A method for preparing a fruiting body mixed fermentation broth and a product and application thereof

Abstract

The application discloses a kind of fruit body mixed fermentation liquor preparation method and product and its application, including, ganoderma lucidum fruit body, Sanghuang fruit body, birch fungus fruit, respectively after drying, grinding, pass through 60-80 mesh screen, prepare ganoderma lucidum fruit body powder, Sanghuang fruit body powder, birch fungus fruit powder;Ganoderma lucidum fruit body powder, Sanghuang fruit body powder, birch fungus fruit powder are mixed to obtain fungal fruit body powder composition, add distilled water, stir to prepare fruit body mother liquor;After sterilization of fruit body mother liquor, inoculate saccharomyces cerevisiae, at 37-40 DEG C under fermentation 48-72h after, after sterilization treatment filtration, fruit body mixed fermentation liquor is obtained, the stimulating property of microorganism fermentation process is eliminated, antioxidant while removing free radical, safe and non-irritating, belong to a kind of mild, friendly cosmetic raw materials.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological fermentation, and particularly relates to a preparation method and product of a fruiting body mixed fermentation liquid and application thereof. BACKGROUND

[0002] Aging senescence, also known as aging, generally refers to an irreversible phenomenon that, under normal conditions, after a biological organism matures, its function declines with age, the stability and stress resistance of the internal environment decrease, the structure and components gradually degenerate, and the organism tends to die. The free radical theory of aging was proposed by Harman D in 1956. The theory holds that free radicals are constantly generated in the body, but there is also an effective free radical scavenging system (such as GSH and SOD) to maintain the free radicals in the body at a normal level. However, with age, this balance is gradually disrupted, resulting in an excess of free radicals. Excessive oxygen free radicals can damage cell membranes and other important components, causing protein and enzyme denaturation. When the accumulation of damage caused by free radicals overcomes the body's repair capacity, it can lead to changes in cell differentiation status, even loss, thereby accelerating aging.

[0003] With the improvement of living standards, people pay more and more attention to the aging problem of the skin. At present, synthetic antioxidants are widely used in functional foods and drugs. However, long-term use of antioxidants can have toxic effects, so people are increasingly pursuing more natural and safer antioxidants and anti-aging drugs.

[0004] Polysaccharides are essential components of all living organisms, and they are closely related to maintaining various physiological functions of life. In recent years, polysaccharides from fungi have emerged as an important type of biologically active natural products. The polysaccharides can enhance the free radical scavenging capacity and reduce the damage of free radicals to the body, thereby achieving the effect of anti-aging. At the same time, triterpenoids, as one of the main active ingredients of fungal substances, also have the effects of anti-tumor, anti-oxidation, anti-inflammatory, etc.

[0005] At present, there are fungal extracts and fungal fermentation liquid products on the market that have certain free radical scavenging capacity, but the active substances contained in the products are few, and the anti-aging effect is very limited. At the same time, there are problems such as low extraction efficiency and unverified safety. SUMMARY

[0006] This section is intended to summarize some aspects of the embodiments of the present application and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the abstract and title of the specification to avoid obscuring the purpose of this section, the abstract and the title, and such simplifications or omissions cannot be used to limit the scope of the present application.

[0007] In view of the above and / or problems existing in the prior art, the present application is proposed.

[0008] Therefore, the purpose of the present application is to overcome the deficiencies in the prior art, and provide a preparation method of fruiting body mixed fermentation liquor.

[0009] To solve the above technical problems, the present application provides the following technical scheme: a preparation method of fruiting body mixed fermentation liquor, comprising,

[0010] The ganoderma lucidum fruiting body, the phellinus igniarius fruiting body and the betulae fruiting body are dried and then crushed, and then passed through a 60-80 mesh sieve to obtain ganoderma lucidum fruiting body powder, phellinus igniarius fruiting body powder and betulae fruiting body powder;

[0011] The ganoderma lucidum fruiting body powder, the phellinus igniarius fruiting body powder and the betulae fruiting body powder are mixed to obtain a fungal fruiting body powder composition, and then distilled water is added and stirred to obtain a fruiting body mother liquor;

[0012] After the fruiting body mother liquor is sterilized, saccharomyces cerevisiae is inoculated, and then the mixture is fermented at 37-40°C for 48-72h, and then filtered after sterilization to obtain the fruiting body mixed fermentation liquor;

[0013] The mixture ratio of the ganoderma lucidum fruiting body powder, the phellinus igniarius fruiting body powder and the betulae fruiting body powder is 6:3:1.

