A firming and anti-wrinkle composition and its preparation method and application

Abstract

The present application belongs to the technical field of cosmetic production, and discloses a firming and anti-wrinkle composition, which is prepared by drying, crushing, enzymolysis, fermentation and filtering of purslane, emblica and tremella, wherein the mass ratio of purslane, emblica and tremella is 5-30:1-10:1-10. The present application obtains a firming and anti-wrinkle composition by fermenting the above plants, and the composition is obtained by fermenting and screening the functional ingredients in the above plants with yeast to obtain a composition with good anti-wrinkle, firming and soothing effects. In addition, the present application also discloses a preparation method of the firming and anti-wrinkle composition and the use of the firming and anti-wrinkle composition.

Description

Technical Field

[0001] The present invention relates to the technical field of cosmetic production, and in particular to a firming and anti-wrinkle composition and a preparation method and application thereof. Background Art

[0002] Antioxidation is an important mechanism that can protect the skin from free radical damage and is of great significance for delaying skin aging. Before understanding the significance of antioxidation, we first need to understand what free radicals are and how they cause skin aging. Free radicals are highly active molecules or atoms with unpaired electrons. They will constantly react with surrounding molecules, causing oxidative damage. When there are too many free radicals, it will cause oxidative stress and accelerate the aging process of the skin, which will manifest as wrinkles, spots, sagging and other skin problems.

[0003] The role of anti-oxidation is to neutralize free radicals and prevent them from damaging the skin. By using ingredients with antioxidant effects, the production of free radicals can be reduced, the skin's self-repair ability can be enhanced, and the skin can be protected from external factors such as environmental pollution and ultraviolet radiation. Therefore, anti-oxidation has the following important significance for delaying skin aging:

[0004] 1. Prevent oxidative stress: Antioxidant ingredients can neutralize free radicals, prevent the occurrence of oxidative stress, protect cells from damage, and slow down skin aging.

[0005] 2. Promote collagen synthesis: Oxidative stress can destroy collagen in the skin, causing skin sagging and deepening wrinkles. Antioxidants can promote collagen synthesis, improve skin elasticity, and slow down the aging process.

[0006] 3. Reduce pigmentation: Free radicals can increase melanin production and promote the formation of pigmentation. Antioxidants can reduce the production of melanin, reduce pigmentation, and brighten the skin.

[0007] 4. Maintain the water-oil balance of the skin: Oxidative stress can damage the skin's protective barrier and cause an imbalance of water and oil. Antioxidants help maintain the water-oil balance of the skin and keep the skin healthy.

[0008] Prior art 1: Chinese patent application 202010260346.0 discloses a cosmetic composition with anti-glycation and anti-aging effects and its application, which belongs to the technical field of functional compositions, including 0.5-4 parts of vitamin C, 0.5-4 parts of SOD yeast powder, 0.2-5 parts of Haematococcus pluvialis, 0.5-10 parts of thread-leaved broom, 0.5-8 parts of licorice extract, 0.1-6 parts of bayberry extract, 0.5-8 parts of longan shell extract, 0.1-8 parts of burdock polyphenols, 0.5-8 parts of cinnamon extract, 0.1-5 parts of ginseng extract, 0.5-8 parts of kudzu root extract, 1-10 parts of purslane extract, 2-40 parts of carbohydrates, 2-30 parts of proteins and 0.5-5 parts of lipids. This solution can improve skin aging caused by skin oxidation, ultraviolet damage, skin glycation, melanin deposition, etc. The solution can prevent aging and darkening of the skin, make the skin in good condition, and achieve the effect of delaying aging and whitening. In addition, the solution also rationally mixes various nutritional components that can be used as both medicine and food, such as vitamin C, yeast powder containing SOD, Haematococcus pluvialis, broom, licorice extract, bayberry extract, longan shell extract, burdock polyphenols, and cinnamon extract, to make them synergistic with each other. SOD yeast powder is used as the first line of defense against free radicals, and Haematococcus pluvialis that can penetrate the cell membrane is used in combination with broom to effectively remove free radicals, thereby enhancing the skin's antioxidant defense ability and fundamentally preventing the accumulation of oxidative stress that causes skin damage and aging.

[0009] Prior art 2: Chinese patent application 202110238566.8 discloses a whitening, moisturizing, anti-glycation soft candy and a preparation method thereof. The whitening, moisturizing, anti-glycation soft candy includes the following components by weight: 500-900 parts of sweetener; 10-60 parts of gelling agent; 50-200 parts of functional components; 1-20 parts of acidity regulator;

[0010] In the scheme, after the Tremella is ground into pulp, pectinase and hemicellulase are added. The hemicellulase can enzymatically hydrolyze Tremella polysaccharide into small-molecule Tremella polysaccharide, which is convenient for human body absorption. At the same time, the addition of acidic regulator can form glucuronic acid with small-molecule Tremella polysaccharide. Glucuronic acid can effectively remove free radicals. Tremella polysaccharide itself has gelling properties. When combined with gelling agent, the amount of gelling agent can be reduced, so that the obtained soft candy has good elasticity and hardness.

