CN114085271B - A medicine or antioxidant whitening cosmetic prepared from microorganism fermentation broth - Google Patents

Abstract

The invention relates to a medicament or an antioxidant whitening cosmetic prepared according to microbial fermentation liquor. According to the invention, the fermentation broth with high oxidation resistance activity is obtained by fermenting the medlar through lactobacillus plantarum and aspergillus niger, three polypeptides with oxidation resistance and melanin synthesis inhibition are separated and identified from the fermentation broth, the polypeptides can enhance the proliferation of skin fibroblasts on one hand and inhibit the synthesis of melanin and improve the oxidation resistance of mice on the other hand, and the polypeptides have good application prospects in the preparation of medicines and whitening cosmetics.

Description

A medicine or antioxidant whitening cosmetic prepared from microorganism fermentation liquid

Technical Field

The invention relates to the field of medicines, and particularly relates to a medicine or an antioxidant cosmetic prepared from microbial fermentation liquor.

Background

The aging process of the body results from damage to cells and tissues by free radicals. Free ions are by-products produced by the human body during metabolism. With the age, the body's own redox system is out of balance, and the ability to scavenge free ions in the body is gradually weakened, so that free radicals can invade our body tissues. Antioxidant factors are natural ingredients, and can neutralize oxygen "free radicals" and protect your cells from damage. Poor dietary habits, heavy living pressure, and environmental pollution all consume a large amount of nutrients, and disturb the immune system of a human.

Oxidation is a necessary process for living organisms and can also cause damage to biological macromolecules in the organism. Although the body itself has antioxidant defense systems, these systems are not completely effective in defending or repairing damage from oxidation. At this time, some antioxidant is added to help balance oxygen metabolism in vivo. Because of the safety issues associated with many chemically synthesized antioxidants, natural products having antioxidant activity are highly desirable. Lactic acid bacteria (Lactcacid bacteria) are probiotics existing in human body, can tolerate digestive tract environment, and have the functions of regulating normal flora of gastrointestinal tract, preventing and treating diarrhea, eliminating endotoxin, improving immunity, etc. The research of some scholars at home and abroad shows that part of lactic acid bacteria have antioxidant activity. The research shows that animal experiments prove that part of lactobacillus acidophilus (L.acidophilus) has the capacity of resisting lipid peroxidation and eliminating hydroxyl radical. However, the antioxidant activity and mechanism of these small fractions of lactic acid bacteria have not yet been clarified. It is believed that a small fraction of lactic acid bacteria may have some antioxidant enzyme substances (e.g., superoxide dismutase (SOD), NADH oxidase (NADH oxidase), Catalase (CAT), etc.) that make them antioxidant.

Moreover, with the progress of society and the improvement of living standard of people, the pursuit of health happiness and beauty becomes a common appeal of people. The oriental people want to get white, ruddy, fine and smooth skin, and the European consumers want to eliminate or reduce chloasma and age spots, so that the use of antioxidant cosmetics becomes a necessity of daily life of people. However, because the antioxidant cosmetics added with chemical whitening components have cytotoxicity, allergenicity and some potential unsafe factors, the search and development of antioxidant cosmetics which are safe and efficient and have nutrition functions and are added with natural antioxidant components become hot spots of research in the field of cosmetics. China has abundant natural plant resources, extracts antioxidant components in natural products, and further develops medicaments or cosmetics with better antioxidant property, thereby having unique resource advantages.

However, at present, research on antioxidant drugs and corresponding cosmetics developed by fermentation of natural components is few, and the research is not enough.

Disclosure of Invention

The invention adopts specific microorganism to ferment medlar to obtain fermentation liquor, and the fermentation liquor is screened to obtain polypeptide which has the functions of resisting oxidation and inhibiting melanin synthesis.

In one aspect, the invention provides a method for preparing a fermentation broth with high antioxidant activity.

