Preparation method and application of cordyceps sinensis fermentation product

Abstract

The invention relates to the technical field of daily chemicals, in particular to a preparation method and application of a cordyceps sinensis fermentation product. The cordyceps sinensis fermentation product is obtained by fermenting cordyceps sinensis flowers serving as a solid culture medium and lactic acid bacteria and saccharomyces cerevisiae serving as strains. According to the cordyceps sinensis probiotic fermentation composition provided by the invention, a composite solid-liquid two-stage fermentation system taking the cordyceps sinensis flowers as the raw material is constructed, and the lactic acid bacteria and the saccharomycetes are cooperatively applied for metabolic conversion, so that the content and the stability of active substances such as cordycepin, cordyceps polysaccharide, adenosine, small molecule peptide and amino acid in a fermentation product are remarkably improved; meanwhile, good skin adaptability and a skin care effect are endowed.

Description

Technical Field

[0001] The present invention relates to the field of daily chemical technology, and particularly to a preparation method and application of a cordyceps fermentation product. Background Art

[0002] Cordyceps is a complex that combines the properties of insects and fungi and is regarded as a rare traditional Chinese medicine with various effects such as nourishing the body, strengthening the lungs, relieving cough, and enhancing immunity in traditional Chinese medicine. In recent years, the artificial cultivation technology of cordyceps has gradually matured. In particular, artificially cultivated cordyceps represented by cordyceps flower (i.e., the fruiting body of cordyceps militaris) has been widely used in the fields of health care, food, and cosmetics due to its high nutritional value and rich active ingredients.

[0003] Cordyceps flower is rich in various natural bioactive components such as cordycepin, cordyceps polysaccharide, adenosine, mannitol, and amino acids. These components have significant antioxidant, anti-inflammatory, antibacterial, and immunomodulatory activities, and can improve the skin condition and enhance the skin barrier function to a certain extent. However, the natural content of active ingredients in cordyceps is limited, and traditional extraction processes often have problems such as low efficiency, poor stability, and insufficient efficacy expression, making it difficult to meet the requirements of high-efficiency functional skin care products for the content and stability of active substances.

[0004] As a natural and controllable biotransformation method, probiotic fermentation technology has been widely used in recent years to enhance the biological activity of plant or animal-derived extracts. Through the directional fermentation of specific strains, not only can the closed active ingredients in raw materials be released, but also functional metabolites such as bioactive peptides, short-chain fatty acids, hyaluronic acid, and lactic acid can be produced, thereby synergistically enhancing the anti-inflammatory, repair, and barrier-enhancing abilities of raw materials.

[0005] Based on this, researchers have tried to apply functional probiotics such as lactic acid bacteria and yeasts to the fermentation system of cordyceps flower, with the expectation of enriching its key active substances and endowing it with new functional characteristics through two-stage, multi-strain complex fermentation. Further, combined with plant enzyme hydrolysates or traditional Chinese medicine plants (such as wolfberry, astragalus, etc.) for compound fermentation, it helps to introduce auxiliary components such as natural polysaccharides, small molecule peptides, and amino acids, thereby achieving synergistic effects and improving the skin adaptability and activity performance of cordyceps fermentation products.

[0006] Therefore, developing a cordyceps probiotic composite fermentation composition and its preparation method, by optimizing the fermentation process and component formulation, to improve the content and stability of effective active substances from the source, has important application value, especially showing broad application prospects in the research and development of high-functional skin care products. Summary of the Invention

[0007] In order to solve the above technical problems, the present invention provides a method for preparing a Cordyceps fermentation product, wherein the Cordyceps fermentation product is obtained by fermenting Cordyceps flowers as a solid culture medium with lactic acid bacteria and Saccharomyces cerevisiae as strains; the steps of the preparation method are as follows: (1) Crush the dried Cordyceps flowers to a solid-liquid ratio of (1.6-2.5) ml: (0.5-1.5) g, mix the crushed Cordyceps residue with the solution, put it into a container and sterilize it, and the total volume of the solid culture medium occupies 10%-15% of the entire container; (2) Inoculating lactic acid bacteria into cordyceps solid culture medium: inoculate lactic acid bacteria into cordyceps solid culture medium at an addition rate of 3wt% to 5wt%, stir the bacterial liquid and solid culture medium evenly, and culture anaerobically at 37°C for 48 to 72 hours; (3) solid fermentation: adding a sterile liquid culture medium to the above Cordyceps solid culture medium, adding 3wt% to 5wt% of Saccharomyces cerevisiae according to the total volume, and culturing under microaerobic conditions at 28°C for 24 to 48 hours; The formula of the sterile liquid culture medium is: 0.6 parts of peptone, 85-90 parts of water, sterilized at 115°C for 20 minutes, and added to the cordyceps solid culture medium after cooling; (4) Liquid fermentation: After the fermentation is completed, the solution is quickly heated to 72-75°C using a pasteurization method for 15-30 minutes, and then immediately cooled to 4-5°C. The fermentation solution is then filtered using an 8-10 μm filter plate to obtain Cordyceps fermentation liquid.

