Sea rice fermentation filtrate, preparation method thereof and whitening cosmetic

Abstract

The invention relates to the field of cosmetics, in particular to sea rice fermentation filtrate, a preparation method thereof and a whitening cosmetic. Inoculating the first fermentation seed solution and the second fermentation seed solution into a fermentation culture medium, fermenting at 25-30 DEG C to obtain a fermentation product, and removing thalli in the fermentation product; the fermentation culture medium is prepared from the following components in percentage by mass: 5 to 10 percent of sea rice powder, 1 to 5 percent of saccharides and the balance of water; the pH value of the fermentation culture medium is 4-5; the first fermentation seed solution is a saccharomyces cerevisiae strain fermentation seed solution; the second fermentation seed solution is a lactobacillus plantarum strain fermentation seed solution. The method can promote the growth of saccharomyces cerevisiae and lactobacillus plantarum while promoting the release of an active component phytic acid in the sea rice, and has a remarkable effect of removing excessive free radicals and inhibiting the activity of related enzymes for synthesizing melanin; therefore, the whitening effect of the cosmetic can be effectively improved.

Description

Technical Field

[0001] The present application relates to the field of cosmetics, and more specifically, to a sea rice fermentation filtrate, a preparation method thereof, and whitening cosmetics. Background Art

[0002] Currently, the whitening ingredients in whitening cosmetics primarily include vitamin C, glutathione, arbutin, niacinamide, tranexamic acid, hyaluronic acid, and other chemical ingredients. However, as the concept of green skincare becomes increasingly popular, the use of these chemical whitening ingredients has been limited. The market demand for green and natural whitening ingredients is growing. Summary of the Invention

[0003] The purpose of the embodiments of the present application is to provide a sea rice fermentation filtrate and a preparation method thereof, and a whitening cosmetic.

[0004] In a first aspect, the present application provides a method for preparing sea rice fermentation filtrate, comprising: Inoculating the first fermentation seed liquid and the second fermentation seed liquid into a fermentation medium, fermenting at 25° C.-30° C. to obtain a fermentation product, and removing bacteria from the fermentation product; The fermentation medium consists of the following components in percentage by mass: 5%-10% sea rice powder, 1%-5% sugar, and the balance water; the pH of the fermentation medium is 4-5; The first fermentation seed liquid is fermentation seed liquid of Saccharomyces cerevisiae; the second fermentation seed liquid is fermentation seed liquid of Lactobacillus plantarum.

[0005] The above technical solution adopts a mixed strain of saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice. While promoting the release of phytic acid, an active ingredient in sea rice, it can promote the growth of saccharomyces cerevisiae and Lactobacillus plantarum. The saccharomyces cerevisiae fermentation product and the Lactobacillus plantarum fermentation product have excellent antioxidant effects, which are conducive to improving the whitening effect. Phytic acid has a significant effect on scavenging excess free radicals and inhibiting the activity of enzymes related to melanin synthesis. Therefore, this fermentation product is used in cosmetics to effectively improve the antioxidant and whitening effects of cosmetics. Moreover, the fermentation product prepared by this method is green and natural. Not only are the raw materials pure and natural, but the process is green and environmentally friendly, and no other chemical solvents are introduced. Therefore, it is of great significance for developing green and natural whitening efficacy ingredients. This fermentation product is applied to whitening cosmetics to effectively improve the whitening effect of cosmetics.

[0006] In other embodiments of the present application, the fermentation medium is composed of the following components in percentage by mass: 6%-9% sea rice powder, 2%-4% sugars, and the balance water; the pH of the fermentation medium is adjusted to 4.5.

[0007] In other embodiments of the present application, inoculating the first fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, 1%-5% of the first fermentation seed liquid is added to the fermentation medium in volume percentage.

[0008] In other embodiments of the present application, inoculating the second fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, 2%-6% of the cerevisiae strain is inoculated into the fermentation medium in terms of volume percentage.

[0009] In other embodiments of the present application, inoculating the first fermentation seed liquid and the second fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, in terms of volume percentage, a total amount of the first fermentation seed liquid and the second fermentation seed liquid greater than 0 and less than or equal to 10% is inoculated into the fermentation medium.

[0010] In other embodiments of the present application, inoculating the first fermentation seed liquid and the second fermentation seed liquid into the fermentation medium comprises: The volume ratio of the added amount of the first fermentation seed liquid and the second fermentation seed liquid is: (1-5): (2-6).

[0011] In other embodiments of the present application, the sugar includes at least one of glucose or sucrose.

[0012] In a second aspect, the present application provides a sea rice fermentation filtrate, which is prepared using the preparation method of the sea rice fermentation filtrate provided in the first aspect.