Specifically, the method comprises the steps of pouring dry medlar into a 500mL triangular flask, adding water for soaking until the medlar is full of water, adding water for pulping according to the material-liquid ratio of 1: 6 (m: V), weighing sucrose and yeast extract, adding into the prepared medlar pulp, and adjusting the pH value to 6.0. The concentration of the cells was adjusted to 10% by 2% inoculation ⁷ Mixing cfu/mL Lactobacillus plantarum CGMCC NO.1258 into fructus Lycii pulp, sealing with gauze, and placing into shaking table (130 r.min) ^-1 ) Fermenting at 37 deg.C for 48 h.

And (3) activating purchased Aspergillus niger CGMCC 3.3927 on a PDA slant culture medium for 3d, and inoculating the activated Aspergillus niger spores in the PDA culture medium for 5d at the constant temperature of 28 ℃. Taking a certain amount of Aspergillus niger spores from PDA culture medium, adding into physiological saline, and adjusting thallus concentration to 10 ⁶ cfu/mL. Inoculating according to 6% of inoculum size, adding into lactobacillus plantarum fermentation liquor fermented for 48h, uniformly mixing, fermenting for 80h at 30 ℃ at 100r/min, centrifuging at 4000r/min for 10min after fermentation, and taking supernatant to obtain the fermentation liquor.

Furthermore, specific polypeptides with high antioxidant activity for inhibiting melanin synthesis are obtained by screening according to the activity of each component in the fermentation liquor, wherein the specific polypeptides are YX-1 polypeptide, YX-2 polypeptide and YX-3 polypeptide. The three polypeptides have good safety for resisting erythrolysis, and the mouse test in the embodiment also proves that the polypeptides have no animal toxicity and good safety effect.

Furthermore, the invention also provides a pharmaceutical composition with antioxidant activity.

The pharmaceutical composition contains SEQ ID NO: 1 and a pharmaceutically acceptable carrier.

Wherein, further, provided herein is a pharmaceutical composition comprising a therapeutically effective amount of the above peptide product or an acceptable salt thereof, and at least one pharmaceutically acceptable carrier or excipient. In some embodiments, the carrier is an aqueous-based carrier. In some embodiments, the carrier is a non-aqueous based carrier. In some embodiments, the non-aqueous carrier is a hydrofluoroalkane solvent containing submicron anhydrous-lactose or other excipients.

As used herein, "carrier" includes any solvent, dispersion medium, carrier, coating, diluent, isotonic agent, absorption delaying agent, buffer, carrier solution, suspension, colloid, and the like. The use of such media and/or agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients may also be incorporated into the composition. As used herein, "pharmaceutically acceptable" refers to a material that is not biologically or otherwise undesirable, i.e., the material may be administered to an individual along with a polypeptide without causing any undesirable biological effects or interacting in a deleterious manner with any of the other components of the pharmaceutical composition in which it is contained.

The pharmaceutical composition may be formulated in various forms suitable for the preferred route of administration. Thus, the compositions may be administered by known routes, including, for example, orally, parenterally (e.g., intradermally, transdermally, subcutaneously, intramuscularly, intravenously, intraperitoneally, etc.) or topically (e.g., intranasally, intrapulmonary, intramammary, intravaginally, intrauterine, intradermally, transdermally, rectally, etc.). A pharmaceutical composition may be administered to a mucosal surface, for example by administration to, for example, the nasal or respiratory mucosa (e.g., by a spray or aerosol). A composition may also be administered by sustained or delayed release.