[0014] As a preferred scheme of the preparation method, the saccharomyces cerevisiae includes saccharomyces cerevisiae Rayting-P2 (Saccharomyces cerevisiae Rayting-P2), which is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO.M 2021849.

[0015] As a preferred scheme of the preparation method, the mass ratio of the fungal fruiting body powder composition to distilled water is 1:20-1:25.

[0016] As a preferred scheme of the preparation method, the sterilization of the fruiting body mother liquor is performed at 121°C, 0.1Mpa for 30min.

[0017] As a preferred scheme of the preparation method, the inoculation of saccharomyces cerevisiae includes the following steps:

[0018] The strain is first inoculated in YPD culture medium and cultured at 37°C, 200rpm for 18-24h, and then the concentration of the bacteria is adjusted to 10 7 ~10 8 CFU / mL to obtain the activated saccharomyces cerevisiae.

[0019] As a preferred scheme of the preparation method, the YPD culture medium comprises 20.0 g of peptone, 10.0 g of yeast powder, 20.0 g of glucose, 20.0 g of agar, 1.0 L of distilled water, and pH 6.8.

[0020] As a preferred scheme of the preparation method, the Saccharomyces cerevisiae is inoculated, and the inoculation amount of the Saccharomyces cerevisiae is 10% to 15% relative to the mass percentage of the fruiting body mother liquor.

[0021] As a preferred scheme of the preparation method, the filtering comprises removing solid insoluble substances through 200 to 300 mesh gauze, and the filtrate is subjected to secondary filtration through a ceramic membrane with a filter diameter of 200 nm.

[0022] Still another object of the present application is to provide a fermentation liquid prepared by the preparation method of the fruiting body mixed fermentation liquid.

[0023] Still another object of the present application is to provide an application of the fruiting body mixed fermentation liquid as an anti-aging component in cosmetics.

[0024] The present application has the following beneficial effects:

[0025] (1) The present application provides a preparation method of a fruiting body combined fermentation liquid with anti-aging effect. After liquid submerged fermentation of the fungal fruiting body, the active ingredient content is increased, the molecular weight is reduced, and the effect is obviously enhanced. At the same time, the microbial fermentation process eliminates the irritation of the fungus, and the antioxidant effect of free radical clearance is safe and non-irritating, which is a kind of mild and friendly cosmetic raw material for the skin.

[0026] (2) The present application provides a fungal fruiting body combined fermentation liquid, which takes ganoderma lucidum fruiting body as the main raw material component, is supplemented with phellinus igniarius fruiting body and betulae fungus fruiting body, and is fermented by lactobacillus plantarum. The active components synergistically enhance the effect of free radical clearance and antioxidant, so as to achieve the purpose of anti-aging. The mass ratio of the raw materials is ganoderma lucidum fruiting body: phellinus igniarius fruiting body: betulae fungus fruiting body = 6:3:1, the polysaccharide content in the fermentation liquid is 4.84 mg / mL, the total triterpene content is 317.95 ppm, and the effect of free radical clearance and lipid peroxidation is best. BRIEF DESCRIPTION OF DRAWINGS

[0027] In order to more clearly illustrate the technical solutions of the embodiments of the present application, the drawings needed to be used in the embodiments will be briefly introduced as follows. Obviously, the drawings in the following description only constitute some embodiments of the present application, and other drawings can be obtained by those skilled in the art without any creative effort on the basis of these drawings. Among them:

