A galactose yeast fermented oil and its preparation method, application, and cosmetics

Abstract

This application belongs to the technical field of cosmetic production, and discloses a galactomyces fermented oil, which is obtained by fermenting macadamia nut oil and phyllanthus emblica fruits with a fermentation broth containing live galactomyces; the fermentation broth containing live galactomyces is obtained by fermenting galactomyces with a first cereal extract; the first cereal extract is selected from at least one of malt extract, soybean extract, and corn extract; the mass ratio of macadamia nut oil to phyllanthus emblica fruits is 16:1 to 6; in addition, this application also discloses a preparation method for preparing the above-mentioned galactomyces fermented oil, the use of the galactomyces fermented oil, and a cosmetic.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetics production, and particularly relates to a galactomyces fermented oil and its preparation method, application, and cosmetics. Background Art

[0002] For centuries, people have been using oils to care for their skin because of their conditioning and nourishing properties. With the development of social economy and the improvement of living quality, people's consumption concepts are gradually changing. With the rapid development of the cosmetics industry, people's demand for green, safe, and healthy cosmetics is increasing day by day. People pay more and more attention to whether the washing and care products contain their favorite ingredients and effects. However, some ingredients are harmful to skin health. Therefore, developing oils with obvious effects, safety, and high efficiency has a broad market prospect.

[0003] Fats and oils extracted from plant seeds, pulp, and other parts are collectively called vegetable oils. Vegetable oils are natural macromolecular compounds formed by the combination of fatty acids and glycerol, and are widely distributed in nature. More than 95% of vegetable oils are fatty acid triglycerides and are accompanied by a small amount of lipid substances. Lipid substances include fatty acids, monoglycerides, triglycerides, phospholipids, sterols, vitamins, pigments, terpenes, fatty alcohols, hydrocarbons, etc. Vegetable oils have functions such as storing and releasing energy, providing essential fatty acids, and serving as components of the organism structure. However, there are a small number of macromolecular free groups in vegetable oils, which results in poor spreading property of vegetable oils, and thus affects the use feeling. Through fermentation, the macromolecular groups in the oil can be decomposed into small molecular groups, and beneficial metabolites can be produced, making the use feeling of vegetable oils better.

[0004] Fermentation technology is a biochemical reaction process in which organic matter is decomposed through the growth metabolism and life activities of microorganisms based on a suitable environment and culture medium. Enzymes secreted by microorganisms in fermentation change some substances in the raw materials. For example, macromolecular substances can be decomposed into small molecular substances by fermentation, and the cell walls of tissue cells can also be broken, thereby releasing more nutrient elements. At present, galactomyces-like fungi are mainly applied in the fermentation of sugars such as galactose and lactose, and the products are mainly yeast-like fungus fermentation products such as galactomyces-like fungus fermentation product filtrate. However, its application in plant fermentation production is still blank. There has been no report on the exploration of using galactomyces-like fungi for plant fermentation research on macadamia nut oil and Phyllanthus emblica fruit, as well as the efficacy test analysis of the prepared cosmetics.

[0005] Prior Art 1: Chinese Patent Application No. 201410310229.5 discloses a natural composition and a natural cosmetic with whitening and freckle-reducing effects. The composition mainly consists of the following components in parts by weight: 0.05 - 0.2 parts of Glycyrrhiza glabra root extract, 0.1 - 2 parts of Phyllanthus emblica fruit extract, 0.1 - 2 parts of Ascorbyl glucoside, 0.1 - 0.5 parts of Bisabolol, 0.1 - 0.4 parts of Piper nigrum extract, and 1 - 15 parts of yeast extract. It also discloses a cosmetic, which includes the above composition and vegetable oil, where the vegetable oil is selected from at least one of grape seed oil, rose hip oil, evening primrose oil, wheat germ oil, corn oil, rice bran oil, sweet almond oil, olive oil, jojoba oil, nut oil, shea butter, or camellia oil;

[0006] Through the compounding of six components, namely Glycyrrhiza glabra root extract, Phyllanthus emblica fruit extract, Ascorbyl glucoside, Bisabolol, Piper nigrum extract, and yeast extract, this solution enables the composition to have a significant whitening and freckle-reducing effect. It can quickly penetrate through the epidermis to reach the deep skin tissue, mainly inhibit the activity of tyrosinase, inhibit melanin formation at all stages, inhibit inflammation, and provide sufficient nutrients for the skin to promote its metabolism, overall achieving a synergistic effect of freckle reduction from the inside out. Additionally, it can be compounded with white tea extract, raspberry extract, and plant essential oils to further enhance the whitening and freckle-reducing effect;

[0007] It can be seen that this solution uses yeast extract and Phyllanthus emblica fruit extract to prepare a natural composition with whitening and freckle-reducing effects and uses the above composition and vegetable oil to prepare a cosmetic, but this solution does not disclose specifically which yeast is selected.

[0008] Problem to be solved by this solution: How to develop a galactomyces ferment oil with good free radical resistance and antioxidant ability. Summary of the Invention

[0009] The purpose of this application is to provide a galactomyces ferment oil with good free radical resistance and antioxidant ability. This ferment oil ferments macadamia nut oil and Phyllanthus emblica fruit with a fermentation broth containing galactomyces viable bacteria, thereby obtaining a fermentation product with good free radical resistance ability, and further enhancing the antioxidant ability of the ferment oil.

[0010] A galactomyces ferment oil, which is obtained by fermenting macadamia nut oil and Phyllanthus emblica fruit with a fermentation broth containing galactomyces viable bacteria;

[0011] The fermentation broth containing galactomyces viable bacteria is obtained by fermenting galactomyces with a first cereal extract;

[0012] The first cereal extract is selected from at least one of malt extract, soybean extract, and corn extract;

[0013] The mass ratio of the fermentation broth containing live Galactomyces yeasts to the total mass of macadamia nut oil and Phyllanthus emblica fruits is 3-15:8-20;

[0014] The mass ratio of the macadamia nut oil to the Phyllanthus emblica fruits is 16:1-6.

[0015] It should be noted that in this application, macadamia nut oil contains a large amount of antioxidants, has a very low saturated fatty acid content, is rich in a large amount of monounsaturated fatty acids, has sufficient polyunsaturated fatty acids but is not excessive, and at the same time, the ratio of the essential fatty acids ω3 and ω6 it contains is very ideal. It can soothe the skin, immediately repair skin sensitivity, sunburn, and windburn caused by the environment, protect cell membranes, moisturize and hydrate, has good ductility, doubles the moisturizing effect on the skin, and makes the skin soft and energetic.

[0016] Phyllanthus emblica, also known as Indian pepper, is a common herbaceous plant widely distributed in India, China, and other Asian countries. The extract of Phyllanthus emblica is obtained by extracting the fruits of Phyllanthus emblica. The extract of Phyllanthus emblica fruits can promote the production of body fluid and relieve cough, clear heat and detoxify, reduce fat and lose weight, resist fatigue and aging, and beautify the skin. The superoxide dismutase in it can protect cells, resist aging, and also scavenge free radicals, having a good antioxidant effect.

[0017] Galactomyces is a special yeast called Galactomyces, which can secrete small molecules beneficial to skin care such as vitamin B group, minerals, amino acids, organic acids, and inorganic acids during the fermentation process. Applying it in fermentation, the fermentation product has excellent skin care effects and is an excellent skin conditioner;

[0018] Preferably, the preservation number of the Galactomyces is GDMCC No: 64235, the preservation date is January 2, 2024, the preservation unit is the Guangdong Provincial Culture Collection Center of Microorganisms, and the taxonomic name is Geotrichum candidum.

