CN111617025A

CN111617025A - Fermentation product cosmetic for whitening, removing freckles and resisting saccharification

Info

Publication number: CN111617025A
Application number: CN202010540446.9A
Authority: CN
Inventors: 罗春梅
Original assignee: Yangpu Jishang Biological Technology Co ltd
Current assignee: GUANGZHOU QIANBANG COSMETICS Co.,Ltd.
Priority date: 2020-06-12
Filing date: 2020-06-12
Publication date: 2020-09-04
Anticipated expiration: 2040-06-12
Also published as: CN111617025B

Abstract

The invention relates to a fermented product cosmetic for whitening, removing freckles and resisting saccharification, which comprises the following steps: (1) inoculating, fermenting and culturing the preserved lactobacillus crispatus CGMCC No.6364 into a seed solution under the aseptic condition; (2) inoculating 10 v/v% of lactobacillus crispatus CGMCC No.6364 seed solution to a liquid culture medium containing cortex mori extract and curcumin for fermentation culture; (3) centrifuging the fermentation liquor obtained in the step (2) at a centrifugal rotation speed of 8000r/min for 30min, collecting supernatant to obtain fermentation filtrate, filtering with a 0.22um sterilizing filter, and further making into facial mask, essence, cosmetic water, lotion, and cream.

Description

Fermentation product cosmetic for whitening, removing freckles and resisting saccharification

Technical Field

The invention relates to the technical field of cosmetic formulas, in particular to a plant fermentation composition for whitening, removing freckles and resisting saccharification and an application thereof in a fermentation product cosmetic for whitening, removing freckles and resisting saccharification.

Background

The color of human skin can therefore take on different shades, not depending on the number of melanocytes, but on the amount of various melanosomes produced, depending on the number, size, distribution and degree of melanosomes. The formation and deposition of normal melanin in melanocytes not only has the functions of protecting skin, resisting ultraviolet radiation, preventing internal tissues from overheating, and the like, but also has important significance on the health and beauty of skin. When abnormal changes such as hyperpigmentation, hypopigmentation or hypopigmentation occur in the skin, different types of dyschromatosis skin diseases such as vitiligo, albinism, chloasma, etc. are caused, and a corresponding unpleasant psychological sensation is produced.

Senescence-associated glycation is in fact a abbreviation, known collectively as non-enzymatic glycosylation, and in brief, the glycation reaction is the binding of sugars to proteins in the absence of enzymatic action, causing the proteins to lose normal structure and color, become brittle, and yellow. The protein may be of various types, including hemoglobin, elastin, collagen, and the like. The dermis layer of the skin contains a lot of collagen, so the skin is tight and elastic. After the reaction between sugars (glucose and fructose) and collagen, reversible primary glycosylation products are formed, followed by the formation of irreversible advanced glycosylation end products (AGEs). After glycation of collagen, in addition to further aging of the skin, the skin also looks yellow and therefore loses its fair texture. Not only does glycation occur in the dermis, new studies have found that proteins in the epidermis (particularly keratin) can also be glycated. In addition, AGEs can stimulate AGEs receptors and promote the production of melanin.

The whitening product in the prior art is usually only researched for inhibiting the activity of tyrosinase, and the invention aims to provide a plant fermentation composition for whitening, removing freckles and resisting saccharification and an application thereof in cosmetics.

Disclosure of Invention

In view of the above problems, the present invention aims to provide a plant fermentation composition for whitening, removing spots and resisting saccharification, and its application in cosmetics. The pure plant culture medium is safe and reliable, and is matched with probiotic fermentation, so that the inhibitory activity of the neuraminidase can be improved unexpectedly, and a good anti-saccharification effect and an anti-oxidation effect can be obtained, so that the effects of whitening, removing freckles and resisting aging are achieved.

Lactobacillus, lactic acid bacteria, are well-established probiotics and their fermentation products (usually lysates) have a certain cosmetic effect. It is known to those skilled in the art that the metabolism of probiotics is regulated in many ways, and the metabolites include many bioactive substances, such as small peptides, exopolysaccharides, saponins, amino acids, minerals, phenols, etc. The specific structure of exopolysaccharide greatly influences the physiological activity. In contrast, the team of the invention carries out a great amount of screening on the strains and improvement on the culture process so as to obtain the fermentation process with outstanding beauty effect.