[0011] Further observation of the scheme shows that the functional components in the scheme also include yeast extract and emblica extract, but the embodiments of the scheme do not disclose a case in which the above-mentioned Tremella polysaccharide, yeast extract and emblica extract are used together.

[0012] The problem that this program needs to solve is: How to develop a composite plant fermentation product with good anti-wrinkle, firming and soothing abilities. Summary of the invention

[0013] The purpose of the present application is to provide a composite plant fermentation product with good anti-wrinkle, firming and soothing abilities. In addition, the present application also provides a preparation method and application of the above-mentioned composite plant fermentation product.

[0014] To achieve the above-mentioned purpose, the present application discloses a firming and anti-wrinkle composition, which is prepared by drying, crushing, enzymolysis, fermentation and filtering of purslane, emblica, and tremella, wherein the mass ratio of purslane, emblica, and tremella is 5-30:1-10:1-10;

[0015] Among them, purslane has anti-inflammatory effect, can prevent and treat skin diseases such as eczema, allergic dermatitis and contact dermatitis, and it has good free radical scavenging ability and antioxidant effect. At the same time, purslane has good skin moisturizing properties.

[0016] Tremella can bind more than 500 times its own weight of water, helping the skin to replenish more water and increase the water content of the stratum corneum. At the same time, the active ingredients in Tremella have good film-forming properties, and the unique multi-side chain spatial network structure and complex sugar chain structure can firmly bind water molecules, showing a strong ability to lock in moisture and retain moisture.

[0017] At the same time, Tremella also has the ability to scavenge hydroxyl free radicals, can inhibit lipid peroxidation, has obvious antioxidant effects, can delay skin aging, has a certain soothing effect on skin allergies, and can be used to inhibit skin sensitivity;

[0018] Emblica officinalis has good antioxidant activity and can delay skin aging.

[0019] Preferably, the enzyme used in the enzymatic hydrolysis process is selected from at least one of alkaline protease, cellulase, trypsin and neutral protease.

[0020] The mass ratio of the enzyme to purslane, emblica fruit and tremella is 0.1-5:5-30:1-10:1-10.

[0021] Preferably, the yeast used in the fermentation process is selected from at least one of galactosaccharomyces, bifidosaccharomyces and Pichia pastoris.

[0022] Preferably, the yeast used in the fermentation process comprises at least galactosaccharomyces galactosidase.

[0023] Preferably, the Latin name of the galactosaccharomyces galactosidum is Geotrichum candidum, the strain collection number is GDMCC: 64235, the collection date is January 2, 2024, the collection unit is Guangdong Microbiological Culture Collection Center, and the collection address is 5th Floor, Experimental Building, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.

[0024] Preferably, the yeast used in the fermentation process is a mixture of galactose yeast and Pichia pastoris, and the mass ratio of galactose yeast to Pichia pastoris is 2 to 4:1.

[0025] Preferably, the enzyme used in the enzymatic hydrolysis process is selected from a mixture of alkaline protease, cellulase and trypsin, and the mass ratio of the alkaline protease, cellulase and trypsin is 1-3:1-3:1-3.

[0026] In addition, the present application also discloses a method for preparing the above-mentioned firming and anti-wrinkle composition, comprising the following steps:

[0027] Step 1: taking, drying and crushing purslane, emblica fruit and tremella in a mass ratio of 5-30:1-10:1-10 to obtain powder;

[0028] Step 2: performing enzymatic hydrolysis on the powder obtained in step 1 to obtain an enzymatic hydrolyzate;

[0029] Step 3: The enzymatic hydrolysate obtained in step 2 is made into a culture medium and inoculated with yeast for fermentation and filtration to obtain a firming and anti-wrinkle composition.

[0030] In addition, the present application also discloses the use of the firming and anti-wrinkle composition as an active substance in cosmetics.

[0031] The beneficial effects of the present application are as follows: the present application uses purslane, tremella, and emblica fruit as main ingredients, and the above ingredients are subjected to enzymatic hydrolysis and fermentation to obtain a composition with good antioxidant capacity, wherein purslane has an anti-inflammatory effect, can prevent and treat skin diseases such as skin eczema, allergic dermatitis, contact dermatitis, and has good free radical scavenging ability and antioxidant effect, and purslane has good skin moisturizing property;

[0032] Tremella can bind more than 500 times its own weight of water, helping the skin to replenish more water and increase the water content of the stratum corneum. At the same time, the active ingredients in Tremella have good film-forming properties, and the unique multi-side chain spatial network structure and complex sugar chain structure can firmly bind water molecules, showing a strong ability to lock in moisture and retain moisture.

