A fermentation composition with antioxidant effect and its application

Abstract

The present application provides a fermented composition with antioxidant efficacy; through component optimization, the composition has a special active ingredient and peptide molecular weight distribution, the composition has high bioavailability, rich functional ingredients, and excellent stability, has obvious antioxidant, cell vitality enhancement, skin brightening, moisturizing and repairing effects, and can be used in skin care products and cosmetics.

Description

Technical Field

[0001] The present invention relates to the technical field of skin antioxidant, A61K8 / 64, and in particular to a fermentation composition with antioxidant effect and application thereof. Background Art

[0002] Affected by internal and external environments such as oxidative stress, ultraviolet light, non-enzymatic glycation, etc., the skin is prone to a series of adverse manifestations such as wrinkles, pigmentation, darkening, sagging, roughness and aging. The reason is that the DNA, RNA, lipids, collagen, elastic fibers, etc. inside the skin are damaged or destroyed; currently, a large number of cosmetics have appeared on the market to improve the skin condition, but most of them cannot fundamentally improve the skin's antioxidant capacity, and people's abuse of these cosmetics may have destroyed the microbial flora on the surface of the skin, aggravating the above adverse reactions and affecting the skin's absorption of other active substances. In view of the above situation, it is necessary to develop an antioxidant composition that is less irritating to the skin, can be fully absorbed, deeply stimulates skin cells, and brightens the skin color.

[0003] Chinese patent CN114621881A discloses a fermented liquor of brewer's yeast and a skin care product comprising the fermented liquor of brewer's yeast. By using the inhibitory effect of brewer's yeast on pathogenic bacteria such as Staphylococcus aureus and Enterococcus pyogenes, the skin brightening effect of the skin care product is improved. Chinese patent CN114948803A discloses an anti-aging essence and a preparation method thereof, in which the essence uses a filtrate of a yeast fermentation lysate to regulate the oil-water balance of the skin, inhibit the activity of tyrosinase, eliminate dark complexion, and reduce fine lines. However, in these technologies, the effective ingredients absorbed by the skin are limited, and there is still much room for improvement in their antioxidant effect on the skin. Summary of the invention

[0004] In order to solve the above technical problems, the present invention first provides a fermentation composition with antioxidant effect, which comprises at least two of the fermentation product or its filtrate, the lysate extract, and the fermentation lysate filtrate.

[0005] The fermentation product or its filtrate, lysate extract, fermentation lysate filtrate contains multiple B vitamins, amino acids, peptides, minerals and other active skin care substances, which, as prebiotics, have excellent oil balancing, anti-aging and moisturizing and repairing effects on the skin. By compounding different components and utilizing their combined effects, the best anti-oxidation, cell vitality enhancement and repair effects are achieved.

[0006] Furthermore, the fermentation product or the filtrate thereof is fermented by at least one of bacteria, yeast, algae and plants.

[0007] Furthermore, the bacteria include but are not limited to at least one of acetobacillus, lactobacillus, lactobacillus, and brevibacterium.

[0008] Furthermore, the yeast includes but is not limited to at least one of Galactomyces cerevisiae, Saccharomyces cerevisiae, Candida, Pichia pastoris, and Schizosaccharomyces pombe.

[0009] Furthermore, the algae is at least one of Chlorella, Spirulina, Chlamydomonas, Salmonella, Chromococcus, Microcystis, Closterium, and Scenedesmus.

[0010] Furthermore, the plant includes but is not limited to at least one of lotus leaf, ganoderma lucidum, sea buckthorn, ginseng, dendrobium officinale, and barley seed.

[0011] Further, the composition includes at least two of a fermentation product, a fermentation product filtrate, and a fermentation lysate filtrate.

[0012] Furthermore, the DPPH clearance rates of the fermentation product, the fermentation product filtrate and the fermentation lysate filtrate are all greater than 20%.

[0013] Preferably, the fermentation product is derived from algae fermentation, including Chlorella fermentation product.