Chinese patent 2013100043553 discloses a strain of Lactobacillus crispatus, which is a strain of Lactobacillus crispatus, characterized in that the preservation number of the strain is CGMCC No. 6364. The known function of the strain is to effectively realize antagonism to various pathogenic bacteria, and in addition, the strain has higher adhesion capability to vaginal epithelial cells and higher lactic acid production capability. The group of the invention finds that the lactobacillus strain has great potential as a strain of lactobacillus which is not commonly used in cosmetics, and the active extract of the conventional fermentation product has good antioxidant effect and certain activity of inhibiting the tyrosinase. The plant fermentation product is characterized by plasticity, and the fermentation process is tried to be changed, so that the addition of some plant components can obviously improve various cosmetic effects, including anti-saccharification performance, whitening performance and the like. While we observed that other strains of Lactobacillus crispatus did not have such outstanding plasticity properties.

Curcumin is a chemical component extracted from the rhizome of some plants in the families of Zingiberaceae and Araceae, wherein the Curcuma rhizome contains about 3-6% of pigment with diketone, which is a rare pigment with diketone in the plant world. Curcumin is orange yellow crystal powder, slightly bitter in taste and insoluble in water, and is mainly used for coloring products such as sausage products, cans, sauced and marinated products and the like in food production. The medical research shows that curcumin has the effects of reducing blood fat, resisting tumor, resisting inflammation, benefiting gallbladder, resisting oxidation and the like. The team of the invention tries to add a certain amount of curcumin into the culture medium, and finds that the performance of the fermentation product can be greatly changed.

The cortex mori radicis a traditional Chinese medicinal material, is root bark of mulberry, contains a large amount of nutrient substances, can promote metabolism of a body, and has a repairing effect on cells, so that the cortex mori radicis has a certain removing effect on scars, can enable the skin to become finer and smoother, and has good beautifying effect. The invention adds the cortex mori aqueous extract into the culture medium, and finds that the cortex mori aqueous extract and the curcumin can synergistically improve the beauty effect of the lactobacillus crispatus CGMCC No.6364 fermentation product. In addition, the cortex mori radicis also has certain beautifying effect and has double functions of regulating and controlling the growth of microorganisms and directly acting on the skin for beautifying.

In order to achieve the purpose, the invention is realized by the following technical scheme:

the plant fermentation composition for whitening, removing freckles and resisting saccharification is characterized by comprising the following preparation methods:

(1) inoculating the preserved Lactobacillus crispatus CGMCC No.6364 into MRS culture medium under aseptic condition, culturing at 35 deg.C and 200rpm for 12 hr to obtain culture as seed liquid for fermentation culture with final concentration of 6 × 10⁷cfu/ml；

(2) Inoculating lactobacillus crispatus CGMCC No.6364 seed liquid into a liquid culture medium according to the inoculation amount of 10 v/v%, and performing shake table culture at 30 ℃ and 200rpm for 24 hours;

the preparation method of the liquid culture medium comprises the following steps: mixing 0.3g curcumin, 50mL cortex Mori extractive solution, 25g glucose, 8g peptone, 7g yeast extract, 8g beef extract, 5g sodium acetate, 2g ammonium citrate, 2mL tween-80, 0.6g MgSO₄·7H₂O, 0.3g of MnSO₄·4H₂O, 2g of K₂HPO₄Mixing with 1000mL of distilled water, adjusting the pH value to 6.2-6.4, and sterilizing by high pressure steam to obtain a sterile culture medium;

(3) and (3) centrifuging the fermentation liquor obtained in the step (2) at a centrifugal rotation speed of 8000r/min for 30min, collecting supernatant to obtain fermentation filtrate, and filtering the fermentation filtrate by using a 0.22um sterilizing filter to obtain a sterile fermentation composition.

The invention also provides a cosmetic which contains the plant fermentation composition for whitening, removing freckles and resisting saccharification.

Preferably, the cosmetic formulation may be a mask, a serum, a cream, etc.

Compared with the prior art, the invention has the following beneficial effects:

the active ingredient of the cosmetic is prepared by fermenting natural and safe substrates with screened probiotics, is wider in applicable population, has no obvious toxic or side effect, has no irritation, has obvious anti-saccharification and whitening effects, and also has good antioxidant activity. As a preferred embodiment, the facial mask can be used for facial masks, is convenient to use and has an excellent effect.

Detailed Description

The present invention will be further described in detail with reference to the following specific examples, which are provided for illustration only and are not intended to limit the scope of the present invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.

The cortex mori radicis extract of the present example is commercially available from shanxi kang jumping biotechnology limited.

