4-6, indicating that the sophorolipid preparation process has a significant impact on the final oil-controlling effect of the green biosurfactant. Comparative Examples 4 and 5 replaced the fermented oil of *Candida baccata*, and Comparative Example 6 changed the post-purification method of sophorolipids. These steps all change the proportion of various sophorolipids in the sophorolipid, thereby altering the synergistic effect between sophorolipids and tea saponins. In this invention, the carbon source used for fermenting sophorolipids is preferably rapeseed oil and / or sunflower seed oil, and the purification step is preferably a purification step involving sequential anion exchange resin, cation exchange resin, and macroporous adsorption resin. The oil-controlling effect of Examples 1-10 is better than that of Comparative Examples 7-9, indicating that the bacterial strain used for extracting tea saponins has a significant impact on the oil-controlling effect of the final green biosurfactant. This invention replaces the bacterial strain used for extracting tea saponins with similar strains such as *Lactobacillus fermentum* and / or *Lactobacillus casei*. Different bacterial strains result in different metabolic products, leading to differences in the structure of the final purified tea saponins. This weakens the synergistic effect between sophorolipids and tea saponins, thus reducing the oil-controlling effect of the green biosurfactant. The oil-controlling effect of Examples 1-10 is better than that of Comparative Examples 10-12, indicating that the fermentation method, the purification steps of tea saponins, and the solvent used in subcritical cycle extraction all affect the oil-controlling effect of the green biosurfactant. Comparative Example 10 used n-butanol to extract tea saponins from the crude fermentation broth; Comparative Example 11 used Lactobacillus fermentation seed liquid to directly ferment Camellia oleifera residue powder; Comparative Example 12 changed the subcritical circulating extraction method. These steps resulted in differences in the structure and glycosides of the finally extracted tea saponins, leading to a poorer synergistic effect between sophorolipids and tea saponins. In this invention, tea saponins should be extracted by adding Camellia oleifera residue powder after further fermentation of Lactobacillus fermentation seed liquid, and the crude fermentation broth containing tea saponins is preferably extracted by subcritical fluid extraction, with methanol and / or ethanol being the preferred extractant. The oil control effect of Examples 1-10 is better than that of Comparative Examples 13 and 14, indicating that the synergistic effect between commercially available sophorolipids or tea saponins and the tea saponins and sophorolipids of this invention is poor, resulting in a poorer oil control effect in the final product.

[0173] Test Example 3: Dandruff Removal Effect Test Test Principle: Malassezia is a resident fungus on the human scalp. It feeds on triglycerides in sebum and breaks down lipids using lipases (such as LIP1) to produce free fatty acids (such as oleic acid). When the scalp's sebaceous glands are overactive, producing large amounts of sebum (mainly composed of triglycerides, free fatty acids, squalene, etc.), Malassezia will overproduce. Simultaneously, Malassezia uses triglycerides in sebum as a nutrient source, breaking down lipids using lipases (such as LIP1) to produce free fatty acids (such as oleic acid). These fatty acids damage the scalp's barrier function, triggering an inflammatory response, stimulating excessive proliferation of keratinocytes, and causing a large amount of immature keratinocytes to shed, forming dandruff. This can be simply summarized as: excessive sebum on the scalp → excessive proliferation of Malassezia → decomposition of sebum to produce irritating oleic acid → scalp inflammation and stratum corneum dysfunction → dandruff formation. Therefore, inhibiting the excessive proliferation of Malassezia can effectively reduce dandruff production, thereby achieving a dandruff-removing effect.

[0174] Test samples: Green biosurfactants of Examples 1-10 and Comparative Examples 1-14 were prepared into solutions with a mass concentration of 2% using deionized water.

[0175] Test procedure: The Malassezia inhibition rate test was conducted according to the procedure in QBT2738-2012(7.3) "Evaluation method for antibacterial and bacteriostatic effects of daily chemical products". The Malassezia used was ATCC44344.

