Fermented product of houttuynia cordata and preparation method thereof, cosmetic

Abstract

This application relates to the field of cosmetics, specifically a fermented product of Houttuynia cordata, its preparation method, and cosmetics. A brewer's yeast strain is inoculated into a culture medium and fermented at a temperature below 25°C. The culture medium consists of Houttuynia cordata homogenate and sugars; the Houttuynia cordata homogenate serves as the substrate; the sugars are added to the substrate. By inoculating the brewer's yeast strain into the culture medium and fermenting at a temperature below 25°C, the culture medium includes Houttuynia cordata powder and sugars. Using brewer's yeast as the starting strain, low-temperature fermentation of Houttuynia cordata not only promotes the growth of the brewer's yeast strain but also allows for the extraction of more active ingredients such as flavonoids, polyphenols, polysaccharides, and volatile oils from Houttuynia cordata, thereby effectively improving the utilization of active substances in Houttuynia cordata. Applying the Houttuynia cordata fermented product obtained using this method to cosmetics can effectively enhance the anti-aging effects of the cosmetics.

Description

Technical Field

[0001] This application relates to the field of cosmetics, specifically to a Houttuynia cordata fermentation product, its preparation method, and cosmetics. Background Technology

[0002] Houttuynia cordata contains active ingredients such as flavonoids, polyphenols, polysaccharides, and volatile oils, which can be used in cosmetics. However, current methods for utilizing these active ingredients typically employ physical extraction methods, such as using common extractants like water and alcohol. However, this method not only leaves solvent residues but also fails to fully extract the active ingredients, resulting in poor utilization rates. Consequently, the efficacy of Houttuynia cordata's active ingredients in cosmetics is difficult to fully realize. Summary of the Invention

[0003] The purpose of this application is to provide a Houttuynia cordata fermentation product and its preparation method, as well as a cosmetic, with the aim of improving the utilization effect of the active ingredients in Houttuynia cordata so as to truly exert the efficacy of the active ingredients in cosmetics.

[0004] In a first aspect, this application provides a method for preparing a fermentation product of Houttuynia cordata, comprising: The brewer's yeast strain was inoculated into the culture medium and fermented at a temperature below 25°C; The culture medium consists of Houttuynia cordata homogenate and sugars; the culture medium uses Houttuynia cordata homogenate as a matrix; the sugars are added to the matrix. The preparation of the houttuynia cordata homogenate includes: Add water to the houttuynia cordata powder and stir evenly; by mass percentage, the amount of houttuynia cordata powder added to the houttuynia cordata homogenate is 5%-15% of the mass of water.

[0005] Currently, brewing yeast is typically fermented at 28-30℃; however, the inventors have found that this higher fermentation temperature is not conducive to obtaining more active ingredients from Houttuynia cordata. In the above-mentioned technical solution of this application, brewing yeast strains are inoculated into a culture medium and fermented at a temperature below 25℃. The culture medium includes Houttuynia cordata powder and sugars. Using brewing yeast strains as the starting strain, low-temperature fermentation of Houttuynia cordata not only promotes the growth of brewing yeast strains but also extracts more active ingredients such as flavonoids, polyphenols, polysaccharides, and volatile oils from Houttuynia cordata, thereby effectively improving the utilization of active substances in Houttuynia cordata. The Houttuynia cordata fermentation product obtained using this method is applied to cosmetics. This fermentation product contains active ingredients from Houttuynia cordata and metabolites from brewing yeast. By scavenging excess free radicals and inhibiting the activity of related enzymes, it achieves an anti-aging effect without solvent residue, thus effectively improving the anti-aging effect of cosmetics. This truly allows the active ingredients of Houttuynia cordata to exert their efficacy in cosmetics.

[0006] In other embodiments of this application, the above-mentioned brewing yeast strain is inoculated into the culture medium and fermented at a temperature of 16°C to 20°C.

[0007] In other embodiments of this application, the amount of sugar added to the culture medium is 5%-15% of the mass of water, by weight percentage.

[0008] In other embodiments of this application, inoculating the Saccharomyces cerevisiae strain into the culture medium includes: Inoculate 5%-10% of the Saccharomyces cerevisiae culture medium with the total volume of the culture medium after inoculation, based on the Saccharomyces cerevisiae culture strain / total volume of the culture medium after inoculation.