[0033] At the same time, Tremella also has the ability to scavenge hydroxyl free radicals, can inhibit lipid peroxidation, has obvious antioxidant effects, can delay skin aging, has a certain soothing effect on skin allergies, and can be used to inhibit skin sensitivity;

[0034] Phyllanthus emblica has good antioxidant activity and can delay skin aging;

[0035] Moreover, after the above ingredients undergo enzymatic hydrolysis and fermentation, the antioxidant capacity of the obtained products is further improved. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a schematic diagram of a chicken embryo before adding the solution prepared by the firming and anti-wrinkle composition of Example 1;

[0037] Figure 2 This is a schematic diagram of a chicken embryo after adding the solution prepared from the firming and anti-wrinkle composition of Example 1. DETAILED DESCRIPTION

[0038] The present invention will be described clearly and completely below in conjunction with the embodiments of the present invention. In the description of the present invention, it should be noted that, if no specific conditions are specified in the embodiments, the conditions are carried out according to conventional conditions or conditions recommended by the manufacturer. If the manufacturer of the reagents or instruments used is not specified, they are all conventional products that can be purchased commercially.

[0039] Before describing the embodiments and comparative examples, the raw material information involved in the embodiments and comparative examples is first described as follows, as shown in Table 1:

[0040] Table 1

[0041]

[0042] Example 1

[0043] A firming and anti-wrinkle composition, the preparation method is as follows:

[0044] Step 1: Take purslane, emblica fruit and tremella in a mass ratio of 30:10:10, dry and crush to obtain powder;

[0045] Step 2: performing enzymatic hydrolysis on the powder obtained in step 1 to obtain an enzymatic hydrolyzate;

[0046] The enzyme used in the enzymatic hydrolysis process is alkaline protease, and the mass ratio of the enzyme to purslane, emblica fructus, and tremella is 5:30:10:10;

[0047] Step 3: Mix the enzymatic hydrolysate prepared in step 2 with peptone, glucose, sodium dihydrogen phosphate and pure water to prepare a culture medium, wherein the mass fraction of the enzymatic hydrolysate is 80%, the mass fraction of peptone is 2%, the mass fraction of glucose is 2%, the mass fraction of sodium dihydrogen phosphate is 1.5%, and pure water is added to a total mass fraction of 100%;

[0048] Then, the yeast is inoculated, fermented, and filtered to obtain a firming and anti-wrinkle composition;

[0049] The yeast used in the fermentation process is selected from galactose yeast, the galactose yeast is the galactose yeast with the Latin name Geotrichum candidum, the strain collection number GDMCC: 64235, the preservation date January 2, 2024, and the galactose yeast preserved in the Guangdong Provincial Microbiological Culture Collection Center, and the mass ratio of yeast to purslane, emblica, and tremella is 3:30:10:10.

[0050] Example 2

[0051] A firming and anti-wrinkle composition, the preparation method is as follows:

[0052] Step 1: taking purslane, emblica fruit and tremella in a mass ratio of 5:1:1, drying and crushing to obtain powder;

[0053] Step 2: performing enzymatic hydrolysis on the powder obtained in step 1 to obtain an enzymatic hydrolyzate;

[0054] The enzyme used in the enzymatic hydrolysis process is cellulase, and the mass ratio of the enzyme to purslane, emblica fructus, and tremella is 0.1:5:1:1.

[0055] Step 3: Mix the enzymatic hydrolysate prepared in step 2 with peptone, glucose, sodium dihydrogen phosphate and pure water to prepare a culture medium, wherein the mass fraction of the enzymatic hydrolysate is 80%, the mass fraction of peptone is 2%, the mass fraction of glucose is 2%, the mass fraction of sodium dihydrogen phosphate is 1.5%, and pure water is added to a total mass fraction of 100%;

[0056] Then, the yeast is inoculated, fermented, and filtered to obtain a firming and anti-wrinkle composition;

[0057] The yeast used in the fermentation process is Pichia pastoris

[0058] The mass ratio of Pichia pastoris to Purslane, Emblica officinalis and Tremella fuciformis is 0.1:5:1:1.

[0059] Example 3

[0060] A firming and anti-wrinkle composition, the preparation method is as follows:

[0061] Step 1: Take purslane, emblica fruit and tremella in a mass ratio of 15:7:8, dry and crush to obtain powder;

[0062] Step 2: performing enzymatic hydrolysis on the powder obtained in step 1 to obtain an enzymatic hydrolyzate;

[0063] The enzyme used in the enzymatic hydrolysis process is trypsin, and the mass ratio of the enzyme to purslane, emblica fructus, and tremella is 1:15:7:8;

[0064] Step 3: Mix the enzymatic hydrolysate prepared in step 2 with peptone, glucose, sodium dihydrogen phosphate and pure water to prepare a culture medium, wherein the mass fraction of the enzymatic hydrolysate is 80%, the mass fraction of peptone is 2%, the mass fraction of glucose is 2%, the mass fraction of sodium dihydrogen phosphate is 1.5%, and pure water is added to a total mass fraction of 100%;

[0065] Then, the yeast is inoculated, fermented, and filtered to obtain a firming and anti-wrinkle composition;

[0066] The yeast used in the fermentation process is bifid yeast, and the mass ratio of yeast to purslane, emblica fructus and tremella is 1:15:7:8.

