CN116370446A - Composition, method and application for realizing anti-inflammatory effect by regulating specific skin microorganisms - Google Patents

Abstract

The invention discloses a composition for realizing anti-inflammatory effect by regulating and controlling specific skin microorganisms, which comprises a carbon source and a nitrogen source; the carbon source is selected from one or more of saccharide isomer, sphingomonas fermentation product extract, glycerol, sodium hyaluronate and trehalose; the nitrogen source is one or more selected from recombinant type I collagen, recombinant type III collagen, recombinant type XVII collagen, arginine, cocoyl glutamic acid TEA salt and ceramide. The invention also discloses a method for realizing anti-inflammatory effect by regulating specific skin microorganisms and application of the composition. The invention can effectively help the stabilization of skin microecology, resist the colonization of conditional pathogenic bacteria and reduce the risk of skin diseases.

Description

A composition and method for achieving anti-inflammatory effects by regulating specific skin microorganisms and application

technical field

The invention belongs to the field of biotechnology, and in particular relates to a composition, method and application for realizing anti-inflammatory effects by regulating specific skin microorganisms.

Background technique

The existing super organism composed of human body and symbiotic microorganisms can be regarded as a "walking" bioreactor. The part of the food ingested by the human body that cannot be digested and absorbed by the human body and the substances secreted by the human body into the intestinal cavity, such as mucin, can be regarded as the "culture medium" of the intestinal flora, changing the quality of the medium entering the reactor. Nutrient composition is an effective way to alter the microbial composition within them. For human skin, the skin microbiome is a similar situation and logic. Both cosmetics and substances secreted by the skin can be fermented by skin microorganisms, thereby affecting the physiological functions of the skin. More and more studies have shown that skin microbes and their metabolites play a very important role in skin barrier and function, and can even participate in the polarization of some immune cells in the basal layer of the skin, thereby affecting the physiological and immune responses of the skin. For example, the lipopoly-peptide of Staphylococcus epidermidis can also stimulate keratinocytes to secrete antimicrobial peptides to resist the colonization and reproduction of pathogenic bacteria in the skin. Therefore, in-depth research on the skin microbiome and the development of new technologies based on the research on the skin microbiome have huge opportunities in the development of cosmetic formulations and functional raw materials.

A large number of studies on the intestinal microbiome have revealed that short-chain fatty acids are involved in many physiological processes in the human body, such as increasing the insulin sensitivity of fat cells, providing nutrition for intestinal epithelial cells, maintaining the health of the intestinal barrier, and so on. Studies have also shown that the introduction of short-chain fatty acids into cosmetic formulations can also help repair the skin barrier and play a soothing role. By introducing short-chain fatty acids into the formula, the biological functions of short-chain fatty acids can be directly used to help repair the skin barrier, but there are also some limitations, which are reflected in: short-chain fatty acids have small molecular weight and strong skin permeability, so they have Irritation risk; short-chain fatty acids are highly volatile, and have high requirements for formulation technology; at the same time, because of their good water solubility, they may not stay on the skin for a long time after use due to penetration or sweat, and play a long-lasting role. Effect: short-chain fatty acids have a certain special smell, which puts forward new requirements for the fragrance of the formula.

Contents of the invention

In view of the various limitations of the application of the above-mentioned short-chain fatty acids to skin repair, in the first aspect, the present invention discloses a composition that achieves an anti-inflammatory effect by regulating specific skin microorganisms, including a carbon source and a nitrogen source; the carbon source is selected from One or more of sugar isomers, Sphingomonas fermentation product extract, glycerin, sodium hyaluronate, and trehalose; the nitrogen source is selected from recombinant type I collagen, recombinant III One or more of type collagen, recombinant type XVII collagen, arginine, cocoyl glutamic acid TEA salt, and ceramide.

In some embodiments, the carbon source is carbohydrate; the nitrogen source is protein.

In some embodiments, the carbohydrate is a carbohydrate of human origin; the protein is a protein of human origin.