[0008] The second aspect of the present invention provides a Cordyceps probiotic fermentation composition, which is prepared by the preparation method.

[0009] The third aspect of the present invention provides a soothing and moisturizing facial cream, wherein the raw materials for preparing the facial cream include the Cordyceps probiotic fermentation composition.

[0010] As an embodiment of the present invention, the raw materials for preparing the facial cream include, in parts by mass: Phase A: Water 62.77; Allantoin 0.20; Lubrezia gum 1.00; Water-soluble silicone wax 2.00; Glycerin 4.00; Sodium hyaluronate 0.03; Phase B: ammonium acryloyldimethyltaurate / VP copolymer 0.50; sodium stearoyl glutamate 0.20; M68 0.50; ARLACEL 165 0.80; Phase C: OLIVEM1000 2.00; Cetearyl Alcohol 2.00; Behenyl Alcohol 0.50; Ethylhexyl Palmitate 4.00; Dimethicone 1.00; Shea Butter 1.00; Caprylic / Capric Triglyceride 4.00; Phase D: Vaseline 0.30; Tocopheryl Acetate 0.20; Phytosteryl Isostearate 2.00; Cyclomethicone 2.00; DC1403 1.00; 1,3-Butanediol 3.00; p-Hydroxyacetophenone 0.50; 1,2-Hexanediol 0.50; β-Glucan 2.00; the Cordyceps probiotic fermentation composition 2.00.

[0011] As an embodiment of the present invention, the steps of the preparation method of a soothing and moisturizing cream are as follows: ①Disperse Phase A well, heat it to about 85°C, and homogenize or stir to dissolve evenly; ②Heat Phase B to about 85°C, and homogenize or stir to dissolve evenly; ③Add Phase A to Phase B, homogenize for about 2 minutes, add Phase C and Phase D, continue to homogenize for about 5 minutes, and stir to cool down; ④Cool down to about 50°C, add Phase E and Phase F, stir evenly, and discharge.

[0012] Adopting the above technical solution, the present invention has the following beneficial effects: The Cordyceps probiotic fermentation composition provided by the present invention constructs a composite solid-liquid two-stage fermentation system with Cordyceps flower as the raw material, and synergistically applies lactic acid bacteria and yeast for metabolic transformation, significantly improving the content and stability of active substances such as cordycepin, cordyceps polysaccharide, adenosine, small molecule peptides, and amino acids in the fermentation product, and at the same time endowing it with good skin adaptability and skin care efficacy.

[0013] The technical solution of the present invention has the following beneficial effects: High enrichment efficiency of active ingredients: Through the metabolic action of probiotics and the synergistic enzymatic hydrolysis of plant coenzymes, the release rate and conversion efficiency of functional factors such as cordycepin, adenosine, and polysaccharide are significantly improved. Compared with traditional water extraction or single-strain fermentation methods, the increase in key components can reach 1.3 - 3.5 times.

[0014] Synergistic enhancement of efficacy: Lactic acid bacteria and yeast produce active metabolites (such as lactic acid, hyaluronic acid, glutathione) during the fermentation process, and synergistically play comprehensive skin care effects such as soothing, repairing, anti-inflammatory, antioxidant, and enhancing the barrier with the active ingredients of Cordyceps itself.

[0015] Better system stability: This fermentation system combines the advantages of solid-state pre-fermentation and liquid-state subsequent fermentation, can effectively control the risk of contamination by miscellaneous bacteria, and forms a microcolloid structure containing natural polysaccharides and mucopolysaccharide polymers, making the final product easier to emulsify and having higher stability, suitable for application in skin care systems such as creams and essences.

[0016] Wide application adaptability: The fermentation product is mild, safe, and non-irritating. The pH is stably maintained between 5.0 and 6.0, with good skin tolerance. It is especially suitable for sensitive skin, post-operative repair, barrier-damaged skin, etc., expanding its application scope in functional cosmetics. Brief Description of the Drawings

[0017] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0018] Figure 1 It is the cordyceps fermentation broth provided by Example 1 of the present invention. Detailed Embodiments

[0019] The present invention provides a cordyceps probiotic fermentation composition, which is obtained by fermenting cordyceps flowers as a solid medium with lactic acid bacteria and Saccharomyces cerevisiae as strains.