[0067] Example 4

[0068] It is basically the same as Example 1, except that the commercially available galactose yeast in Example 1 is replaced by the galactose yeast whose Latin name is Geotrichum candidum, the strain collection number is GDMCC: 64235, the collection date is January 2, 2024, the collection unit is Guangdong Microbiological Culture Collection Center, and the collection address is 5th Floor, Experimental Building, No. 100, Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.

[0069] Example 5

[0070] The method is basically the same as Example 1, except that the yeast used in the fermentation process is a mixture of galactosaccharomyces, bifid yeast and Pichia pastoris; and the mass ratio of galactosaccharomyces, bifid yeast and Pichia pastoris is 3:0.5:0.5;

[0071] The Latin name of galactosidase is Geotrichum candidum, the strain collection number is GDMCC: 64235, the collection date is January 2, 2024, the collection unit is Guangdong Microbiological Culture Collection Center, and the collection address is galactosidase on the 5th floor of the laboratory building at No. 100 Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.

[0072] Example 6

[0073] The method is basically the same as Example 1, except that the yeast used in the fermentation process is a mixture of galactosaccharomyces and Pichia pastoris; and the mass ratio of galactosaccharomyces to Pichia pastoris is 3:1;

[0074] The Latin name of galactosidase is Geotrichum candidum, the strain collection number is GDMCC: 64235, the collection date is January 2, 2024, the collection unit is Guangdong Microbiological Culture Collection Center, and the collection address is galactosidase on the 5th floor of the laboratory building at No. 100 Xianlie Middle Road, Yuexiu District, Guangzhou City, Guangdong Province.

[0075] Example 7

[0076] The method is basically the same as Example 1, except that the yeast used in the fermentation process is a mixture of galactosaccharomyces and Pichia pastoris; and the mass ratio of galactosaccharomyces to Pichia pastoris is 3:1;

[0077] The galactosidase is commercially available galactosidase.

[0078] Example 8

[0079] The method is basically the same as Example 1, except that the yeast used in the fermentation process is a mixture of galactosyl yeast and bifid yeast; and the mass ratio of galactosyl yeast to bifid yeast is 3:1.

[0080] Example 9

[0081] The method is basically the same as Example 1, except that the enzyme used in the enzymatic hydrolysis process is a mixture of alkaline protease, cellulolytic protease and trypsin, and the mass ratio of the alkaline protease, cellulolytic protease and trypsin is 1:1:1.

[0082] Example 10

[0083] The method is basically the same as Example 6, except that the enzyme used in the enzymatic hydrolysis process is a mixture of alkaline protease, cellulolytic protease and trypsin, and the mass ratio of the alkaline protease, cellulolytic protease and trypsin is 1:1:1.

[0084] Comparative Example 1

[0085] The method is basically the same as Example 1, except that step 1 specifically comprises: taking, drying and crushing the emblica oleracea and tremella fuciformis having the same total mass as that of purslane, emblica oleracea and tremella fuciformis in Example 1 in a mass ratio of 10:10 to obtain powder.

[0086] Comparative Example 2

[0087] The method is basically the same as Example 1, except that step 1 specifically comprises: taking, drying and crushing purslane and tremella having the same total mass as that of purslane, emblica and tremella in Example 1 in a mass ratio of 30:10 to obtain powder.

[0088] Comparative Example 3

[0089] The method is basically the same as Example 1, except that step 1 specifically comprises: taking, drying and crushing purslane and emblica of the same total mass as that of purslane, emblica and tremella in Example 1 in a mass ratio of 30:10 to obtain powder.

[0090] Comparative Example 4

[0091] The method is basically the same as Example 1, except that step 1 specifically comprises: taking Pueraria root, Emblica fructus and Tremella fuciformis having the same total mass as that of Purslane, Emblica fructus and Tremella fuciformis in Example 1 in a mass ratio of 30:10:10, drying and crushing to obtain powder.

[0092] Comparative Example 5

[0093] The method is basically the same as Example 1, except that step 1 specifically comprises: taking purslane, double red rose and tremella with the same total mass as that of purslane, emblica and tremella in Example 1 in a mass ratio of 30:10:10, drying and crushing to obtain powder.

[0094] Comparative Example 6

[0095] The method is basically the same as Example 1, except that step 1 specifically comprises: taking Pueraria root, Emblica emblica, and grape seed having the same total mass as that of Purslane, Emblica emblica, and Tremella fuciformis in Example 1 in a mass ratio of 30:10:10, drying, and crushing to obtain powder.

[0096] Comparative Example 7

[0097] A composition, the preparation method is as follows:

[0098] Step 1: Take purslane, emblica fruit and tremella in a mass ratio of 30:10:10, dry and crush to obtain powder;

[0099] Step 2: enzymatically hydrolyzing the powder obtained in step 1 to obtain a composition;

[0100] The enzyme used in the enzymatic hydrolysis process is alkaline protease, and the mass ratio of the enzyme to purslane, emblica fructus, and tremella is 5:30:10:10.

