A sargassum thunbergii fermentation liquor, a preparation method thereof, and a cosmetic

Abstract

The application provides a sargassum fusiforme fermentation liquor, a preparation method thereof and a cosmetic, wherein the preparation method of the sargassum fusiforme fermentation liquor comprises the following steps: S10: sterilizing a mixture of sargassum fusiforme, a carbon source and water to obtain an initial fermentation system; S20: inoculating lactobacillus liquid into the initial fermentation system and then performing fermentation for a certain time; and S30: sterilizing and centrifuging the fermentation system to obtain supernatant, which is the sargassum fusiforme fermentation liquor. Through the fermentation of lactobacillus, the active ingredients in the sargassum fusiforme are released to a large extent, and the metabolic products produced by the lactobacillus can synergize with the active ingredients in the sargassum fusiforme to exert a good antioxidant effect. The preparation method is simple, raw materials are easy to obtain, and the cost is low, and the method can be applied to the production of cosmetics.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of cosmetics, in particular to a Sargassum foliaceum fermentation liquor, a preparation method thereof and a cosmetic. BACKGROUND

[0002] Sargassum foliaceum, a kind of algae, is also known as antler tip, sea vegetable bud, goat milk, sea barley and so on. The algal body is yellow-brown and thick and juicy, and the thallus varies greatly in shape. It grows on rocks in the low tide zone. Research has found that Sargassum foliaceum contains 18 important amino acids required by the human body (including 8 essential amino acids that the human body cannot synthesize) and 14 important trace elements, and has high nutritional and health care value.

[0003] Sargassum foliaceum contains polysaccharides, polyphenols, proteins and other antioxidant active ingredients, which can prevent free radical damage caused by skin aging, increase the ability of the skin to resist photoaging, and resist inflammatory reactions of the skin, reduce the occurrence of skin problems such as pigmentation, redness and sensitivity.

[0004] In the prior art, organic solvents are generally used to extract active ingredients from Sargassum foliaceum, which not only causes environmental pollution, but also has low extraction efficiency and high cost, and the extract may contain residual organic solvents, which poses a safety hazard and is not conducive to industrialization. SUMMARY

[0005] The present application provides a Sargassum foliaceum fermentation liquor, a preparation method thereof and a cosmetic. The Sargassum foliaceum fermentation liquor is prepared by fermenting Sargassum foliaceum using lactobacillus. The fermentation of lactobacillus releases active ingredients in Sargassum foliaceum to a large extent, and the metabolites produced by lactobacillus can synergize with the active ingredients in Sargassum foliaceum to produce good antioxidant effects.

[0006] In a first aspect, the present application provides a preparation method of a Sargassum foliaceum fermentation liquor, comprising the following steps:

[0007] S10: sterilizing the mixture of Sargassum foliaceum raw materials, carbon source and water to obtain an initial fermentation system;

[0008] S20: inoculating lactobacillus liquid into the initial fermentation system to obtain a mixed fermentation liquor;

[0009] S30: separating the supernatant from the mixed fermentation liquor to obtain the Sargassum foliaceum fermentation liquor.

[0010] In the technical solution of the present application, Sargassum foliaceum is fermented using lactobacillus, which promotes the dissolution of active ingredients in Sargassum foliaceum under the action of extracellular enzymes produced by lactobacillus. Therefore, the preparation method of the Sargassum foliaceum fermentation liquor provided by the present application is simple, has high extraction efficiency, short production cycle and good safety.

[0011] In some embodiments of the present application, in the step S10, the raw material of the Sargassum fusiforme is dried Sargassum fusiforme powder with a particle size corresponding to a 50-150 mesh sieve.

[0012] In some embodiments of the present application, in the step S10, the carbon source is one or more of glucose, sucrose, fructose, and maltose.

[0013] In some embodiments of the present application, in the step S10, the mass ratio of the dried Sargassum fusiforme powder, the carbon source, and water is 1:0.1-2:10-300.

[0014] In some embodiments of the present application, in the step S20, the inoculation amount of the Lactobacillus liquid is 1%-5%, and the viable bacterial count of the Lactobacillus liquid is 10 6 11 CFU / mL.