[0014] Chlorella fermentation products are fermented from Chlorella and are rich in protein, multiple vitamins, unsaturated fatty acids and other ingredients. They can promote skin metabolism from deep inside, activate skin cells, repair damaged genes, promote the growth of skin cell tissue, eliminate free radicals and delay aging.

[0015] Furthermore, the nitrogen content of the fermentation product is 0.05-0.08 g / 100 g, preferably 0.06-0.075 g / 100 g.

[0016] Furthermore, based on the total peptide mass of the fermentation product, peptides with a molecular weight (Mw) of 200-2000Da account for 10-90% of the total peptide molecule mass; peptides with Mw greater than 2000Da account for 8-25%, preferably 8-13% of the total peptide molecule mass.

[0017] Furthermore, based on the total peptide mass of the fermentation product, in the fermentation product, peptides with a Mw of 200-600Da account for 15-40%, and peptides with a Mw of 600-2000Da account for 30-70%; more preferably, peptides with a Mw of 200-600Da account for 20-40%, peptides with a Mw of 600-1000Da account for 15-25%, and peptides with a Mw of 1000-2000Da account for 30-50%.

[0018] Preferably, the fermentation product filtrate is obtained by yeast fermentation, including at least one of Galactomyces cerevisiae, Saccharomyces cerevisiae, Pichia pastoris, and Schizosaccharomyces pombe.

[0019] Furthermore, the nitrogen content of the fermentation product filtrate is 0.04-0.06 g / 100 g; preferably 0.05-0.06 g / 100 g.

[0020] Furthermore, for the fermentation product filtrate, the peptide molecular weight is less than 1200Da, preferably 200-1200Da.

[0021] Furthermore, in the fermentation product filtrate, based on the total mass of peptide molecules, peptides with Mw of 200-800Da account for more than 99%.

[0022] Preferably, in the fermentation product filtrate, based on the total mass of peptide molecules, peptides with a Mw of 300-600 Da account for 85-96%, and peptides with a Mw of 600-800 Da account for 0.5-5%.

[0023] More preferably, the fermentation product filtrate comprises at least one of a galactosaccharomyces-like bacteria fermentation product filtrate, a cerevisiae fermentation product filtrate, and a Pichia pastoris fermentation product filtrate.

[0024] In a preferred embodiment, the fermentation product filtrate comprises a galactosidase-like yeast fermentation product filtrate.

[0025] The filtrate of galactosyl yeast fermentation product is the product of the fermentation process of coated yeast. It is rich in small molecules of active ingredients such as minerals, free amino acids, vitamin B group, inorganic acids, organic acids, etc. It has multiple functions such as whitening, moisturizing, and anti-aging. It can significantly balance oil and increase skin transparency.

[0026] Preferably, the fermentation lysate filtrate is a yeast fermentation lysate filtrate.

[0027] Yeast fermentation lysate filtrate is a product of probiotic metabolism, transformation or release, which contains a large amount of short-chain fatty acids, various amino acids, peptides and minerals. It can significantly improve the distribution of bacteria on the skin surface, promote the stratum corneum to better lock in moisture, inhibit tyrosinase activity, eliminate the dull skin tone after oxidation, and show good anti-aging, moisturizing, repair, water and oil balance and other functions.

[0028] Furthermore, the nitrogen content of the fermentation lysate filtrate is 0.06-0.09 g / 100 g, preferably 0.07-0.09 g / 100 g.

[0029] Furthermore, in the fermentation lysate filtrate, based on the total mass of peptide molecules, peptides with Mw of 200-600Da account for 50-70%, peptides with Mw of 600-1000Da account for 20-35%, peptides with Mw of 1000-2000Da account for 5-15%, and peptides above 2000Da account for less than 1%.

[0030] Preferably, in the fermentation lysate filtrate, based on the total mass of peptide molecules, peptides with Mw of 200-600Da account for 60-70%, peptides with Mw of 600-1000Da account for 20-30%, peptides with Mw of 1000-2000Da account for 5-9%, and peptides above 2000Da account for less than 1%.

[0031] In a preferred embodiment, the composition comprises a fermentation product, a fermentation product filtrate and a fermentation lysate filtrate.