[0020] As an embodiment of the present invention, the preparation method of the cordyceps probiotic fermentation composition is as follows: 1. Preparation of cordyceps solid medium: Crush the dried cordyceps flowers, and mix the crushed cordyceps residues with a solution at a solid-liquid ratio of (1.6 - 2.5):(0.5 - 1.5) ml / g. Load them into a flask or fermenter for sterilization. The total volume of the solid medium accounts for 10% - 15% of the entire container.

[0021] Among them, the solution is a plant enzyme hydrolysate, and the specific preparation method of the plant enzyme hydrolysate is as follows: (1) Crush Codonopsis pilosula, and add deionized water at a solid-liquid ratio of 1:12 g / ml; pre-soak in a water bath at 60 - 70 °C for 30 min to pre-soften the cell wall.

[0022] (2) Cool down to 45 - 50 °C, adjust the pH to 5.5 - 6.0 with citric acid buffer, and then add neutral protease: 0.2% (w / w), cellulase: 0.2% (w / w), pectinase: 0.1% (w / w), and gently stir and react for 6 hours.

[0023] (3) Heat in a water bath at 85 °C for 10 minutes to terminate the enzyme activity; filter and take the liquid, and cool it to 35 - 37 °C for standby.

[0024] 2. Inoculate lactic acid bacteria into the cordyceps solid medium: Inoculate Lactobacillus plantarum into the Cordyceps solid medium at an addition amount of 3 wt% - 5 wt%, stir evenly and anaerobically culture at 37 °C for 48 - 72 h.

[0025] Add the above Cordyceps solid medium into the sterile liquid medium, add 3 wt% - 5 wt% of Saccharomyces cerevisiae according to the total volume and culture under microaerobic conditions at 28 °C for 24 - 48 h.

[0026] The formula of the sterile liquid medium is: 0.6 parts of peptone, 85 - 90 parts of plant enzyme hydrolysate, sterilize at 115 °C for 20 min, and add it to the Cordyceps solid medium after cooling.

[0027] After the fermentation is completed, quickly heat to 72 - 75 °C using pasteurization for 15 - 30 min, then immediately cool to 4 - 5 °C, then filter the fermentation solution using an 8 - 10 μm filter plate, and then filter with a 0.22 μm PES membrane to obtain the Cordyceps fermentation broth.

[0028] The present invention also provides a soothing and moisturizing cream. According to mass parts, the raw materials for preparing the cream include: Phase A: 62.77 of water; 0.20 of allantoin; 1.00 of lubrajel; 2.00 of water-soluble silicon wax; 4.00 of glycerol; 0.03 of sodium hyaluronate; Phase B: 0.50 of acryloyldimethyltaurine ammonium / VP copolymer; 0.20 of sodium stearoyl glutamate; 0.50 of M68; 0.80 of ARLACEL165; Phase C: 2.00 of OLIVEM1000; 2.00 of cetearyl alcohol; 0.50 of behenyl alcohol; 4.00 of ethylhexyl palmitate; 1.00 of polydimethylsiloxane; 1.00 of shea butter; 4.00 of caprylic / capric triglyceride; Phase D: 0.30 of petrolatum; 0.20 of tocopheryl acetate; 2.00 of phytosterol isostearate; 2.00 of cyclic dimethyl silicone oil; 1.00 of DC1403; 3.00 of 1,3 - butanediol; 0.50 of p - hydroxyacetophenone; 0.50 of 1,2 - hexanediol; 2.00 of β - glucan; 2.00 of the Cordyceps probiotic fermentation composition.

[0029] As a more preferred embodiment of the present invention, according to mass parts, the raw materials for preparing the cream include: Phase A: 62.77 of water; 0.20 of allantoin; 1.00 of lubrajel; 2.00 of water-soluble silicon wax; 4.00 of glycerol; 0.03 of sodium hyaluronate; Phase B: Ammonium acryloyldimethyltaurate / VP copolymer 0.30; Sodium stearoyl glutamate 0.20; M68 0.50; ARLACEL 165 0.40; C14-22 alcohols 0.50; C12-20 alkyl glucoside 0.50; PEG-20 stearate 0.40; Polyglycerol-6 stearate 0.20; Phase C: OLIVEM 1000 1.00; Cetearyl alcohol 2.00; Behenyl alcohol 0.50; Ethylhexyl palmitate 4.00; Polydimethylsiloxane 1.00; Shea butter 1.00; Caprylic / capric triglyceride 4.00; Phase D: Petrolatum 0.30; Tocopheryl acetate 0.20; Phytosteryl isostearate 2.00; Cyclomethicone 2.00; DC 1403 1.00; 1,3-Butanediol 3.00; p-Hydroxyacetophenone 0.50; 1,2-Hexanediol 0.50; β-Glucan 2.00; The cordyceps probiotic fermentation composition 2.00.