[0019] Preferably, the Galactomyces is activated Galactomyces, and the activation method of the activated Galactomyces includes the following steps:

[0020] Step 1: Mix the second cereal extract, glucose, yeast extract, and pure water, sterilize to obtain an activation medium, and then add Galactomyces to the activation medium to obtain a Galactomyces activation solution;

[0021] Step 2: Mix the third cereal extract, glucose, yeast extract, and pure water, and sterilize to obtain a seed medium. Subsequently, add a galactose yeast activation solution to the seed medium to obtain a galactose yeast seed solution, i.e., activated galactose yeast.

[0022] In the activation medium, the mass fraction of glucose is 0.1wt% - 10wt%, the mass fraction of the second cereal extract is 0.1wt% - 20wt%, the mass fraction of yeast extract is 0.1wt% - 5wt%, and the balance is water.

[0023] In the seed medium, the mass fraction of glucose is 0.1wt% - 10wt%, the mass fraction of the third cereal extract is 0.1wt% - 20wt%, the mass fraction of yeast extract is 0.1wt% - 5wt%, and the balance is water.

[0024] The second cereal extract is selected from at least one of malt extract, soybean extract, corn extract, sorghum extract, jujube extract, and sugarcane extract.

[0025] The third cereal extract is selected from at least one of malt extract, soybean extract, corn extract, sorghum extract, jujube extract, and sugarcane extract.

[0026] Preferably, the viable cell concentration of galactose yeast in the galactose yeast seed solution is 1x10 6 - 2x10 8 CFU / g.

[0027] Preferably, the mass ratio of the galactose yeast seed solution to the first cereal extract is 5 - 10:2 - 30.

[0028] In addition, the present application also discloses a preparation method for preparing the above-mentioned galactose yeast fermented oil, comprising the following steps:

[0029] Step A: Inoculate galactose yeast into a fermentation medium to obtain a fermentation broth containing viable galactose yeast.

[0030] Step B: Crush the Phyllanthus emblica fruit to obtain Phyllanthus emblica fruit powder, and mix the macadamia nut oil and the Phyllanthus emblica fruit powder in a mass ratio of 16:1 - 6 to obtain an intermediate.

[0031] Step C: Mix and ferment the fermentation broth containing viable galactose yeast prepared in Step A with the intermediate prepared in Step B to obtain galactose yeast fermented oil.

[0032] The fermentation medium is prepared by mixing a first cereal extract, glucose, yeast extract, and pure water, and the mass fraction of glucose in the fermentation medium is 0.1 wt% to 10 wt%, the mass fraction of the first cereal extract is 0.1 wt% to 20 wt%, the mass fraction of yeast extract is 0.1 wt% to 5 wt%, and the balance is water;

[0033] The mass ratio of the fermentation broth containing viable Saccharomyces galactomyces obtained in step A to the intermediate obtained in step B is 3 to 15:8 to 20.

[0034] Preferably, step B is specifically: mixing macadamia nut oil and Phyllanthus emblica fruits in a mass ratio of 16:1 to 6 and stirring in an environment at 70 °C for 2 to 4 h to obtain an intermediate.

[0035] Preferably, step C is specifically: mixing the fermentation broth containing viable Saccharomyces galactomyces obtained in step A with the intermediate obtained in step B, and fermenting in an environment at 27 to 30 °C for 20 to 26 h to obtain Saccharomyces galactomyces fermentation oil.

[0036] In addition, the present application also discloses the use of the above-mentioned Saccharomyces galactomyces fermentation oil in the preparation of cosmetics.

[0037] In addition, the present application also discloses a cosmetic containing 0.00001 to 30 wt% of the above-mentioned Saccharomyces galactomyces fermentation oil.

[0038] The beneficial effects of the present application are: The macadamia nut oil used in the present application contains a large amount of antioxidants, has a very low saturated fatty acid content, is rich in a large amount of monounsaturated fatty acids, has sufficient polyunsaturated fatty acids but does not exceed the limit, and at the same time, the ratio of the essential fatty acids ω3 and ω6 for the human body is very ideal, can soothe the skin, immediately repair skin sensitivity, sunburn and windburn caused by the environment, protect cell membranes and moisturize, has good ductility, doubles the moisturizing of the skin, and makes the skin soft and energetic.

[0039] Phyllanthus emblica, also known as Indian pepper, is a common herbaceous plant widely distributed in India, China and other Asian countries. The Phyllanthus emblica extract is obtained by extracting Phyllanthus emblica fruits. The Phyllanthus emblica fruit extract can promote the production of body fluid and relieve cough, clear heat and detoxify, reduce fat and lose weight, resist fatigue and anti-aging, and beautify the skin. The superoxide dismutase in it can protect cells, resist aging, and can also scavenge free radicals, and has good antioxidant effects.

[0040] Saccharomyces galactomyces-like is a special yeast called Saccharomycopsis, which can secrete small molecules beneficial to skin care such as vitamin B group, minerals, amino acids, organic acids, and inorganic acids during the fermentation process. Applying it in fermentation, the fermentation product has excellent skin care effects and is an excellent skin conditioner. Description of the Drawings

[0041] Figure 1 Schematic diagram before adding the galactose yeast fermentation oil prepared in Example 1 to the chicken embryo;

[0042] Figure 2 Schematic diagram after adding the galactose yeast fermentation oil prepared in Example 1 to the chicken embryo;

[0043] Figure 3 Schematic diagram of the comparison of hair friction before and after using the galactose yeast fermentation oil prepared in Example 1 in the repair efficacy test;

[0044] Figure 4 Schematic diagram of the comparison of hair friction before and after using the galactose yeast fermentation oil prepared in Example 1 in the heat protection performance test;

[0045] Figure 5 Schematic diagram of hair before using the galactose yeast fermentation oil prepared in Example 1;

[0046] Figure 6 Schematic diagram of hair after using the galactose yeast fermentation oil prepared in Example 1;

[0047] Figure 7 Schematic diagram of the fluorescence analysis of the negative control group in the ROS scavenging rate test;

[0048] Figure 8 Schematic diagram of the fluorescence analysis of the positive control group in the ROS scavenging rate test;

[0049] Figure 9 Schematic diagram of the fluorescence analysis of the galactose yeast fermentation oil prepared in Example 1 in the ROS scavenging rate test. Detailed implementation manners

[0050] Next, the embodiments of the present invention will be combined to clearly and completely describe the present invention. In the description of the present invention, it should be noted that for those embodiments not specified with specific conditions, they are carried out according to conventional conditions or conditions recommended by the manufacturer. For reagents or instruments not specified with the manufacturer, they are all conventional products that can be obtained through commercial purchase.

[0051] Supplier information and / or product information

[0052] Macadamia nut oil was purchased from the Australian company FICCECODE AUSTRALIA PTY LTD;

[0053] Phyllanthus emblica fruits were purchased from Bozhou Yuanshengtang Pharmaceutical Co., Ltd.;

[0054] Malt extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0055] The soybean extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0056] The corn extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0057] The sorghum extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0058] The red date extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0059] The sugarcane extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd.;

[0060] The yeast extract was purchased from Guangzhou Dingguo Biotechnology Co., Ltd., brand OXOID, model LP0021B;

[0061] Galactose yeast-like fungus HM.GY.168: The preservation number is GDMCC No: 64235, the preservation date is January 2, 2024, the preservation unit is the Guangdong Provincial Microbial Culture Collection Center, and the taxonomic name is Geotrichum candidum. The galactose yeast used in the examples and comparative examples is this strain except as otherwise specified.