[0015] In some embodiments of the present application, the Lactobacillus is one or more of Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus acidophilus, and Lactobacillus bulgaricus.

[0016] In some embodiments of the present application, the Lactobacillus is a mixture of Lactobacillus helveticus and Lactobacillus plantarum with a viable bacterial count ratio of 1:0.2-5.

[0017] In some embodiments of the present application, in the step S20, the fermentation conditions are a fermentation temperature of 30-45°C and a fermentation time of 12-48 h.

[0018] In some embodiments of the present application, the method for preparing the Sargassum fusiforme fermentation liquor further comprises:

[0019] Step S40: decolorizing the supernatant using a decolorizing agent and then sterilizing again to obtain the Sargassum fusiforme fermentation liquor.

[0020] In a second aspect, the present application provides a Sargassum fusiforme fermentation liquor prepared according to the preparation method of any one of the above embodiments.

[0021] In the technical solution of the present application, the Sargassum fusiforme fermentation liquor prepared by fermenting Sargassum fusiforme with Lactobacillus has a high dissolution rate of active ingredients after Lactobacillus fermentation, and the small molecule monosaccharides in Sargassum fusiforme can be utilized by Lactobacillus to produce metabolites. Therefore, the Sargassum fusiforme fermentation liquor provided by the present application has good antioxidant activity.

[0022] In a third aspect, the present application provides a cosmetic product comprising the Sargassum fusiforme fermentation liquor prepared according to the preparation method of any one of the above embodiments or the Sargassum fusiforme fermentation liquor according to any one of the above embodiments.

[0023] ​In the technical solution of the present application, under the action of the enzyme secreted by the lactobacillus, the macromolecular active ingredients in the Sargassum fusiforme are decomposed into small molecular active ingredients, so that they are more easily absorbed by the skin. Meanwhile, the metabolites produced by the lactobacillus are used in combination, and the Sargassum fusiforme fermentation liquor is directly used as a cosmetic or added to a cosmetic base formula, which has good antioxidant, anti-aging and skin nourishing effects. BRIEF DESCRIPTION OF DRAWINGS

[0024] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and serve to explain the principles of the present application, together with the description.

[0025] Figure 1 The DPPH free radical scavenging test results of Examples 1-4 and Comparative Examples 1-2.

[0026] Figure 2 The hydroxyl radical scavenging test results of Examples 1-4 and Comparative Examples 1-2.

[0027] Figure 3 The protein content test results of Examples 1-4 and Comparative Examples 1-2.

[0028] Figure 4 The total sugar content test results of Examples 1-4 and Comparative Examples 1-2.

[0029] It should be noted that in each figure, * represents the difference analysis results of Examples 2-4, Comparative Examples 1-2 and the corresponding examples, wherein one * represents a significant difference, two * represents a very significant difference, and three * represents an extremely significant difference.

[0030] Through the above drawings, the specific embodiments of the present application have been shown, and more detailed descriptions will be given hereinafter. These drawings and textual descriptions are not intended to limit the scope of the concept of the present application in any way, but to illustrate the concept of the present application to those skilled in the art by referring to specific embodiments. DETAILED DESCRIPTION

[0031] Each embodiment or implementation in the present specification is described in a progressive manner, and each embodiment focuses on the difference from other embodiments.

[0032] In the description of the present specification, the description referring to the terms "one embodiment", "some embodiments", "exemplary embodiment", "example", "specific example", or "some examples" and the like means that the specific features, structures, materials or characteristics described in connection with the implementation or example are included in at least one implementation or example of the present application. In the present specification, the exemplary description of the above terms does not necessarily mean the same implementation or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more implementations or examples as appropriate.

[0033] In addition, the terms "first", "second" are only for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined with "first", "second" can explicitly or implicitly include at least one of the features. In the description of the present application, the meaning of "a plurality of" is at least two, for example, two, three, etc., unless otherwise explicitly specified.