In some embodiments, the carbon source and the nitrogen source are used in combination for microbial fermentation, so that the content of butyric acid produced by the microorganism is the same as that produced by the microbial fermentation without the composition under the same conditions. 1.2 to 7.9 times that of butyric acid. Furthermore, it is 1.5 to 7.8 times.

In some embodiments, butyric acid is produced by microorganisms as a component of the fermentation medium; preferably, the concentration of the carbon source in the fermentation medium is 0.1% to 3%, preferably 0.2% to 2%, More preferably 0.5%-1%; the concentration of the nitrogen source in the fermentation medium is 0.1%-3%, preferably 0.2%-2%, more preferably 0.25%-1%.

In some embodiments, the concentration ratio of the carbon source to the nitrogen source is 1:(0.1-3), preferably 1:(0.25-2.5), more preferably 1:(0.5-2).

In some embodiments, the composition is a raw material with skin anti-inflammatory effect screened by an evaluation method for rapid screening of raw materials with skin anti-inflammatory effect;

The evaluation method for the rapid screening of raw materials with skin anti-inflammatory effect comprises the following steps:

S1. Obtain the draft genome data of the target strain through NGS sequencing or public database retrieval;

S2. Using the gene prediction software to predict genes from the draft genome data of the target strain;

S3. By collecting the nucleic acid sequences of key genes in the short-chain fatty acid synthesis pathway, use multiple sequence alignment software to perform multiple sequence alignments, output the sequence alignment in STOCKHOLM format, and then use hmmer software to construct the key genes in the short-chain fatty acid synthesis pathway Hidden Markov model; each key gene corresponds to a hidden Markov model, so there are n key genes on the short-chain fatty acid synthesis pathway, which corresponds to the hidden Markov model of n key genes on the short-chain fatty acid synthesis pathway , n>0; use the hidden Markov model of key genes on the short-chain fatty acid synthesis pathway to determine whether each gene predicted in step S2 is a key gene on the short-chain fatty acid synthesis pathway: if the target strain has key genes on the short-chain fatty acid synthesis pathway, the possibility of the target strain producing short-chain fatty acids is large; otherwise, it is small; determine the target strain according to the key gene types on the short-chain fatty acid synthesis pathway possessed by the target strain Synthetic pathway for producing short-chain fatty acids, so as to delineate the selection range of raw materials and construct the target strain library;

S4. Ferment and co-culture the target strains selected from the target strain library and the raw materials to be screened within the defined range according to the set ratio and conditions to obtain a fermentation co-culture solution;

S5. Determining the content of short-chain fatty acids in the fermentation co-culture solution by using a targeted GC-MS analysis and determination method;

S6. It is used to irradiate keratinocytes with UVB to trigger the expression of IL-6 in the keratinocytes, and then add the fermentation co-culture solution to detect the decrease of IL-6 expression to obtain the IL-6 inhibitory effect of the fermentation co-culture solution, The purpose of evaluating the skin anti-inflammatory effect of the raw material to be screened is realized; the evaluation standard is that the better the IL-6 inhibitory effect is, the better the skin anti-inflammatory effect of the raw material to be screened is.

Further, the gene prediction software is prokka software; the multiple sequence alignment software is clustalo software.

In some embodiments, the short-chain fatty acid is one or more of acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and caproic acid. Butyric acid is preferred.

In some embodiments, the target strain is a microorganism derived from skin or other tissues of the human body. Further, the draft genome is a draft genome that has been published in the prior art, or is obtained through sequencing and assembly.