[0032] Furthermore, based on the total weight of the composition, the composition comprises: 20-40% of fermentation product, 5-20% of fermentation product filtrate, and the balance of 100% is made up of fermentation lysate filtrate.

[0033] In a preferred embodiment, the composition comprises, by total weight of the composition: 20-40% of Chlorella fermentation product, 5-20% of Galactomyces fermentation product filtrate, and the balance of 100% is yeast fermentation lysate filtrate.

[0034] The present application utilizes the rich skin-benefitting active substances in the chlorella fermentation product, the filtrate of the galactose yeast-like fermentation product, and the filtrate of the yeast fermentation lysate to regulate the microbial flora of the skin epidermal environment; most importantly, the present application found in the experiment that the smaller the molecular weight is, the better the penetration and promotion effect of the composition on the skin. When the nitrogen content and the peptide molecular weight distribution of the fermentation product, the fermentation product filtrate, and the lysate filtrate are further optimized to make the system have a special peptide molecular weight distribution range, the composition at this time can better promote the penetration of active peptide molecules to the surface of the skin, fundamentally stimulating the regeneration of skin tissue cells; in addition, due to the presence of a large number of polar bonds such as hydroxyl, amino, ester, ether bonds, etc. in the molecules of the peptide molecular chains, organic acids and proteins in the system, intermolecular forces such as hydrogen bonds and electrostatic effects can make active substances with different chain lengths and polarities have a suitable degree of cross-linking in the system, maintain the stable state of the system, and accelerate each other's penetration into the skin.

[0035] Furthermore, the composition described in the present application has the effects of anti-oxidation, improving cell vitality, brightening skin, etc., and can be used in skin care products and cosmetics with the effects of anti-oxidation, whitening, brightening skin tone, repairing cells, etc.

[0036] Furthermore, the skin care products and cosmetics include but are not limited to any one or more of water-based, emulsion-based, gel-based, and cream-based types, and may be any one or more of water, emulsion, essence, gel, cream, ointment, and the like.

[0037] Beneficial Effects

[0038] The present application provides a fermentation composition with antioxidant effect. By optimizing the ingredients of the composition, the composition has a good skin care effect when it includes a chlorella fermentation product, a galactosaccharomyces fermentation product filtrate and a yeast fermentation lysate filtrate; the nitrogen content and peptide molecular weight are further specified, and the relative contents of the three are regulated so that the composition system maintains an optimal peptide molecule distribution range, and utilizes the intermolecular effects and chain entanglement of peptide molecules, inorganic acids, organic acids, minerals, proteins, etc. in the composition so that the components can exert excellent synergistic effects, which not only promotes the stability of each other, but also promotes its penetration and absorption into the surface layer of the skin, fundamentally stimulates the regeneration of skin cells, and regulates the skin surface microenvironment; the composition of the present application has high bioavailability, rich functional ingredients, and has obvious antioxidant, cell vitality-enhancing, skin-brightening, and moisturizing and repairing effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] Figure 1 : Experimental phenotype diagram of the antioxidant efficacy of the composition of Example 1 at 1% concentration;

[0040] Figure 2 : Fibroblast survival rate results of the composition of Example 1 at 1% concentration;

[0041] Figure 3 : Fibroblast growth number curve of the composition of Example 1 at 1% concentration;

[0042] Figure 4 : Cell growth cycle diagram of the composition of Example 1 at 1% concentration;

[0043] Figure 5 : Pictures of the stability effect of the composition of Example 1 placed in different environments for one week (from left to right: -20°C, 4°C, 25°C, 37°C, 50°C, cycle, light);

[0044] Figure 6 : Pictures of the stability effect of the composition of Example 1 placed in different environments for two weeks (from left to right: -20°C, 4°C, 25°C, 37°C, 50°C, cycle, light);

[0045] Figure 7 : Pictures of the stability effect of the composition of Example 1 placed in different environments for 8 weeks (from left to right: -20°C, 4°C, 25°C, 37°C, 50°C, cycle, light). DETAILED DESCRIPTION