[0030] As an embodiment of the present invention, the steps of a preparation method of a soothing and moisturizing cream are as follows: ①Disperse Phase A well and heat it to about 85°C, homogenize or stir to dissolve evenly; ②Heat Phase B to about 85°C, homogenize or stir to dissolve evenly; ③Add Phase A to Phase B, homogenize for about 2 minutes, add Phase C and Phase D, continue to homogenize for about 5 minutes, and stir to cool down; ④Cool down to about 50°C, add Phase E (pre-melted) and Phase F, stir evenly, and discharge.

[0031] The present invention selects cordyceps flowers as the fermentation substrate, which can utilize the combined action of hydration and fermentation enzymes, facilitating the loosening of cell structures, making it easier for lactic acid bacteria and yeast to invade and secrete enzymes for the substrate; thus achieving more sufficient release; at the same time, using Saccharomyces cerevisiae and an acidic environment can increase the free degree of cordycepin. If a commercial solid medium is used, there will be exogenous nutrients to assist in decomposition, but it will form competitive substrates, and there is no need to introduce impurities such as rice flour / bran, which is cleaner and more focused on the release of the active product, but instead inhibits the release of cordycepin; at the same time, it avoids focusing the strains on growth but rather on metabolizing non-cordyceps source substrates.

[0032] Meanwhile, when preparing the solid medium, the added enzymolysis solution contains small molecules such as oligosaccharides, monosaccharides, polyphenols, and amino acids; these components can be preferentially utilized by lactic acid bacteria or yeast, activating metabolic pathways (glycolysis, TCA cycle); thereby promoting the release and transformation of endogenous polysaccharides and cordycepin in Cordyceps flowers. The enzymolysis solution often contains residues of plant proteases and polysaccharidases; it can assist in degrading the Cordyceps cell wall and increasing the dissolution rate of the inclusion. The plant enzymolysis solution is rich in antioxidant factors such as polyphenols, flavonoids, and VC; it can reduce the degradation of easily oxidized components such as cordycepin during fermentation and increase the retention rate. It contains a high proportion of water-soluble polysaccharides; after fermentation, these polysaccharides are restructured (oligosaccharification, branch chain breakage), the total yield and functionality are enhanced; at the same time, Cordyceps polysaccharides are co-extracted, and the overall polysaccharide content increases significantly.

[0033] The following further explains and illustrates the present invention in combination with specific embodiments.

[0034] Example 1 This example provides a Cordyceps probiotic fermentation composition, and the preparation method of the Cordyceps probiotic fermentation composition is as follows: 1. Prepare the Cordyceps solid medium: Crush the dried Cordyceps flowers, mix the crushed Cordyceps residues with a solution at a material-liquid ratio of 2:1 ml / g, load them into a flask or fermenter for sterilization, and the total volume of the solid medium accounts for 15% of the entire container.

[0035] The solution is a plant enzymolysis solution, and the specific preparation method of the plant enzymolysis solution is as follows: (1) Crush Codonopsis pilosula, add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a 60°C water bath for 30 min to pre-soften the cell wall.

[0036] (2) Cool down to 45°C, adjust the pH to 6.0 with citric acid buffer, and then add neutral protease: 0.2% (w / w), cellulase: 0.2% (w / w), pectinase: 0.1% (w / w), and stir gently for 6 hours.

[0037] (3) Heat in an 85°C water bath for 10 minutes to terminate the enzyme activity; filter to obtain the liquid and cool it to 35 for standby.

[0038] 2. Inoculate lactic acid bacteria into the Cordyceps solid medium: Inoculate Lactobacillus plantarum into the Cordyceps solid medium at an addition amount of 3 wt%, stir evenly and anaerobically culture at 37°C for 72 h.

[0039] 3. Add the above Cordyceps solid medium to the sterile liquid medium (volume ratio 9:1), and add 5 wt% Saccharomyces cerevisiae according to the total volume and culture at 28°C under microaerobic conditions for 36 h.

[0040] The formula of the sterile liquid culture medium is as follows: 0.6 parts of peptone, 85 parts of plant enzyme hydrolysate, sterilized at 115°C for 20 minutes, and added to the Cordyceps solid culture medium after cooling.

[0041] 4. After the fermentation is completed, use pasteurization to quickly heat to 72°C and keep it for 15 minutes, then immediately cool to 4°C, then filter the fermentation solution with an 8μm filter plate, and then filter it with a 0.22μm PES membrane to obtain the Cordyceps fermentation broth.