[0013] In other embodiments of the present application, the phytic acid content in the sea rice fermentation filtrate is greater than 413.33 mg / mL.

[0014] In a third aspect, the present application provides a whitening cosmetic comprising the sea rice fermentation filtrate provided in the second aspect. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the following is a brief introduction to the drawings required for use in the embodiments. It should be understood that the following drawings only show certain embodiments of the present application and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other relevant drawings can be obtained based on these drawings without creative work.

[0016] Figure 1 The standard curve of phytic acid. DETAILED DESCRIPTION

[0017] In order to make the purpose, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be clearly and completely described below. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments.

[0018] Therefore, the following detailed description of the embodiments of the present application is not intended to limit the scope of the present application for protection, but merely represents selected embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0019] The present application provides a method for preparing sea rice fermentation filtrate, which comprises: inoculating the first bacterial strain and the second bacterial strain into a fermentation medium, fermenting at 25° C. to 30° C. to obtain a fermentation product, and removing bacterial cells from the fermentation product; The fermentation medium consists of the following components in percentage by mass: 5%-10% sea rice powder, 1%-5% sugar, and the balance water; the pH of the fermentation medium is 4-5; The first bacterial species is a Saccharomyces cerevisiae bacterial species; the second bacterial species is a Lactobacillus plantarum bacterial species.

[0020] In this technical solution, sea rice is fermented with yeast and Lactobacillus plantarum as the starting strains. This not only promotes the growth of these two strains but also fully extracts the phytic acid present in the sea rice. The fermentation product prepared using this method contains active ingredients from sea rice and metabolites of yeast and Lactobacillus plantarum. By scavenging excess free radicals and inhibiting the activity of enzymes involved in melanin synthesis, it achieves both antioxidant and whitening effects.

[0021] Furthermore, in some embodiments of the present application, the method for preparing sea rice fermentation filtrate comprises the following steps: Step S1, preparation of sea rice powder.

[0022] Furthermore, in some embodiments of the present application, the steps of preparing sea rice powder include: The dried sea rice is crushed and passed through a sieve with mesh size of 40 or above, and the residue under the sieve is taken.

[0023] Illustratively, in some embodiments of the present application, the dried sea rice is crushed and then passed through a 40-mesh, 45-mesh, 50-mesh, 55-mesh, 60-mesh, 65-mesh, 70-mesh, 75-mesh or 80-mesh sieve.

[0024] Further optionally, in some embodiments of the present application, the step of preparing sea rice powder includes: The sea rice is dried, crushed, and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0025] Step S2: preparing a first fermentation seed liquid.

[0026] In some embodiments of the present application, the first fermentation seed liquid contains a first bacterial species.

[0027] In some embodiments of the present application, the first bacterial species is a Saccharomyces cerevisiae species.

[0028] In some embodiments of the present application, the above-mentioned Saccharomyces cerevisiae strain was purchased from Guangdong Provincial Microbiological Culture Collection Center with the strain number GDMCC 2.90.

[0029] In some embodiments of the present application, preparing a first fermentation seed liquid comprises: The cerevisiae yeast strain is transferred to a solid culture medium and allowed to stand at 25°C to 30°C for activation until a single colony grows; the single colony on the solid culture medium is then transferred to a liquid culture medium and cultured at 25°C to 30°C for 20h to 48h to obtain a first fermentation seed liquid.

[0030] Illustratively, in some embodiments of the present application, the brewer's yeast strain is transferred to a solid culture medium and allowed to stand and activate at a temperature of 25°C, 26°C, 27°C, 28°C, 29°C, 30°C or a range between any two of the foregoing values until a single colony grows; then the single colony strain on the solid culture medium is transferred to a liquid culture medium, and the culture is continued to be expanded at a temperature of 25°C, 26°C, 27°C, 28°C, 29°C, 30°C or a range between any two of the foregoing values for 20h, 22h, 24h, 26h, 28h, 30h, 32h, 34h, 38h, 40h, 42h, 45h, 46h, 48h or a range between any two of the foregoing values to obtain a first fermentation seed liquid.

[0031] In some embodiments of the present application, the solid culture medium comprises: YPD medium and agar, wherein the added mass of agar is 2 wt % of the mass of water.

[0032] In some embodiments of the present application, the liquid culture medium is YPD culture medium.

[0033] Optionally, in some embodiments of the present application, the above-mentioned YPD culture medium is purchased from hopebio brand 250g YPD liquid culture medium (product number: HB5193-1).

[0034] Step S3: preparing a second fermentation seed liquid.

[0035] In some embodiments of the present application, the second fermentation seed liquid contains a second bacterial species.

[0036] In some embodiments of the present application, the second bacterial species is Lactobacillus plantarum.