[0009] In other embodiments of this application, fermentation is carried out at a temperature below 25°C, including: Ferment at a temperature below 25°C for 3 to 7 days.

[0010] In other embodiments of this application, the pH of the culture medium is less than 7.

[0011] In other embodiments of this application, the pH of the culture medium is 6 to 6.5.

[0012] In other embodiments of this application, sugars include at least one of glucose or sucrose.

[0013] Secondly, this application provides a Houttuynia cordata fermentation product, which is prepared by the method for preparing Houttuynia cordata fermentation product provided in any of the first aspects above.

[0014] Thirdly, this application provides a cosmetic product, including the Houttuynia cordata fermentation product provided in the second aspect above. Attached Figure Description

[0015] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 The standard curve for rutin; Figure 2 This is the standard curve for gallic acid. Detailed Implementation

[0017] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are some embodiments of this application, but not all embodiments.

[0018] Therefore, the following detailed description of the embodiments of this application is not intended to limit the scope of the claimed application, but merely to illustrate selected embodiments of the application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without inventive effort are within the scope of protection of this application.

[0019] This application provides a method for preparing a fermentation product of Houttuynia cordata, comprising: The brewer's yeast strain was inoculated into the culture medium and fermented at a temperature below 25°C; The culture medium comprises a houttuynia cordata homogenate and sugars; the culture medium uses the houttuynia cordata homogenate as a matrix; and the sugars are added to the matrix. The preparation of the houttuynia cordata homogenate includes: Add water to the houttuynia cordata powder and stir evenly; by mass percentage, the amount of houttuynia cordata powder added to the houttuynia cordata homogenate is 5%-15% of the mass of water.

[0020] In the above technical solution, brewer's yeast strains are inoculated into a culture medium and fermented at a temperature below 25°C. The culture medium includes Houttuynia cordata powder and sugars. Using brewer's yeast strains as the starting strain, low-temperature fermentation of Houttuynia cordata not only promotes the growth of brewer's yeast strains but also extracts more active ingredients such as flavonoids, polyphenols, polysaccharides, and volatile oils from Houttuynia cordata. This effectively improves the utilization of active substances in Houttuynia cordata. The Houttuynia cordata fermentation product obtained by this method is applied to cosmetics. This fermentation product contains active ingredients of Houttuynia cordata and metabolites of brewer's yeast. By scavenging excess free radicals and inhibiting the activity of related enzymes, it achieves an anti-aging effect, thereby effectively improving the anti-aging effect of cosmetics.

[0021] Further optionally, and exemplary, in some embodiments of this application, the brewing yeast strain is inoculated into a culture medium and fermented at a temperature of 24°C, 23°C, 22°C, 21°C, 20°C, 19°C, 18°C, 17°C, 16°C, 15°C, or any two of the aforementioned values.

[0022] Existing technologies typically employ fermentation at temperatures above 25°C, such as 28°C to 30°C, for example, to ferment brewer's yeast. However, the inventors discovered that within this temperature range, the active ingredients in houttuynia cordata decompose, significantly reducing their effectiveness. Houttuynia cordata contains abundant flavonoids, polyphenols, polysaccharides, volatile oils, and other active ingredients; their application in cosmetics can effectively enhance the anti-aging effects. The inventors creatively discovered that using brewer's yeast as the starting strain for low-temperature fermentation of houttuynia cordata (fermentation at temperatures below 25°C) not only promotes the growth of brewer's yeast but also effectively extracts flavonoids, polyphenols, polysaccharides, volatile oils, and other active ingredients from houttuynia cordata, while greatly reducing the decomposition of these active ingredients and significantly protecting their effectiveness.

[0023] Furthermore, in some embodiments of this application, the method for preparing the Houttuynia cordata fermentation product includes the following steps: Step S1: Prepare houttuynia cordata powder.

[0024] In some embodiments of this application, Houttuynia cordata is dried, pulverized, and passed through a sieve of 40 mesh or larger to obtain Houttuynia cordata powder.

[0025] For example, in some embodiments of this application, Houttuynia cordata is dried, pulverized, and passed through a 40-mesh sieve, a 41-mesh sieve, a 42-mesh sieve, a 45-mesh sieve, a 46-mesh sieve, a 48-mesh sieve, and a 50-mesh sieve to obtain Houttuynia cordata powder.