In some embodiments, the method may comprise administering to the subject sufficient polypeptide to provide a dose of, for example, about 10ng/kg to about 10mg/kg, although in some embodiments, the method may be performed by administering a dose of the scFv antibody that is outside of this range. Thus, in some embodiments, the method can comprise administering the polypeptide in an amount effective to provide a minimum dose of at least 100ng/kg, e.g., at least 500ng/kg, at least 5mg/kg, at least 1mg/kg, at least 2mg/kg, at least 5mg/kg, at least 7mg/kg, at least 10mg/kg, at least 250mg/kg, at least 500mg/kg, at least 1mg/kg, at least 5mg/kg, or at least 10 mg/kg. In some embodiments, the method can include administering the polypeptide in an amount effective to provide a maximum dose of no more than 10mg/kg, e.g., no more than 10mg/kg, no more than 50g/kg, no more than 2mg/kg, no more than 1mg/kg, no more than 0.1 mg/kg. In some embodiments, the method can comprise administering the polypeptide in an amount effective to provide a dose that falls within a range having an endpoint defined by any of the minimum doses listed above and any of the maximum doses listed above that are greater than the minimum dose. In some of these embodiments, the method comprises administering to the subject sufficient polypeptide to provide a dose of about 1mg/kg to about 5 mg/kg.

The pharmaceutical composition of the present invention can be used in the form of a dosage form in the form of an external solvent such as an acid agent, granules, a purified capsule, a suspension, a syrup, an aerosol, or a sterile injectable solution according to a conventional method, and can be used for the above-mentioned composition. Possible carrier excipients and diluents are: lactose, dckstros, tomasotson, xylosylmannitol, locust bean gum phospholipid, calcium gelatin, calcium silica gel, cellulose methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone; water; methyl oxybenzone; an oxybenzone; magnesium removal; a stearate ester; and mineral oil; when formulated, the composition is formulated with diluents or excipients such as fillers, reinforcing agents, binders, wetting agents, disintegrators, surfactants, etc., which are generally used. Solid preparations for oral administration include a granular preparation of a refined epoxy preparation, a capsule preparation of a capsule preparation, etc., which contains at least one excipient among the above-mentioned compounds, such as calcium carbonate, Sucrose or lactose gel. And mixing the components. In addition to the simple excipient, a lubricant such as magnesium foam is used. Liquid phase formulations for hard balls include: the suspending agent is liquid, emulsion, syrup, etc. and may include various kinds of excipient, such as wetting agent, sweetener, aromatic, preservative, etc. besides water cubic paraffin as common diluent. The preparation for non-oral administration comprises sterile aqueous solution achiral solvent suspending agent emulsion freeze-drying preparation adjuvant. Achiral solvent suspensions can be used in injectable esters such as propylene glycol (glycol) olive oil. The mechanism of the adjuvant may be gastrin (witepsol), Macro Goltwin (tween), 61-Carkol lauryl triglyceride, etc.

The invention further provides an antioxidant whitening cosmetic with antioxidant and melanin synthesis inhibiting functions, which contains the amino acid sequence shown in SEQ ID No: 1.

The cosmetic composition of the present invention contains the effective ingredient, and may contain, in addition to the above-mentioned extract, ingredients generally used in cosmetic compositions, such as stabilizer-dissolving agents, vitamin pigments, perfumes, and other conventional adjuvants, and a carrier.

The cosmetic raw material composition of the present invention can be prepared into any dosage form generally manufactured in the industry, such as emulsion cream, lotion, foundation emulsion, beauty lotion, hair cosmetic, and the like.

Advantageous effects

According to the invention, the fermentation broth with high oxidation resistance activity is obtained by fermenting the medlar through lactobacillus plantarum and aspergillus niger, three polypeptides with oxidation resistance and melanin synthesis inhibition are separated and identified from the fermentation broth, the polypeptides can enhance the proliferation of skin fibroblasts on one hand and inhibit the synthesis of melanin and improve the oxidation resistance of mice on the other hand, and the polypeptides have good application prospects in the preparation of medicines and whitening cosmetics.

Drawings

FIG. 1 is a graph showing the results of the detection of antioxidant activity of the polypeptide

Detailed Description

The following examples and experimental examples are provided to facilitate understanding of the present invention. However, the following examples and experimental examples are provided only to make the present invention more easily understood, and the contents of the present invention are not limited by the following examples and experimental examples.