[0028] Figure 1 It is a polysaccharide content comparison chart of single fermentation liquor of fungal fruiting bodies and fermentation liquor of fungal fruiting bodies in different mass ratios. Fermentation liquor 1 is Ganoderma lucidum fruiting body 100%; fermentation liquor 2 is Phellinus igniarius fruiting body 100%; fermentation liquor 3 is Inonotus obliquus fruiting body 100%; fermentation liquor 4 is Ganoderma lucidum fruiting body 50%, Phellinus igniarius fruiting body 50%; fermentation liquor 5 is Phellinus igniarius fruiting body 50%, Inonotus obliquus fruiting body 50%; fermentation liquor 6 is Ganoderma lucidum fruiting body 50%, Inonotus obliquus fruiting body 50%; fermentation liquor 7 is Ganoderma lucidum fruiting body: Phellinus igniarius fruiting body: Inonotus obliquus fruiting body = 60%: 30%: 10%; fermentation liquor 8 is Ganoderma lucidum fruiting body: Phellinus igniarius fruiting body: Inonotus obliquus fruiting body = 50%: 30%: 20%; fermentation liquor 9 is Ganoderma lucidum fruiting body: Phellinus igniarius fruiting body: Inonotus obliquus fruiting body = 40%: 20%: 40%.

[0029] Figure 2 It is a total triterpenoid content comparison chart of single fermentation liquor of fungal fruiting bodies and fermentation liquor of fungal fruiting bodies in different mass ratios.

[0030] Figure 3 It is a free radical scavenging rate comparison chart of single fermentation liquor of fungal fruiting bodies and fermentation liquor of fungal fruiting bodies in different mass ratios. A is a DPPH free radical scavenging rate comparison chart, and B is an ABTS free radical scavenging rate comparison chart.

[0031] Figure 4 It is a lipid peroxidation inhibition rate comparison chart of single fermentation liquor of fungal fruiting bodies and fermentation liquor of fungal fruiting bodies in different mass ratios.

[0032] Figure 5 It is a stability comparison chart of single fermentation liquor of fungal fruiting bodies and fermentation liquor of fungal fruiting bodies in different mass ratios after being placed at 50°C. From left to right, they are fermentation liquor 1-9.

[0033] Figure 6 It is a transdermal water loss rate test result of skin (cheek) of the essence formula of fermentation liquor 7.

[0034] Figure 7 It is a wrinkle area test result of skin (eye corner) of the essence formula of fermentation liquor 7.

[0035] Figure 8 It is a wrinkle area comparison chart of skin (eye corner) of the essence formula of fermentation liquor 7. DETAILED DESCRIPTION

[0036] In order to make the above objectives, characteristics and advantages of the present application more obvious and easy to understand, the specific embodiments of the present application are described in detail below with reference to the description examples.

[0037] In the following description, a large number of specific details are set forth in order to facilitate a thorough understanding of the present application, but the present application can also be implemented in other ways different from those described herein, and those skilled in the art can make similar generalizations without departing from the connotation of the present application, therefore the present application is not limited by the specific examples disclosed below.

[0038] Secondly, the "one embodiment" or "embodiment" referred to herein means that the specific features, structures or characteristics can be included in at least one implementation of the present application. "In one embodiment" appearing in different places in the specification does not mean the same embodiment, nor is it an embodiment that is separate or alternative to other embodiments.

[0039] In the embodiments of the present application, Ganoderma lucidum fruiting bodies, Phellinus igniarius fruiting bodies and Inonotus obliquus fruiting bodies are all ordinary commercially available products; in the embodiments of the present application, the Saccharomyces cerevisiae is Saccharomyces cerevisiae Rayting-P2 (CCTCC NO. M2021849), which is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO. M2021849, which is the same as the Saccharomyces cerevisiae CCTCC NO. M2021849 in Chinese Patent Application No. 2022117362409.

[0040] In the embodiments of the present application, the activation method of Saccharomyces cerevisiae is as follows: the glycerol frozen tube of Saccharomyces cerevisiae is taken out from the-80℃ refrigerator, and is streaked on YPD solid culture medium (yeast powder 1%, peptone 2%, glucose 2%, agar 2%, sterilized at 121℃ for 20min) under sterile operation, and is placed in a 30-degree incubator for 24h of inverted culture to grow single colonies.