Example 1

(1) inoculating the preserved Lactobacillus crispatus CGMCC No.6364 into MRS culture medium at 1 v/v%, culturing at 35 deg.C and 200rpm for 12 hr to obtain culture as seed liquid for fermentation culture with final concentration of 6 x 10⁷cfu/ml；

(2) (2) inoculating lactobacillus crispatus CGMCC No.6364 seed liquid into a liquid culture medium according to the inoculation amount of 10 v/v%, and performing shake table culture at 30 ℃ and 200rpm for 24 hours;

Example 2 comparative example

The research and development team finds that curcumin and the cortex mori radicis have the functions of synergistically increasing the oxidation resistance, whitening and anti-saccharification performance of the fermented extract.

Comparative example 1

The experimental method is the same as that of example 1, but the fermentation medium is not added with curcumin and cortex mori radicis extract, and the rest is unchanged.

Comparative example 2:

the experimental procedure was the same as in example 1, except that no curcumin was added to the fermentation medium, and the rest was unchanged.

Comparative example 3:

the experimental procedure was the same as in example 1, except that the fermentation medium was not supplemented with the cortex Mori extract, and the rest was unchanged.

Comparative example 4:

the experimental method is the same as that of example 1, but the Lactobacillus crispatus CGMCC No.6364 is replaced by the Lactobacillus crispatus CGMCC1.2743, and the others are not changed.

Example 3

And (3) testing the cytotoxicity:

the plant fermentation composition obtained in example 1 is dissolved in serum-free DMEM culture solution, 4 samples are prepared respectively, the mass fractions of the samples are 0.25%, 0.5%, 1% and 2%, MTT detection is performed by taking the serum-free DMEM culture solution as a negative control sample group, and the cytotoxicity of the whitening, freckle-removing and anti-saccharification plant fermentation composition is evaluated.

L929 cells were seeded at 100. mu.L/well in 96-well plates at 1 × 10⁵one/mL, 5% CO at 37 ℃₂And after 24 hours of culture in serum-free DMEM culture solution for adherence, removing the culture solution by suction, adding 100 mu L of sample solution with different mass fractions into each hole, and continuously culturing the negative control sample group in serum-free DMEM for 48 hours by a conventional method. Then 20. mu.L of 5mg/mL MTT solution was added, and the mixture was incubated in an incubator for 4 hours in the absence of light. The supernatant was discarded and added to DMSO in an amount of 100. mu.L, and the mixture was shaken on a shaker at a low speed for 20min to measure the absorbance at a wavelength of 570 nm. The other groups of cells were calculated as followsRelative survival (β) results are shown in table 1.

β＝An/A₀

In the formula, An is the absorbance of the experimental group, A₀Absorbance of negative control group

TABLE 1 cytotoxicity assays

Relative cell survival (%)	0.25％	0.5％	1％	2％
					EXAMPLE 1 group	96.37±8.56	93.43±7.55	91.55±9.27	90.72±6.51

According to ISO 10993-5: 2009, the cell viability was greater than 70% and was considered non-toxic. As can be seen from the above results, the plant fermentation composition of the present invention has a higher cell viability than 90% at the selected mass concentration, considering its non-cytotoxicity.

Example 4

And (3) evaluating the hydroxyl radical scavenging capacity of the plant fermentation composition for whitening, removing freckles and resisting saccharification.

Antioxidation refers to the abbreviation of antioxidant free radical, Anti-Oxidant. The human body continuously generates free radicals in the human body due to continuous contact with the outside, including respiration (oxidation reaction), external pollution, radiation irradiation and other factors. Scientific studies have shown that cancer, aging or other diseases are mostly associated with the production of excess free radicals. Research on antioxidation can effectively overcome the harm caused by the antioxidation, so the antioxidation is listed as one of the main research and development directions by health-care products and cosmetic enterprises, and is also one of the most important functional requirements of the market. Antioxidant effects are well recognized as indicative of anti-aging effects.