[0101] Performance Testing

[0102] 1. Irritation test

[0103] 1.1 Instruments and Equipment

[0104] Fully automatic incubator, stereo microscope, SPF chicken embryos;

[0105] 1.2 Reagents

[0106] Sodium chloride, sodium dodecyl sulfate (SDS);

[0107] 1.3 Incubation conditions

[0108] Room temperature is 20℃~25℃, relative humidity is 45%~70%, incubation temperature is 37.5℃±0.5℃, relative humidity is 55%~70%, turntable is 3 times / h~6 times / h, and it is not necessary to rotate when incubating 9-day-old chicken embryos.

[0109] 1.4 Test methods

[0110] (1) Test operation steps

[0111] In this test, 6 embryos were selected for each group, and the conditions of the chorioallantoic membrane were recorded with a camera, and a polytetrafluoroethylene resin ring was placed on the chorioallantoic membrane of the chicken embryo, and photographed and recorded. The composite plant fermentation product of the embodiment and the comparative example was diluted with pure water to a solution with a mass fraction of 1% of the composite plant fermentation product and used as a sample to be tested, and then the sample to be tested was added to the polytetrafluoroethylene resin ring, the time of adding the sample was recorded, and the air chamber was covered with a wetted plastic wrap, and the chicken embryo was moved to a constant temperature and humidity chamber for culture, and the degree of change of each toxic effect was observed.

[0112] (2) Results observation

[0113] Observe and record bleeding, coagulation, and vascular melting, and score them according to their severity

[0114] The scoring criteria for bleeding, coagulation, and vascular thaw are shown in Table 2:

[0115] Table 2

[0116]

[0117]

[0118] Rating criteria: ES≤12: no / mild irritation; 12<ES<16: moderate irritation; ES≥16: strong irritation.

[0119] According to the test, the embodiments and comparative examples did not produce irritation. Figure 1-2 , which is a comparison of the states of chicken embryos before and after the solution prepared from the composition of Example 1 is added dropwise to chicken embryos. According to statistics, the ES value of the solution prepared from the composition of Example 1 is 4.00.

[0120] 2.DPPH free radical scavenging rate test

[0121] 2.1 Instruments and Equipment

[0122] BSA224S analytical balance, L6s UV spectrophotometer

[0123] 2.2 Reagents

[0124] DPPH (1,1-diphenyl-2-picrylhydrazyl), 98%

[0125] 2.3 Test methods

[0126] (1) Treatment of control and test samples

[0127] Sample group: the compositions prepared in Examples 1 to 10 and Comparative Examples 1 to 7 at a concentration of 100%;

[0128] Negative control: pure water.

[0129] (2) Test operation steps

[0130] Set up sample group and negative control group. Set up 3 parallel tubes for each group. Add the corresponding reagent solution to each group, shake gently, and let stand at room temperature for 5 minutes. Transfer the reaction solution of each group into a 1cm

[0131] The absorbance was measured at 517 nm in a cuvette.

[0132] (3) The calculation formula is shown in Formula 3:

[0133]

[0134] Where: T—absorbance of sample tube, i.e. absorbance of solution after sample reacts with DPPH;

[0135] T0—sample background absorbance;

[0136] C—the average value of the absorbance of the DPPH tube three times, that is, the absorbance of the DPPH solution when no sample is added;

[0137] C0—Solvent background absorbance.

[0138] (4) Data analysis

[0139] The statistical analysis software was SPSS, and the comparison between the DPPH free radical scavenging rate of the test sample group and the negative control was performed using the independent sample t test. The above statistical analyses were all two-tailed tests, and the significance level was α = 0.05. P>0.05 indicated that there was no significant difference between the two groups; P<0.05 indicated that there was a significant difference between the two groups.

[0140] The test results are shown in Table 3:

[0141] Table 3

[0142]

[0143]

[0144] Result analysis:

[0145] 1. It can be seen from Examples 1-3 that after adjusting the mass ratios of the plant components and the mass ratios of the enzymes and yeast to the plant components, there are certain fluctuations in the DPPH radical scavenging rates between Examples 1-3, and the DPPH radical scavenging rate of Example 3 is higher than that of Examples 1 and 2. It can be seen that the composition prepared by the parameters of Example 3 has better free radical scavenging ability;

[0146] 2. It can be seen from Examples 1 and 4-6 that when Example 4 uses the same type of commercially available galactose yeast to replace Geotrichum candidum, the free radical scavenging ability of Example 4 shows a significant downward trend. It can be seen that even if the same type of galactose yeast is used to replace it, it is still impossible to achieve an effect similar to that of Geotrichum candidum.