[0041] The strain is first inoculated in the YPD culture medium under sterile operation, and is cultured at 37℃, 200rpm for 18-24h, and when the bacterial concentration reaches 10 7 ~10 8 CFU / mL, the activated Saccharomyces cerevisiae is obtained.

[0042] In the embodiments of the present application, filtration is carried out by removing solid insoluble substances through 200-300 mesh gauze, and the clear liquid is subjected to secondary filtration through a ceramic membrane with a filter diameter of 200nm.

[0043] Example 1

[0044] The present embodiment provides a preparation method of fruiting body single fermentation liquid, and the main steps are as follows:

[0045] (1) Take the dried fruiting bodies of Ganoderma lucidum, Phellinus igniarius and Inonotus obliquus respectively, and crush them with a crusher, then pass them through a 60-80 mesh sieve;

[0046] (2) Take the sieved fruiting body powder and add distilled water to stir it into a fruiting body mother liquor, wherein the mass ratio of the fungal fruiting body powder to distilled water is 1:20;

[0047] (3) After the mother liquor is sterilized and inoculated with microorganisms, it is fermented at 37°C for 48h, and the inoculation amount of the activated Saccharomyces cerevisiae relative to the mass percentage of the fruiting body mother liquor is 10%.

[0048] (4) After sterilization, filter to obtain fruiting body single fermentation liquor 1-3;

[0049] Among them, taking the mass of the raw materials as 100%, the fermentation liquor 1 is 100% Ganoderma lucidum fruiting body, the fermentation liquor 2 is 100% Phellinus igniarius fruiting body, and the fermentation liquor 3 is 100% Inonotus obliquus fruiting body.

[0050] Take the obtained fruiting body single fermentation liquor respectively, and determine the polysaccharide and triterpene contents.

[0051] Example 2

[0052] The present embodiment provides a fruiting body combined fermentation liquor preparation method, and the main steps are:

[0053] (1) Take the dried fruiting bodies of Ganoderma lucidum, Phellinus igniarius and Inonotus obliquus respectively, and crush them with a crusher, then pass them through a 60-80 mesh sieve;

[0054] (2) Take the sieved fruiting body powder and mix it uniformly according to a certain proportion, then add distilled water to stir it into a fruiting body mother liquor, wherein the mass ratio of the fungal fruiting body powder to distilled water is 1:20;

[0055] (3) After the mother liquor is sterilized and inoculated with microorganisms, it is fermented at 37°C for 48h, and the inoculation amount of the activated Saccharomyces cerevisiae relative to the mass percentage of the fruiting body mother liquor is 10%.

[0056] (4) After sterilization, filter to obtain fruiting body combined fermentation liquor 4-6;

[0057] Among them, taking the mass of the raw materials as 100%, the fermentation liquor 4 is 50% Ganoderma lucidum fruiting body and 50% Phellinus igniarius fruiting body; the fermentation liquor 5 is 50% Phellinus igniarius fruiting body and 50% Inonotus obliquus fruiting body; and the fermentation liquor 6 is 50% Ganoderma lucidum fruiting body and 50% Inonotus obliquus fruiting body.

[0058] Take the obtained fruiting body combined fermentation liquor respectively, and determine the polysaccharide and triterpene contents.

[0059] Example 3

[0060] The embodiment provides a preparation method of a combined fruiting body fermentation liquor, and main steps are as follows:

[0061] (1) dried ganoderma lucidum fruiting bodies, phellinus igniarius fruiting bodies and betulae fruiting bodies are broken by a pulverizer, and then sieved through a 60-80 mesh screen;

[0062] (2) sieved fruiting body powders are uniformly mixed according to a certain proportion, distilled water is added, and fruiting body mother liquor is prepared by stirring, wherein the mass ratio of the fungal fruiting body powder to the distilled water is 1:20;

[0063] (3) the mother liquor is sterilized after being divided into portions, inoculated with microorganisms, and fermented at 37 DEG C for 48 h, wherein the inoculation amount of the activated saccharomyces cerevisiae relative to the mass percentage of the fruiting body mother liquor is 10 %;

[0064] (4) the three kinds of fruiting body mixed fermentation liquor 7 is obtained after sterilization and filtration;

[0065] wherein, the ganoderma lucidum fruiting bodies account for 60 % of the total raw material, the phellinus igniarius fruiting bodies account for 30 % of the total raw material, and the betulae fruiting bodies account for 10 % of the total raw material.