In some embodiments, the synthesis pathway of butyric acid includes 4 pathways, respectively the first pathway, the second pathway, the third pathway and the fourth pathway, and these 4 synthesis pathways involve 3 common genes; the 3 common The hidden Markov models of the key genes corresponding to the genes are bcd.hmm, etfA.hmm, etfB.hmm; the hidden Markov models of the key genes corresponding to the first pathway are thl.hmm, bhbd.hmm, cro .hmm, but.hmm, buk.hmm; the hidden Markov models of key genes corresponding to the second pathway include gctA.hmm, gctB.hmm, hgCoAdA.hmm, hgCoAdB.hmm, hgCoAdC.hmm, gcdA.hmm , gcdB.hmm; the hidden Markov model of the key gene corresponding to the third pathway has 4hbt.hmm, abfD.hmm, abfH.hmm; the hidden Markov model of the key gene corresponding to the third pathway has kamA .hmm, kamD.hmm, kamE.hmm, kdd.hmm, kce.hmm, kal.hmm, atoA.hmm, AtoD.hmm;

bcd.hmm, etfA.hmm, etfB.hmm, thl.hmm, bhbd.hmm, cro.hmm, but.hmm, buk.hmm, gctA.hmm, gctB.hmm, hgCoAdA.hmm, hgCoAdB.hmm listed above , hgCoAdC.hmm, gcdA.hmm, gcdB.hmm, 4hbt.hmm, abfD.hmm, abfH.hmm, kamA.hmm, kamD.hmm, kamE.hmm, kdd.hmm, kce.hmm, kal.hmm, atoA .hmm and AtoD.hmm are hidden Markov models of key genes in the short-chain fatty acid synthesis pathway.

In some embodiments, the target strain is a microorganism derived from skin or other tissues of the human body.

In some embodiments, the target bacterial strain is selected from Staphylococcus epidermidis; further, the Staphylococcus epidermidis is Staphylococcus epidermidis ATCC 12228, Staphylococcus epidermidis ATCC 14990, Staphylococcus epidermidis ATCC 35984, Staphylococcus epidermidis ATCC 700926 and/or or Staphylococcus epidermidis ATCC 51625.

In the second aspect, the present invention also discloses the application of the above-mentioned composition in the preparation of skin anti-inflammatory preparations. Further, the skin anti-inflammatory preparation is skin care product, cosmeceutical preparation, cosmetic and/or medicine.

In the third aspect, the present invention also discloses a method for achieving an anti-inflammatory effect by regulating specific skin microorganisms, providing a fermentation medium to the microorganisms on the skin for producing short-chain fatty acids to achieve an anti-inflammatory effect; the fermentation medium contains the above said composition.

In some embodiments, the short chain fatty acid is acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid, and/or caproic acid.

In some embodiments, modulation of specific skin microbial flora is achieved by altering the composition of the fermentation medium.

The beneficial effects are:

1) Skin micro-ecological friendliness: Using selected skin strains, fermentation raw materials, and raw materials as the fermentation medium of the selected strains can provide nutrients for the selected strains, and then provide the growth of the strains on the skin, ecologically Stability provides a good support, which can effectively help the stability of skin microecology, resist the colonization of opportunistic pathogens, and reduce the risk of skin diseases;

2) Long-term exertion of the function of short-chain fatty acids: Compared with the method of directly adding short-chain fatty acids, the method of continuously producing short-chain fatty acids by using the raw material itself to ferment selected strains can provide a relatively stable and continuous low-dose short-chain fatty acid. Chain fatty acid stimulation, long-term maintenance of skin biological effects of short-chain fatty acids, and can effectively avoid problems such as irritation and odor caused by direct addition due to high concentration;

3) Based on the evaluation method and database system of selected strains and raw materials, it is possible to carry out personalized raw material combinations and formula combinations for specific strains, so as to realize the development of precise personalized customization methods and effects. The evaluation database system of selected strains-raw materials is an extension based on the present invention, and is used to establish the corresponding relationship between skin microbial strains and raw materials, that is, which strains can be effectively acted on by the fermentation medium containing which raw materials to generate short-chain fatty acids, And these corresponding relationships are collected into a database to form a system.

The idea, specific structure and technical effects of the present invention will be further described below in conjunction with the accompanying drawings, so as to fully understand the purpose, features and effects of the present invention.

Description of drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following will briefly introduce the accompanying drawings that need to be used in the description of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only some of the present invention. Embodiments, for those of ordinary skill in the art, other drawings can also be obtained according to these drawings without paying creative labor.