[0147] 3. Further observation of Examples 5-6 shows that when Example 5 uses a mixture of galactose yeast, bifid yeast, and Pichia pastoris to replace galactose yeast, its DPPH free radical scavenging rate is slightly improved, but when galactose yeast and Pichia pastoris are used together in Example 6, the free radical scavenging ability of the composition is significantly improved. It can be seen that galactose yeast and Pichia pastoris can cooperate with each other to ferment more fermentation products with free radical scavenging ability, and the reason why the free radical scavenging ability of Example 5 is not high may be that bifid yeast has occupied the fermentation raw materials or intermediates that should have been fermented by galactose yeast and Pichia pastoris to a certain extent;

[0148] Further observation of Examples 7-8 shows that when Example 7 and Example 6 are compounded with Galacto-saccharomyces and Pichia pastoris, but commercially available Galacto-saccharomyces is used, the free radical scavenging ability of Example 7 is close to that of Example 4, and is lower than that of Example 1 and significantly lower than that of Example 6. It can be seen that the synergistic ability between commercially available Galacto-saccharomyces and Pichia pastoris is weak.

[0149] The effect of the combined use of galactosyl yeast and bifid yeast in Example 8 is also weaker than that in Example 1, which shows that bifid yeast does occupy part of the fermentation resources originally belonging to galactosyl yeast.

[0150] 4. It can be seen from Examples 1 and 9-10 that when the enzymes are compounded, the free radical scavenging rate of Example 9 is improved to a certain extent, and when Example 10, i.e., the enzymes are further compounded on the basis of Example 6, the free radical scavenging rate of Example 10 is significantly improved, and this improvement trend is greater than the sum of the improvements of Example 6 and Example 9 relative to Example 1. It can be seen that the compounding of enzymes can improve the fermentation capacity of the system with Galactomyces galactosidase and Pichia pastoris as fermentation bacteria, thereby producing more fermentation products with free radical scavenging ability.

[0151] 5. It can be seen from Example 1 and Comparative Examples 1-6 that when any one of Purslane, Emblica emblica, and Tremella fuciformis is missing, the free radical scavenging ability of Comparative Examples 1-3 has a significant downward trend. It can be seen that any one of Purslane, Emblica emblica, and Tremella fuciformis is indispensable;

[0152] When other plant ingredients are used to replace any of the ingredients in Purslane, Emblica fructus, and Tremella fuciformis, the free radical scavenging ability of Examples 4-6 shows a significant downward trend. It can be seen that any of Purslane, Emblica fructus, and Tremella fuciformis is irreplaceable.

[0153] 3. Elastase inhibition rate test

[0154] 3.1 Instruments and Equipment

[0155] BSA224S analytical balance, RT-6100 ELISA analyzer;

[0156] 3.2 Reagents

[0157] Elastase (porcine pancreas), BR, N-succinyl-L-alanyl-L-alanyl-L-alanine, 98%;

[0158] 3.3 Test methods

[0159] (1) Treatment of control and test samples

[0160] Sample group: the compositions prepared in Examples 1 to 10 and Comparative Examples 1 to 7 at a concentration of 100%;

[0161] Negative control: pure water.

[0162] (2) Test operation steps

[0163] Set up a sample group and a negative control group, with 3 parallels for each group. Add different reagent solutions to a 96-well plate, shake gently, incubate at 25°C for 15 minutes, place it in an enzyme reader, and measure the absorbance at 410 nm.

[0164] (3) The calculation formula is shown in Formula 2:

[0165]

[0166] Where: A—is the absorbance of the reaction solution without sample;

[0167] B—is the absorbance of the reaction solution without sample and enzyme;

[0168] C—is the absorbance of the reaction solution containing sample and enzyme;

[0169] D—is the absorbance of the reaction solution containing sample and without enzyme.

[0170] (4) Data analysis

[0171] The statistical analysis software was SPSS, and the comparison between the elastase inhibition rate of the test sample and the negative control was performed using the independent sample t test. The above statistical analyses were all two-tailed tests, and the significance level was α = 0.05. P>0.05 indicated that there was no significant difference between the two groups; P<0.05 indicated that there was a significant difference between the two groups.

[0172] The test results are shown in Table 4:

[0173] Table 4

[0174]

[0175]

[0176] Result analysis:

[0177] 1. It can be seen from Examples 1-3 that after adjusting the mass ratio of the plant components and the mass ratio of the enzyme and yeast to the plant components, there is a certain fluctuation in the elastase inhibition rate between Examples 1-3, and the elastase inhibition rate of Example 3 is higher than that of Examples 1 and 2. It can be seen that the composition prepared by the parameters of Example 3 has a better elastase inhibition ability;

[0178] 2. It can be seen from Examples 1 and 4-6 that when Example 4 uses the same type of commercially available galactose yeast to replace Geotrichum candidum, the elastase inhibition ability of Example 4 shows a significant downward trend. It can be seen that even if the same type of galactose yeast is used to replace it, it is still impossible to achieve an effect similar to that of Geotrichum candidum.