[0066] The obtained fruiting body mixed fermentation liquor is taken, and the polysaccharide and triterpene contents are determined.

[0067] Embodiment 4

[0068] The embodiment provides a preparation method of a combined fruiting body fermentation liquor, and main steps are as follows:

[0069] (1) dried ganoderma lucidum fruiting bodies, phellinus igniarius fruiting bodies and betulae fruiting bodies are broken by a pulverizer, and then sieved through a 60-80 mesh screen;

[0070] (2) sieved fruiting body powders are uniformly mixed according to a certain proportion, distilled water is added, and fruiting body mother liquor is prepared by stirring, wherein the mass ratio of the fungal fruiting body powder to the distilled water is 1:20;

[0071] (3) the mother liquor is sterilized after being divided into portions, inoculated with microorganisms, and fermented at 37 DEG C for 48 h, wherein the inoculation amount of the activated saccharomyces cerevisiae relative to the mass percentage of the fruiting body mother liquor is 10 %;

[0072] the three kinds of fruiting body mixed fermentation liquor 8 is obtained after sterilization and filtration;

[0073] wherein, the ganoderma lucidum fruiting bodies account for 50 % of the total raw material, the phellinus igniarius fruiting bodies account for 30 % of the total raw material, and the betulae fruiting bodies account for 20 % of the total raw material.

[0074] The obtained fruiting body mixed fermentation liquor is taken, and the polysaccharide and triterpene contents are determined.

[0075] Embodiment 5

[0076] The embodiment provides a preparation method of a fruiting body combination fermentation liquor, and main steps are as follows:

[0077] (1) The ganoderma lucidum fruiting body, the phellinus igniarius fruiting body and the inonotus obliquus fruiting body are respectively dried, then are broken by a pulverizer, and are passed through a 60-80 mesh screen.

[0078] (2) The screened fruiting body powder is uniformly mixed according to a certain proportion, distilled water is added, and stirring is performed to prepare a fruiting body mother liquor, wherein the mass ratio of the fungus fruiting body powder to the distilled water is 1:20.

[0079] (3) The mother liquor is sterilized after being divided, is inoculated with microorganisms, is fermented at 37 DEG C for 48 h, and the inoculation amount of the activated saccharomyces cerevisiae is 10% relative to the mass percentage of the fruiting body mother liquor.

[0080] (4) After sterilization treatment, the three kinds of fruiting body mixed fermentation liquor 9 are obtained by filtration, wherein, taking the raw material mass as 100%, the ganoderma lucidum fruiting body is 40%, the phellinus igniarius fruiting body is 20%, and the inonotus obliquus fruiting body is 40%.

[0081] The obtained fruiting body mixed fermentation liquor is taken, and the polysaccharide and triterpene contents are determined.

[0082] The raw materials and mass ratios of the fermentation liquors in the embodiment 1 to the embodiment 5 are shown in table 1.

[0083] Table 1

[0084]

[0085] The polysaccharide content is determined by using a phenol-sulfuric acid method:

[0086] 1.0 mL of the fermentation liquor obtained in the above embodiment 1 to 5 is taken, 1.0 mL of 5% phenol is added, 5 mL of ice water is slowly added in an ice water bath, is shaken uniformly, is placed in a boiling water bath for 25 min, and the absorbance value is determined at 490 nm, and three parallel samples are taken for each sample.

[0087] The polysaccharide content in the fermentation liquor obtained in the embodiment 1 to 5 is determined and calculated by taking glucose as a standard (linear range 20-160 ppm), and the results are shown in Figure 1 .

[0088] From Figure 1 It can be seen that the polysaccharide content of the fermentation liquor 7 obtained in the embodiment 3 is the most.