[0046] Example

[0047] Example 1

[0048] This embodiment provides a fermentation composition with antioxidant effect; based on the total weight of the composition, the composition comprises: 30% of Chlorella fermentation product, 10% of Galactomyces galactosidase fermentation product filtrate, and 60% of yeast fermentation lysate product filtrate;

[0049] The nitrogen content of the chlorella fermentation product is 0.063g / 100g, and the peptide molecular weight distribution is as follows: based on the total weight of the peptide molecules of the chlorella fermentation product, Mw (200-400Da) accounts for 21.6%, Mw (400-600Da) accounts for 13.6%, Mw (600-1000Da) accounts for 19.3%, Mw (1000-2000Da) accounts for 36.2%, and Mw (above 2000Da) accounts for 9.3%.

[0050] The nitrogen content of the filtrate of the galactose yeast-like bacteria fermentation product is 0.054g / 100g, and the peptide molecular weight distribution is as follows: based on the total mass of the peptide molecules in the filtrate of the galactose yeast-like bacteria fermentation product, Mw (200-300Da) accounts for 4.4%, Mw (300-400Da) accounts for 39.7%, Mw (400-600Da) accounts for 54.4%, Mw (600-800Da) accounts for 1.4%, and Mw (800-1200Da) accounts for 0.1%.

[0051] The nitrogen content of the yeast fermentation lysate filtrate is 0.075g / 100g, and the peptide molecular weight distribution is as follows: based on the total mass of peptide molecules in the yeast fermentation lysate filtrate, Mw (200-400Da) accounts for 14%, Mw (400-600Da) accounts for 51.6%, Mw (600-1000Da) accounts for 26.1%, Mw (1000-2000Da) accounts for 7.6%, and Mw (above 2000Da) accounts for 0.7%.

[0052] Example 2

[0053] This embodiment provides a fermentation composition with antioxidant effect; based on the total weight of the composition, the composition comprises: 40% of Chlorella fermentation product, 5% of Galactomyces galactosidase fermentation product filtrate, and 55% of yeast fermentation lysate filtrate;

[0054] The nitrogen content of the chlorella fermentation product is 0.078g / 100g, and the peptide molecular weight distribution is as follows: based on the total weight of the peptide molecules of the chlorella fermentation product, Mw (200-400Da) accounts for 23.4%, Mw (400-600Da) accounts for 16.6%, Mw (600-1000Da) accounts for 17.2%, Mw (1000-2000Da) accounts for 39.3%, and Mw (above 2000Da) accounts for 3.5%.

[0055] The nitrogen content of the filtrate of the galactose yeast-like bacteria fermentation product is 0.058g / 100g, and the peptide molecular weight distribution is: based on the total mass of the peptide molecules in the filtrate of the galactose yeast-like bacteria fermentation product, Mw (200-300Da) accounts for 6.9%, Mw (300-400Da) accounts for 34.2%, Mw (400-600Da) accounts for 58.4%, Mw (600-800Da) accounts for 3.5%, and Mw (800-1200Da) accounts for 0.5%.

[0056] The nitrogen content of the yeast fermentation lysate filtrate is 0.086g / 100g, and the peptide molecular weight distribution is as follows: based on the total mass of peptide molecules in the yeast fermentation lysate filtrate, Mw (200-400Da) accounts for 14.8%, Mw (400-600Da) accounts for 45.2%, Mw (600-1000Da) accounts for 30.8%, Mw (1000-2000Da) accounts for 8.8%, and Mw (above 2000Da) accounts for 0.4%.

[0057] Example 3

[0058] This embodiment provides a fermentation composition with antioxidant effect; it is consistent with Example 1, except that: based on the total weight of the composition, the composition includes: 20% of Chlorella fermentation product, 20% of Galactomyces fermentation product filtrate, and the balance of 60% of yeast fermentation lysate filtrate.