Vitamin C is selected as a positive control group for administration, and the groups of example 1, comparative examples 1-4 and V are set_CAnd (4) grouping. Dissolving the above components with purified water to obtain sample solutions with concentration of 15ug/ml, 25ug/ml, 35ug/ml, 45ug/ml and 55ug/ml, adding sample, FeSO 4.7H2O solution (4.5mmol/L), salicylic acid-ethanol solution and H2 according to the preparation method of A, A1 and A2₂O₂Mixing the solutions (4.4mmol/L), warm bathing in a constant temperature water bath at 37 deg.C in dark for 30min, and measuring absorbance at 510 nm;

hydroxyl radical scavenging rate (%) [1- (A1-A2)/A0] X100%

A0: 1.0ml of FeSO4 & 7H2O +1.0ml of salicylic acid +1.0ml of hydrogen peroxide +1.0mLH2O

A1: 1.0mL FeSO4 & 7H2O +1.0mL salicylic acid +1.0mL hydrogen peroxide +1.0mL sample

A2: 1.0mL FeSO4 & 7H2O +1.0mL salicylic acid +1.0mL H2O +1.0mL sample

The relevant test data are shown in table 2:

TABLE 2 various groups of different concentrations of scavenging ability for hydroxyl radical

As can be seen from the test data in table 2, example 1 has a higher hydroxyl radical scavenging ability as compared with the positive test group using Vc; the lactobacillus crispatus CGMCC No.6364 is contained, but the clearance rate of the culture solution fermented by the extract containing curcumin or the white mulberry root-bark is better than that of the simple lactobacillus crispatus CGMCC No.6364 (the clearance rate of the group of comparative example 2 and the group of comparative example 3 is more than that of the comparative example 1); the lactobacillus crispatus CGMCC No.6364 is replaced by the lactobacillus crispatus CGMCC1.2743 for fermentation (other culture conditions are not changed), the effect of removing free radicals is the worst, even the effect is inferior to the effect of simple lactobacillus crispatus CGMCC No.6364 fermentation, which shows that the active extract of the conventional fermentation product of the lactobacillus crispatus CGMCC No.6364 has better hydroxyl free radical removing capability, and the effect can be obviously improved by adding curcumin or a cortex mori extract, while the lactobacillus crispatus of other strains has no outstanding effect.

Example 5

Inhibition of non-enzymatic glycosylation assays

The test simulates the non-enzymatic glycosylation reaction in human body by mixing and incubating bovine serum albumin and fructose, the inhibitor of the non-enzymatic glycosylation reaction is added to inhibit the reaction, the fluorescence intensity of the system with the added inhibitor is weaker than that of the control without the added inhibitor, and the inhibition rate is used for reflecting the inhibition condition of the sample on the non-enzymatic glycosylation reaction. A certain amount of a mixed solution containing 10mg/mL bovine serum albumin and 0.25mol/L fructose was taken as a reaction solution. PBS (1% by mass) solution of example 1 and comparative examples 1-4 was prepared as a treatment group, and a positive control group (1.00% by mass of Vc ethyl ether), a negative control group (PBS instead of sample) and a blank group (PBS instead of reaction solution) were set, each group having 3 parallel tubes. Incubate at 37 ℃ in the dark. The Fluorescence intensity (RFU) of each test group was measured at 370nm (absorption wavelength)/440 nm (excitation wavelength) using a Fluorescence microplate reader selected from items 1, 3, 5 and 7d, and the inhibition ratio was calculated. The results are shown in Table 3.

Inhibition (%) [1- (RFU inhibition group-RFU blank group)/RFU negative control group ] × 100%

TABLE 3 results of inhibition of non-enzymatic glycosylation in each experimental group

Relative inhibition rate	D1	D3	D5	D7
					Positive group (%)	40.3±2.3	50.6±4.7	57.0±4.1	62.8±7.7
Example 1 group (%)	47.6±5.1	58.8±6.7	70.3±5.1	73.8±4.3
					Comparative example 1 group (%)	12.2±1.3	15.6±1.4	16.2±2.6	18.4±2.0
Comparative example 2 group (%)	14.3±1.8	18.4±1.9	18.5±1.7	21.7±1.8
					Comparative example 3 group (%)	15.4±1.5	17.4±1.7	20.7±1.5	22.8±3.1
Comparative example 4 group (%)	5.3±0.63	6.4±0.9	7.3±1.1	9.3±2.3

As can be seen from Table 4, the inhibition rate of the non-enzymatic glycosylation reaction was gradually increased by the respective experimental groups with the increase of the incubation time.

Example 1 compared with the positive test group, the inhibition rate of the non-enzymatic glycosylation reaction is higher; the lactobacillus crispatus CGMCC No.6364 is contained, but the inhibition rate of the culture solution containing curcumin or cortex mori radicis extract fermentation is better than that of the simple lactobacillus crispatus CGMCC No.6364 fermentation culture (the inhibition rates of the comparative example 2 group and the comparative example 3 group are higher than that of the comparative example 1); and the lactobacillus crispatus CGMCC No.6364 is replaced by the lactobacillus crispatus CGMCC1.2743 for fermentation (other culture conditions are not changed), and the inhibition rate is lowest. The active extract of the conventional fermentation product of lactobacillus crispatus CGMCC No.6364 has better inhibition rate of non-enzymatic glycosylation, and has good synergistic effect by adding curcumin or cortex mori extract, so that the inhibition rate of non-enzymatic glycosylation can be improved. Other strains of Lactobacillus crispatus did not have such outstanding efficacy.