[0026] Further, optionally, in some embodiments of this application, after obtaining houttuynia cordata powder by pulverizing, the houttuynia cordata powder is further packaged and sterilized; the sterilized houttuynia cordata powder is then used for subsequent fermentation.

[0027] Step S2: Activation of brewing yeast strain.

[0028] In some embodiments of this application, the brewing yeast is transferred to a solid culture medium and activated by static incubation at 28°C until a single colony grows; then the single colony on the solid culture medium is transferred to a liquid culture medium and cultured at 30°C for 48 hours to obtain a fermentation seed liquid.

[0029] In some embodiments of this application, the above-mentioned brewing yeast was purchased from the Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90.

[0030] In some embodiments of this application, the above-mentioned solid culture medium includes YPD culture medium and agar, wherein the mass of agar added is 2 wt% of the mass of water.

[0031] In some embodiments of this application, the liquid culture medium described above is YPD culture medium.

[0032] Optionally, in some embodiments of this application, the YPD culture medium described above was purchased from Hopebio brand 250g YPD liquid culture medium (product number: HB5193-1).

[0033] In other optional embodiments of this application, the above-mentioned activation of the Saccharomyces cerevisiae strain can also be carried out using other common solid and liquid culture media in the art.

[0034] Step S3: Prepare the fermentation medium.

[0035] In some embodiments of this application, the culture medium includes: houttuynia cordata powder and sugars.

[0036] Further, optionally, in some embodiments of this application, the preparation of the houttuynia cordata homogenate includes: Add water to the houttuynia cordata powder and stir evenly; by mass percentage, the amount of houttuynia cordata powder added to the houttuynia cordata homogenate is 5%-15% of the mass of water.

[0037] For example, in some embodiments of this application, the amount of houttuynia cordata powder added to the houttuynia cordata homogenate is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15% of the mass of water, or the amount of houttuynia cordata powder within any two of the aforementioned values.

[0038] In some embodiments of this application, the amount of sugar added to the culture medium is 5%-15% of the mass of water, by weight percentage.

[0039] For example, in some embodiments of this application, the amount of sugar added is 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, or any two of the aforementioned values.

[0040] Further, optionally, in some embodiments of this application, the sugars include at least one of glucose or sucrose.

[0041] For example, in some embodiments of this application, the sugar selection is either glucose or sucrose; or in some embodiments of this application, the sugar selection is a mixture of glucose and sucrose; the raw materials in the mixture can be mixed in any proportion.

[0042] Further, alternatively, in some embodiments of this application, the pH of the culture medium is less than 7.

[0043] For example, in some embodiments of this application, the pH of the culture medium is 6.9, 6.8, 6.7, 6.6, 6.5, 6.4, 6.3, 6.2, 6.1, 6.0 or a range between any two of the aforementioned values.

[0044] Further alternatively, in some embodiments of this application, the pH of the culture medium is 6 to 6.5.

[0045] Step S4: Fermentation of Houttuynia cordata by brewing yeast.

[0046] Furthermore, in some embodiments of this application, the brewing yeast strain is inoculated into the culture medium and fermented at a temperature below 25°C.

[0047] Alternatively, in some embodiments of this application, the brewing yeast strain is inoculated into the culture medium and fermented at a temperature of 16°C to 20°C.

[0048] For example, in some embodiments of this application, the brewer's yeast strain is inoculated into a culture medium and fermented at a temperature of 16°C, 16.5°C, 17°C, 17.5°C, 18°C, 18.5°C, 19°C, 19.5°C, 20°C or any two of the aforementioned values.

[0049] Furthermore, in some embodiments of this application, inoculating the Saccharomyces cerevisiae strain into the culture medium includes: Inoculate 5%-10% of the Saccharomyces cerevisiae culture medium with the total volume of the culture medium after inoculation, based on the Saccharomyces cerevisiae culture strain / total volume of the culture medium after inoculation.

[0050] For example, inoculating a culture medium with a strain of *Saccharomyces cerevisiae* includes: Based on the total volume of the culture medium after inoculation with Saccharomyces cerevisiae, inoculate the culture medium with 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, or any two of the aforementioned values.

[0051] Furthermore, in some embodiments of this application, fermentation is carried out at a temperature below 25°C, including: Ferment at a temperature below 25°C for 3 to 7 days.