[0089] The total triterpene content is determined by using a vanillin-glacial acetic acid-perchloric acid spectrophotometric method:

[0090] The fermentation liquor obtained in the above Examples 1-5 was respectively taken 0.5 mL into a volumetric flask, 0.4 mL of pre-cooled 5% vanillin-glacial acetic acid solution was added, and after shaking, 1 mL of perchloric acid solution was added, and after shaking, it was placed in a water bath at 60°C for 30 min, and after cooling in an ice water bath, it was diluted to 10 mL with glacial acetic acid, and after shaking, the absorbance value was measured at 550 nm by ultraviolet spectrophotometry, and 3 parallel samples were taken for each sample.

[0091] The total triterpenoid content in the fermentation liquor obtained in Examples 1-5 was determined and calculated by using oleanolic acid-methanol solution as a standard (linear range 0-20 ppm), and the results are shown in Table 1. Figure 2 .

[0092] From Figure 2 It can be seen that the total triterpenoid content of the fermentation liquor 7 obtained in Example 3 is the highest.

[0093] Example 6

[0094] The free radical scavenging ability of the fruiting body fermentation liquor in Examples 1-5 was detected:

[0095] The DPPH free radical scavenging experiment of the present application: DPPH (1,1-diphenyl-2-trinitrobenzene hydrazine) method is one of the most commonly used methods for evaluating natural antioxidants, which is fast, sensitive and simple. DPPH as a stable free radical has a maximum absorption wavelength at 517 nm, and it is scavenged after receiving electrons and protons, thereby reducing the absorbance at 517 nm.

[0096] The fermentation liquor obtained in the above Examples 1-5 was respectively dissolved into a 10% solution with deionized water, and the antioxidant activity was determined with 8 ppm of VC as a positive control. The DPPH free radical scavenging rate was calculated.

[0097] DPPH scavenging rate % = [1-(A s -A sb ) / (A c -A cb )] x 100%

[0098] Wherein A s is the absorbance of the sample group; A sb为 is the absorbance of the sample control group; A c is the absorbance of the blank group, i.e. using purified water instead of the sample; and A cb is the absorbance of the blank control group.

[0099] The results are shown in Table 2. Figure 3 From Figure 3 A can be seen, the DPPH free radical inhibition rate of the fermentation liquor 7 of the fungal fruiting body combination obtained in Example 3 is higher, which indicates that the antioxidant effect of the fermentation liquor 7 of the fungal fruiting body combination obtained in Example 3 is better.

[0100] ABTS radical scavenging experiment of the present application: ABTS [2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] method is first used to determine the total antioxidant capacity of biological samples, which is time-saving, low-cost and highly relevant to antioxidants. The fermentation broth obtained in Examples 1-5 above is dissolved in deionized water to form a 5% solution, and 10 ppm VC is used as a positive control for antioxidant activity determination. The ABTS radical scavenging rate is calculated.

[0101] ABTS radical scavenging rate % = (1-A s / A c ) x 100%

[0102] Wherein A s is the absorbance of the sample group; A c is the absorbance of the blank group, i.e. using purified water instead of the sample.

[0103] The results are shown in Figure 3 , from Figure 3 B, it can be seen that the ABTS radical inhibition rate of the fermentation broth 7 of the fungal fruiting body combination obtained in Example 3 is higher, indicating that the antioxidant effect of the fermentation broth 7 of the fungal fruiting body combination obtained in Example 3 is better.

[0104] Example 7

[0105] The fruiting body fermentation broth in Examples 1-5 is used for anti-lipid peroxidation capacity detection.

[0106] The anti-lipid peroxidation capacity detection of the present application uses the thiobarbituric acid method: Harman D's free radical aging theory believes that aging is caused by the accumulation of damage to cells, especially biological membranes and genetic molecules, caused by free radicals and free radicals in the body. The sebaceous glands secrete oil, and the oil is easily oxidized by external smog, ultraviolet rays, etc., stimulates the skin, causes inflammation, acne, etc., and long-term inflammation can also cause aging. The thiobarbituric acid method is based on the oxidation of unsaturated fatty acids by free radical reaction to form oxidized radicals to form epoxy compounds, which decompose to form malondialdehyde (MDA), which reacts with thiobarbituric acid (TBA) to form a TBA dye complex. This compound has a maximum absorption value at 532 nm. The test results are usually expressed as the product of the absorbance at 532 nm and the malondialdehyde absorbance coefficient, i.e. the commonly used TBA value.