[0034] Applicants found that Sargassum fusiforme contains a variety of antioxidant active ingredients that can be extracted and used, but in the prior art, the extraction of active ingredients in Sargassum fusiforme is mainly through the method of organic solvent extraction, and the extraction efficiency of this method is low, which is not suitable for industrial production, and the residual organic solvent in the extract will also affect the further use of Sargassum fusiforme extract.

[0035] Therefore, the applicant uses the method of microbial fermentation to extract the active ingredients in Sargassum fusiforme, and a large number of experiments on the types and methods of fermentation microorganisms are carried out, thereby providing a technical scheme.

[0036] In a first aspect, the present application provides a preparation method of Sargassum fusiforme fermentation liquor, comprising the following steps:

[0037] S10: sterilizing the mixture of Sargassum fusiforme raw materials, carbon source and water to obtain an initial fermentation system;

[0038] S20: inoculating Lactobacillus liquid into the initial fermentation system to obtain a mixed fermentation liquor by fermentation;

[0039] S30: separating the supernatant from the mixed fermentation liquor to obtain the Sargassum fusiforme fermentation liquor.

[0040] In the technical scheme of the present application, the Sargassum fusiforme is fermented by using Lactobacillus, and under the action of the extracellular enzymes produced by Lactobacillus, the active ingredients of Sargassum fusiforme are fully dissolved, so that the preparation method of Sargassum fusiforme fermentation liquor provided by the present application is simple, has high extraction efficiency, short production cycle and good safety.

[0041] In the technical solution of the present application, the applicant finds that the antioxidant performance of the fermentation liquor obtained by using Lactobacillus to ferment Sargassum fusiforme is significantly higher than that of other commonly used fermentation microorganisms, such as yeast; the possible reason is that most of the active ingredients of Sargassum fusiforme are distributed in the cells and intercellular layers, and need to pass through multiple cell walls to be utilized, the extracellular enzymes produced by Lactobacillus in the fermentation process have higher efficiency in degrading the cell wall, so that the active ingredients are more easily dissolved, in addition, there is a conversion of active ingredients in the fermentation process, the dissolved active ingredients of Sargassum fusiforme, such as flavonoids, polysaccharides, saponins, etc., can be utilized by Lactobacillus as prebiotics, on the one hand, promoting the growth and metabolism of Lactobacillus, on the other hand, various enzymes produced by Lactobacillus in the metabolism process will cause the original active ingredients of Sargassum fusiforme to be converted to obtain new active ingredients, or be degraded into more low-polarity and small-molecular-weight metabolites, thereby improving the antioxidant activity of the Sargassum fusiforme fermentation liquor.

[0042] In some embodiments of the present application, in step S10, the Sargassum fusiforme raw material is dry Sargassum fusiforme powder with a particle size corresponding to a 50-150 mesh sieve.

[0043] In the above some embodiments, the dry Sargassum fusiforme powder destroys the original tissue structure of Sargassum fusiforme, thereby improving the dissolution efficiency of the active ingredients of Sargassum fusiforme from a physical level, which is beneficial to the fermentation and utilization of Lactobacillus. Generally speaking, the smaller the particle size, the more sufficient the fermentation extraction, but considering the cost problem, therefore, the particle size of the dry Sargassum fusiforme powder is controlled to correspond to a 50-150 mesh sieve.

[0044] In some embodiments of the present application, in step S10, the carbon source is one or more of glucose, sucrose, fructose, and maltose.

[0045] In the above some embodiments, it is necessary to add a carbon source in the fermentation system to provide nutrients required for the early growth and reproduction of Lactobacillus, thereby improving the fermentation activity of Lactobacillus, and making the effective components of Sargassum fusiforme be fully dissolved and utilized by fermentation. Therefore, the selected carbon source is a common monosaccharide or disaccharide that is easily utilized by Lactobacillus, and more preferably, the carbon source is glucose.

[0046] In some embodiments of the present application, in step S10, the mass ratio of dry Sargassum fusiforme powder, carbon source, and water is 1:0.1-2:10-300.