Example 1 preparation of a fermentation broth of a Lycium barbarum polypeptide

Pouring 20g of dry medlar into a 500mL triangular flask, adding water to soak until the medlar is full of water, and thenAdding water and pulping at a ratio of 1: 6 (m: V), adding sucrose 1g and yeast extract 0.5g into the prepared fructus Lycii pulp, and adjusting pH to 6.0. The bacterial concentration was adjusted to 10% by 2% inoculation ⁷ Mixing cfu/mL Lactobacillus plantarum CGMCC NO.1258 into fructus Lycii pulp, sealing with gauze, and placing into a shaking table (130 r.min.) ^－1 ) Fermenting at 37 deg.C for 48 h.

And (3) activating purchased Aspergillus niger CGMCC 3.3927 on a PDA slant culture medium for 3d, and inoculating the activated Aspergillus niger spores in the PDA culture medium for 5d at the constant temperature of 28 ℃. Taking a certain amount of Aspergillus niger spores from PDA culture medium, adding into physiological saline, and adjusting thallus concentration to 10 ⁶ cfu/mL. Inoculating according to 6% inoculum size, adding into lactobacillus plantarum fermentation broth fermented for 48h, mixing well, fermenting at 30 deg.C for 80h at 100r/min, centrifuging at 4000r/min for 10min after fermentation, collecting supernatant, and storing at 4 deg.C for use.

Example 2 preparation of a fermentation broth of Lycium barbarum polypeptide comparative example 1

Pouring 20g of dried medlar into a 500mL triangular flask, adding water for soaking until the medlar is full of water, adding water for pulping according to the material-liquid ratio of 1: 6 (m: V), weighing 1g of sucrose and 0.5g of yeast extract, adding into the prepared medlar pulp, and adjusting the pH value to 6.0. The bacterial concentration was adjusted to 10% by 2% inoculation ⁷ Mixing cfu/mL Lactobacillus plantarum CGMCC NO.1258 into fructus Lycii pulp, sealing with gauze, and placing into shaking table (130 r.min) ^－1 ) Fermenting at 37 deg.C for 48 h. Centrifuging at 4000r/min for 10min after fermentation, collecting supernatant, and storing at 4 deg.C for use.

Example 3 preparation of a fermentation broth of Lycium barbarum polypeptide control example 2

And (3) activating purchased Aspergillus niger CGMCC 3.3927 on a PDA slant culture medium for 3d, and inoculating the activated Aspergillus niger spores in the PDA culture medium for 5d at the constant temperature of 28 ℃. Taking a certain amount of Aspergillus niger spores from PDA culture medium, adding into physiological saline, and adjusting thallus concentration to 10 ⁶ cfu/mL. Inoculating according to 6% inoculation amount, adding into culture medium, and its preparation method comprises pouring 20g dry fructus Lycii into 500mL triangular flask, soaking in water until fructus Lycii is full, pulping with water at material-to-liquid ratio of 1: 6 (m: V), weighing 1g sucrose and 0.5g yeast extract, adding into the prepared fructus Lycii pulpIn (3), the pH was adjusted to 6.0. Uniformly mixing, fermenting at 30 ℃ for 80h at 100r/min, centrifuging at 4000r/min for 10min after fermentation, collecting supernatant, and storing at 4 ℃ for later use.

Example 4 antioxidant Capacity assay of Lycium barbarum polypeptide fermentation broth

Measurement of DPPH scavenging ability: preparing 0.2mmol/L DPPH by 95% ethanol, taking 100uL of the fermentation liquor of the embodiment 1-3 and 100uL of DPPH solution to be uniformly mixed in a 96-hole enzyme label plate, reacting for 30min at normal temperature in a dark place, and measuring the light absorption value A1 at 517 nm. A blank control (A0) was prepared by reacting 100uL of 95% ethanol instead of DPPH with 100uL of deionized water, and a negative control (A2) was prepared by reacting 100uL of DPPH with 100uL of deionized water. The DPPH clearance calculation formula is as follows: DPPH clearance (%) (a2-a1)/(a2-a0) × 100%, the results are shown in table 1 below.