Figure 1 is a schematic diagram of the structure of the fermentation co-cultivation process.

Fig. 2 is the morphological diagram of keratinocytes under the action of different concentrations of samples C3, wherein Solvent Control is the control group without adding C3 solvent.

Fig. 3 is a graph of cell viability of keratinocytes under the action of different concentrations of sample C3.

Fig. 4 is a histogram of IL-6 content results of sample C3.

Fig. 5 is a histogram of IL-6 content results of samples C17 and C19.

Fig. 6 is a histogram of IL-6 content results of samples B1 and B10.

Figure 7 is a schematic diagram of butyrate synthesis pathway and key genes.

Detailed ways

In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments It is a part of embodiments of the present invention, but not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

Example 1

Take Staphylococcus epidermidis ATCC 12228 as an example for illustration. Figure 1 shows a fermentation co-cultivation model or approach. The raw material formula is pre-treated (including mixing and sterilization, etc.); the Staphylococcus epidermidis ATCC 12228 (S.epidemidisATCC 12228) is subjected to strain pre-treatment (including strain activation, enrichment, etc.); and then the pre-treated raw material formula is combined with After strain pretreatment, Staphylococcus epidermidis ATCC 12228 was co-cultured for 24 hours (aerobic, 37°C, 220rpm), and the metabolites in the bacterial liquid were detected by GC-MS (gas chromatography-mass spectrometry) to detect short-chain fatty acids (SCFAs).

raw material:

Carbohydrate isomers (from DSM), recombinant type I collagen (from Jiangsu Create Medical Technology Co., Ltd.), recombinant III collagen (from Jiangsu Create Medical Technology Co., Ltd.), recombinant type XVII collagen (from Jiangsu Create Medical Technology Co., Ltd.) From Jiangsu Create Medical Technology Co., Ltd.), Trehalose (from Hayashibara Co., Ltd.), Glycerin (from Indonesia Spring Gold), Sphingomonas Ferment Extract (from Lubrizol), Sodium Hyaluronate (from Bloomage), cocoyl glutamic acid TEA salt (from Ajinomoto), ceramide (from Shenzhen Dickman).

Specific experimental steps:

1. Prepare the fermentation medium

Take a 50ml centrifuge tube, add the corresponding amount of substrate sample (such as the fermentation system except the strains in Table 1 to Table 3), add the corresponding amount of phosphate buffer saline (PBS), mix well, and for high temperature sterilization samples, wrapped in aluminum foil, sterilized under high temperature and high pressure at 121°C for 30-60min, and temporarily stored at 4°C after sterilization. For samples that cannot be sterilized by high temperature, use a 0.22 μm filter membrane to filter and sterilize, and store them temporarily at 4°C after sterilization.

2. Fermentation

S. epidermidis ATCC 12228 was cultured overnight on a tryptic soy broth (TSB) agar plate at 37°C. A single colony was inoculated in TSB medium and cultured at 37°C for about 24h. Centrifuge at 4000rpm for 10 minutes (20°C) to obtain bacterial microspheres, and resuspend in PBS.

Add 1ml of Staphylococcus epidermidis (concentration about 10 ⁸ CFU/mL) to each tube (15ml) of the fermentation system. After mixing, incubate at 37° C. under aerobic conditions (220 rpm shaking) for 24 hours. The PBS system containing bacteria and no fermentation medium is used as a control. Since the bacteria in this example refers to S. epidermidis ATCC 12228, it is referred to as epidermidis+medium blank for short, and the medium blank refers to PBS.

3. Sampling

Take out the fermented 50ml centrifuge tube, use a disposable syringe to absorb 2ml, filter it into a 2ml centrifuge tube with a 0.22 μm filter, and store it in a -80°C refrigerator for subsequent determination of short-chain fatty acids by gas chromatography-mass spectrometry. Use a disposable syringe to draw 8-10ml, filter it into a 15ml centrifuge tube with a 0.22μm filter, and store it in a -80°C refrigerator for subsequent evaluation of the inhibitory effect of UVB-induced IL-6 expression in skin keratinocytes. The remaining samples were filtered through a 0.22 μm filter into a 15ml centrifuge tube, measured with a pH meter, and frozen in a -80°C refrigerator for later use.