[0047] In some of the above embodiments, since the initial fermentation system in the technical scheme of the present application mainly comprises Sargassum fusiforme, a carbon source and water, and Lactobacillus is sensitive to the fermentation conditions, if the content of the carbon source is too low, it cannot meet the requirements of the early growth and reproduction of Lactobacillus, and if the content of the carbon source is too high, it will affect the fermentation of Lactobacillus on Sargassum fusiforme; in addition, the content of Sargassum fusiforme will also affect the water activity and oxygen concentration and other parameters of the initial fermentation system, which will lead to differences in the fermentation process and the product, and therefore the content of Sargassum fusiforme will significantly affect the composition of the fermentation broth. The applicant determines through a large number of experiments that the mass ratio of dried Sargassum fusiforme powder, carbon source and water in the range of 1:0.1-2:10-300 is beneficial to obtaining Sargassum fusiforme fermentation broth with high antioxidant activity; preferably, the mass ratio of dried Sargassum fusiforme powder, carbon source and water is 1:0.1-1:20-150; most preferably, the mass ratio of dried Sargassum fusiforme powder, carbon source and water is 1:0.2:100.

[0048] In some embodiments of the present application, in step S20, the inoculation amount of the Lactobacillus bacterial solution is 1%-5%, and the viable bacterial count of the Lactobacillus bacterial solution is 10 6 ~ 10 11 CFU / mL.

[0049] In some of the above embodiments, the inoculation amount and the viable bacterial count of the Lactobacillus bacterial solution are further limited, and the inoculation amount and the viable bacterial count of the Lactobacillus bacterial solution have a certain influence on the final fermentation broth. The applicant determines through experiments that the better Lactobacillus bacterial solution inoculation amount is 1%-5%, and the viable bacterial count of the Lactobacillus bacterial solution is 10 6 ~ 10 11 CFU / mL; preferably, the viable bacterial count of the Lactobacillus bacterial solution is 10 8 ~ 10 10 CFU / mL.

[0050] In some embodiments of the present application, the Lactobacillus is one or more of Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus bulgaricus.

[0051] In some of the above embodiments, the applicant finds that the Sargassum fusiforme fermentation broth obtained by fermenting Sargassum fusiforme with Lactobacillus helveticus, Lactobacillus plantarum, Lactobacillus acidophilus and Lactobacillus bulgaricus in the Lactobacillus genus all have good antioxidant activity, and the antioxidant activity of the Sargassum fusiforme fermentation broth obtained by fermenting Sargassum fusiforme with Lactobacillus helveticus and Lactobacillus plantarum is the highest. Therefore, the applicant further studies the fermentation of Sargassum fusiforme with Lactobacillus helveticus and Lactobacillus plantarum in subsequent embodiments.

[0052] In some embodiments of the present application, the Lactobacillus is a mixture of Lactobacillus helveticus and Lactobacillus plantarum with a viable bacterial count ratio of 1:0.2-5.

[0053] In some of the above embodiments, the applicants use Lactobacillus helveticus and Lactobacillus plantarum which have good effects on fermenting Sargassum siliqustrum to carry out further tests. The applicants find that the antioxidant activity of the Sargassum siliqustrum fermentation liquid obtained by mixed fermentation is higher than that of the Sargassum siliqustrum fermentation liquid obtained by single-strain fermentation. The possible reason is that there is a certain difference between the two strains in the way of improving the antioxidant activity of the Sargassum siliqustrum fermentation liquid, and there is a certain synergistic effect between the corresponding ways, so that the antioxidant activity of the mixed Sargassum siliqustrum fermentation liquid is further improved.

[0054] In some embodiments of the present application, the fermentation conditions in step S20 are as follows: the fermentation temperature is 30-45℃, and the fermentation time is 12-48h.

[0055] In some of the above embodiments, the applicants find that the fermentation temperature and the fermentation time determined according to the fermentation strain have a great influence on the fermentation product. If the time is too short, the active ingredients of Sargassum siliqustrum are not fully dissolved and there are few Lactobacillus metabolic products. If the time is too long, the microorganisms may use too much active ingredient, resulting in a decrease in effective components in the fermentation liquid. Therefore, the fermentation conditions are set as follows: the fermentation temperature is 30-45℃, and the fermentation time is 12-48h; more preferably, the fermentation temperature is 35-40℃, and the fermentation time is 12-24h.