[0052] Exemplarily, in some embodiments of this application, fermentation is carried out at a temperature below 25°C, including: Fermentation is carried out at a temperature below 25°C for 3, 4, 5, 6, or 7 days, or any two of the aforementioned values.

[0053] In some embodiments of this application, the fermentation product obtained above is further centrifuged to remove the cells and obtain the fermentation product (fermentation broth).

[0054] Some embodiments of this application provide a Houttuynia cordata fermentation product, which is prepared by the preparation method of Houttuynia cordata fermentation product provided in any of the foregoing embodiments.

[0055] This application uses Saccharomyces cerevisiae as the starting strain to conduct low-temperature fermentation of Houttuynia cordata, which not only promotes the growth of Saccharomyces cerevisiae but also effectively extracts active ingredients such as flavonoids, polyphenols, polysaccharides, and volatile oils from Houttuynia cordata.

[0056] Furthermore, the fermentation product prepared in this application contains Houttuynia cordata active ingredients and Saccharomyces cerevisiae metabolites, which achieve anti-aging effects by scavenging excess free radicals and inhibiting the activity of related enzymes.

[0057] Some embodiments of this application provide a cosmetic product, including the houttuynia cordata fermentation product provided in any of the foregoing embodiments.

[0058] The features and performance of this application will be further described in detail below with reference to embodiments: Example 1 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0059] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2; to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0060] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0061] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After fermentation at 18℃ for 5 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0062] Example 2 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0063] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0064] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0065] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 3 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0066] Example 3 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0067] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0068] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0069] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 7.5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 7 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0070] Example 4 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0071] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 15% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0072] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0073] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 20℃ for 5 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0074] Example 5 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0075] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 15% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0076] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0077] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 20℃ for 3 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0078] Example 6 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0079] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 15% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0080] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0081] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 7.5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 20℃ for 7 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0082] Example 7 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0083] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 5% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2; to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0084] (3) The brewing yeast purchased from Guangdong Provincial Microbial Culture Collection Center (culture number GDMCC 2.90) was transferred to a solid culture medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28℃ until single colonies grew; then the single colony culture on the solid culture medium was transferred to a liquid culture medium (YPD medium) and cultured at 30℃ for 48h to obtain brewing yeast fermentation seed liquid.

[0085] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 16℃ for 5 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0086] Example 8 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0087] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 5% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2; to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0088] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0089] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After fermentation at 16℃ for 3 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0090] Example 9 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0091] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 5% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0092] (3) The brewing yeast (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 2.90) was transferred to a solid medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to a liquid medium (YPD medium) and cultured at 30°C for 48 hours to obtain the brewing yeast fermentation seed liquid.

[0093] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 7.5% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 16℃ for 7 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0094] Comparative Example 1 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0095] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0096] (3) The brewing yeast purchased from Guangdong Provincial Microbial Culture Collection Center (culture number GDMCC 2.90) was transferred to a solid culture medium (composed of YPD medium and agar, wherein the mass of agar added was 2wt% of the mass of water) and statically activated at 28℃ until single colonies grew; then the single colony culture on the solid culture medium was transferred to a liquid culture medium (YPD medium) and cultured at 30℃ for 48h to obtain brewing yeast fermentation seed liquid.

[0097] (4) The yeast fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, yeast fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 30℃ for 5 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0098] Comparative Example 2 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0099] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0100] (3) Bacillus subtilis (purchased from China Culture Collection Center, strain number CGMCC 1.15792) was transferred to solid nutrient broth medium and activated by static incubation at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to liquid nutrient broth medium and cultured at 30°C for 48 hours to obtain Bacillus subtilis fermentation seed liquid.

[0101] (4) The Bacillus subtilis fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, Bacillus subtilis fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 5 days, the fermentation product is centrifuged to remove the bacterial cells and obtain the fermentation product (fermentation broth).

[0102] Comparative Example 3 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0103] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0104] (3) Bacillus coagulans (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.645) was transferred to solid LB medium and activated at 28°C until single colonies grew. Then, the single colony strain on the solid medium was transferred to liquid LB medium and cultured at 30°C for 48 hours to obtain Bacillus coagulans fermentation seed liquid.

[0105] (4) The Bacillus coagulans fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, Bacillus coagulans fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 5 days, the fermentation product is centrifuged to remove the bacterial cells and obtain the fermentation product (fermentation liquid).