[0042] This example also provides a soothing and moisturizing cream. According to mass parts, the raw materials for preparing the cream include: Phase A: 62.77 of water; 0.20 of allantoin; 1.00 of lubrajel; 2.00 of water-soluble silicone wax; 4.00 of glycerol; 0.03 of sodium hyaluronate; Phase B: 0.30 of acryloyldimethyltaurate / VP copolymer; 0.20 of sodium stearoyl glutamate; 0.50 of M68; 0.40 of ARLACEL165; 0.50 of C14-22 alcohol; 0.50 of C12-20 alkyl glucoside; 0.40 of PEG-20 stearate; 0.20 of polyglyceryl-6 stearate; Phase C: 1.00 of OLIVEM1000; 2.00 of cetearyl alcohol; 0.50 of behenyl alcohol; 4.00 of ethylhexyl palmitate; 1.00 of polydimethylsiloxane; 1.00 of shea butter; 4.00 of caprylic / capric triglyceride; Phase D: 0.30 of petrolatum; 0.20 of tocopheryl acetate; 2.00 of phytosteryl isostearate; 2.00 of cyclic dimethyl silicone oil; 1.00 of DC1403; 3.00 of 1,3-butanediol; 0.50 of p-hydroxyacetophenone; 0.50 of 1,2-hexanediol; 2.00 of β-glucan; 2.00 of the Cordyceps probiotic fermentation composition described above.

[0043] The steps for a method of preparing a soothing and moisturizing cream are as follows: ① Disperse Phase A well, heat to about 85°C, and homogenize or stir to dissolve evenly; ② Heat Phase B to about 85°C, and homogenize or stir to dissolve evenly; ③ Add Phase A to Phase B, homogenize for about 2 minutes, add Phase C and Phase D, continue to homogenize for about 5 minutes, and stir to cool down; ④ Cool down to about 50°C, add Phase E (pre-melted) and Phase F, stir evenly, and discharge.

[0044] Figure 1 This is the Cordyceps fermentation broth provided by the embodiment of the present invention.

[0045] Example 2 The difference between this embodiment and Embodiment 1 is that this embodiment provides a cordyceps probiotic fermentation composition, and the preparation method of the cordyceps probiotic fermentation composition is as follows: 1. Prepare the cordyceps solid medium: Crush the dried cordyceps flowers, mix the crushed cordyceps residue with a solution at a material-liquid ratio of 2:1 ml / g, put it into a flask or fermenter for sterilization, and the total volume of the solid medium accounts for 15% of the whole container.

[0046] The solution is distilled water.

[0047] 2. Inoculate lactic acid bacteria into the cordyceps solid medium: Inoculate Lactobacillus plantarum into the cordyceps solid medium at an addition amount of 3 wt%, stir evenly and anaerobically culture at 37°C for 72 h.

[0048] 3. Add the sterile liquid medium to the above cordyceps solid medium (volume ratio 9:1), add 5 wt% of Saccharomyces cerevisiae according to the total volume and culture at 28°C under microaerobic conditions for 36 h.

[0049] The formula of the sterile liquid medium is: 0.6 parts of peptone, 85 parts of distilled water, sterilize at 115°C for 20 min, and add it to the cordyceps solid medium after cooling.

[0050] 4. After the fermentation is completed, quickly heat to 72°C using the pasteurization method, keep it for 15 min, then immediately cool to 4°C, then filter the fermentation solution with an 8-μm filter plate, and then filter with a 0.22-μm PES membrane to obtain the cordyceps fermentation broth.

[0051] Embodiment 3 The difference between this embodiment and Embodiment 1 is that this embodiment provides a cordyceps probiotic fermentation composition, and the preparation method of the cordyceps probiotic fermentation composition is as follows: 1. Prepare the cordyceps solid medium: Crush the dried cordyceps flowers, mix them with wheat bran at a ratio of 1:1, mix the crushed cordyceps residue with a solution at a material-liquid ratio of 2:1 ml / g, put it into a flask or fermenter for sterilization, and the total volume of the solid medium accounts for 15% of the whole container.

[0052] The solution is a plant enzyme hydrolysate, and the specific preparation method of the plant enzyme hydrolysate is as follows: (1) Crush Codonopsis pilosula, add deionized water at a material-liquid ratio of 1:12 g / ml; pre-soak in a water bath at 60°C for 30 min to pre-soften the cell wall.

[0053] (2)Cool down to 45°C, adjust the pH to 6.0 with citric acid buffer, and then add neutral protease: 0.2% (w / w), cellulase: 0.2% (w / w), pectinase: 0.1% (w / w), and stir gently for 6 hours.