[0176] The test results are shown in Table 4 below: Table 4

[0177] Test results show that the green biosurfactants of the present invention have a good inhibitory effect on Malassezia, thus exhibiting potential anti-dandruff efficacy. Specifically, Examples 1-10 show better inhibitory effects on Malassezia than Comparative Examples 1-3, indicating a synergistic effect between sophorolipids and tea saponins in this invention. The absence of either component, or the use of rhamnolipin (which has similar efficacy to sophorolipids) as a component, results in poorer inhibitory effects of the green biosurfactants on Malassezia. Examples 1-10 also show better inhibitory effects on Malassezia than Comparative Examples 4-6, indicating that the sophorolipid preparation process significantly affects the final inhibitory effect of the green biosurfactants on Malassezia. Comparative Examples 4 and 5 replaced the fermented oil of *Candida albicans* from bumblebees, and Comparative Example 6 changed the post-purification method of sophorolipids. These steps all alter the proportions of various sophorolipids in the sophorolipid, thereby changing the synergistic effect between sophorolipids and tea saponins. In this invention, the carbon source used for fermenting sophorolipids is preferably rapeseed oil and / or sunflower oil, and the purification step is preferably a purification step involving sequential anion exchange resin, cation exchange resin, and macroporous adsorption resin. The inhibitory effect of Examples 1-10 on Malassezia is better than that of Comparative Examples 7-9, indicating that the bacterial strain used for extracting tea saponins has a significant impact on the oil-controlling effect of the final green biosurfactant. In this invention, the bacterial strain used for extracting tea saponins is replaced from *Lactobacillus fermentum* with similar strains such as *Lactobacillus plantarum* and / or *Lactobacillus casei*. Different bacterial strains result in different metabolic products, leading to differences in the structure of the final purified tea saponins. This weakens the synergistic effect between sophorolipids and tea saponins, thus reducing the inhibitory effect of the green biosurfactant on Malassezia. The inhibitory effect of Examples 1-10 on Malassezia is better than that of Comparative Examples 10-12, indicating that the fermentation method, the purification steps of tea saponins, and the solvent used in subcritical circulating extraction all affect the oil-controlling effect of the green biosurfactant on Malassezia. Comparative Example 10 used n-butanol to extract tea saponins from the crude fermentation broth; Comparative Example 11 used a method of directly fermenting camellia oleifera residue powder with Lactobacillus fermentation seed liquid; Comparative Example 12 changed the subcritical circulating extraction method. These steps resulted in differences in the structure and glycosides of the finally extracted tea saponins, leading to a poorer synergistic effect between sophorolipids and tea saponins. In this invention, tea saponins should be extracted by adding camellia oleifera residue powder after further fermentation of Lactobacillus fermentation seed liquid, and the crude fermentation broth containing tea saponins is preferably extracted by subcritical fluid extraction, with methanol and / or ethanol being the preferred extractant. The inhibitory effect of Examples 1-10 on Malassezia is better than that of Comparative Examples 13 and 14, indicating that the synergistic effect of commercially available sophorolipids or tea saponins with the tea saponins and sophorolipids of this invention is poor, resulting in a poor inhibitory effect of the final product on Malassezia.

[0178] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A green biosurfactant with oil-controlling and dandruff-removing effects, characterized in that... By weight, it includes 30-60 parts sophorolipid, 10-40 parts tea saponin, and water to make up to a total weight of 100.

2. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 1, characterized in that... By weight, it comprises 40 parts sophorolipid, 25 parts tea saponin and 35 parts water.

3. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 1 or 2, characterized in that: The method for preparing the sophorolipid includes: B1. Inoculate the seed culture of *Candida bacillus* into a fermentation medium and ferment to obtain a crude fermentation broth; B2. The crude fermentation broth is sterilized, filtered, purified, and dried to obtain the sophorolipid; And / or, the method for preparing the tea saponin includes: S1. Inoculate the Lactobacillus fermentation seed liquid into the fermentation substrate and culture to obtain a fermentation broth containing bacteria; S2. Add camellia oleifera residue powder to the fermentation broth containing bacteria, ferment and extract, and then heat treat to obtain crude fermentation broth containing tea saponins; S3. Add auxiliary materials to the crude fermentation liquid containing tea saponin, and obtain the tea saponin after subcritical cycle extraction and drying.

4. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 3, characterized in that: In step B1, the method for preparing the bumblebee Candida seed liquid includes: inoculating bumblebee Candida colonies into YM medium, and obtaining the bumblebee Candida seed liquid after expansion culture; the expansion culture temperature is 28-33℃, the expansion culture time is 36-60h, and the fermentation shake flask rotation speed is 120-180rpm; In step S1, the method for preparing the Lactobacillus fermentation seed liquid includes: inoculating Lactobacillus fermentation colonies into MRS medium, and obtaining the Lactobacillus fermentation seed liquid after expansion culture. The expansion culture temperature is 33-40℃, the expansion culture time is 36-60h, and the rotation speed of the fermentation shake flask is 50-120rpm.

5. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 3, characterized in that: In step B1, the fermentation medium comprises oil, carbon source, nitrogen source, inorganic salts, and water; the oil comprises rapeseed oil and / or sunflower oil; the amount of oil added is 2-10% of the total mass of the fermentation medium; the carbon source comprises at least one of glucose, fructose, and sucrose; the amount of carbon source added is 1-5% of the total mass of the fermentation medium; the nitrogen source comprises at least one of yeast extract, urea, soybean peptone, tryptone, and ammonium sulfate; the amount of nitrogen source added is 0.2-1% of the total mass of the fermentation medium; the inorganic salt comprises at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate, and the inorganic salts adjust the pH of the fermentation medium to 4.5-5.5; the amount of water added is to bring the total mass of the fermentation medium to 100%. In step B1, the inoculation amount of the bumblebee Candida seed liquid is 2-6% (v / v); the fermentation culture conditions are: fermentation temperature of 28-33℃, fermentation time of 3-7 days, stirring speed of 350-450 rpm, and sterile air ventilation rate of 0.3-0.6 vvm.

6. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 3, characterized in that: In step S1, the inoculation amount of the Lactobacillus fermentation seed liquid is 1-3% (v / v). In step S1, the fermentation substrate includes a nitrogen source, a carbon source, inorganic salts, and water; the nitrogen source includes at least one of soybean peptone, tryptone, peanut meal, and soybean meal; the amount of nitrogen source added is 1-3% of the total mass of the fermentation substrate; the carbon source includes at least one of glucose, maltose, and sucrose; the amount of carbon source added is 1-3% of the total mass of the fermentation substrate; the inorganic salt includes at least one of sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, and dipotassium hydrogen phosphate, and the inorganic salt adjusts the pH of the fermentation substrate to 3.5-5.5; In step S1, the culture temperature is 33-40℃, the culture time is 18-30h, and the sterile air ventilation rate is 0.05-0.2 vvm.

7. The green biosurfactant with oil-controlling and dandruff-removing effects according to claim 3, characterized in that: In step S2, the mass ratio of the fermentation broth containing bacteria to the camellia oleifera residue powder is (1-3):1; In step S2, the fermentation extraction temperature is 33-40℃, the fermentation extraction time is 18-30h, and the sterile air ventilation rate is 0.05-0.2vvm. In step S2, the heat treatment temperature is 85-95℃, and the heat treatment time is 1-3 hours. In step S3, the auxiliary material includes silica granulation, and the mass ratio of the auxiliary material to the crude fermentation liquid containing tea saponin is (2-4):

1. In step S3, the conditions for the subcritical cyclic extraction include: the extractant includes methanol and / or ethanol; the temperature is 40-50℃; and the pressure is 0.4-0.6 MPa.

8. A method for preparing a green biosurfactant with oil-controlling and dandruff-removing effects according to any one of claims 1-7, characterized in that... Includes the following steps: Sophorolipids and tea saponins are added to water and mixed evenly to obtain a green biosurfactant with oil-controlling and dandruff-removing effects.

9. The application of the green biosurfactant with oil-controlling and dandruff-removing effects according to any one of claims 1-7 in the preparation of daily chemical products.

10. The application of the green biosurfactant with oil-controlling and dandruff-removing effects according to claim 9 in the preparation of daily chemical products, characterized in that... The amount of the green biosurfactant with oil-controlling and dandruff-removing effects added to the daily chemical product is 0.1-20 wt%.

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