[0179] 3. Further observation of Examples 5-6 shows that when Example 5 uses a mixture of galactose yeast, bifid yeast, and Pichia pastoris to replace galactose yeast, its elastase inhibition rate is slightly improved, but when galactose yeast and Pichia pastoris are used together in Example 6, the elastase inhibition ability of the composition is significantly improved. It can be seen that galactose yeast and Pichia pastoris can cooperate with each other to ferment more fermentation products with elastase inhibition ability, and the reason why the elastase inhibition ability of Example 5 is not high may be that bifid yeast has occupied the fermentation raw materials or intermediates that should have been fermented by galactose yeast and Pichia pastoris to a certain extent;

[0180] Further observation of Examples 7-8 shows that when Galacto-saccharomyces and Pichia pastoris are compounded in Example 7 and Example 6, but commercially available Galacto-saccharomyces is used, the elastase inhibition ability of Example 7 is close to that of Example 1 and significantly lower than that of Example 6. It can be seen that the synergistic ability between commercially available Galacto-saccharomyces and Pichia pastoris is weak.

[0181] The effect of the combined use of galactosaccharomyces and bifid yeast in Example 8 is close to that in Example 1, which shows that bifid yeast does occupy part of the fermentation resources originally belonging to galactosaccharomyces, and the fermentation capacity of the two for similar raw materials (referring to the fermentation capacity of products with elastase inhibition ability) is relatively close, but there is a synergistic effect between galactosaccharomyces and Pichia pastoris, while the synergistic effect between bifid yeast and Pichia pastoris is not high or even does not exist.

[0182] 4. It can be seen from Examples 1 and 9-10 that when the enzymes are compounded, the elastase inhibition rate of Example 9 is improved to a certain extent, and when Example 10, that is, the enzymes are further compounded on the basis of Example 6, the elastase inhibition rate of Example 10 is significantly improved, and this improvement trend is greater than the sum of the improvement amounts of Example 6 and Example 9 relative to Example 1. It can be seen that the compounding of enzymes can improve the fermentation capacity of the system with Galactomyces and Pichia pastoris as fermentation bacteria, thereby producing more fermentation products with elastase inhibition ability.

[0183] 5. It can be seen from Example 1 and Comparative Examples 1-6 that when any one of Purslane, Emblica emblica, and Tremella fuciformis is missing, the elastase inhibition ability of Comparative Examples 1-3 has a significant downward trend. It can be seen that any one of Purslane, Emblica emblica, and Tremella fuciformis is indispensable;

[0184] When any of purslane, emblica, and tremella is replaced by other plant ingredients, the elastase inhibition ability of Examples 4-6 shows a significant downward trend. It can be seen that any of purslane, emblica, and tremella is irreplaceable.

[0185] 4. Hyaluronidase inhibition rate test

[0186] 4.1 Instruments and Equipment

[0187] BSA224S analytical balance, L6s UV spectrophotometer

[0188] 4.2 Reagents

[0189] Hyaluronidase, BR, Sodium Hyaluronate, BR

[0190] 4.3 Test methods

[0191] (1) Treatment of control and test samples

[0192] Sample group: the compositions prepared in Examples 1 to 10 and Comparative Examples 1 to 7 at a concentration of 100%;

[0193] Negative control: pure water.

[0194] (2) Test operation steps

[0195] Set up a sample group and a negative control group. Each group needs to have 3 parallels. Add the corresponding reagent solution to each group, shake well, and leave it at room temperature for 30 minutes for color development. Use an ultraviolet spectrophotometer to measure the absorbance value at a wavelength of 528nm.

[0196] (3) The calculation formula is shown in Formula 1:

[0197]

[0198] Where: A—is the absorbance of the reaction solution without sample;

[0199] B—is the absorbance of the reaction solution without sample and enzyme;

[0200] C—is the absorbance of the reaction solution containing sample and enzyme;

[0201] D—is the absorbance of the reaction solution containing sample and without enzyme.

[0202] (4) Data analysis

[0203] The statistical analysis software was SPSS, and the comparison between the hyaluronidase inhibition rate of the test sample and the negative control was performed using an independent sample t-test. The above statistical analyses were all two-tailed tests, with a significance level of a = 0.05. P ≥ 0.05 indicated that there was no significant difference between the two groups; P < 0.05 indicated that there was a significant difference between the two groups.

[0204] The test results are shown in Table 5:

[0205] Table 5

[0206]

[0207]

[0208] Results Analysis

[0209] 1. It can be seen from Examples 1-3 that after adjusting the mass ratios of the plant components and the mass ratios of the enzymes and yeast to the plant components, the hyaluronidase inhibition rates of Examples 1-3 show certain fluctuations, but the fluctuation range is relatively small;

[0210] 2. It can be seen from Examples 1 and 4-6 that when Example 4 uses the same type of commercially available galactose yeast to replace Geotrichum candidum, the hyaluronidase inhibition ability of Example 4 shows a significant downward trend. It can be seen that even if the same type of galactose yeast is used to replace Geotrichum candidum, the effect similar to that of Geotrichum candidum cannot be achieved;

[0211] 3. Further observation of Examples 5-6 shows that when Example 5 uses a mixture of galactose yeast, bifid yeast, and Pichia pastoris to replace galactose yeast, its hyaluronidase inhibition rate decreases slightly, but when galactose yeast and Pichia pastoris are used together in Example 6, the hyaluronidase inhibition ability of the composition is significantly improved. It can be seen that galactose yeast and Pichia pastoris can cooperate with each other to ferment more fermentation products with hyaluronidase inhibition ability, and the reason why the hyaluronidase inhibition ability of Example 5 is not high may be that bifid yeast has occupied the fermentation raw materials or intermediates that should have been fermented by galactose yeast and Pichia pastoris to a certain extent;