[0054] (3)Heat in a water bath at 85°C for 10 minutes to terminate the enzyme activity; filter to obtain the liquid and cool it to 35 for standby.

[0055] 2. Inoculate the cordyceps solid medium with lactic acid bacteria: Inoculate Lactobacillus plantarum into the cordyceps solid medium at an addition amount of 3 wt%, stir evenly, and anaerobically culture at 37°C for 72 h.

[0056] 3. Add the above cordyceps solid medium to the sterile liquid medium (volume ratio 9:1), and add 5 wt% Saccharomyces cerevisiae according to the total volume and culture at 28°C under microaerobic conditions for 36 h.

[0057] The formula of the sterile liquid medium is: 0.6 parts of peptone, 85 parts of plant enzymolysis solution, sterilize at 115°C for 20 min, and add it to the cordyceps solid medium after cooling.

[0058] 4. After the fermentation is completed, quickly heat to 72°C using the pasteurization method for 15 min, then immediately cool to 4°C, then filter the fermentation solution using an 8μm filter plate, and then filter with a 0.22μm PES membrane to obtain the cordyceps fermentation broth.

[0059] Example 4 The difference between this example and Example 1 is that this example provides a cordyceps probiotic fermentation composition, and the preparation method of the cordyceps probiotic fermentation composition is as follows: 1. Crush the dried cordyceps flowers, inoculate Lactobacillus plantarum into the liquid medium at an addition amount of 3 wt% (the mass ratio of the liquid medium to the cordyceps flowers is 10:1), stir evenly, anaerobically culture at 37°C for 72 h, and add 5 wt% Saccharomyces cerevisiae according to the total volume and culture at 28°C under microaerobic conditions for 36 h.

[0060] The formula of the liquid medium is: 0.6 parts of peptone, 85 parts of plant enzymolysis solution, sterilize at 115°C for 20 min, and use it in the medium after cooling.

[0061] 2. After the fermentation is completed, quickly heat to 72°C using the pasteurization method for 15 min, then immediately cool to 4°C, then filter the fermentation solution using an 8μm filter plate, and then filter with a 0.22μm PES membrane to obtain the cordyceps fermentation broth.

[0062] Example 5 The difference between this embodiment and Embodiment 1 is that this embodiment provides a cordyceps probiotic fermentation composition, and the preparation method of the cordyceps probiotic fermentation composition is as follows: 1. Prepare the cordyceps solid medium: Crush the dried cordyceps flowers, with a material-liquid ratio of 2:1 ml / g, and then add codonopsis powder, and the mass ratio of the codonopsis powder to the cordyceps flower powder is 1:24; mix the crushed cordyceps residue with a solution, put it into a flask or fermenter for sterilization, and the total volume of the solid medium accounts for 15% of the whole container. The solution is deionized water.

[0063] 2. Inoculate lactic acid bacteria into the cordyceps solid medium: Inoculate Lactobacillus plantarum into the cordyceps solid medium at an addition amount of 3 wt%, stir evenly and anaerobically culture at 37 °C for 72 h.

[0064] 3. Add the above-mentioned cordyceps solid medium to the sterile liquid medium (volume ratio 9:1), and add 5 wt% of Saccharomyces cerevisiae according to the total volume and culture at 28 °C under microaerobic conditions for 36 h.

[0065] The formula of the sterile liquid medium is: 0.6 parts of peptone, 85 parts of deionized water, sterilize at 115 °C for 20 min, and add it to the cordyceps solid medium after cooling.

[0066] 4. After the fermentation is completed, quickly heat to 72 °C using the pasteurization method, keep it for 15 min, then immediately cool to 4 °C, then filter the fermentation solution with an 8 μm filter plate, and then filter with a 0.22 μm PES membrane to obtain the cordyceps fermentation broth.

[0067] Embodiment 6 The difference between this embodiment and Embodiment 1 is that this embodiment provides a soothing and moisturizing cream, and the preparation raw materials of the cream include: Phase A: water 62.77; allantoin 0.20; lubrajel 1.00; water-soluble silicone wax 2.00; glycerol 4.00; sodium hyaluronate 0.03; Phase B: acryloyldimethyltauramide / VP copolymer 0.50; sodium stearoyl glutamate 0.20; M68 0.50; ARLACEL165 0.80; Phase C: OLIVEM1000 2.00; cetearyl alcohol 2.00; behenyl alcohol 0.50; ethylhexyl palmitate 4.00; polydimethylsiloxane 1.00; shea butter 1.00; caprylic / capric triglyceride 4.00; Phase D: Vaseline 0.30; Tocopheryl Acetate 0.20; Phytosteryl Isostearate 2.00; Cyclic Dimethyl Silicone 2.00; DC1403 1.00; 1,3-Butanediol 3.00; p-Hydroxyacetophenone 0.50; 1,2-Hexanediol 0.50; β-Glucan 2.00; the Cordyceps probiotic fermentation composition 2.00.