[0037] In some embodiments of the present application, the aforementioned Lactobacillus plantarum was purchased from Guangdong Provincial Microbiological Culture Collection Center with the strain number GDMCC 1.1797.

[0038] In some embodiments of the present application, preparing the second fermentation seed liquid includes: transferring Lactobacillus plantarum to a solid culture medium and letting it stand at 30°C to 40°C for activation until a single colony grows; then transferring the single colony on the solid culture medium to a liquid culture medium, and continuing to expand the culture at 30°C to 40°C for 20h to 48h to obtain the second fermentation seed liquid.

[0039] Illustratively, in some embodiments of the present application, the plant lactobacillus is transferred to a solid culture medium and allowed to stand and activate at a temperature of 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C or a range between any two of the foregoing values until a single colony grows; then the single colony on the solid culture medium is transferred to a liquid culture medium, and the culture is expanded for 20h, 22h, 24h, 26h, 28h, 30h, 32h, 34h, 38h, 40h, 42h, 45h, 46h, 48h or a range between any two of the foregoing values at a temperature of 30°C, 31°C, 32°C, 33°C, 34°C, 35°C, 36°C, 37°C, 38°C, 39°C, 40°C or a range between any two of the foregoing values to obtain a second fermentation seed liquid.

[0040] In some embodiments of the present application, the solid culture medium comprises: MRS culture medium, agar and water, wherein the added mass of agar is 2 wt % of the mass of water.

[0041] In some embodiments of the present application, the liquid culture medium is MRS culture medium.

[0042] Optionally, in some embodiments of the present application, the above-mentioned MRS culture medium is purchased from Huankai Microbiology brand 250g MRS broth.

[0043] Step S4: preparing fermentation medium.

[0044] Furthermore, in some embodiments of the present application, the fermentation medium is composed of the following components in percentage by mass: 5%-10% sea rice powder, 1%-5% sugar and the balance water; the pH of the fermentation medium is 4~5.

[0045] In some embodiments of the present application, the sea rice powder is prepared using the sea rice powder prepared in step S1.

[0046] For example, in some embodiments of the present application, the fermentation medium is composed of the following components in percentage by mass: 5%, 6%, 7%, 8%, 9%, 10% or a range between any two of the foregoing sea rice powder, 1%, 2%, 3%, 4%, 5% or a range between any two of the foregoing sugars, and the remainder water; the pH of the fermentation medium is 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.

[0047] Further optionally, in some embodiments of the present application, the fermentation medium is composed of the following components in percentage by mass: 6%-9% sea rice powder, 2%-4% sugars, and the balance water; the pH of the fermentation medium is adjusted to 4.5.

[0048] Furthermore, in some embodiments of the present application, the sugar includes at least one of glucose or sucrose.

[0049] For example, in some embodiments of the present application, the sugar is selected from either glucose or sucrose. Alternatively, in some embodiments of the present application, the sugar is selected from a mixture of glucose and sucrose; the raw materials in the mixture can be mixed in any proportion.

[0050] Step S5: inoculating the first fermentation seed liquid containing the first bacterial species and the second fermentation seed liquid containing the second bacterial species into a fermentation medium, fermenting at 25° C.-30° C. to obtain a fermentation product, and removing the bacterial cells in the fermentation product.

[0051] Illustratively, in some embodiments of the present application, fermentation is carried out at 25°C, 26°C, 27°C, 28°C, 29°C or 30°C.

[0052] Furthermore, in some embodiments of the present application, the first fermentation seed liquid prepared in the aforementioned step S2; and the second fermentation seed liquid prepared in the aforementioned step S3 are inoculated into the fermentation medium prepared in the aforementioned step S4.

[0053] Furthermore, in some embodiments of the present application, inoculating the first fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, 1%-5% of the first fermentation seed liquid is added to the fermentation medium in volume percentage.

[0054] For example, in some embodiments of the present application, inoculating the first fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, the first fermentation seed liquid is added to the fermentation medium in an amount of 1%, 1.1%, 1.2%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5% or a range between any two of the foregoing values in volume percentage.

[0055] Furthermore, in some embodiments of the present application, inoculating the second fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, 2%-6% of the cerevisiae strain is inoculated into the fermentation medium in terms of volume percentage.

[0056] Illustratively, in some embodiments of the present application, based on the total volume of the fermentation medium, the Saccharomyces cerevisiae strain is inoculated into the fermentation medium at a content of 2%, 2.5%, 3%, 4%, 4.5%, 5.5%, 6% or a range between any two of the foregoing values, as a percentage by volume.

[0057] Furthermore, in some embodiments of the present application, inoculating the first fermentation seed liquid and the second fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, in terms of volume percentage, a total amount of the first fermentation seed liquid and the second fermentation seed liquid greater than 0 and less than or equal to 10% is inoculated into the fermentation medium.