[0106] Comparative Example 4 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0107] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0108] (3) Sphingosine monocytogenes (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC1.587) was transferred to solid TSA medium and activated by static incubation at 28°C until single colonies grew; then the single colony strain on the solid medium was transferred to liquid TSA medium and cultured at 30°C for 48 hours to obtain Sphingosine monocytogenes fermentation seed liquid.

[0109] (4) The Sphingomonas sphingosine monocytogenes fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, Sphingomonas sphingosine monocytogenes fermentation seed liquid / total volume of culture medium after inoculation with seed liquid). After fermentation at 18℃ for 5 days, the fermentation product is centrifuged to remove the bacterial cells and obtain the fermentation product (fermentation broth).

[0110] Comparative Example 5 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0111] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0112] (3) Aspergillus niger (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 3.25) was transferred to solid PDA medium and activated at 28°C until single colonies grew. Then, the single colony strain on the solid medium was transferred to liquid PDA medium and cultured at 30°C for 48 hours to obtain Aspergillus niger fermentation seed liquid.

[0113] (4) The Aspergillus niger fermentation seed liquid obtained in step (3) is inoculated into the culture medium obtained in step (2) at a volume percentage of 10% (v / v, Aspergillus niger fermentation seed liquid / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 5 days, the fermentation product is centrifuged to remove the bacterial cells and obtain the fermentation product (fermentation clear liquid).

[0114] Comparative Example 6 A method for preparing Houttuynia cordata fermentation product is provided, comprising the following steps: (1) After drying the houttuynia cordata, crush it and pass it through a 40-mesh sieve to obtain houttuynia cordata powder. Then, package and sterilize it for later use.

[0115] (2) Take the Houttuynia cordata powder obtained in step (1), add water and stir evenly to make Houttuynia cordata homogenate; wherein, by mass percentage, the amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 10% of the mass of water. Using the Houttuynia cordata homogenate as the culture medium matrix, add glucose and adjust the pH to 6.2 to obtain the culture medium; wherein, by mass percentage, the amount of glucose added to the culture medium is 10% of the mass of water.

[0116] (3) The fermenting Lactobacillus (purchased from Guangdong Provincial Microbial Culture Collection Center, strain number GDMCC 1.1796) was transferred to solid MRS medium and activated at 28°C until single colonies grew. Then, the single colony strain on the solid medium was transferred to liquid MRS medium and cultured at 30°C for 48 hours to obtain fermenting Lactobacillus fermentation seed liquid.

[0117] (4) The fermentation seed liquid of Lactobacillus fermentation prepared in step (3) is inoculated into the culture medium prepared in step (2) at a volume percentage of 10% (v / v, fermentation seed liquid of Lactobacillus fermentation / total volume of culture medium after inoculation of seed liquid). After standing at 18℃ for 5 days, the fermentation product is centrifuged to remove the cells and obtain the fermentation product (fermentation liquid).

[0118] The performance of the fermentation products obtained in each embodiment and comparative example was tested as follows: Experimental Example 1: Determination of Total Flavonoid Content by Spectrophotometry Under neutral or weakly alkaline conditions and in the presence of sodium nitrite, flavonoids form chelates with aluminum salts, which turn red-orange upon the addition of sodium hydroxide solution, with maximum absorption at 510 nm. Using rutin as a reference standard and aluminum nitrate as the colorimetric reagent for flavonoid determination, the absorbance shows a linear relationship with the concentration of rutin. The total flavonoid content is determined spectrophotometrically. (Instructions for use attached) Figure 1 The standard curve for rutin is shown.

[0119] The specific experimental steps are as follows: Transfer 2.4 mL each of the standard stock solution, water (for blank control), and the test solution from each example or comparative example into stoppered test tubes. Add 0.4 mL of 5% sodium nitrite solution to each test tube, shake well, and react for 6 min. Add 0.4 mL of 10% aluminum nitrate solution, shake well, and react for 6 min. Add 1.0 mL of 4% sodium hydroxide solution and 0.7 mL of water, shake well, and react for 15 min. Measure the absorbance using a spectrophotometer at 510 nm using a 10 mm cuvette.

[0120] The test results of flavonoid content in the fermentation products of each embodiment and comparative example are shown in Table 1.