[0062] Example 1

[0063] Preparation of a galactose yeast fermented oil

[0064] Step 1: Mix according to the ratio of 5 g of malt extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance being water, adjust the pH value of the solution to 6.0, and sterilize the above medium at 121 °C for 20 min to obtain an activation medium. After cooling, add 0.5 g of frozen and preserved galactose yeast solution (concentration 1x10 6 CFU / g) to the activation medium for primary culture. Set the constant temperature culture temperature at 28 °C and culture for 24 h to obtain a galactose yeast activation solution. After detection, the viable bacteria concentration in the galactose yeast activation solution is 0.6x10 8 CFU / g.

[0065] Step 2: Mix according to the ratio of 5 g of malt extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance being water, adjust the pH value of the solution to 6.0, and sterilize the above medium at 121 °C for 20 min to obtain a seed medium;

[0066] Subsequently, inoculate the galactose yeast activation solution into the seed medium and culture at a constant temperature of 28 °C for 24 h to obtain a galactose yeast seed solution (i.e., activated galactose yeast). After detection, the viable bacteria concentration in the galactose yeast seed solution is 1.8x10 8 CFU / g;

[0067] The weight of the galactose yeast activation solution added to every 1 kg of the seed medium is 0.5 g.

[0068] Step A: Prepare a fermentation medium by mixing in the ratio of 5 g of malt extract, 15 g of glucose, 5 g of yeast extract per 1 kg, with the balance being water.

[0069] Subsequently, inoculate the fermentation medium with the galactose yeast seed solution. Inoculate 5 g of the galactose yeast-like bacteria seed solution per 1 kg of the fermentation medium, and set the dissolved oxygen content value in the fermenter to 25%. The stirring speed is associated with the dissolved oxygen content and automatically compensates and adjusts the speed. Ferment and culture at 28°C for 18 h to obtain a fermentation broth containing live galactose yeast (after testing, in the fermentation broth containing live galactose yeast, the content of the oil-soluble components is about 13.5 ± 0.5%).

[0070] Step B: Crush the Phyllanthus emblica fruits to obtain Phyllanthus emblica fruit powder. According to the ratio of adding 250 g of Phyllanthus emblica fruit powder per 1 kg of macadamia nut oil (after testing, in the Phyllanthus emblica fruit powder, the oil content is about 26 ± 1%), add the materials to another fermenter, set the fermenter rotation speed at 50 rpm, and cook at 70°C for 3 h to prepare an intermediate.

[0071] Step C: Pump the fermentation broth containing live galactose yeast into the fermenter containing the intermediate according to the mass ratio of the fermentation broth containing live galactose yeast:intermediate being 11:10. Set the fermenter rotation speed at 50 rpm and ferment and culture at 28°C for 24 h.

[0072] After fermentation, separate the bacteria and the mixed fermentation culture broth at 12000 rpm, and collect the oil phase part after centrifugation to prepare the galactose yeast fermentation oil.

[0073] Example 2

[0074] It is basically the same as Example 1, except that the fermentation medium is prepared by mixing in the ratio of 10 g of malt extract, 25 g of glucose, 12.5 g of lactose, 20 g of yeast extract per 1 kg, with the balance being water.

[0075] Prepare the activation medium and the seed medium by mixing in the ratio of 10 g of malt extract, 25 g of glucose, 12.5 g of lactose, 20 g of yeast extract per 1 kg, with the balance being water.

[0076] Example 3

[0077] It is basically the same as Example 1, except that the fermentation medium is prepared by mixing in the ratio of 30 g of malt extract, 37.5 g of glucose, 2 g of yeast extract per 1 kg, with the balance being water.

[0078] Prepare the activation medium and the seed medium by mixing in the ratio of 30 g of malt extract, 37.5 g of glucose, 2 g of yeast extract per 1 kg, with the balance being water.

[0079] Example 4

[0080] It is basically the same as Example 1, except that the fermentation medium is prepared by mixing in the ratio of 2 g of malt extract, 40 g of glucose, 15 g of yeast extract per 1 kg, with the balance being water.

[0081] Prepare the activation medium and the seed medium by mixing in the ratio of 2 g of malt extract, 40 g of glucose, 15 g of yeast extract per 1 kg, with the balance being water.

[0082] Example 5

[0083] It is basically the same as Example 1, except that 8 g of galactose yeast seed liquid is inoculated into every 1 kg of the fermentation medium.

[0084] Example 6

[0085] It is basically the same as Example 1, except that 10 g of galactose yeast seed liquid is inoculated into every 1 kg of the fermentation medium.

[0086] Example 7

[0087] It is basically the same as Example 1, except that in step B: according to the ratio of adding 62.5 g of Phyllanthus emblica fruit powder per 1 kg of macadamia nut oil, add the materials into another fermentation tank, set the rotation speed of the fermentation tank at 50 r / min, and cook at 70 °C for 3 h to obtain the intermediate.

[0088] Step C: Pump the fermentation broth containing live galactose yeast into the fermentation tank according to the mass ratio of fermentation broth: intermediate of 3:20, set the rotation speed of the fermentation tank at 50 r / min, and ferment and culture at 28 °C for 24 h.

[0089] Example 8

[0090] It is basically the same as Example 1, except that in step B: according to the ratio of adding 375 g of Phyllanthus emblica fruit powder per 1 kg of macadamia nut oil, add the materials into another fermentation tank, set the rotation speed of the fermentation tank at 50 r / min, and cook at 70 °C for 3 h to obtain the intermediate.

[0091] Step C: Pump the fermentation broth containing live galactose yeast into the fermentation tank according to the mass ratio of fermentation broth: intermediate of 15:8, set the rotation speed of the fermentation tank at 50 r / min, and ferment and culture at 28 °C for 24 h.

[0092] Example 9

[0093] Basically the same as Example 1, except that the activation medium and the seed medium in Steps 1 and 2 are prepared according to the ratio of 2 g of malt extract, 2 g of red date extract, 1 g of corn extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance is water.

[0094] Example 10

[0095] Basically the same as Example 1, except that the activation medium and the seed medium in Steps 1 and 2 are prepared according to the ratio of 2 g of malt extract, 1 g of soybean extract, 1 g of corn extract, 1 g of sorghum extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance is water.

[0096] Example 11

[0097] Basically the same as Example 1, except that the activation medium and the seed medium in Steps 1 and 2 are prepared according to the ratio of 2 g of malt extract, 1 g of red date extract, 1 g of corn extract, 1 g of sorghum extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance is water.

[0098] Example 12

[0099] Basically the same as Example 1, except that in Step A: the fermentation medium is prepared according to the ratio of 3 g of soybean extract, 2 g of corn extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance is water.

[0100] Example 13

[0101] Basically the same as Example 1, except that in Step A: the fermentation medium is prepared according to the ratio of 2.5 g of malt extract, 1.5 g of soybean extract, 1 g of corn extract, 15 g of glucose, 5 g of yeast extract per 1 kg, and the balance is water.