TABLE 1 DPPH clearance of various groups of fermentation broths

Group of	DPPH clearance (%)
		Example 1 fermentation broth	98.5±6.6
Example 2 fermentation broth	79.3±4.5
		Example 3 fermentation broth	82.5±2.4

From the results in table 1, it can be seen that the fermentation broth obtained by fermenting lycium barbarum with specific lactobacillus plantarum and aspergillus niger has a higher DPPH clearance rate (98.5 ± 6.6)%, which is better than the fermentation effect obtained by fermenting with lactobacillus plantarum or aspergillus niger alone.

EXAMPLE 5 purification and isolation of the antioxidant Polypeptides in fermentation broths

The fermentation liquor of example 1 is centrifuged at 12000r/min for 10min, and after ultrafiltration and concentration, DEAE-Sepharose FF anion exchange chromatography is carried out, and then linear gradient elution is carried out by using a buffer solution containing 0-1.0 mol/L NaC1, and the antioxidant activity is detected. Collecting 5 groups of active components with strong antioxidant activity, and respectively ultrafiltering and concentrating. Then loading a SephadexG-15 molecular sieve gel filtration chromatographic column, collecting 4 groups of active components, loading the active components on a C18 column balanced by 0.1% TFA double distilled water, performing 0.8mL/min linear gradient elution by using 2% -100% acetonitrile (0.1% TFA), collecting samples in parts, performing ultrafiltration concentration on the active 3 groups of collected peaks again, performing RT-HPLC separation, freeze-drying the samples, hydrolyzing the freeze-dried samples at 110 ℃ by using 6mol/L HC1, drying the hydrolysate by distillation, dissolving the hydrolysate by adding 5mL of 0.02mol/L HCl, and performing amino acid composition analysis on the samples by using an L-8800 full-automatic amino acid analyzer to obtain 3 polypeptides with better antioxidant activity, which are respectively named as YX-1, YX-2 and YX-3 polypeptides, wherein the YX-1 sequence is as SEQ ID NO: 1 is shown. And (3) preparing the polypeptide for later use after the polypeptide is consigned to Shanghai for synthesis.

Example 6 detection of antioxidant Activity of YX-1 polypeptide

The hydroxyl radical (OH) scavenging capacity is measured by adopting an o-diazaphenanthrene-Fe 2+ oxidation method, taking 1mL of 0.75mmol/L o-diazaphenanthrene solution in a test tube, sequentially adding 2mL of phosphate buffer solution (0.2mol/L, pH7.4) and 1mL of distilled water and 1mL of 0.75mmol/L ferrous sulfate solution, fully and uniformly mixing, adding 0.01% H by volume fraction ₂ O ₂ 1mL, reacting at 37 ℃ for 60min, and measuring the absorbance at 536nm to obtain Ap. The formula for the hydroxyl radical scavenging capacity is as follows: clearance (%) ═ As-Ap/Ao-Ap × 100 formula: replacement of H by 1mL polypeptide solution ₂ O ₂ The absorbance measured was taken As; 1mL of distilled water was used in place of H ₂ O ₂ The absorbance measured was designated as Ao. VC was used as a positive control. The results are shown in FIG. 1.

As can be seen from figure 1, the clearance of polypeptide to hydroxyl free radical also increases rapidly along with the increase of the concentration of polypeptide and VC, and when the concentration is 250ug/mL, the clearance of polypeptide reaches about 76 percent and is basically similar to the clearance of Vc. Has better effect.