[0058] For example, in some embodiments of the present application, inoculating the first fermentation seed liquid and the second fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, the total amount of the first fermentation seed liquid and the second fermentation seed liquid in an amount of 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10% or a range between any two of the foregoing values is inoculated into the fermentation medium in terms of volume percentage.

[0059] Furthermore, in some embodiments of the present application, inoculating the first fermentation seed liquid into the fermentation medium comprises: The volume ratio of the first fermentation seed liquid and the added amount of the first fermentation seed liquid is: (1-5): (2-6).

[0060] For example, in some embodiments of the present application, inoculating the first fermentation seed liquid into the fermentation medium comprises: The volume ratio of the first fermentation seed liquid and the added amount of the first fermentation seed liquid is: (1, 2, 3, 4 or 5): (2, 3, 4, 5 or 6).

[0061] Furthermore, in some embodiments of the present application, removing the bacterial cells in the fermentation product comprises: The fermentation product obtained above is centrifuged to remove the bacterial cells and obtain the fermentation filtrate.

[0062] Some embodiments of the present application provide a sea rice fermentation filtrate, which is prepared using the preparation method of the sea rice fermentation filtrate provided by any of the aforementioned embodiments.

[0063] Furthermore, in some embodiments of the present application, the phytic acid content in the sea rice fermentation filtrate is greater than 413.33 mg / mL.

[0064] Illustratively, in some embodiments of the present application, the phytic acid content in the sea rice fermentation filtrate is 413.33 mg / mL, 415 mg / mL, 420 mg / mL, 450 mg / mL, 480 mg / mL, 500 mg / mL, 550 mg / mL, 600 mg / mL, 650 mg / mL or a range between any two of the foregoing values.

[0065] Some embodiments of the present application provide a whitening cosmetic comprising the sea rice fermentation filtrate provided in any of the aforementioned embodiments.

[0066] The features and performance of the present invention are further described in detail below with reference to the embodiments: Example 1 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0067] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0068] Example 2 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0069] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 6% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 12 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0070] Example 3 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0071] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 4% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 36 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0072] Example 4 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0073] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 5% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 12 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0074] Example 5 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0075] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 3% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 36 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0076] Example 6 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0077] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 4% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0078] Example 7 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0079] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 4% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0080] Example 8 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0081] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0082] Example 9 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0083] (2) Preparation of the first fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid culture medium (YPD medium and agar, wherein the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain the first fermentation seed liquid; (3) Preparation of the second fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to a solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 37°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to a liquid culture medium (MRS culture medium) and continued to expand and culture at 37°C for 24 h to obtain the second fermentation seed liquid; (4) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 5; (5) Fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum: 4% of the first fermentation seed liquid prepared in step (2) and 5% of the second fermentation seed liquid prepared in step (3) (v / v, seed liquid / total volume of the culture medium after seed liquid inoculation) were inoculated into the fermentation medium prepared in step (4), and the mixture was allowed to stand at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0084] Comparative Example 1 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0085] (2) Preparation of fermentation seed liquid: Saccharomyces cerevisiae (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to solid culture medium (YPD medium and agar, where the added mass of agar was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to liquid culture medium (YPD medium) and continued to expand and culture at 28°C for 24 h to obtain fermentation seed liquid; (3) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (4) Fermentation of sea rice with brewer's yeast: 4% by volume of the fermentation seed liquid prepared in step (2) was added to the fermentation medium prepared in step (3), and the mixture was allowed to ferment at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0086] Comparative Example 2 Provided is a method for preparing sea rice fermentation filtrate, which is prepared by the following steps: (1) Preparation of sea rice powder: After the sea rice is dried, it is crushed and passed through a 40-mesh sieve to obtain sea rice powder, which is then packaged and sterilized for later use.

[0087] (2) Preparation of fermentation seed liquid: Lactobacillus plantarum (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1797) was transferred to solid culture medium (MRS culture medium, agar and water, where the mass of agar added was 2 wt% of the mass of water) and left to stand at 28°C for activation until a single colony grew; then the single colony on the solid culture medium was transferred to liquid culture medium (MRS culture medium) and continued to expand and culture at 28°C for 24 h to obtain fermentation seed liquid; (3) Prepare fermentation medium: prepare the medium according to the following mass percentages: 6% sea rice powder prepared in step (1), 3% glucose, and the balance water; adjust the pH to 4.5; (4) Fermentation of sea rice with Lactobacillus plantarum: 4% by volume of the fermentation seed liquid prepared in step (2) was added to the fermentation medium prepared in step (3), and the mixture was allowed to ferment at 28°C for 24 h. The fermentation product was then centrifuged to obtain a fermentation clear liquid.