[0102] Example 14

[0103] Basically the same as Example 1, except that the galactose yeast used in Example 1 is replaced with galactose yeast (commercially available).

[0104] Comparative Example 1

[0105] Basically the same as Example 1, except that the Saccharomyces cerevisiae is used to replace the galactose yeast.

[0106] Comparative Example 2

[0107] Basically the same as Example 1, except that the Lactobacillus plantarum is used to replace the galactose yeast.

[0108] Comparative Example 3

[0109] It is basically the same as Example 1, except that in Step 4, under the condition of constant total mass, macadamia nut oil is used to replace macadamia nut oil and Phyllanthus emblica fruit powder to prepare the intermediate.

[0110] Comparative Example 4

[0111] It is basically the same as Example 1, except that in Step 4, under the condition of constant total mass, Phyllanthus emblica fruit powder is used to replace macadamia nut oil and Phyllanthus emblica fruit powder to prepare the intermediate.

[0112] Comparative Example 5

[0113] It is basically the same as Example 1, except that in Step 4, according to the ratio of adding 195 g of Saussurea involucrata powder to every 1 kg of macadamia nut oil, the materials are added into another fermentation tank, and the rotation speed of the fermentation tank is set at 50 r / min, and the steaming time at 70 °C is 3 h to prepare the intermediate.

[0114] Comparative Example 6

[0115] It is basically the same as Example 1, except that in Step 4, according to the ratio of adding 195 g of Phyllanthus emblica fruit powder to every 1 kg of sunflower oil, the materials are added into another fermentation tank, and the rotation speed of the fermentation tank is set at 50 r / min, and the steaming time at 70 °C is 3 h to prepare the intermediate.

[0116] Comparative Example 7

[0117] It is basically the same as Example 1, except that the fermentation medium is prepared by mixing according to the ratio of containing 5 g of sugarcane extract, 15 g of glucose, 5 g of yeast extract, and the balance being water per 1 kg.

[0118] Comparative Example 8

[0119] It is basically the same as Example 1, except that the fermentation medium is prepared by mixing according to the ratio of containing 5 g of red date extract, 15 g of glucose, 5 g of yeast extract, and the balance being water per 1 kg

[0120] I. Performance Test:

[0121] 1. Irritation Test

[0122] 1.1 Instrument and Equipment

[0123] Fully automatic incubator, stereomicroscope, SPF chicken embryos;

[0124] 1.2 Reagents

[0125] Sodium chloride, sodium dodecyl sulfate (SDS);

[0126] 1.3 Incubation Conditions

[0127] Room temperature is 20°C to 25°C, relative humidity is 45% to 70%, incubation temperature is 37.5°C ± 0.5°C, relative humidity is 55% to 70%, the turntable rotates 3 times / h to 6 times / h, and chicken embryos at 9 days old do not need to be rotated during incubation.

[0128] 1.4 Test method

[0129] (1) Test operation steps

[0130] For each group in this test, 6 embryos were selected. The situation of the chorioallantoic membrane was recorded with a photographing device. A polytetrafluoroethylene resin ring was placed on the chorioallantoic membrane of the chicken embryo and photographed. The fermentation oil prepared in the examples and comparative examples was diluted with peanut oil to a solution with a mass fraction of 2% of the fermentation oil and used as the sample to be tested. Subsequently, the sample to be tested was added into the polytetrafluoroethylene resin ring, the time of adding the sample was recorded, and the air chamber was covered with a moistened plastic wrap. The chicken embryo was moved to an incubator with constant temperature and humidity for cultivation, and the degree of change in each toxic effect was observed.

[0131] (2) Result observation

[0132] Observe and record the manifestations of bleeding, blood coagulation and vascular lysis, and score according to their severity

[0133] The scoring criteria for bleeding, blood coagulation and vascular lysis are shown in Table 1:

[0134] Table 1

[0135]

[0136]

[0137] Scoring criteria: ES ≤ 12: no / mild irritation; 12 < ES < 16: moderate irritation; ES ≥ 16: strong irritation.

[0138] 1.5 Test results

[0139] Table 2

[0140] Name Test Result (ES) Negative Control (Peanut Oil) 0.00 Positive Control (1% SDS) 18.00 Example 1 3.00

[0141] For the negative control (peanut oil), ES = 0.00, meeting the standard of the negative control sample; for the positive control (1% SDS), ES = 18.00, meeting the standard of the positive control sample;

[0142] The ES value of Example 1 was 3.00, indicating no irritation, and no irritation was shown in the remaining examples and comparative examples.

[0143] 2. Nourishing efficacy test

[0144] Treatment and reagents for test groups

[0145] Sample: Dilute the galactose yeast fermentation oil prepared in Example 1 with absolute ethanol to a solution with a mass fraction of 10% of galactose yeast fermentation oil.

[0146] Blank control: Absolute ethanol

[0147] Test procedure:

[0148] Select in vitro human hair bundles that meet the conditions, wash the hair bundles with a cleaning solution, and after rinsing thoroughly, label the hair bundles with serial numbers and place them in a thermo-hygrostat for equilibration. The number of effective hair bundles in each group should meet the statistical requirements.

[0149] The hair bundles to be tested are fully wetted with flowing constant-temperature clear water (38 ± 1 °C), and 100 μL of the sample is evenly applied to the surface of the hair bundles at a rate of 100 μL per gram of hair bundle. The application time is about 30 seconds, and then left standing for about 1 minute, and then rinsed for about 30 seconds. The hair bundles after the wet hair test are placed in a thermo-hygrostat for more than 4 hours to stabilize, taken out and fixed at the test position, and a combing cycle test (10 cycles of combing) is carried out.

[0150] Result calculation

[0151] Calculate the initial value of the measured parameters and the values between other groups, and use professional statistical software for analysis. If the data is normally distributed, t-test or analysis of variance methods are used for data analysis and statistics; if the data is non-normally distributed, the rank sum test method is used for statistics.

[0152] Result judgment criteria

[0153] Compared with before using the sample, the hair combing force after using the sample is significantly reduced, indicating that the sample has the effect of improving hair combability; compared with before using the sample, the hair frizziness after using the sample is significantly reduced, indicating that the sample has the effect of improving hair frizziness; when the hair gloss and frizziness after using the sample are improved simultaneously, and the difference compared with the control group or before using the sample is statistically significant, it can be considered that the sample has the effect of nourishing hair.

[0154] 3. Repair efficacy test

[0155] Treatment and reagents for test groups

[0156] Sample: Dilute the galactose yeast fermentation oil prepared in Example 1 with absolute ethanol to a solution with a mass fraction of 10% of galactose yeast fermentation oil.

[0157] Blank control: Absolute ethanol

[0158] Test procedure:

[0159] Select qualified ex vivo human hair bundles, wash the hair bundles with a cleaning solution, and after rinsing thoroughly, attach serial number labels to the hair bundles and place them in a thermo-hygrostat for equilibration before use. The number of effective hair bundles in each group should meet the statistical requirements.

[0160] The hair bundles to be tested are fully wetted with flowing thermostatic water (38 ± 1 °C). At a rate of 100 μL of the sample per gram of hair bundle, apply it evenly on the surface of the hair bundle for about 30 seconds, let it stand for about 3 - 5 minutes, and then rinse for about 30 seconds. Place the hair bundles after applying the sample in a thermo-hygrostat for at least 4 hours, take them out and fix them in the test position, and conduct a friction cycle test (10 cycles of friction).