[0212] Further observation of Examples 7-8 shows that when Galacto-saccharomyces and Pichia pastoris are compounded in Example 7 and Example 6, but commercially available Galacto-saccharomyces is used, the hyaluronidase inhibition ability of Example 7 is significantly lower than that of Example 6, which shows that the synergistic ability between commercially available Galacto-saccharomyces and Pichia pastoris is weak;

[0213] The effect of the combined use of galactose yeast and bifid yeast in Example 8 is close to that in Example 1, which shows that bifid yeast does occupy part of the fermentation resources originally belonging to galactose yeast, and the fermentation capacity of the two for similar raw materials (referring to the fermentation capacity of products with hyaluronidase inhibition ability) is relatively close, but there is a synergistic effect between galactose yeast and Pichia pastoris, while the synergistic effect between bifid yeast and Pichia pastoris is not high or even does not exist.

[0214] 4. It can be seen from Examples 1 and 9-10 that when the enzyme is compounded for use, the hyaluronidase inhibition rate of Example 9 is improved to a certain extent, and when Example 10, that is, the enzyme is further compounded on the basis of Example 6, the hyaluronidase inhibition rate of Example 10 is significantly improved, and this improvement trend is greater than the sum of the improvement amounts of Example 6 and Example 9 relative to Example 1. It can be seen that the compounding of enzymes can improve the fermentation capacity of the system with Galacto-saccharomyces and Pichia pastoris as fermentation bacteria, thereby producing more fermentation products with hyaluronidase inhibition ability.

[0215] 5. It can be seen from Example 1 and Comparative Examples 1-6 that when any one of Purslane, Emblica emblica, and Tremella fuciformis is missing, the hyaluronidase inhibition ability of Comparative Examples 1-3 has a significant downward trend. It can be seen that any one of Purslane, Emblica emblica, and Tremella fuciformis is indispensable;

[0216] When any of purslane, emblica fruit, and tremella is replaced by other plant ingredients, the hyaluronidase inhibition ability of Examples 4-6 shows a significant downward trend. It can be seen that any of purslane, emblica fruit, and tremella is irreplaceable.

[0217] Application Examples

[0218] A firming and anti-wrinkle composition essence, the formula ratio of which is shown in Table 6.

[0219] Table 6

[0220]

[0221]

[0222] The embodiments presented herein are only embodiments selected according to the combination of all possible embodiments. The appended claims should not be limited by the embodiments describing the present invention. Some numerical ranges used in the claims include sub-ranges therein, and the changes in these ranges should also be covered by the appended claims.

Claims

1. A firming and anti-wrinkle composition, characterized in that: The purslane, the emblica fruit and the tremella are dried, crushed, enzymolyzed, fermented and then filtered to prepare the product, wherein the mass ratio of the purslane, the emblica fruit and the tremella is 5-30:1-10:1-10; The enzyme used in the enzymatic hydrolysis process is selected from at least one of alkaline protease, cellulase, trypsin, and neutral protease; The mass ratio of the enzyme to purslane, emblica fructus, and tremella is 0.1-5:5-30:1-10:1-10; The yeast used in the fermentation process is selected from at least one of galactosaccharomyces, bifid yeast, and Pichia pastoris; The mass ratio of yeast to purslane, emblica fructus, and tremella is 0.1-3:5-30:1-10:1-10; The Latin name of the galactose yeast is Geotrichum candidum The strain deposit number is GDMCC No.64235.

2. The firming and anti-wrinkle composition according to claim 1, characterized in that: The yeast used in the fermentation process at least comprises galactosyl yeast.

3. The firming and anti-wrinkle composition according to claim 1, characterized in that: The yeast used in the fermentation process is a mixture of galactose yeast and Pichia pastoris, and the mass ratio of galactose yeast to Pichia pastoris is 2-4:

1.

4. The firming and anti-wrinkle composition according to claim 1, characterized in that: The enzyme used in the enzymatic hydrolysis process is selected from a mixture of alkaline protease, cellulose protease and trypsin, and the mass ratio of the alkaline protease, cellulose protease and trypsin is 1-3:1-3:1-3.

5. A method for preparing the firming and anti-wrinkle composition according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step 1: taking, drying and crushing purslane, emblica fruit and tremella in a mass ratio of 5-30:1-10:1-10 to obtain powder; Step 2: performing enzymatic hydrolysis on the powder obtained in step 1 to obtain an enzymatic hydrolyzate; Step 3: The enzymatic hydrolysate obtained in step 2 is made into a culture medium and inoculated with yeast for fermentation and filtration to obtain a firming and anti-wrinkle composition.

6. Use of the firming and anti-wrinkle composition according to any one of claims 1 to 4 in preparing cosmetics.

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