[0121] Table 1 Flavonoid content of fermentation products in various examples and comparative examples

[0122] As shown in Table 1, the flavonoid content in the fermentation products of each embodiment is higher than that in the fermentation products of each comparative example; this indicates that the various solutions in the embodiments of this application can effectively increase the flavonoid content in the fermentation products. The solutions of this application can not only promote the release of active ingredients in Houttuynia cordata, but also simultaneously promote the growth of Saccharomyces cerevisiae and increase the production of Saccharomyces cerevisiae metabolites.

[0123] Experimental Example 2: Determination of Total Content by Folin-f Phenol Method Polyphenols undergo a specific reaction with Folin-Ciocalteu reagent, and the reaction product exhibits maximum absorption at a specific wavelength. Furthermore, the absorbance is linearly related to the amount of polyphenol within a certain concentration range. A standard curve is first established using standard phenolic substances, and then the absorbance of the sample is measured. Based on this, the polyphenol content in the sample can be determined. (Instructions for use are attached.) Figure 2 The standard curve for gallic acid is shown.

[0124] The specific experimental steps are as follows: Transfer 1.0 mL each of the standard working liquid, water (for blank control), and the test solution from each example or comparative example into a stoppered test tube. Add 5.0 mL of 10% Folin-Ciocalteu reagent to each test tube and shake well. React for 5 min, then add 4.0 mL of 7.5% sodium carbonate solution and shake well. Incubate at room temperature for 60 min. Measure the absorbance using a spectrophotometer at 765 nm using a 10 mm cuvette. The polyphenol content test results of the fermentation products of each example and comparative example are shown in Table 2.

[0125] Table 2. Polyphenol content of fermentation products from various examples and comparative examples.

[0126] As shown in Table 2, the polyphenol content in the fermentation products of each embodiment is higher than that in the fermentation products of each comparative example; this indicates that the various solutions in the embodiments of this application can effectively increase the polyphenol content in the fermentation products. The solutions of this application can not only promote the release of active ingredients in Houttuynia cordata, but also simultaneously promote the growth of Saccharomyces cerevisiae and increase the production of Saccharomyces cerevisiae metabolites.

[0127] Experimental Example 3: One of the antioxidant properties 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, preventing the unpaired electrons on the central nitrogen atom from performing their proper electron pairing function. Its anhydrous ethanol solution is purple, exhibiting maximum absorption at 517 nm, with absorbance showing a linear relationship with concentration. Adding free radical scavengers can bind to or replace DPPH·, reducing the number of free radicals, decreasing absorbance, and lightening the solution color, thus allowing evaluation of its free radical scavenging ability.

[0128] The specific experimental steps are as follows: Pipette 2.0 mL of each example or comparative sample solution (or anhydrous ethanol) and 2.0 mL of DPPH solution into a stoppered test tube, mix well, react in the dark for 30 min, and measure the absorbance at a wavelength of 517 nm.

[0129]

[0130] In the formula: A0: No sample added, DPPH added; A1: Add sample or anhydrous ethanol, then add DPPH; A2: Added sample or anhydrous ethanol, no DPPH added.

[0131] The test results of the DPPH free radical scavenging rate of the fermentation products of each embodiment and comparative example are shown in Table 3.

[0132] Table 3. DPPH radical scavenging rates of fermentation products from various examples and comparative studies.

[0133] As can be seen from Table 3, the DPPH radical scavenging rate of the fermentation products of each embodiment is higher than that of the fermentation products of each comparative example.

[0134] This demonstrates that the various solutions in the embodiments of this application can effectively improve the DPPH free radical scavenging rate of fermentation products.

[0135] Experimental Example 4: Antioxidant Capacity II The experiment investigated the scavenging ability of the fermentation products of various examples and comparative examples on ABTS+·. ABTS is oxidized to green ABTS+· under the action of 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.

[0136] 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) Take 190 μL of ABTS working solution (or PBS), add 10 μL of sample solution (or PBS) of each example or comparative example, shake for 10 s, let stand at 30 °C for 6 min, and measure the absorbance value A at 734 nm.

[0137]

[0138] In the formula: A0: No sample added, ABTS added; A1: Add sample or PBS, add ABTS; A2: Added sample or PBS, no ABTS added.

[0139] The test results of the scavenging ability of the fermentation products of each embodiment and comparative example against ABTS+ are shown in Table 4.

[0140] Table 4. Scavenging ability of fermentation products from various examples and comparative studies against ABTS+·

[0141] As can be seen from Table 4, the ABTS+ scavenging ability of the fermentation products of each embodiment is higher than that of the fermentation products of each comparative example.