[0161] Result calculation

[0162] Calculate the initial value of the measured parameters and the values among other groups, and use professional statistical software for analysis. If the data is normally distributed, use the t-test or analysis of variance method for data analysis and statistics; if the data is non-normally distributed, use the rank sum test method for statistics.

[0163] Result judgment criteria

[0164] If the friction force result of the test group after using the product shows a significant improvement compared to the hair bundle before using the product (P < 0.05), it indicates that the sample has the ability to improve hair softness and has a hair repair effect.

[0165] 4. Thermal protection performance test

[0166] Treatment and reagents for test groups

[0167] Sample: Dilute the galactose yeast fermentation oil prepared in Example 1 with absolute ethanol to a solution with a mass fraction of galactose yeast fermentation oil of 10%.

[0168] Blank control: Absolute ethanol

[0169] Test steps:

[0170] Select qualified ex vivo human hair bundles, wash the hair bundles with a cleaning solution, and after rinsing thoroughly, attach serial number labels to the hair bundles and place them in a thermo-hygrostat for equilibration before use. The number of effective hair bundles in each group should meet the statistical requirements (3 hair bundles for each sample).

[0171] For each hair bundle, use 100 μL of the sample evenly on the surface of the hair bundle for about 30 seconds. Immediately use a straightening hair iron at 180 °C to heat for 5 minutes. Each time, use it uniformly from top to bottom for 30 seconds and repeat 10 times. Fix the hair bundle at the test position and conduct a friction cycle test (friction cycle 10 times). Without adding the sample to the hair bundle, use a straightening hair iron at 180 °C to heat for 5 minutes. Each time, use it uniformly from top to bottom for 30 seconds and repeat 10 times. Fix the hair bundle at the test position and conduct a friction cycle test (friction cycle 10 times).

[0172] Result calculation

[0173] Calculate the initial value of the measured parameter / the value between the blank group and other measurement time points / other groups, and use professional statistical software for analysis. If the data is normally distributed, use the t-test or analysis of variance method for data analysis and statistics; if the data is non-normally distributed, use the rank sum test method for statistics.

[0174] Result judgment criteria

[0175] If the friction force result of the test group after using the product shows a significant improvement compared to the hair bundles of the blank control group (P < 0.05), it indicates that the sample has the ability to improve the hair softness and has a hair repair effect.

[0176] 5. SEM scanning electron microscope test

[0177] Treatment and reagents for test groups

[0178] Sample: Dilute the galactose yeast fermentation oil prepared in Example 1 with absolute ethanol to a solution with a mass fraction of galactose yeast fermentation oil of 10%.

[0179] Blank control: Absolute ethanol

[0180] Test steps

[0181] Select in vitro human hair bundles that meet the conditions, clean the hair bundles with a cleaning solution, and after rinsing thoroughly, label the hair bundles with serial numbers and place them in a constant temperature and humidity chamber for equilibration. The number of effective hair bundles in each group should meet the statistical requirements (use 3 hair bundles for each sample).

[0182] Fully moisten the hair bundles to be tested with flowing constant-temperature clear water (38 ± 1 °C). For each gram of hair bundle, use 100 μL of the sample evenly on the surface of the hair bundle for about 30 seconds, let it stand for about 3 - 5 minutes, and then rinse for about 30 seconds. Place the hair bundles after using the sample in a constant temperature and humidity chamber for more than 4 hours, take out 5 hairs, cut the middle segments, and obtain images through an SEM scanning electron microscope.

[0183] Result judgment criteria

[0184] If some hair cuticles are slightly closed after the test group uses the product, it indicates that the sample has a certain repair effect on hair cuticles; otherwise, it has no effect.

[0185] 6. Elastic protease inhibition rate test

[0186] Treatment of test groups and reagents

[0187] Sample: Dilute the fermentation oils prepared in Examples 1-14 and Comparative Examples 1-6 to a solution with a mass fraction of 10% of the fermentation oil using absolute ethanol.

[0188] Positive control: 0.1% epigallocatechin gallate (EGCG, 98%) aqueous solution.

[0189] Negative control: Absolute ethanol.

[0190] Reagents: Elastic protease (porcine pancreas), BR; N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%; EGCG, 98%.

[0191] Test procedure

[0192] Set up a sample group, a sample background group, a solvent group, and a solvent background group. Each group needs to have 3 parallels. Add different reagent solutions to a 96-well plate, shake gently, and after incubating at 25 °C for 15 min, place it in an enzyme-linked immunosorbent assay (ELISA) reader and measure the absorbance at 410 nm.

[0193] Result calculation

[0194] Calculate the elastic protease inhibition rate % according to Equation 1.

[0195]

[0196] In the formula:

[0197] A—Absorbance of the reaction solution without the sample;

[0198] B—Absorbance of the reaction solution without the sample and the enzyme;

[0199] C—Absorbance of the reaction solution containing the sample and the enzyme;

[0200] D—Absorbance of the reaction solution containing the sample and without the enzyme.

[0201] Data analysis

[0202] The statistical analysis software was SPSS. The independent-samples t-test was used to compare the elastase inhibition rates among the test samples, positive control substances, and negative control substances. All of the above statistical analyses were two-tailed tests, and the significance level was α = 0.05. P ≥ 0.05 indicated no significant difference between the two groups; P < 0.05 indicated a significant difference between the two groups.

[0203] Result judgment criteria

[0204] If the elastase inhibition rate of the sample was higher than that of the negative control and there was a significant difference (P < 0.05), it could be considered that the test sample had a certain firming effect.

[0205] 7. Hyaluronidase inhibition rate test

[0206] Treatment of test groups and reagents

[0207] Sample group: Dilute the fermentation oils prepared in Examples 1 to 14 and Comparative Examples 1 to 6 with absolute ethanol to a solution with a mass fraction of the fermentation oil of 10%.

[0208] Positive control group: 3% aqueous solution of dipotassium glycyrrhizinate (dipotassium glycyrrhizinate, purity ≥ 98%).

[0209] Negative control group: Absolute ethanol.

[0210] Reagents: Hyaluronidase, BR; Sodium hyaluronate, BR.

[0211] Test steps

[0212] Set up a sample group, a sample background group, a solvent group, and a solvent background group. Three parallels should be set up for each group. Add different reagent solutions to the four groups respectively, shake well, place at room temperature for 30 min for color development, and measure the absorbance value at a wavelength of 528 nm with a UV spectrophotometer.

[0213] Result calculation: Calculate the hyaluronidase inhibition rate % according to Equation 2.

[0214]

[0215] In the formula:

[0216] A—Absorbance of the reaction solution without the sample;

[0217] B—Absorbance of the reaction solution without the sample and the enzyme;

[0218] C—Absorbance of the reaction solution containing the sample and the enzyme;

[0219] D—Absorbance of the reaction solution containing the sample and without the enzyme.

[0220] Data analysis

[0221] The statistical analysis software was SPSS. The independent samples t-test was used to compare the hyaluronidase inhibition rates among the test samples, positive control substances, and negative control substances. All the above statistical analyses were two-tailed tests, and the significance level was α = 0.05. P ≥ 0.05 indicates no significant difference between the two groups; P < 0.05 indicates a significant difference between the two groups.