[0056] In some embodiments of the present application, step S30 specifically comprises sterilizing the mixed fermentation liquid and centrifuging to obtain the supernatant, i.e., the Sargassum siliqustrum fermentation liquid.

[0057] In the technical solutions of the present application, the specific conditions for sterilization are not further limited. The sterilization process is mainly to inactivate the microorganisms in the system, so the sterilization techniques commonly used in the prior art can be used, such as high-temperature sterilization at 110-125℃, 0.1-0.14MPa for 15-35min.

[0058] In the technical solutions of the present application, the centrifugation conditions are also not further limited. The purpose of centrifugation is to separate the residues after fermentation from the fermentation liquid. Generally, under the conditions of a centrifugal speed of 3000-9000r / min and a centrifugal radius of 8-15cm, centrifugation for 10-40min can separate the two.

[0059] In some embodiments of the present application, the method for preparing the Sargassum siliqustrum fermentation liquid further comprises:

[0060] Step S40: decolorizing the supernatant with a decolorizing agent and then sterilizing to obtain the Sargassum siliqustrum fermentation liquid.

[0061] In some of the above embodiments, since there are more pigments in Sargassum fusiforme, the pigments cannot be separated from the fermentation broth by centrifugation. Therefore, in order to expand the application range of the Sargassum fusiforme fermentation broth, the supernatant after centrifugation needs to be decolorized. The Sargassum fusiforme fermentation broth obtained after sterilization after decolorization is colorless, and can be used as a cosmetic or a cosmetic raw material.

[0062] In some of the above embodiments, the decolorization method is not further limited. Generally, a decolorizing agent commonly used in the art is added to the supernatant for decolorization, and then filtration is performed to obtain the Sargassum fusiforme fermentation broth. Specifically, the decolorizing agent can be selected from one or more of activated carbon, bentonite and diatomite, the mass / volume ratio of the decolorizing agent to the supernatant can be 10-20 g / L, and the decolorization time can be 50-150 min.

[0063] In a second aspect, the present application provides a Sargassum fusiforme fermentation broth prepared according to the preparation method of any one of the above embodiments.

[0064] In the technical solution of the present application, the Sargassum fusiforme fermentation broth prepared by fermenting Sargassum fusiforme with Lactobacillus has a high dissolution rate of active ingredients of Sargassum fusiforme after fermentation, and small molecule monosaccharides in Sargassum fusiforme can be utilized by Lactobacillus to produce metabolites. Therefore, the Sargassum fusiforme fermentation broth provided by the present application has good antioxidant activity.

[0065] In a third aspect, the present application provides a cosmetic containing the Sargassum fusiforme fermentation broth prepared according to the preparation method of any one of the above embodiments or the Sargassum fusiforme fermentation broth of any one of the above embodiments.

[0066] In the technical solution of the present application, under the action of enzymes secreted by Lactobacillus, the macromolecular active ingredients in Sargassum fusiforme are decomposed into small molecule active ingredients, which are more easily absorbed by the skin. In addition, the metabolites produced by Lactobacillus are used as a cosmetic or added to a cosmetic base formula, which has good antioxidant, anti-aging and skin nourishing effects.

[0067] In the technical solution of the present application, a preservative commonly used in the cosmetic field can be added to the Sargassum fusiforme fermentation broth to be used directly as a cosmetic. Specifically, the preservative can be selected from p-hydroxyacetophenone and / or 1,2-hexanediol, and the amount of the preservative added is 0.5% to 2% of the total mass of the cosmetic.

[0068] In the technical solution of the present application, the Sargassum fusiforme fermentation broth can also be added to a cosmetic base formula for use. Specifically, the mass percentage of the Sargassum fusiforme fermentation broth in the skin external agent cosmetic is 5% to 99%; more preferably, the mass percentage of the Sargassum fusiforme fermentation broth in the skin external agent cosmetic is 60% to 99%.