Example 7 Activity assay of YX-1 Polypeptides on fibroblasts

Fibroblast proliferation assay: selecting fibroblasts with good logarithmic growth phase state, digesting with 0.25% trypsin, centrifuging, suspending in DMEM medium containing 10% fetal calf serum, and counting; fibroblasts were seeded in 96-well culture plates at about 5X 10 cells per well ³ And a blank control without added cells was set, and the marginal wells were filled with sterile Phosphate Buffered Saline (PBS) at 37 ℃ with 5% CO ₂ Culturing overnight in the environment; adding polypeptide solution to be tested according to the respective final volume fraction, using untreated group as cell control group, setting 3 multiple wells in each group, and culturing at 37 deg.C under 5% CO ₂ Culturing for 48h in the environment. Adding 5g/L MTI solution 4h before the culture is finished, and terminating the culture after continuing the culture for 4 h; the liquid in the hole is sucked and removed, washed by PBS for 1 time, and added with DMSO to dissolve the crystal; the absorbance of each well was measured at a wavelength of 570nm using an M3 plate reader. Cell viability ═ (assay well absorbance value — blank control absorbance value)/(cell control absorbance value-blank control absorbance value) X100%. The results are shown in Table 2.

TABLE 2 cell viability after treatment with different concentrations of polypeptide

Concentration group	Cell survival rate (%)
		0	100±2.4
10μg/mL	118.4±4.8
		100μg/mL	134.5±6.7
200μg/mL	155.9±8.6
		1mg/mL	159.4±9.5

Fibroblasts have an important role in maintaining the elastic and elastic properties of the skin and aging. As can be seen from Table 2, the polypeptide has a certain promoting effect on the proliferation of fibroblasts, and the highest promoting effect of 159.4 +/-9.5% can be achieved at the concentration of 1 mg/mL.

Example 8 YX-1 whitening ability test

Intracellular tyrosinase activity inhibition assay: b16 cells in the logarithmic growth phase were seeded in 6-well cell culture plates and cultured overnight. The final volume fraction of 200. mu.g/mL of the polypeptide was added separately, and the untreated group was used as a cell control group, with 2 duplicate wells per group. After 48h of culture, the cells were washed 1 time with PBS, 100. mu.L of lysate was added to each well, the cells were scraped off and collected, and the supernatant was centrifuged. The 5OL cell supernatant was transferred to a 96-well plate, followed by addition of 50. mu.L of 1% L-dopa solution and incubation at 37 ℃ for 1 hour. The M3 plate reader reads the absorbance at 475 nm. Relative tyrosinase activity ═ (assay well absorbance value — blank absorbance value)/(cell control absorbance value — blank absorbance value) x 100%. The result shows that the relative tyrosinase activity in the cell is only (45.6 +/-5.3)%, which shows that the relative tyrosinase activity of B16 is obviously reduced under the action of the polypeptide, and the polypeptide has an inhibition effect on the tyrosinase activity.

Example 9 antioxidant assay of YX-1 Polypeptides in mice

40 male Kunming mice with the weight of 20-25 g are randomly divided into four groups, namely a normal control group, a D-galactose oxidative damage model group, a polypeptide treatment group and a Vc positive control group. Except for the normal control group, the mice of other groups are injected with D-galactose 400mg/kg bw subcutaneously every day for 32 days, the mice of the control group are injected with physiological saline with the same amount every day subcutaneously, the polypeptide group and the Vc group are respectively irrigated with stomach at the dosage of 1mg/kg bw every day, the normal control group and the model group are irrigated with distilled water with the same amount, and the mice are weighed for 1 time every 3 days. After injecting D-galactose and gavage polypeptide for 2h at 32D, dissecting the mouse, taking 0.2g of liver tissue, cutting into pieces, placing in a glass homogenizer, adding 9 times of cold normal saline to prepare 10% (W/V) liver tissue homogenate, centrifuging at 3000r/min for 10min, and taking the supernatant to measure the activity of the liver cell SOD and GSH-Px and the content of MDA. Determining the MDA content of the liver by adopting a thiobarbituric acid (TBA) colorimetric method; SOD is measured by xanthine oxidase; GSH-Px activity was determined by the DTNB method. The results are shown in Table 3.