[0222] Result determination criteria

[0223] If the hyaluronidase inhibition rate of the sample is higher than that of the negative control and there is a significant difference (P < 0.05), it can be considered that the test sample has a certain soothing effect.

[0224] 8. DPPH free radical scavenging rate test

[0225] Treatment and reagents for test groups

[0226] Sample: Dilute the fermentation oils prepared in Examples 1 to 14 and Comparative Examples 1 to 6 with absolute ethanol to a solution with a mass fraction of the fermentation oil of 10%.

[0227] Positive control: Ethanol (95%) solution of 0.1% vitamin E (purity ≥ 95%).

[0228] Negative control: Absolute ethanol.

[0229] Test procedure:

[0230] Set up sample tubes, sample background tubes, DPPH tubes, and solvent background tubes. Three parallel tubes need to be set up for each group. Add different reagent solutions to the four groups respectively, shake gently, and let stand at room temperature for 5 min. Transfer the reaction solutions of each group into 1 cm cuvettes and measure the absorbance at 517 nm.

[0231] Result calculation

[0232] Calculate the DPPH free radical scavenging rate (%) according to Equation 3.

[0233]

[0234] In the formula:

[0235] T — is the absorbance of the sample tube, that is, the absorbance of the solution after the sample reacts with DPPH;

[0236] T0 — is the absorbance of the sample background;

[0237] C — is the average absorbance of the DPPH tube in 3 parallel measurements, that is, the absorbance of the DPPH solution without adding the sample;

[0238] C0 — is the absorbance of the solution containing the background.

[0239] Data analysis

[0240] The statistical analysis software was SPSS. The independent samples t-test was used to compare the DPPH radical scavenging rates among the test samples, positive control substances, and negative control substances. The above statistical analyses were all two-tailed tests, and the significance level was a = 0.05. P ≥ 0.05 indicates no significant difference between the two groups; P < 0.05 indicates a significant difference between the two groups.

[0241] Result judgment criteria

[0242] If the DPPH radical scavenging rate of the sample is higher than that of the negative control and there is a significant difference (P < 0.05), it can be considered that the test sample has a certain anti-wrinkle effect.

[0243] 9. Test for the ROS scavenging rate of keratinocytes

[0244] Treatment and reagents for each test group

[0245] Blank group: Cells (+) H2O2 (-) DCFH-DA (+) Sample (-);

[0246] Negative group: Cells (+) H2O2 (+) DCFH-DA (+) Sample (-);

[0247] Positive group: Cells (+) H2O2 (+) DCFH-DA (+) Vitamin E (+);

[0248] Sample group: Cells (+) H2O2 (+) DCFH-DA (+) Sample (+).

[0249] Sample: The fermentation oil prepared in Example 1 was diluted with absolute ethanol to a solution with a mass fraction of 10% of the fermentation oil.

[0250] Negative control: Cell culture medium

[0251] Positive control: Vitamin E

[0252] Test steps

[0253] Take cells in the logarithmic growth phase, prepare cell suspensions, inoculate them in 6-well plates / 24-well plates at 1 mL / well, and culture for 24 h. Discard the culture medium in each well of the plate. Add the culture medium containing a certain concentration of the test substance to the test substance wells, add normal culture medium to the negative control, blank control, and naked cell wells, and add the culture medium containing the corresponding concentration of the positive control to the positive control well. After administration, place the plate in a CO2 incubator for 4 h ± 2 h. After incubation, without discarding the liquid, except for the blank control well, the other wells need to be induced with H2O2. The induction time of H2O2 is overnight, and the induction dose is 300 μM. Discard the culture medium in the wells, wash twice with serum-free DMEM, and add 1 mL of DCFH-DA working solution to each well (the DCFH-DA working solution is prepared according to a dilution ratio of 1:2000, and DCFH-DA is diluted with serum-free DMEM to a final concentration of 5 μmol / L). Incubate in a CO2 incubator for 30 min. After incubation, wash twice with serum-free DMEM. Place the plate after the operation on the microscope stage, use the B channel, excitation light wavelength 488 nm, objective lens 20 times, set the same exposure time, and take pictures. For the collected pictures, use Image Pro Plus software to analyze the fluorescence intensity of the pictures.

[0254] Result calculation

[0255] The calculation of the ROS inhibition rate is shown in Equation 4:

[0256]

[0257] In the formula: T—the average value of the fluorescence intensity of the test sample for 3 times;

[0258] C—the average value of the fluorescence intensity of the negative control for 3 times.

[0259] C0—the average value of the fluorescence intensity of the blank group for 3 times.

[0260] Result judgment criteria

[0261] To evaluate the ability of the test substance to inhibit ROS, the inhibition rate value of the test substance needs to be positive, and there is a significant difference between the fluorescence intensity value and the fluorescence intensity value of the negative control (P < 0.05).

[0262] II. Test results:

[0263] (1) Test results of nourishing efficacy

[0264] Table 3

[0265]

[0266] Result analysis:

[0267] As shown in Table 3, the galactose yeast fermented oil prepared in Example 1 has a dry combing improvement rate of 6.490% and a glossiness improvement rate of 9.153% after the hair bundles are treated with the sample compared with those before the treatment. There is a significant difference between the two groups of data after the hair bundles are treated with the sample and before the treatment (significance factor P<0.050), indicating that the sample has a nourishing effect.

[0268] (2) Repair efficacy test results

[0269] Table 4

[0270]

[0271] Result analysis:

[0272] As shown in Table 4, the galactose yeast fermented oil prepared in Example 1, after being treated with the sample, the friction force of the hair bundle after being treated was reduced by 5.647% compared with that before being treated. There was a significant difference between the two groups of data after being treated with the sample and before being treated (significant factor P<0.050), indicating that this sample has a repairing effect.

[0273] (3) Thermal protection performance test results

[0274] Table 5

[0275]

[0276]

[0277] Result analysis:

[0278] As shown in Table 5, the galactose yeast fermented oil prepared in Example 1 was heated to 180 degrees Celsius by a direct current heating rod, and the friction of the hair bundle treated with the sample decreased by 6.171%, while that of the blank hair bundle increased by 6.004%; the friction change rate of the hair bundle treated with the sample and the blank hair bundle was 12.175%, and there was a significant difference between the two groups of data (significant factor P<0.050). This indicates that the sample can reduce the heat damage to the hair bundle caused by heating and has a repairing effect.

[0279] (4) Scanning electron microscope test results

[0280] Result analysis:

[0281] refer to Figures 5 - 6 It can be seen that the galactose yeast fermented oil prepared in Example 1, the scanning electron microscope image of the hair before using the sample, can be seen that the hair scales are curled and partially peeled off and uneven before using the sample; after using the sample, it can be seen that some hair scales are slightly closed, indicating that the sample has a certain repair effect on the hair scales.

[0282] (5) The test results of elastase inhibition rate are shown in Table 6 as follows:

[0283] Table 6

[0284]

[0285]

[0286] Result analysis:

[0287] 1. As can be seen from Table 6, the galactose yeast fermentation oil prepared in Examples 1-4 has a significant inhibitory effect on elastase.