[0068] Performance Test Test 1: The skin care products prepared in Examples 1-6 were placed at 0 °C, 25 °C, and 40 °C for 90 days, and the product properties and colors were examined. The test results are shown in Table 1.

[0069] Thermal cycling experiment: The skin care products prepared in Examples 1-6 were placed in a sealed container to ensure complete sealing and prevent interference from external humidity or gas. The samples were placed in an incubator set at 45 °C for 12 hours. After taking out the samples, they were quickly transferred to a low-temperature incubator at -5 °C for 12 hours. After completing 1 high-low temperature cycle, repeat 6 times; after the cycle, check the physical properties of the samples such as appearance, color, odor, viscosity, layering, and particle precipitation.

[0070] Table 1 Test results of Performance Test 1

[0071] Test 2: Hyaluronidase inhibition rate Hyaluronidase is a specific lyase of hyaluronic acid and is a participant in allergic reactions, with a strong correlation with the release of histamine by mast cells. Whether the test sample has a soothing effect can be judged by the hyaluronidase inhibition rate. The higher the hyaluronidase inhibition rate, the stronger the soothing effect of the substance, and vice versa.

[0072] This test refers to the laboratory method (HMC-WI-029 Hyaluronidase Inhibition Rate), and the hyaluronidase inhibition rate test results of the test sample are compared with those of the negative control. If the inhibition rate of the test sample is higher than that of the negative control and has a significant difference (P < 0.05), it can be considered that the test sample has a certain soothing effect. This test method is an in vitro method and is applicable to evaluating cosmetics that claim to achieve a soothing effect by inhibiting hyaluronidase.

[0073] Instrumentation: BSA224S analytical balance, L6s ultraviolet spectrophotometer.

[0074] Reagents: Hyaluronidase, BR; Sodium Hyaluronate, BR.

[0075] Test method (1) Treatment of control and test samples Sample group: Dilute with pure water until the sample concentration is 10%; Positive control (dipotassium glycyrrhizinate, purity ≥ 98%): Dilute with pure water until the positive control concentration is 3%; Negative control: Pure water.

[0076] (2) Test operation steps Set up a sample group, a sample background group, a solvent group, and a solvent background group. Three parallels should be set up for each group. Add different reagent solutions to the four groups respectively, shake well, let it stand at room temperature for 30 min for color development, and measure the absorbance value at a wavelength of 528 nm with an ultraviolet spectrophotometer.

[0077] Calculation formula Hyaluronidase inhibition rate (%) = (1 - (C - D) / (A - B)) * 100% Where: A - Absorbance of the reaction solution without the sample; B - Absorbance of the reaction solution without the sample and enzyme; C - Absorbance of the reaction solution containing the sample and enzyme; D - Absorbance of the reaction solution containing the sample and without enzyme.

[0078] Table 2 Results table of Test 2

[0079] The hyaluronidase inhibition rate is retained to three decimal places. P < 0.05 indicates that there is a significant difference between the sample and the positive control compared with the negative control.

[0080] Test 3: Firming and anti-wrinkle test The anti-wrinkle and firming effects are mainly characterized by evaluating the inhibition rate of the test sample on elastase. Elastase is mainly synthesized and secreted by fibroblasts and can degrade elastin in the skin, leading to skin aging. The experimental principle of inhibiting elastase is that porcine pancreatic elastase undergoes a catalytic reaction with the enzyme substrate, and the absorbance changes after adding the active substance. The inhibition rate of the elastase inhibitor is reflected by the magnitude of the absorbance change. In this test, referring to the laboratory method (HMC-WI-028 elastase inhibition rate), the inhibition rate test results of the test sample and the negative control are compared. If the inhibition rate of the test sample is higher than that of the negative control and there is a significant difference (P < 0.05), it can be considered that the test sample has anti-wrinkle and firming effects.

[0081] This test method is an in vitro method and is applicable to evaluating cosmetics that claim to achieve anti-wrinkle and firming effects by inhibiting elastase.

[0082] Instrument and equipment: BSA224S analytical balance, RT-6100 microplate reader.

[0083] Reagents: Elastase (porcine pancreas), BR, N-Succinyl-L-alanyl-L-alanyl-L-alanine, 98%, Epigallocatechin gallate (EGCG), 98%.