[0059] Example 4

[0060] This embodiment provides a fermentation composition with antioxidant effect; based on the total weight of the composition, the composition comprises: 25% of Chlorella fermentation product, 15% of Galactomyces galactosidase fermentation product filtrate, and 60% of yeast fermentation lysate filtrate;

[0061] The nitrogen content of the chlorella fermentation product is 0.073g / 100g, and the peptide molecular weight distribution is as follows: based on the total weight of the peptide molecules of the chlorella fermentation product, Mw (200-400Da) accounts for 12.8%, Mw (400-600Da) accounts for 6.5%, Mw (600-1000Da) accounts for 16.8%, Mw (1000-2000Da) accounts for 45.2%, and Mw (above 2000Da) accounts for 18.7%.

[0062] The nitrogen content of the filtrate of the galactose yeast-like bacteria fermentation product is 0.054g / 100g, and the peptide molecular weight distribution is as follows: based on the total mass of the peptide molecules in the filtrate of the galactose yeast-like bacteria fermentation product, Mw (200-300Da) accounts for 4.4%, Mw (300-400Da) accounts for 39.7%, Mw (400-600Da) accounts for 54.4%, Mw (600-800Da) accounts for 1.4%, and Mw (800-1200Da) accounts for 0.1%.

[0063] The nitrogen content of the yeast fermentation lysate filtrate is 0.075g / 100g, and the peptide molecular weight distribution is as follows: based on the total mass of peptide molecules in the yeast fermentation lysate filtrate, Mw (200-400Da) accounts for 14%, Mw (400-600Da) accounts for 51.6%, Mw (600-1000Da) accounts for 26.1%, Mw (1000-2000Da) accounts for 7.6%, and Mw (above 2000Da) accounts for 0.7%.

[0064] Comparative Example 1

[0065] Basically the same as Example 1, except that: the nitrogen content of the filtrate of the galactose yeast-like bacteria fermentation product is 0.058g / 100g, and the peptide molecular weight distribution is: based on the total mass of peptide molecules in the filtrate of the galactose yeast-like bacteria fermentation product, Mw (200-400Da) accounts for 14.3%, Mw (400-600Da) accounts for 24.2%, Mw (600-1000Da) accounts for 44.69%, Mw (1000-2000Da) accounts for 11.34%, and Mw (above 2000Da) accounts for 5.47%.

[0066] Comparative Example 2

[0067] The method is basically the same as Example 1, except that the nitrogen content of the yeast fermentation lysate filtrate is 0.028 g / 100 g, and the peptide molecular weight distribution is as follows: based on the total mass of peptide molecules in the yeast fermentation lysate filtrate, Mw (500-600 Da) accounts for 4%, Mw (600-800 Da) accounts for 71.2%, Mw (800-1300 Da) accounts for 15.8%, Mw (1300-2000 Da) accounts for 8%, and Mw (above 2000 Da) accounts for 1%.

[0068] Comparative Example 3

[0069] The method is basically the same as Example 1, except that, based on the total weight of the composition, the composition comprises: 10% of the chlorella fermentation product, 30% of the filtrate of the galactosaccharomyces fermentation product, and 60% of the filtrate of the yeast fermentation lysate product.

[0070] Comparative Example 4

[0071] The method is basically the same as Example 1, except that, based on the total weight of the composition, the composition comprises: 30% of Chlorella fermentation product, 60% of Galactomyces fermentation product filtrate, and 10% of yeast fermentation lysate filtrate.

[0072] The Chlorella fermentation product was purchased from Weidan Biotechnology Co., Ltd., Jiangsu Ruiting Biotechnology Co., Ltd., and Dezhou Anlida Biotechnology Co., Ltd.; the galactosaccharomyces fermentation product filtrate was purchased from Angel Yeast Co., Ltd., Japan Katakura Industry Co., Ltd., and Dimei New Materials Technology Co., Ltd.; the yeast fermentation lysate filtrate was purchased from Jiangsu Ruiting Biotechnology Co., Ltd., Angel Yeast Co., Ltd., and Germany CLR.