[0107] The specific test method of lipid peroxidation is as follows: the fermentation broth obtained in Examples 1-5 is dissolved into a 5% solution with deionized water, 200 μL of phosphate buffer solution (50 mM, pH 7.4), 200 μL of 2% lecithin solution, 200 μL of 5% fermentation broth, and 1 mL of 5 mM EDTA-Fe (II) are sequentially added into a sample tube, and the sample tube is incubated in a 37°C water bath for 45 min. Then, 400 μL of TBA mixture is added into the solution, and after mixing, the solution is heated in boiling water for 15 min, and immediately cooled with an ice water bath after being taken out. After being fully cooled, centrifugation is performed, and the supernatant is taken, and the absorbance is measured at 535 nm, with 3 parallel samples. The positive control is 2500 μmol / L of VC.

[0108] Inhibition rate % = [1-(A-B) / (C-B)] x 100%

[0109] Wherein A is the absorbance of the sample group; C is the absorbance of the sample control group, i.e. the sample is replaced with its solvent; and B is the absorbance of the blank group, i.e. without adding EDTA-Fe (II) based on the control group.

[0110] The results are shown in Table 1. Figure 4 From Table 1, it can be seen that the fermentation broth of the fungus fruiting body combination of Example 3 has a higher lipid peroxidation inhibition rate. Figure 4

[0111] The fermentation broth of the fungus fruiting body, the active substance content, and the anti-aging efficacy of Examples 1-5 are shown in Table 2.

[0112] Table 2

[0113]

[0114]

[0115] Example 8

[0116] The present application provides a fungus fruiting body combination fermentation broth in the application of essence:

[0117] The essence formula is shown in Table 3.

[0118] Table 3

[0119] Raw material name Addition amount (%) Water 82.68 Fermented liquid 7 of mushroom fruit body combination 10.00 Acrylamidodimethylammonium taurate / VP copolymer 0.30 Glycerin 3.00 Butylene glycol 3.00 Sodium hyaluronate 0.02 Trehalose 0.50 Phenoxyethanol, ethylhexylglycerin 0.50

[0120] Example 9

[0121] The essence in Example 8 is taken for anti-aging efficacy evaluation, including human testing and self-evaluation of the subjects.

[0122] The human testing of the anti-aging efficacy of the present application is the influence of the essence in Example 8 on the relative change rate of transdermal water loss of the skin (cheek) and the wrinkle area of the skin (eye corner). ​

[0123] Ten subjects, 5 males and 5 females, who met the test conditions were recruited. After facial cleansing, the subjects evenly applied the same amount of product to the face. The test was conducted on the 7th, 14th, 21st and 28th day. The test environment was controlled at a temperature of 21±1℃ and a relative humidity of 50±5%.

[0124] The relative change rate of transdermal water loss was detected by a skin moisture tester, and the wrinkle area was detected by a facial image analyzer. During the test, a questionnaire was issued to complete the self-evaluation of the subjects.

[0125] The test results of the effect of the essence in Example 8 on the transdermal water loss rate of the skin (cheek) and the wrinkle area reduction rate of the skin (crows' feet) are shown in Table 3. Figures 6-8 During the 28-day test, the relative change rate of transdermal water loss of the skin showed a downward trend, indicating that the essence can repair the skin barrier; at the same time, the wrinkle area of the skin (crows' feet) showed a decreasing trend and was lower than that before using the essence, indicating that the essence can effectively improve the skin wrinkle condition. The results of the consumer subjective evaluation showed that 97% of the subjects believed that the skin looked more radiant, 90% of the subjects believed that the skin felt more elastic, and 94% of the subjects believed that the skin looked more youthful. Most of the subjects believed that the product set helped to even out the skin tone, tighten the skin and refine the pores, and were satisfied with the effect of the test sample.