Example 5

Inhibition of tyrosinase in vitro experiments:

PBS solution is used for preparing 1mg/ml tyrosinase solution and 200U/ml tyrosinase solution. Example 1 and comparative examples 1 to 4 were designed as treatment groups, and vitamin C ethyl ether solution was a positive control group. The fermentation composition and the vitamin C ethyl ether solution of the treatment group were prepared using PBS to 4 experimental concentrations with mass fractions of 0.25%, 0.5%, 1%, 2%, respectively.

And (3) enzyme activity detection: transferring four groups of sample solutions a, b, c and d according to the table 3 by using a micropipette, placing the sample solutions in a 1.5ml EP tube (the reaction system is prepared into 1000u1), and carrying out water bath at 35 ℃ for 10 min; adding 200ul tyrosinase solution, carrying out water bath at 35 ℃ for 30min, respectively taking 150ul tyrosinase solution from each reaction tube, adding the solution into a 96-well plate, and measuring the absorbance A value at 475nm by using an enzyme-labeling instrument. Each group was tested in duplicate 3 times.

TABLE 4

Relative inhibition rate	a	b	c	d
					Tyrosine solution	0	200ul	0	200ul
Tyrosinase solution	200ul	200ul	200ul	200ul
					Aqueous solution	200ul	200ul	0	0
PBS	800ul	600ul	800ul	600ul
					Experimental group	0	0	200ul	200ul

Tyrosinase inhibition (I) was calculated for each experimental group, I ═ 100% of [1- (Ad-Ac)/(Ab-Aa) ]. The test results are shown in table 4.

TABLE 5 tyrosinase inhibition at different concentrations in each experimental group

As can be seen from the above table, the tyrosinase inhibition rate of each experimental group gradually increased with the increase of the concentration.

Example 1 had a higher tyrosinase inhibition rate compared to the positive test group; the lactobacillus crispatus CGMCC No.6364 is contained, but the inhibition rate of the culture solution containing curcumin or cortex mori radicis extract fermentation is better than that of the simple lactobacillus crispatus CGMCC No.6364 fermentation culture (the inhibition rates of the comparative example 2 group and the comparative example 3 group are higher than that of the comparative example 1); the lactobacillus crispatus CGMCC No.6364 is replaced by the lactobacillus crispatus CGMCC1.2743 for fermentation (other culture conditions are not changed), the inhibition rate is lowest and even worse than that of the single lactobacillus crispatus CGMCC No.6364 for fermentation, which shows that the active extract of the conventional fermentation product of the lactobacillus crispatus CGMCC No.6364 has better tyrosinase inhibition rate, and the added curcumin or the mulberry bark extract has good synergistic effect, so that the tyrosinase can be greatly inhibited. Other strains of Lactobacillus crispatus did not have such outstanding efficacy.

Example 7 an exemplary embodiment of a facial mask

The whitening, freckle-removing and anti-saccharification plant fermentation composition in the embodiment 1 is added with the following raw materials to prepare a corresponding facial mask according to the parts by weight:

5 parts of a plant fermentation composition for whitening, removing freckles and resisting saccharification, 0.1 part of micromolecular hyaluronic acid, 3 parts of silk protein powder, 1 part of sodium carboxymethylcellulose, 2 parts of sodium alginate, 10 parts of cetostearyl alcohol, 5 parts of a rose extract and 200 parts of deionized water.

The preparation method of the mask comprises the following steps:

s1, respectively weighing sodium carboxymethylcellulose, cetostearyl alcohol and the like according to parts by weight, uniformly mixing, simultaneously adding a proper amount of deionized water, heating to 85-95 ℃, continuously stirring in the heating process until the deionized water is completely dissolved, and standing in a beaker for later use (component A);

s2, respectively weighing micromolecule hyaluronic acid, silk protein powder, sodium alginate and an aloe extracting solution according to parts by weight, uniformly mixing the micromolecule hyaluronic acid, the silk protein powder, the sodium alginate and the aloe extracting solution, simultaneously adding a proper amount of deionized water, heating the mixture to 70-85 ℃ until the mixture is completely dissolved, and standing the mixture in a beaker for later use (component B);