TABLE 3 Effect of the Polypeptides on mouse liver MDA and SOD and GSH-Px Activity

Group of	MDA(μmol/g)	SOD(U/mg)	GSH-Px(U/mg)
				Control group	14.6±1.1	512±23	614±35
Model set	23.0±1.2	448±13	420±37
				Polypeptide group	18.5±1.3	525±22	587±29
Vc control group	19.6±1.2	495±31	576±40

As can be seen from Table 3, compared with the control group, the MDA content in the liver of the mice in the oxidation model group is remarkably increased, and the SOD activity and the GSH-Px activity are remarkably reduced. Compared with the model group, the MDA content of the mouse liver of the polypeptide group and the mouse liver of the positive control Vc group is obviously reduced, and the GSH-Px activity and the SOD activity are obviously improved. In particular, the MDA of the polypeptide group reaches 18.5 +/-1.3 mu mol/g, the SOD reaches 525 +/-22U/mg, the GSH-Px reaches 587 +/-29U/mg, and the result has statistical significance compared with the P <0.05 of a control group.

The foregoing description of the present invention is for illustrative purposes, and those skilled in the art to which the present invention pertains will be readily capable of adapting to other specific forms without changing the technical spirit or essential characteristics thereof. As can be understood in the affirmative. Therefore, the above-described examples and experimental examples should be construed as being indicative, but not limiting, of all aspects.

Sequence listing

<110> Beijing Yunshenda Biotechnology development Co., Ltd

<120> a pharmaceutical or antioxidant whitening cosmetic prepared from microbial fermentation broth

<160> 1

<170> SIPOSequenceListing 1.0

<210> 1

<211> 14

<212> PRT

<213> Artificial Sequence (Artificial Sequence)

<400> 1

Tyr Phe Asp Asp Met Cys Gly Asp Ser Arg Arg Glu Gly Trp

1 5 10

Claims (7)

1. A preparation method of fermentation liquor with high antioxidant activity is characterized by comprising the following steps: pouring dry fructus Lycii into 500mL triangular flask, soaking in water until fructus Lycii is full of water, pulping with water at a material-liquid ratio of 1: 6 (m: V), weighing sucrose and yeast extract, adding into the prepared fructus Lycii pulp, and adjusting pH to 6.0; the bacterial concentration was adjusted to 10% by 2% inoculation ⁷ Mixing cfu/mL Lactobacillus plantarum CGMCC NO.1258, adding fructus Lycii pulp, sealing with gauze, and placing in a shaking table for 130r min ^-1 Fermenting at 37 deg.C for 48 h;

aspergillus niger CGMCC 3.3927 is planted on a PDA slant culture medium to be activated for 3d, and the activated Aspergillus niger spores are inoculated on the PDA culture medium to be cultured for 5d at the constant temperature of 28 ℃; taking a certain amount of Aspergillus niger spores from PDA culture medium, adding into physiological saline, and adjusting thallus concentration to 10 ⁶ cfu/mL; inoculating and adding the lactobacillus plantarum fermentation liquor which is fermented for 48 hours according to the inoculation amount of 6%, uniformly mixing, fermenting for 80 hours at the temperature of 30 ℃ at the speed of 100r/min, centrifuging at the speed of 4000r/min for 10min after fermentation, and taking supernate, thus obtaining the fermentation liquor.

2. Use of the fermentation broth of claim 1 for the preparation of a medicament for enhancing antioxidant capacity.

3, SEQ ID NO: 1 in the preparation of antioxidant whitening cosmetics.

4. An antioxidant whitening cosmetic with antioxidant and melanin synthesis inhibiting effects, the cosmetic comprises an amino acid sequence shown in SEQ ID No: 1, or a pharmaceutically acceptable salt thereof.

5. Use according to claim 3 or cosmetic product according to claim 4, characterized in that the cosmetic product comprises stabilizers and solubilizers.

6. Use according to claim 3 or cosmetic product according to claim 4, wherein the cosmetic product is in the form of an emulsion.

7. The use according to claim 3 or the cosmetic according to claim 4, wherein the cosmetic is in the form of a mask.

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