[0088] The properties of the fermentation supernatants obtained in the above-mentioned embodiments and comparative examples were tested.

[0089] Experimental Example 1 Determination of phytic acid content by spectrophotometry: Phytic acid can fade the purple-red color developer of ferric chloride-sulfosalicylic acid, and the phytic acid content is proportional to the degree of fading. The phytic acid content in the sample can be obtained by detecting the decrease in absorbance at 500 nm using a microplate reader.

[0090] The specific experimental steps are as follows: Use a phytic acid content determination kit, prepare the color developer according to the instructions, and perform a gradient dilution of the samples of each embodiment and comparative example, such as 2-fold, 5-fold, and 10-fold dilution. Add the assay tube A (50 μL of sample, 100 μL of color developer, and 50 μL of distilled water) and blank tube A (50 μL of distilled water, 100 μL of color developer, and 50 μL of distilled water) to a 96-well plate in sequence. Each well has 200 μL of the system. The blank tube is only tested once. After mixing, let it stand for 2 minutes and then read the absorbance value A at 500 nm. △A=A blank-A determination. Substitute △A as y into the standard curve (the phytic acid standard curve is attached to the instructions). Figure 1 The phytic acid content can be calculated using the equation shown in Figure 2.

[0091] The test results of the above embodiments and comparative examples are shown in Table 1.

[0092] Table 1

[0093] From the experimental data in Table 1 above, we can see that: The phytic acid content in the sea rice fermentation filtrate prepared in each example was higher than that in each comparative example. This shows that the method of the present application can effectively increase the phytic acid content in the sea rice fermentation filtrate.

[0094] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Saccharomyces cerevisiae, to ferment sea rice, the phytic acid content in the obtained fermentation product is low; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively increase the content of phytic acid in the fermentation product; it can be seen that it is difficult to increase the content of phytic acid in the fermentation product by using a single strain, Saccharomyces cerevisiae, to ferment sea rice. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly increase the release of active ingredients in sea rice, especially phytic acid. Phytic acid has a significant effect on scavenging excess free radicals and inhibiting the activity of enzymes related to the synthesis of melanin; therefore, using this fermentation product in cosmetics can effectively improve the antioxidant and whitening effects of cosmetics.

[0095] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: under the premise of the same fermentation medium and inoculum size, when Comparative Example 2 uses a single strain, Lactobacillus plantarum, to ferment sea rice, the phytic acid content in the fermentation product obtained is low; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively increase the phytic acid content in the fermentation product; thus, it can be seen that it is difficult to increase the phytic acid content in the fermentation product by using a single strain, Lactobacillus plantarum, to ferment sea rice. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly increase the release of active ingredients in sea rice, especially phytic acid. Phytic acid has a significant effect on scavenging excess free radicals and inhibiting the activity of enzymes related to the synthesis of melanin; therefore, using this fermentation product in cosmetics can effectively enhance the antioxidant and whitening effects of cosmetics.

[0096] Experimental Example 2 Antioxidant Energy DPPH·, also known as 1,1-diphenyl-2-trinitrophenylhydrazine, is a very stable nitrogen-centered free radical. Its stability primarily stems from the steric hindrance of the three benzene rings through resonance stabilization, which prevents the unpaired electrons on the central nitrogen atom from performing their proper electron pairing function. Its anhydrous ethanol solution is purple, with maximum absorption at 517nm. The absorbance is linearly related to concentration. Adding a free radical scavenger, either in combination with or in place of DPPH·, reduces the number of free radicals, lowers the absorbance, and lightens the solution color, thereby evaluating its free radical scavenging ability.

[0097] The specific experimental steps are as follows: 2.0 mL of the sample solution (or anhydrous ethanol) of each example or comparative example and 2.0 mL of the DPPH· solution were respectively pipetted into stoppered test tubes and mixed. The mixture was allowed to react for 30 min in the dark, and the absorbance at a wavelength of 517 nm was measured.

[0098]

[0099] Where: A0: no sample added, DPPH added; A1: Add sample or anhydrous ethanol and DPPH; A2: Add sample or anhydrous ethanol, without adding DPPH.

[0100] The test results are shown in Table 2.

[0101] Table 2 DPPH free radical scavenging rate of different embodiments and comparative examples

[0102] From the experimental data in Table 2 above, we can see that: The DPPH free radical scavenging rates of the sea rice fermentation filtrates prepared in each example were higher than those in each comparative example. This indicates that the method of the present application can effectively improve the DPPH free radical scavenging rate of the sea rice fermentation filtrate.