[0069] Hereinafter, the Sargassum fermenting liquor and the preparation method thereof of the present application will be explained in more details through examples, but the present application is by no means limited to these examples.

[0070] It should be noted that the Lactobacillus helveticus and the Lactobacillus plantarum used in the examples are both purchased from the China Industrial Microbial Culture Collection Center, wherein the Lactobacillus helveticus has a preservation number of CICC20243, and the Lactobacillus plantarum has a preservation number of CICC20261; the Saccharomyces cerevisiae is preserved in the China General Microbiological Culture Collection Center, and has a number of CGMCC17452.

[0071] Example 1

[0072] Preparation of the Sargassum fermenting liquor:

[0073] S10: dry Sargassum is crushed and sieved, the powder particle size is 100 mesh, 300 mL of deionized water and 3 g of Sargassum are taken, 0.6 g of glucose is taken as a carbon source, and the mixture is uniformly mixed, and then sterilized in a 121℃ high-pressure sterilization pot for 20 min, and then cooled to room temperature to obtain an initial fermentation system;

[0074] S20: Lactobacillus helveticus and Lactobacillus plantarum are added to the initial fermentation system for fermentation, wherein the viable cell count of the Lactobacillus helveticus and the Lactobacillus plantarum is 10 9 CFU / mL, and the addition amount of the two bacterial liquids is 1.5 mL, the bacterial liquid is shaken after being inoculated, and then placed in a 40℃ constant temperature incubator for culture for 18 h to obtain a mixed fermentation liquor;

[0075] S30: the mixed fermentation liquor is sterilized in a high-pressure steam sterilization pot at 121℃ for 20 min. After sterilization is completed, the fermentation liquor is cooled to room temperature for centrifugation, the centrifugation speed is 5000 r / min, the centrifugation time is 30 min, and the supernatant is collected as the Sargassum fermenting liquor.

[0076] Example 2

[0077] Preparation of the Sargassum fermenting liquor:

[0078] The difference from Example 1 is only that in step S20, Lactobacillus helveticus is added to the initial fermentation system for fermentation, wherein the viable cell count of the Lactobacillus helveticus is 10 9 CFU / mL, and the addition amount is 3 mL, the bacterial liquid is shaken after being inoculated, and then placed in a 40℃ constant temperature incubator for culture for 18 h.

[0079] Example 3

[0080] Preparation of the Sargassum fermenting liquor:

[0081] The difference from Example 1 is that in step S20, Lactobacillus plantarum is added to the initial fermentation system for fermentation, wherein the viable cell count of the Lactobacillus plantarum liquid is 10 9 CFU / mL, the addition amount is 3 mL, and after the bacterial liquid is inoculated, it is uniformly shaken and placed in a 40℃ constant temperature incubator for culture for 18h.

[0082] Example 4

[0083] Preparation of Sargassum fusiforme fermentation liquid:

[0084] The difference from Example 1 is that in step S10, the addition amount of glucose is 0.3g, and the addition amount of Sargassum fusiforme is 5g.

[0085] Comparative Example 1

[0086] Preparation of Sargassum fusiforme fermentation liquid:

[0087] The difference from Example 1 is that in step S20, 3mL of deionized water is added to the initial fermentation system, and it is placed in a 40℃ constant temperature incubator for culture for 18h.

[0088] Comparative Example 2

[0089] Preparation of Sargassum fusiforme fermentation liquid:

[0090] The difference from Example 1 is that in step S20, Saccharomyces cerevisiae is added to the initial fermentation system for fermentation, wherein the viable cell count of the Saccharomyces cerevisiae liquid is 10 9 CFU / mL, the addition amount is 3 mL, and after the bacterial liquid is inoculated, it is uniformly shaken and placed in a 40℃ constant temperature incubator for culture for 18h.