[0142] This demonstrates that the various solutions in the embodiments of this application can effectively improve the ABTS+· scavenging ability of fermentation products.

[0143] Experimental Example 5: Antioxidant Capacity III Hydroxyl radicals are the most chemically reactive free radicals among reactive oxygen species. They can react with almost any biomolecule in living cells at an extremely rapid rate, making them the most harmful free radicals to the body. However, the addition of salicylic acid to the reaction system can effectively capture ·OH radicals and produce the colored product 2,3-dihydroxybenzoic acid. This colored product has a strong absorption peak at 510 nm. If an analyte with ·OH scavenging ability is added to the system, and the analyte's ·OH scavenging effect is greater than that of salicylic acid, ·OH radicals can be promptly removed, thereby reducing the amount of colored product formed and resulting in a decrease in absorbance.

[0144] The specific experimental steps are as follows: Add 20 μL of each example or comparative sample solution (or pure water) and 180 μL of hydroxyl radical reaction colorimetric solution (or hydroxyl radical blank solution) to a 96-well plate and mix well; incubate at 37°C for 15 min and measure the absorbance at 510 nm.

[0145]

[0146] In the formula: A0 is the absorbance value of the reagent blank; AX represents the absorbance of the test solution in the reaction. Axo is the blank absorbance value of the test solution.

[0147] The test results of the hydroxyl radical scavenging ability of each embodiment and comparative example are shown in Table 5.

[0148] Table 5. Hydroxyl radical scavenging ability of fermentation products from various examples and comparative examples.

[0149] As can be seen from Table 5, the hydroxyl radical scavenging ability of the fermentation products of each embodiment is higher than that of the fermentation products of each comparative example.

[0150] This demonstrates that the various solutions in the embodiments of this application can effectively improve the scavenging ability of fermentation products against hydroxyl radicals.

[0151] In summary, the embodiments of this application can effectively improve the utilization of active substances in Houttuynia cordata. When the Houttuynia cordata fermentation product obtained using this method is applied to cosmetics, this fermentation product contains Houttuynia cordata active ingredients and Saccharomyces cerevisiae metabolites. By scavenging excess free radicals and inhibiting the activity of related enzymes, it achieves an anti-aging effect, thereby effectively improving the anti-aging effect of cosmetics. This truly allows the active ingredients of Houttuynia cordata to exert their full efficacy in cosmetics.

[0152] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A method for preparing a fermented product of Houttuynia cordata, characterized in that, include: Saccharomyces cerevisiae inoculated into a culture medium and fermented at 16℃~20℃ for 3 to 7 days. The fermentation product was then centrifuged to remove the cells, yielding a clear fermentation liquid. The culture medium consisted of Houttuynia cordata homogenate and sugars, with Houttuynia cordata homogenate as the substrate. The sugars are added to the matrix; The preparation of the houttuynia cordata homogenate includes: Mix the houttuynia cordata powder with water until smooth; The amount of Houttuynia cordata powder added to the Houttuynia cordata homogenate is 5%-15% of the mass of water, by weight percentage.

2. The method for preparing the Houttuynia cordata fermentation product according to claim 1, characterized in that, In the culture medium, the amount of sugar added is 5%-15% of the mass of water, by weight percentage.

3. The method for preparing the Houttuynia cordata fermentation product according to claim 1, characterized in that, Inoculating the culture medium with Saccharomyces cerevisiae includes: Based on the total volume of the culture medium after inoculation with Saccharomyces cerevisiae, inoculate 5%-10% of the Saccharomyces cerevisiae culture medium.

4. The method for preparing the Houttuynia cordata fermentation product according to any one of claims 1-3, characterized in that, The pH of the culture medium is less than 7.

5. The method for preparing the Houttuynia cordata fermentation product according to any one of claims 1-3, characterized in that, The pH of the culture medium is 6 to 6.

5.

6. The method for preparing the Houttuynia cordata fermentation product according to any one of claims 1-3, characterized in that, The sugars include at least one of glucose and sucrose.

7. A fermented product of Houttuynia cordata, characterized in that, The product was prepared using the method described in any one of claims 1-6 for the preparation of Houttuynia cordata fermentation products.

8. A cosmetic product, characterized in that, Includes the Houttuynia cordata fermentation product as described in claim 7.

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