[0073] Performance testing method:

[0074] 1. DPPH removal rate: 100% stock solution of the test example composition according to GB / T39100-2020;

[0075] 2. Stability: Aseptically pack the samples into colorless and transparent 20mL sample bottles, and place the sample bottles in different environments, such as -20℃, 4℃, 25℃ dark room, 37℃, 50℃, room temperature lighting, high and low temperature cycle (4℃ 24h, 45℃ 24h), and continuously observe the appearance (whether there is turbidity, precipitation, flocculation, etc.), color (whether there is color change), and odor of the product on the 7th, 14th, 30th, 60th, and 90th days, and test its physical and chemical indicators (pH value, viscosity) for abnormalities to determine its stability. If, during the observation and test, a group shows obvious adverse phenomena (such as precipitation of substances, discoloration, or bad odor) or the indicators are significantly different from the last measurement results, the stability of the group is judged to be unqualified, and the stability observation and indicator test tracking of the group are stopped. The stability test picture of the composition in Example 1 is shown in Figure 5-7 .

[0076] 3. Antioxidant efficacy: The composition at a concentration of 1% was tested according to the Standard Operating Procedure for the Evaluation of Antioxidant Efficacy of Zebrafish;

[0077] Experimental system: Translucent Albino strain zebrafish with melanin allele mutation;

[0078] Zebrafish age: 2 days post fertilization (2dpf);

[0079] The sample size of each experimental group was 15 (N=10);

[0080] The results showed that the composition of Example 1 at a concentration of 1% had an antioxidant effect of 28%. This showed a significant antioxidant effect; the efficacy experimental phenotype diagram is shown in Figure 1 .

[0081] 4. Effect of improving cell viability: Fibroblasts were used as the test object, and the third general rule of the 2015 edition of the Chinese Pharmacopoeia, "Biological Activity Assay - Cell Proliferation Method", was used to verify whether the raw materials had an effect of improving cell viability from the three indicators of cell survival rate, cell growth curve and cell cycle. The test results of the composition of Example 1 at a concentration of 1% are shown in Figure 2-4 The experimental results show that compared with the blank control group, it can be found that the composition of Example 1 at a concentration of 1% can significantly improve the cell viability of skin fibroblasts.

[0082] Performance test results:

[0083] The test results are shown in Table 1.

[0084] Table 1

[0085] Sample No. DPPH removal rate / % stability Example 1 43.94 pass Example 2 43.29 pass Example 3 40.21 pass Example 4 35.89 pass Comparative Example 1 26.75 Failure Comparative Example 2 18.70 Failure Comparative Example 3 10.96 pass Comparative Example 4 12.88 pass

Claims

1. A fermented composition with antioxidant effect, characterized in that: Based on the total weight of the composition, the composition comprises: 20-40% of fermentation product, 5-20% of fermentation product filtrate, and the balance of 100% is made up of fermentation lysate filtrate; The fermentation product is a chlorella fermentation product; the nitrogen content of the fermentation product is 0.05-0.08 g / 100 g; based on the total peptide mass of the fermentation product, in the fermentation product, peptides with Mw of 200-600 Da account for 15-40%, peptides with Mw of 600-2000 Da account for 30-70%, and peptides with Mw greater than 2000 Da account for 8-25% of the total peptide molecule mass; The fermentation product filtrate is a galactose yeast-like bacteria fermentation product filtrate, the nitrogen content of the fermentation product filtrate is 0.04-0.06g / 100g; the molecular weight of its peptides is less than 1200Da; in the fermentation product filtrate, based on the total mass of peptide molecules, peptides with Mw of 300-600Da account for 85-96%, and peptides with Mw of 600-800Da account for 0.5-5%; The fermentation lysate filtrate is a yeast fermentation lysate filtrate; the nitrogen content of the fermentation lysate filtrate is 0.06-0.09g / 100g; in the fermentation lysate filtrate, based on the total mass of peptide molecules, peptides with Mw of 200-600Da account for 50-70%, peptides with Mw of 600-1000Da account for 20-35%, peptides with Mw of 1000-2000Da account for 5-15%, and peptides above 2000Da account for less than 1%.

2. Use of the fermented composition according to claim 1 in skin care products and cosmetics.

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