[0091] Effect Example 1

[0092] DPPH free radical scavenging experiment:

[0093] DPPH is an early-synthesized organic free radical, which is often used to evaluate the hydrogen-donating ability of antioxidants. It is very stable in organic solvents, has a purple color, and has a characteristic absorption peak at 517nm. When a free radical scavenger is encountered, the lone pair of electrons of DPPH is paired, which causes it to lose color, that is, the absorbance value at the maximum absorption wavelength decreases. Therefore, the scavenging effect of the sample on DPPH free radicals can be evaluated by measuring the change in absorbance.

[0094] The specific experimental steps of the DPPH free radical scavenging experiment are as follows:

[0095] (1) Mix equal volumes (1mL) of the test liquid and 2×10-4mol / L DPPH solution (A1 tube);

[0096] (2) Take equal volume (1 mL) of anhydrous ethanol (solvent of the sample to be tested) and 2 x 10-4mol / L DPPH solution and mix (A2 tube);

[0097] (3) Take equal volume (1 mL) of anhydrous ethanol and the sample to be tested and mix (A3 tube);

[0098] (4) After 30 min of light-free reaction, measure the absorbance values of A1 tube, A2 tube and A3 tube at 517 nm; the clearance rate calculation formula is: clearance rate = [(A2 + A3) - A1] / A2 x 100%.

[0099] The DPPH free radical clearance experiment of the samples of Examples 1-4 and Comparative Examples 1-2 diluted by 2 times was tested, and the test results are shown in Table 1, Figure 1 .

[0100] Table 1

[0101] DPPH radical scavenging rate (%) Example 1 71.19±2.23 Example 2 62.67±2.54 Example 3 57.12±1.57 Example 4 62.77±2.13 Comparative Example 1 38.37±0.74 Comparative Example 2 44±1.55

[0102] Example 2

[0103] Hydroxyl radical clearance experiment:

[0104] Hydroxyl radical (·OH) was generated by Fenton reaction, salicylic acid was added to the reaction system, ·OH reacted with salicylic acid to generate colored compound 2, 3-dihydroxybenzoic acid which had characteristic absorption at 510 nm. The fixed reaction time method was adopted, the absorbance of the reaction solution containing the measured substance was measured at 510 nm, and compared with the blank solution, the clearance effect of the measured substance on ·OH was determined. Table 2 reagents were added to the test tube in turn, 9 mmol / L FeSO4, 9 mmol / L salicylic acid ethanol solution, sample (product prepared in Examples 1-4 or Comparative Examples 1-4), appropriate amount of deionized water and 8.8 mmol / L H2O2 solution. Shake well, take out after heating at 37℃ water bath for 15 min, measure the absorbance A0 of blank control, the absorbance A of sample addition x and the background absorbance of the solution without H2O2 A x0 . When measuring A0, the reference solution is the system without hydrogen peroxide; calculate the hydroxyl radical clearance rate of each group according to the following formula, and the results are shown in Table 3, Figure 2 .

[0105] Hydroxyl radical clearance rate calculation formula:

[0106] Clearance rate = (A0 - (A x -A x0 )) / A0 x 100%.

[0107] Table 2

[0108]

[0109] Table 3

[0110] Number Hydroxyl radical scavenging rate (%) Example 1 89.07±1.82 Example 2 78.68±2.02 Example 3 79.83±2.03 Example 4 72.76±1.85 Comparative Example 1 56.72±1.74 Comparative Example 2 48.59±1.85

[0111] Effect Example 3

[0112] Protein content test experiment:

[0113] The protein content of the products prepared in Examples 1-4 and Comparative Examples 1-2 was tested using a BCA protein quantitative detection kit produced by Beijing Baiseijibio Technology Co., Ltd. with the product code BN27109, and the results are shown in Table 4. Figure 3

[0114] Table 4

[0115]

[0116]

[0117] Effect Example 4

[0118] Total sugar content test experiment:

[0119] The total sugar content of the products prepared in Examples 1-4 and Comparative Examples 1-2 was tested using a total sugar content detection kit produced by Beijing Solayebao Technology Co., Ltd. with the product code BC2715, and the results are shown in Table 5. Figure 4

[0120] Table 5

[0121] Total sugar content (mg / mL) Example 1 4.35±0.07 Example 2 3.47±0.13 Example 3 3.81±0.05 Example 4 3.15±0.13 Comparative Example 1 2.17±0.08 Comparative Example 2 2.25±0.1