[0288] 2. By comparing Example 1 (5 g), Example 5 (8 g) and Example 6 (10 g), it can be seen that different inoculation amounts of the seed liquid have different inhibitory effects on elastase, but all three have significant inhibitory effects;

[0289] 3. From Examples 1 and 9-11, it can be seen that when the cereal components in the activation medium and the seed medium are malt extract, red date extract, corn extract, and sorghum extract, the elastase inhibitory ability of Example 11 is significantly better than that of Examples 9-10. The difference between Example 10 and Example 11 is only the use of soybean extract and red date extract. It can be seen that although Example 9 contains red date extract, it only lacks sorghum extract compared to Example 11. It can be seen that in the activation medium and the seed medium, the mixed combination of red date extract and sorghum extract can effectively improve the production activity of the elastase inhibitory product of the strain;

[0290] Further observing Examples 1, 12-13, it can be seen that when the fermentation medium is prepared by mixing malt extract, soybean extract, and corn extract, the elastase inhibition rate of Example 13 is significantly better than that of Examples 1 and 12, and the inhibitory ability of elastase obtained in Example 13 far exceeds the simple superposition of Examples 1 and 12. It can be seen that the compound use of malt extract, soybean extract, and corn extract improves the fermentation ability of the elastase inhibitory product of the strain;

[0291] 4. From Examples 1 and Comparative Examples 1-2, it can be seen that when other fermentation strains are used to replace galactose yeast, the elastase inhibition rates of Comparative Examples 1 and 2 both show an obvious downward trend. Further observing Example 14, it can be seen that even when the same kind of galactose yeast is used to replace the galactose yeast used in Example 1, Example 14 still does not obtain good elastase inhibitory ability. It can be seen that the galactose yeast used in Example 1 is difficult to be replaced;

[0292] 5. As can be seen from Example 1 and Comparative Examples 3-6, when any one of macadamia nut oil and Phyllanthus emblica fruit is missing in the fermentation raw materials, the elastase inhibitory ability of Comparative Examples 3 and 4 both shows an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are indispensable;

[0293] Further observing Comparative Examples 5-6, when any one of macadamia nut oil and Phyllanthus emblica fruit is replaced with other similar substances, the elastase inhibitory ability of Comparative Examples 5 and 6 both shows an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are irreplaceable.

[0294] (6) The test results of hyaluronidase inhibition rate are shown in Table 7:

[0295] Table 7

[0296] Name Test Result P - value Example 1 14.407 ＜0.05 Example 2 15.829 ＜0.05 Example 3 16.056 ＜0.05 Example 4 13.843 ＜0.05 Example 5 18.499 ＜0.05 Example 6 18.215 ＜0.05 Example 7 12.386 ＜0.05 Example 8 13.012 ＜0.05 Example 9 13.258 ＜0.05 Example 10 13.153 ＜0.05 Example 11 13.699 ＜0.05 Example 12 14.023 ＜0.05 Example 13 15.543 ＜0.05 Example 14 12.016 ＜0.05 Comparative Example 1 11.935 ＜0.05 Comparative Example 2 10.911 ＜0.05 Comparative Example 3 9.290 ＜0.05 Comparative Example 4 8.887 ＜0.05 Comparative Example 5 10.523 ＜0.05 Comparative Example 6 10.310 ＜0.05 Comparative Example 7 11.874 ＜0.05 Comparative Example 8 12.138 ＜0.05 Negative Control Group -3.131 / Positive Control Group 57.730 ＜0.05

[0297] Result analysis:

[0298] 1. As can be seen from Table 7, the galactose yeast fermentation oil prepared in Examples 1 to 4 all has a significant inhibitory effect on hyaluronidase.

[0299] 2. By comparing Example 1 (5 g), Example 5 (8 g) and Example 6 (10 g), it can be seen that different inoculation amounts of the seed liquid have different inhibitory effects on hyaluronidase, but all three have a significant inhibitory effect;

[0300] 3. As can be seen from Example 1 and Examples 9-11, when the cereal components in the activation medium and the seed medium are malt extract, red date extract, corn extract, and sorghum extract, the hyaluronidase inhibitory ability of Example 11 is significantly better than that of Examples 9-10. The difference between Example 10 and Example 11 is only the use of soybean extract and red date extract. It can be seen that although Example 9 contains red date extract, it only lacks sorghum extract compared with Example 11. It can be seen that in the activation medium and the seed medium, the mixed combination of red date extract and sorghum extract can effectively improve the production activity of the hyaluronidase inhibitory product of the strain;

[0301] Further observing Example 1 and Examples 12-13, when the fermentation medium is prepared by mixing malt extract, soybean extract, and corn extract, the hyaluronidase inhibition rate of Example 13 is significantly better than that of Example 1 and Example 12, and the inhibitory ability of hyaluronidase obtained in Example 13 far exceeds the simple superposition of Example 1 and Example 12. It can be seen that the compound use of malt extract, soybean extract, and corn extract improves the fermentation ability of the hyaluronidase inhibitory product of the strain;

[0302] 4. As can be seen from Example 1 and Comparative Examples 1-2, when using other fermentation strains instead of galactose yeast, the hyaluronidase inhibition rates in Comparative Examples 1 and 2 both showed an obvious downward trend. Further observing Example 14, it can be seen that even when using the same galactose yeast to replace the galactose yeast used in Example 1, Example 14 still did not achieve good hyaluronidase inhibition ability. It can be seen that the galactose yeast used in Example 1 is difficult to be replaced;

[0303] 5. As can be seen from Example 1 and Comparative Examples 3-6, when any one of macadamia nut oil and Phyllanthus emblica fruit is missing in the fermentation raw materials, the hyaluronidase inhibition abilities in Comparative Examples 3 and 4 both showed an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are indispensable;

[0304] Further observing Comparative Examples 5-6, it can be seen that when using other similar substances to replace any one of macadamia nut oil and Phyllanthus emblica fruit, the hyaluronidase inhibition abilities in Comparative Examples 5 and 6 both showed an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are irreplaceable.

[0305] (7) The test results of the DPPH free radical scavenging rate are shown in Table 8:

[0306] Table 8

[0307] Name Test Result P - value Example 1 25.365 ＜0.05 Example 2 24.867 ＜0.05 Example 3 25.671 ＜0.05 Example 4 21.179 ＜0.05 Example 5 29.970 ＜0.05 Example 6 31.376 ＜0.05 Example 7 20.235 ＜0.05 Example 8 21.268 ＜0.05 Example 9 21.384 ＜0.05 Example 10 21.190 ＜0.05 Example 11 22.631 ＜0.05 Example 12 25.108 ＜0.05 Example 13 26.390 ＜0.05 Example 14 21.084 ＜0.05 Comparative Example 1 19.943 ＜0.05 Comparative Example 2 16.695 ＜0.05 Comparative Example 3 17.436 ＜0.05 Comparative Example 4 18.973 ＜0.05 Comparative Example 5 19.888 ＜0.05 Comparative Example 6 19.014 ＜0.05 Comparative Example 7 20.094 ＜0.05 Comparative Example 8 20.357 ＜0.05 Negative Control Group -0.547 / Positive Control Group 92.555 ＜0.05

[0308] Result analysis:

[0309] 1. As can be seen from Table 8, the galactose yeast fermentation oil prepared in Examples 1 to 4 has a significant improvement effect on the DPPH free radical scavenging rate.