[0126] It can be seen that the essence in Example 8 has anti-aging effect. The fungal fruit body combined fermentation liquor provided by the present application has good effect in the essence formula.

[0127] The present application provides a kind of fungal fruit body combined fermentation liquor, with Ganoderma lucidum fruit body as main raw material component, with Morchella esculenta and Fomes fomentarius fruit body, fermented by Lactobacillus plantarum, synergistic effect, can realize multiple target point clearance free radical and antioxidant, to reach the purpose of anti-aging.The present application provides a kind of fungal fruit body combined fermentation liquor, preferably the mass ratio of fruit body in raw material is Ganoderma lucidum fruit body: Morchella esculenta: Fomes fomentarius fruit body = 6:3:1, the polysaccharide content in fermentation liquor is 4.84mg / mL, the total triterpene content is 317.95ppm, the active substance content in the fermentation liquor obtained by this ratio is the most, and the effect of clearing free radicals and resisting lipid peroxidation is the best.

[0128] The fungal fruit body combined fermentation liquor provided by the present application can clear free radicals, is safe and non-irritating, has low cost, and has brownish red color, and is suitable for the field of cosmetics.

[0129] It should be noted that the above examples are only used to illustrate the technical solutions of the present application and are not limiting. Although the present application has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present application can be modified or replaced equivalently without departing from the spirit and scope of the present application, and they should be covered in the scope of the present application.

Claims

1. A method for preparing a fruiting body mixed fermentation broth, characterized by comprising the following steps: Comprising, ​ Ganoderma lucidum fruiting body, Phellinus igniarius fruiting body, Inonotus obliquus fruiting body are dried and then ground into powder through 60-80 mesh sieve to obtain Ganoderma lucidum fruiting body powder, Phellinus igniarius fruiting body powder and Inonotus obliquus fruiting body powder; Ganoderma lucidum fruiting body powder, Phellinus igniarius fruiting body powder and Inonotus obliquus fruiting body powder are mixed to obtain a fungal fruiting body powder composition, and distilled water is added to obtain a fruiting body mother liquor by stirring, wherein the mass ratio of the fungal fruiting body powder composition to distilled water is 1:20-1:25; After sterilization of the fruiting body mother liquor, Saccharomyces cerevisiae is inoculated, and after fermentation at 37-40℃ for 48-72h, the fruiting body mixed fermentation liquor is obtained by sterilization and filtration, wherein the sterilization conditions of the fruiting body mother liquor are 121℃, 0.1Mpa, 30min, the inoculation amount of Saccharomyces cerevisiae is 10%-15% of the mass percentage of the fruiting body mother liquor, and the filtration includes removing solid insoluble substances through 200-300 mesh gauze, and the clear liquid is filtered again through a ceramic membrane with a filter diameter of 200nm; The mixing ratio of Ganoderma lucidum fruiting body powder, Phellinus igniarius fruiting body powder and Inonotus obliquus fruiting body powder is 6:3:

1. The Saccharomyces cerevisiae is Saccharomyces cerevisiae Rayting-P2 (CCTCC NO.M 2021849) preserved in China Center for Type Culture Collection. Saccharomyces cerevisiae Rayting-P2 (CCTCC NO.M 2021849) preserved in China Center for Type Culture Collection. The yeast is activated Saccharomyces cerevisiae, and the activation method comprises the following steps: inoculating the strain in YPD culture medium, culturing at 37℃ and 200 rpm for 18-24 hours, and obtaining the activated Saccharomyces cerevisiae when the bacterial concentration reaches 10 7 ~10 8 CFU / mL. The YPD culture medium comprises the following components: 20.0 g of proteose peptone, 10.0 g of yeast powder, 20.0 g of glucose, 20.0 g of agar, 1.0 L of distilled water, and pH 6.

8.

2. The fruiting body mixed fermentation liquor prepared by the preparation method of claim 1.

3. Use of the fruiting body mixed fermentation liquor of claim 2 as an anti-aging component in cosmetics.

Images