[0103] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Saccharomyces cerevisiae, to ferment sea rice, the resulting fermentation product has a low DPPH radical scavenging rate; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the fermentation product's DPPH radical scavenging rate; it can be seen that using a single strain, Saccharomyces cerevisiae, to ferment sea rice, it is difficult to improve the fermentation product's DPPH radical scavenging rate. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the DPPH radical scavenging rate. This is because the mixed fermentation of sea rice with Saccharomyces cerevisiae and Lactobacillus plantarum not only releases and converts the active ingredients in sea rice efficiently through the complex synergistic effect of these two strains, but also promotes strain growth, obtains more types and quantities of microbial metabolites with antioxidant activity, and thus effectively improves the fermentation product's DPPH radical scavenging rate. And the improvement in DPPH radical scavenging rate is conducive to improving the whitening effect.

[0104] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Lactobacillus plantarum, to ferment sea rice, the obtained fermentation product has a low DPPH radical scavenging rate; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the DPPH radical scavenging rate of the fermentation product; it can be seen that it is difficult to improve the DPPH radical scavenging rate of the fermentation product when a single strain, Lactobacillus plantarum, is used to ferment sea rice. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the DPPH radical scavenging rate. This is because the use of a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice can promote the growth of Saccharomyces cerevisiae and Lactobacillus plantarum while promoting the release of active ingredients in sea rice, and the Saccharomyces cerevisiae fermentation product and Lactobacillus plantarum fermentation product have excellent antioxidant effects, thereby effectively improving the DPPH radical scavenging rate of the fermentation product; and the improvement of the DPPH radical scavenging rate is conducive to improving the whitening effect.

[0105] Experimental Example 3 Antioxidant Capacity II The present invention was experimentally investigated for its ability to scavenge ABTS+·. ABTS is oxidized to green ABTS+· by appropriate oxidants. When antioxidants are added to the reaction, the production of ABTS+· is inhibited. The total antioxidant capacity of the sample can be determined and calculated by measuring the absorbance at 734 nm.

[0106] The specific experimental steps are as follows: (1) Take 7 mmol / L ABTS solution and dilute it 60 times with 10 mmol / L PBS (pH = 7.4) to obtain ABTS working solution; (2) Pipette 190 μL of ABTS working solution (or PBS), add 10 μL of the sample solution (or PBS) of each example or comparative example, shake for 10 seconds, let it stand at 30°C for 6 minutes, and measure the absorbance A at 734 nm.

[0107]

[0108] Where: A0: no sample added, ABTS added; A1: add sample or PBS, add ABTS; A2: Add sample or PBS, without adding ABTS.

[0109] The test results are shown in Table 3.

[0110] Table 3 ABTS+ free radical scavenging ability of different examples

[0111] From the experimental data in Table 3 above, we can see that: The ABTS+· clearance rate of the sea rice fermentation filtrate prepared in each example was higher than that of each comparative example. This shows that the method of the present application can effectively improve the ABTS+· clearance rate of the sea rice fermentation filtrate.

[0112] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum size, when Comparative Example 1 uses a single strain, Saccharomyces cerevisiae, to ferment sea rice, the fermentation product obtained has a low ABTS+ clearance rate; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the fermentation product for ABTS+ clearance rate; it can be seen that using a single strain of Saccharomyces cerevisiae to ferment sea rice, it is difficult to improve the fermentation product for ABTS+ clearance rate. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the ABTS+ clearance rate. This is because the mixed fermentation of sea rice by Saccharomyces cerevisiae and Lactobacillus plantarum not only releases and converts the active ingredients in sea rice efficiently through the complex synergistic effect of these two strains, but also promotes strain growth, obtains more types and quantities of microbial metabolites with antioxidant activity, and thus effectively improves the fermentation product for ABTS+ clearance rate. And the improvement of ABTS+ clearance rate is conducive to improving the whitening effect.

[0113] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: Under the premise of the same fermentation medium and inoculum size, when Comparative Example 1 uses a single strain, Lactobacillus plantarum, to ferment sea rice, the fermentation product obtained has a low ABTS+ clearance rate; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the fermentation product for ABTS+ clearance rate; it can be seen that using a single strain, Lactobacillus plantarum, to ferment sea rice, it is difficult to improve the fermentation product for ABTS+ clearance rate. However, the present application adopts a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the ABTS+ clearance rate. This is because the use of a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice can promote the growth of Saccharomyces cerevisiae and Lactobacillus plantarum while promoting the release of active ingredients in sea rice, and the Saccharomyces cerevisiae fermentation product and the Lactobacillus plantarum fermentation product have excellent antioxidant effects, thereby effectively improving the fermentation product for ABTS+ clearance rate; and improving the ABTS+ clearance rate is conducive to improving the whitening effect.