[0310] 2. By comparing Example 1 (5g), Example 5 (8g) and Example 6 (10g), it can be seen that different inoculation amounts of the seed liquid have different scavenging effects on DPPH free radicals, but all three have a significant improvement effect on the DPPH free radical scavenging rate;

[0311] 3. As can be seen from Example 1 and Examples 9-11, when the cereal components in the activation medium and the seed medium are malt extract, red date extract, corn extract, and sorghum extract, the DPPH free radical scavenging ability of Example 11 is significantly better than that of Examples 9-10. The difference between Example 10 and Example 11 is only the use of soybean extract and red date extract. It can be seen that although Example 9 contains red date extract, it only lacks sorghum extract compared to Example 11. It can be seen that in the activation medium and the seed medium, the mixed combination of red date extract and sorghum extract can effectively improve the production activity of the product with DPPH free radical scavenging ability of the strain;

[0312] Further observation of Example 1 and Examples 12 - 13 shows that when the fermentation medium is prepared by mixing malt extract, soybean extract, and corn extract, the DPPH free radical scavenging rate of Example 13 is significantly better than that of Example 1 and Example 12, and the DPPH free radical scavenging ability obtained in Example 13 far exceeds the simple superposition of Example 1 and Example 12. It can be seen that the combined use of malt extract, soybean extract, and corn extract improves the fermentation ability of the product with the DPPH free radical scavenging ability of the strain;

[0313] 4. It can be seen from Example 1 and Comparative Examples 1 - 2 that when using other fermentation strains to replace galactose yeast, the DPPH free radical scavenging rates of Comparative Examples 1 and 2 both show an obvious downward trend. Further observation of Example 14 shows that even when using the same galactose yeast to replace the galactose yeast used in Example 1, Example 14 still does not achieve good DPPH free radical scavenging ability. It can be seen that the galactose yeast used in Example 1 is difficult to be replaced;

[0314] 5. It can be seen from Example 1 and Comparative Examples 3 - 6 that when any one of macadamia nut oil and Phyllanthus emblica fruit is missing in the fermentation raw materials, the DPPH free radical scavenging abilities of Comparative Examples 3 and 4 both show an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are indispensable;

[0315] Further observation of Comparative Examples 5 - 6 shows that when using other similar substances to replace any one of macadamia nut oil and Phyllanthus emblica fruit, the DPPH free radical scavenging abilities of Comparative Examples 5 and 6 both show an obvious downward trend. It can be seen that macadamia nut oil and Phyllanthus emblica fruit are irreplaceable.

[0316] (8) The test results of the ROS scavenging rate of keratinocytes are shown in Table 9:

[0317] Table 9

[0318] Name ROS Inhibition Rate / % P - value Example 1 36.151±2.336 ＜0.05 Positive Group 52.268±0.585 ＜0.05

[0319] Result analysis:

[0320] As can be seen from Table 9, for the galactose yeast fermentation oil prepared in Example 1, its cell ROS inhibition rate is 36.151%, and there is a significant difference from the negative control (P < 0.05), that is, the test sample has an obvious effect of inhibiting cell ROS, indicating that the test sample has a certain antioxidant effect.

[0321] Application Example

[0322] A hair care essential oil, the formula ratio is shown in Table 10:

[0323] Table 10

[0324]

[0325]

Claims

1. A fermented oil, characterized in that: The fermented oil is obtained by fermenting macadamia seed oil and emblica fruit with a fermentation liquid containing live galactosaccharomyces-like bacteria; The fermentation liquid containing live galactosaccharomyces-like bacteria is obtained by fermenting the first cereal extract with galactosaccharomyces-like bacteria; The first cereal extract is selected from at least one of malt extract, soybean extract and corn extract; The total mass ratio of the fermentation liquid containing live galactosyl yeast-like bacteria to the macadamia seed oil and the emblica fruit is 3-15:8-20; The mass ratio of the macadamia seed oil to the emblica fruit is 16:1-6; The deposit number of the galactose yeast-like bacteria is GDMCC No: 64235, the deposit date is January 2, 2024, the deposit unit is Guangdong Microbiological Culture Collection Center, and the classification name is Geotrichum candidum .

2. The fermented oil according to claim 1, characterized in that The galactose yeast-like bacteria are activated galactose yeast-like bacteria, and the activation method of the activated galactose yeast-like bacteria comprises the following steps: Step 1: Mix the second grain extract, glucose, yeast extract and pure water, sterilize and obtain an activated culture medium, then add galactose yeast-like bacteria to the activated culture medium to obtain a galactose yeast-like bacteria activated liquid; Step 2: Mix the third grain extract, glucose, yeast extract and pure water, sterilize and obtain a seed culture medium, then add a galactose yeast-like bacteria activation liquid to the seed culture medium to obtain a galactose yeast-like bacteria seed liquid, i.e., activated galactose yeast-like bacteria; The activation medium contains 0.1 wt% to 10 wt% glucose, 0.1 wt% to 20 wt% second cereal extract, 0.1 wt% to 5 wt% yeast extract, and the remainder is water; The content of glucose in the seed culture medium is 0.1wt% to 10wt%, the content of the third cereal extract is 0.1wt% to 20wt%, the content of the yeast extract is 0.1wt% to 5wt%, and the balance is water; The second cereal extract is selected from at least one of malt extract, soybean extract, corn extract, sorghum extract, red date extract, and sugarcane extract; The third cereal extract is selected from at least one of malt extract, soybean extract, corn extract, sorghum extract, red date extract and sugarcane extract.

3. The fermented oil according to claim 2, characterized in that The concentration of live galactosaccharomyces cerevisiae in the galactosaccharomyces cerevisiae seed solution is 1×10 6 ~2x10 8 CFU / g.

4. The fermented oil according to claim 2, characterized in that The mass ratio of the galactose yeast-like bacteria seed liquid to the first grain extract is 5-10:2-30.

5. A method for preparing the fermented oil according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step A: inoculating galactosaccharomyces cerevisiae into a fermentation medium to obtain a fermentation liquid containing live galactosaccharomyces cerevisiae; Step B: crushing the emblica fruit to obtain emblica fruit powder, and mixing the macadamia seed oil and the emblica fruit powder in a mass ratio of 16:1-6 to obtain an intermediate; Step C: mixing the fermentation liquid containing live galactosyl yeast-like bacteria obtained in step A with the intermediate obtained in step B, and fermenting them to obtain fermentation oil; The fermentation medium is prepared by mixing a first grain extract, glucose, a yeast extract, and pure water, wherein the content of glucose in the fermentation medium is 0.1wt% to 10wt%, the content of the first grain extract is 0.1wt% to 20wt%, the content of the yeast extract is 0.1wt% to 5wt%, and the balance is water; The mass ratio of the fermentation liquid containing live galactose yeast-like bacteria obtained in step A to the intermediate obtained in step B is 3-15:8-20.

6. The method for preparing fermented oil according to claim 5, characterized in that: The step B specifically comprises: mixing macadamia seed oil and emblica fruit in a mass ratio of 16:1-6 and stirring the mixture in an environment of 70° C. for 2-4 hours to obtain an intermediate.

7. The method for preparing fermented oil according to claim 5, characterized in that: The step C specifically comprises: mixing the fermentation liquid containing live galactose yeast-like bacteria obtained in step A with the intermediate obtained in step B, and fermenting them at 27-30° C. for 20-26 hours to obtain fermented oil.

8. Use of the fermented oil as claimed in any one of claims 1 to 4 in the preparation of cosmetics.

9. A cosmetic, characterized in that: Contains 0.00001 to 30 wt % of the fermented oil as described in any one of claims 1 to 4.

Images