[0084] Test method (1)Treatment of control and test samples Sample group: Dilute with pure water to a sample concentration of 3%; Positive control (EGCG): Dilute with pure water to a positive control concentration of 0.1%; Negative control: Pure water.

[0085] (2)Test operation steps Set up a sample group, a sample background group, a solvent group and a solvent background group. Each group needs to have 3 parallels. Add different reagent solutions to a 96-well plate, shake gently, incubate at 25 °C for 15 min, then place it in an enzyme-linked immunosorbent assay (ELISA) reader and measure the absorbance at 410 nm.

[0086] Elastase inhibition rate (%) = (1 - (C - D) / (A - B)) * 100% Where: A - Absorbance of the reaction solution without the sample; B - Absorbance of the reaction solution without the sample and the enzyme; C - Absorbance of the reaction solution containing the sample and the enzyme; D - Absorbance of the reaction solution containing the sample and without the enzyme.

[0087] Table 3 Results table of Test 3

[0088] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some 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 invention.

Claims

1. A preparation method of a cordyceps fermentation product, characterized in that, The Cordyceps fermentation product is obtained by using Cordyceps flowers as a solid medium and fermenting with lactic acid bacteria and Saccharomyces cerevisiae as strains; the steps of the preparation method are as follows: (1) Crush the dried Cordyceps flowers, with a liquid-to-solid ratio of (1.6 - 2.5) ml:(0.5 - 1.5) g, mix the crushed Cordyceps residue with a solution, put it into a container and sterilize it, and the total volume of the solid medium accounts for 10% - 15% of the entire container; (2) Inoculate lactic acid bacteria into the Cordyceps solid medium: Inoculate lactic acid bacteria into the Cordyceps solid medium at an addition amount of 3wt% - 5wt%, stir the bacterial liquid and the solid medium evenly and anaerobically culture at 37°C for 48 - 72h; (3) Conduct solid fermentation: Add a sterile liquid medium to the above Cordyceps solid medium, add 3wt% - 5wt% Saccharomyces cerevisiae according to the total volume and culture at 28°C under microaerobic conditions for 24 - 48h; The formula of the sterile liquid medium is: 0.6 parts of peptone, 85 - 90 parts of water, sterilize at 115°C for 20 min, and add it to the Cordyceps solid medium after cooling; (4) Conduct liquid fermentation: After the fermentation is completed, quickly heat to 72 - 75°C using the pasteurization method, maintain for 15 - 30 min, then immediately cool to 4 - 5°C, and then filter the fermentation solution using an 8 - 10μm filter plate to obtain the Cordyceps fermentation broth.

2. A cordyceps probiotic fermentation composition, characterized in that, Prepared by the preparation method described in claim 1.

3. A soothing and moisturizing cream, characterized in that, The preparation raw materials of the cream contain the Cordyceps probiotic fermentation composition described in claim 2.

4. A soothing and moisturizing facial cream according to claim 3, characterized in that, By mass, the preparation raw materials of the cream contain: Phase A: 62.77 parts of water; 0.20 parts of allantoin; 1.00 part of lubrajel; 2.00 parts of water-soluble silicone wax; 4.00 parts of glycerol; 0.03 part of sodium hyaluronate; Phase B: 0.50 part of acryloyldimethyltaurine / VP copolymer; 0.20 part of sodium stearoyl glutamate; 0.50 part of M68; 0.80 part of ARLACEL165; Phase C: 2.00 parts of OLIVEM1000; 2.00 parts of cetearyl alcohol; 0.50 part of behenyl alcohol; 4.00 parts of ethylhexyl palmitate; 1.00 part of polydimethylsiloxane; 1.00 part of shea butter; 4.00 parts of caprylic / capric triglyceride; Phase D: 0.30 part of petrolatum; 0.20 part of tocopheryl acetate; 2.00 parts of phytosteryl isostearate; 2.00 parts of cyclic dimethyl silicone oil; 1.00 part of DC1403; 3.00 parts of 1,3 - butanediol; 0.50 part of p - hydroxyacetophenone; 0.50 part of 1,2 - hexanediol; 2.00 parts of β - glucan; 2.00 parts of the Cordyceps probiotic fermentation composition described in claim 2.

5. A method for preparing a soothing and moisturizing cream according to claim 4, characterized in that, The steps are as follows: ① Disperse Phase A, heat to 85°C, and homogenize or stir to dissolve evenly; ② Heat Phase B to 85°C, and homogenize or stir to dissolve evenly; ③ Add Phase A to Phase B, homogenize for 2 min, add Phase C and Phase D, continue to homogenize for 5 min, and stir to cool down; ④ Cool down to 50°C, add Phase E and Phase F, stir evenly, and discharge.

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