[0114] Experimental Example 4 Determination of Melanin Synthesis Inhibition Rate Centrifuge the B16 cell suspension in a centrifuge tube at 4000 rpm for 5 minutes, aspirate the supernatant, add 200 μL of melanin extract (1 mol / mL NaOH containing 10% DMSO) to each tube, shake well, and place in an 80°C water bath for 1 hour. After cooling, centrifuge gently to remove droplets from the tube wall, pipette evenly, and pipette 150 μL of the solution from each tube into a 96-well plate. Measure the absorbance of each well at 405 nm with a microplate reader. Calculate the inhibition rate according to the following formula.

[0115]

[0116] The test results are shown in Table 4.

[0117] Table 4 Melanin synthesis inhibition ability of different examples

[0118] From the experimental data in Table 4 above, we can see that: The sea rice fermentation filtrate obtained in each example has a higher melanin synthesis inhibition rate than that of each comparative example. This shows that the method of the present application can effectively improve the melanin synthesis inhibition rate of the sea rice fermentation filtrate.

[0119] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Saccharomyces cerevisiae, to ferment sea rice, the resulting fermentation product has a low inhibition rate on melanin synthesis; while the present application scheme uses a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the inhibition rate of melanin synthesis of the fermentation product; it can be seen that using a single strain of Saccharomyces cerevisiae to ferment sea rice is difficult to improve the inhibition rate of melanin synthesis of the fermentation product. However, the present application uses a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the inhibition rate on melanin synthesis. Thus, the whitening effect can be effectively improved.

[0120] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Lactobacillus plantarum, to ferment sea rice, the resulting fermentation product has a low inhibition rate on melanin synthesis; while the present application scheme uses a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the inhibition rate of melanin synthesis of the fermentation product; it can be seen that using a single strain, Lactobacillus plantarum, to ferment sea rice, it is difficult to improve the inhibition rate of melanin synthesis of the fermentation product. However, the present application uses a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the inhibition rate on melanin synthesis. Thus, the whitening effect can be effectively improved.

[0121] Example 5 Tyrosinase Activity Inhibition Rate Determination The cell suspension was centrifuged at 1500 rpm for 3 min to obtain a cell pellet. 300 μL of 1% Triton X-100 was added to each tube of cell pellet and ultrasonicated for 5 min. Samples of each embodiment or comparative example were assayed using a BCA kit. 100 μL of the remaining solution was preheated at 37°C in a 96-well plate for 5 min, followed by the addition of 100 μL of 0.25% L-dopa solution. The plate was incubated at 37°C for 1 h, and the OD value was read at 490 nm. The inhibition rate was calculated according to the following formula.

[0122]

[0123] The test results are shown in Table 5.

[0124] Table 5 Tyrosinase activity inhibition rate of different examples

[0125] From the experimental data in Table 5 above, we can see that: The inhibition rate of tyrosinase activity of the sea rice fermentation filtrate prepared in each example was higher than that of each comparative example. This shows that the method of the present application can effectively improve the inhibition rate of tyrosinase activity of the sea rice fermentation filtrate.

[0126] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 used a single strain, Saccharomyces cerevisiae, to ferment sea rice, the resulting fermentation product had a low inhibition rate on tyrosinase activity; while the present application scheme used a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the inhibition rate of the fermentation product on tyrosinase activity; it can be seen that using a single strain, Saccharomyces cerevisiae, to ferment sea rice is difficult to improve the inhibition rate of the fermentation product on tyrosinase activity. However, the present application used a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the inhibition rate on tyrosinase activity. This can effectively enhance the whitening effect.

[0127] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: Under the premise of the same fermentation medium and inoculum size, when comparative example 1 uses a single strain, Lactobacillus plantarum, to ferment sea rice, the resulting fermentation product has a low inhibition rate for tyrosinase activity; while the present application adopts a mixed strain, Saccharomyces cerevisiae and Lactobacillus plantarum, to ferment sea rice, which can effectively improve the inhibition rate of tyrosinase activity of the fermentation product; it can be seen that using a single strain, Lactobacillus plantarum, to ferment sea rice is difficult to improve the inhibition rate of tyrosinase activity of the fermentation product. However, the present application uses a mixed strain of Saccharomyces cerevisiae and Lactobacillus plantarum to ferment sea rice, which can greatly improve the inhibition rate for tyrosinase activity. Thus, the whitening effect can be effectively improved.