[0122] From the test results of the above effect examples, it can be seen that the antioxidant activity, protein content and total sugar content of the Sargassum fusiforme fermentation broth obtained by each example are better than those of the comparative examples. At the same time, the test results of Example 1 compared with other examples and comparative examples have significant differences, which indicates that the fermentation broth obtained by using Lactobacillus helveticus and Lactobacillus plantarum to ferment Sargassum fusiforme in Example 1 has the best activity effect. Through analysis, it can be known that the fermentation broth obtained by fermenting Sargassum fusiforme using Lactobacillus is significantly better than the fermentation broth obtained by fermenting Sargassum fusiforme using yeast in Comparative Example 2.

[0123] ​​Since the mixed fermentation of Lactobacillus helveticus and Lactobacillus plantarum is different from the single fermentation, the possible reason is analyzed by the results, the DPPH free radical scavenging rate of Example 2 is higher than that of Example 3, the hydroxyl radical scavenging rate of Example 3 is slightly higher than that of Example 2, the protein content and the total sugar content are also opposite results compared with Example 2 and Example 3, so it can be found that the fermentation pathways of Lactobacillus helveticus and Lactobacillus plantarum using Sargassum siliqustrum are different, the substances and metabolites used are also different, but they can all improve the dissolution efficiency of active ingredients of Sargassum siliqustrum; and each test index in Example 1 is significantly better than that in Example 2 and 3, which shows that there is interaction between the two strains in the fermentation process, which may change the fermentation pathway by mutual competition, and the metabolites of the two may also have a certain synergistic effect, so that the obtained fermentation liquor is significantly different from the single strain fermentation of Example 2 and 3, the above is only the possible explanation of the mechanism by the applicant through the test results, and the specific mechanism needs to be further studied and explained.

[0124] Finally, it should be noted that: the above examples are only used to illustrate the technical solutions of the present application, but not to limit them; although the present application has been described in detail with reference to the foregoing examples, those skilled in the art should understand that they can still modify the technical solutions recorded in the foregoing examples, or make equivalent replacement for part or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for preparing a Sargassum fusiforme fermentation broth, characterized in that, The method comprises the following steps: S10: sterilizing the mixture of the Sargassum fusiforme raw material, the carbon source and water to obtain an initial fermentation system; S20: inoculating Lactobacillus liquid into the initial fermentation system to perform fermentation to obtain a mixed fermentation liquid; S30: separating the supernatant from the mixed fermentation liquid to obtain the Sargassum fusiforme fermentation liquid; The Lactobacillus is a mixture of Lactobacillus helveticus and Lactobacillus plantarum with a live bacteria quantity ratio of 1:

1. In the step S10, the Sargassum fusiforme raw material is dry Sargassum fusiforme powder, and the mass ratio of the dry Sargassum fusiforme powder, the carbon source and water is 1:0.1-2:10-300.

2. The production method according to claim 1, characterized by, In the step S10, the Sargassum fusiforme raw material is dry Sargassum fusiforme powder with a particle size corresponding to a 50-150 mesh sieve.

3. The preparation method according to claim 1, characterized in that, In the step S10, the carbon source is one or more of glucose, sucrose, fructose and maltose.

4. The production method according to claim 1, characterized by, The inoculation amount of the Lactobacillus liquid in the step S20 is 1% to 5%, and the viable bacterial count of the Lactobacillus liquid is 10 6 ~10 11 CFU / mL.

5. The method of claim 1, wherein, In the step S20, the fermentation conditions are as follows: the fermentation temperature is 30-45 DEG C, and the fermentation time is 12-48 h.

6. A Sargassum fusiforme fermentation broth, characterized in that, The Sargassum fusiforme fermentation liquid prepared by the preparation method according to any one of claims 1-5.

7. A cosmetic product, characterized by The Sargassum fusiforme fermentation liquid prepared by the preparation method according to any one of claims 1-5 or the Sargassum fusiforme fermentation liquid according to claim 6.

Images