s3, weighing the whitening, freckle-removing and anti-saccharification plant fermentation composition according to parts by weight, uniformly mixing, simultaneously adding a proper amount of deionized water, heating to 55-75 ℃ until the deionized water is completely dissolved, and standing in a beaker for later use (component C);

s4, respectively stirring the component A, the component B and the component C for 8-20 min by using a high-speed shearing machine under the condition of 3000-4500 r/min to promote the dissolution of the components, and then mixing and homogenizing the component A, the component B and the component C for 7-15 min; homogenizing for 1-3 times;

s5, after homogenizing, naturally cooling the mixture until the temperature is lower than 25 ℃, and cooling and standing the mixture for 2-3 hours at room temperature to obtain the skin care product.

The prepared mask liquid has moisturizing effect, and all indexes meet relevant national regulations.

Example 8 Patch safety test

The plant fermentation composition for whitening, freckle removing and anti-saccharification is subjected to human safety study, a 1% concentration pure water solution of the plant fermentation composition for whitening, freckle removing and anti-saccharification of the invention in the embodiment 1 is used as a test group 1, and distilled water is used as a control group for testing, 30 persons in each group, and no statistical difference exists between the two groups. According to the regulation in the cosmetic hygiene code (2007 edition), more than 2 persons with skin adverse reactions of grade "++" or any 1 person with skin adverse reactions of grade "+++" or more than grade +++ "are present in 30 subjects, and the subjects are judged to have adverse reactions to human bodies. The results of the relevant tests are shown in the table below.

TABLE 6 safety test for skin patches

As shown in the table, after 0.5 hour, 24 hours and 48 hours of the spot removal tester, no adverse skin reaction is observed in the distilled water control area of 30 subjects, and no 1 case of the tested area of the whitening, spot-removing and anti-saccharification plant fermentation composition has a '++' reaction, which shows that the whitening, spot-removing and anti-saccharification plant fermentation composition has high safety and causes no adverse reaction on human skin.

Finally, it should be noted that the above embodiments are only used for illustrating the technical solutions of the present invention, and not for limiting the protection scope of the present invention, although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that modifications or equivalent substitutions can be made on the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.

Claims

1. A plant fermentation composition for whitening, removing freckles and resisting saccharification is characterized by comprising the following components in parts by weight:

(1) inoculating, fermenting and culturing the preserved lactobacillus crispatus CGMCC No.6364 into a seed solution under the aseptic condition;

(2) inoculating lactobacillus crispatus CGMCC No.6364 seed liquid into a liquid culture medium for fermentation culture;

(3) and (3) centrifuging the fermentation liquor obtained in the step (2), collecting supernatant to obtain fermentation filtrate, and filtering the fermentation filtrate by a 0.22um sterilizing filter.

2. The plant fermentation composition of claim 1, wherein step (1) is:

(1) inoculating the preserved Lactobacillus crispatus CGMCC No.6364 into MRS culture medium under aseptic condition, culturing at 35 deg.C and 200rpm for 12 hr to obtain culture as seed solution for fermentation culture with final concentration of 6 × 10⁷cfu/ml。

3. The plant fermentation composition of claim 1 or 2, wherein step (2) is:

the preparation method of the liquid culture medium comprises the following steps: mixing 0.3g curcumin, 50mL cortex Mori extractive solution, 25g glucose, 8g peptone, 7g yeast extract, 8g beef extract, 5g sodium acetate, 2g ammonium citrate, 2mL tween-80, 0.6g MgSO₄·7H₂O, 0.3g of MnSO₄·4H₂O, 2g of K₂HPO₄And 1000mL of distilled water, adjusting the pH value to 6.2-6.4, and sterilizing with high pressure steam to obtain the sterile culture medium.

4. The phytofermentation composition for whitening, freckle removal and anti-glycation according to any one of claims 1 to 3, which is made into a fermentation product cosmetic for whitening, freckle removal and anti-glycation.

5. The use according to claim 4, characterized in that it is made as a mask.

6. The use of claim 5, wherein the mask formulation comprises, by weight: 5 parts of a plant fermentation composition for whitening, removing freckles and resisting saccharification, 0.1 part of micromolecular hyaluronic acid, 3 parts of silk protein powder, 1 part of sodium carboxymethylcellulose, 2 parts of sodium alginate, 10 parts of cetostearyl alcohol, 5 parts of a rose extract and 200 parts of deionized water.