[0128] Example 6 Determination of relative expression levels of genes related to the whitening pathway The collected cells were centrifuged at 3000 rpm in a low-temperature centrifuge for 5 minutes, and the supernatant was discarded. The RNA was extracted by lysing and rinsing the cell pellet using an RNA extraction kit. The RNA concentration was then measured and normalized using a micro-spectrophotometer. The reaction system was then configured according to the instructions of the reverse transcription kit to prepare the cDNA sample. The cDNA sample and upstream and downstream primers and other reactants, a total reaction system of 20 µL, were mixed into eight tubes according to the operating instructions of the qPCR kit. After gently shaking to mix, the tube was placed in the qPCR instrument. The reaction parameters were set according to the instructions. After the last cycle, the Ct value of each well was obtained, and the 2 of each well was calculated using Excel software. （-ΔΔCt） The values were obtained and the results of the control group and experimental group were compared to analyze the expression levels of TYP, TRP-1 and TRP-2 genes.

[0129] The test results are shown in Tables 6, 7 and 8.

[0130] Table 6 Expression levels of gene TYR in different examples

[0131] Table 7 Expression levels of gene TRP-1 in different examples

[0132] Table 8 Expression levels of gene TRP-2 in different examples

[0133] From the experimental data in Tables 6-8 above, we can see that: The sea rice fermentation filtrates produced in each Example showed lower expression levels for the TYR gene, the TRP-1 gene, and the TRP-2 gene than the control examples. The lower the expression levels for the TYR gene, the TRP-1 gene, and the TRP-2 gene, the better the whitening pathway effect.

[0134] This shows that the method of the present application can effectively enhance the whitening effect of sea rice fermentation filtrate.

[0135] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 1 that: Under the premise of the same fermentation medium and inoculum amount, when the comparative example 1 uses a single strain, Saccharomyces cerevisiae, to ferment sea rice, the whitening pathway effect of the fermentation product obtained is worse than that of Example 1.

[0136] Furthermore, it can be seen from the comparison between Example 1 and Comparative Example 2 that: Under the premise of the same fermentation medium and inoculum amount, when the comparative example 1 used a single strain, Lactobacillus plantarum, to ferment sea rice, the whitening pathway effect of the fermentation product obtained was worse than that of Example 1.

[0137] In summary, the fermentation product prepared by the method of the present application can effectively improve the whitening effect.

[0138] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A method for preparing sea rice fermentation filtrate, characterized in that: include: inoculating the first fermentation seed liquid and the second fermentation seed liquid into a fermentation medium, fermenting at 25° C.-30° C. to obtain a fermentation product, and removing bacteria from the fermentation product; The fermentation medium is composed of the following components in percentage by mass: 5%-10% sea rice powder, 1%-5% sugars and the balance water; the pH of the fermentation medium is 4-5; The first fermentation seed liquid is a fermentation seed liquid of Saccharomyces cerevisiae; The second fermented seed liquid is fermented seed liquid of Lactobacillus plantarum.

2. The method for preparing sea rice fermentation filtrate according to claim 1, characterized in that: The fermentation medium is composed of the following components in percentage by mass: 6%-9% sea rice powder, 2%-4% sugars and the balance water; the pH of the fermentation medium is adjusted to 4.

5.

3. The method for preparing sea rice fermentation filtrate according to claim 1 or 2, characterized in that: The step of inoculating the first fermentation seed liquid into the fermentation medium comprises: Based on the total volume of the fermentation medium, 1%-5% of the Saccharomyces cerevisiae strain is inoculated into the fermentation medium in volume percentage.

4. The method for preparing sea rice fermentation filtrate according to claim 1 or 2, characterized in that: Inoculating the second fermentation seed liquid into a fermentation medium comprises: Based on the total volume of the fermentation medium, 2%-6% of the Saccharomyces cerevisiae strain is inoculated into the fermentation medium in volume percentage.

5. The method for preparing sea rice fermentation filtrate according to claim 1 or 2, characterized in that: Inoculating the first fermentation seed liquid and the second fermentation seed liquid into a fermentation medium comprises: Based on the total volume of the fermentation medium, the total amount of the first fermentation seed liquid and the second fermentation seed liquid, which is greater than 0 and less than or equal to 10%, is inoculated into the fermentation medium in volume percentage.

6. The method for preparing sea rice fermentation filtrate according to claim 1 or 2, characterized in that: Inoculating the first fermentation seed liquid and the second fermentation seed liquid into a fermentation medium comprises: The volume ratio of the added amount of the first fermentation seed liquid to the second fermentation seed liquid is: (1-5): (2-6).

7. The method for preparing sea rice fermentation filtrate according to claim 1, characterized in that: The sugar includes at least one of glucose or sucrose.

8. A sea rice fermentation filtrate, characterized in that: The sea rice fermentation filtrate is prepared by the method for preparing the sea rice fermentation filtrate according to any one of claims 1 to 7.

9. The method for preparing sea rice fermentation filtrate according to claim 8, characterized in that: The phytic acid content in the sea rice fermentation filtrate is greater than 413.33 mg / mL.

10. A whitening cosmetic, characterized in that: The method comprises the sea rice fermentation filtrate according to claim 9.

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