Table 1-Table 3 lists multiple groups of fermentation examples. These fermentation examples all use epiglucose+medium blank as a reference fermentation system to compare the ability to produce short-chain fatty acids, especially the ability to produce butyric acid. As can be seen from Table 1-Table 3, when there is 1% sugar isomers in the fermentation system, the output of acetic acid, propionic acid, isobutyric acid, butyric acid, isovaleric acid, valeric acid and hexanoic acid Compared with the reference fermentation system, there is an improvement, for example, the production of acetic acid is 9.42 times that of the reference fermentation system. When there were 1% sugar isomers and 0.5% recombinant type XVII collagen in the fermentation system (ie, fermentation system C3), the production of butyric acid was 7.80 times that of the reference fermentation system.

Table 1. Single raw material of fermentation example

Fermentation system Fermentation concentration Acetic acid propionic acid Isobutyric acid butyric acid Isovaleric acid Valeric acid caproic acid Epiglucose+medium blank \ 1 1 1 1 1 1 1 sugar isomers 1.00% 9.42 4.69 15.60 2.73 5.62 1.59 1.81 Recombinant Type III Collagen 0.50% 2.81 4.01 51.99 42.57 20.74 2.95 2.57 Recombinant Type XVII Collagen 0.50% 0.09 0.44 0.89 7.25 0.24 1.00 1.25

Table 2. Binary/ternary compound raw material fermentation system and fermentation concentration of fermentation examples

The relative content of short-chain fatty acids corresponding to the fermentation system in table 3 and table 2

The fermentation concentrations in Table 1 and Table 2 correspond to various raw materials in the fermentation system. For example: the fermentation concentration of the recombinant type III collagen in table 1 refers to the fermentation medium containing 0.5% recombinant type III collagen; the fermentation concentration of the C3 fermentation system in table 2 refers to the fermentation medium containing 1% Sugar isomers and 0.5% recombinant collagen type XVII.

Example 2

Materials: Keratinocytes were from Guangdong Boxi Biotechnology Co., Ltd., batch number Ep220707. KC2500 (Guangdong Boxi Biology), PBS (Solebux), dexamethasone (Sigma), IL-6 ELISA kit (Abcam), MTT (Sigma), DMSO (Sigma).

Main equipment: _CO2 incubator (Thermo, 150I), ultra-clean bench (Suzhou Antai, SW-CJ-1F), microplate reader (BioTek, Epoch), UVB irradiation instrument (Philips).

1. Evaluation of the toxicity of the fermentation broth in the sugar isomer + recombinant XVII collagen fermentation system (C3) to keratinocytes

Test Methods:

1) Cell seeding: keratinocytes were seeded into 96-well plates at a seeding density of 1×10 ⁴ cells/well, and incubated overnight in an incubator (37° C., 5% CO ₂ ).

2) Test grouping: The test set up a zero-adjustment group, a solvent control group, a positive control group and a sample group. In the sample group, 8 concentration gradients were set for each sample, and 3 replicate wells were set for each concentration gradient.

3) Dosing solution: prepare sample working solutions with different concentrations: 10%, 5%, 2.5%, 1.25%, 0.625%. That is, the concentration of the fermentation broth in the C3 fermentation system was set as 100%, and then diluted to the above concentrations respectively to obtain the sample working solution.

4) Administration: administration is performed when the keratinocyte plating rate in the 96-well plate reaches 40%-60%. Add 200 μL of culture solution to each well of the solvent control group; add 200 μL of culture solution containing 10% DMSO to each well of the positive control group; 200 μL cell culture medium. After administration, the 96-well plate was placed in an incubator (37° C., 5% CO ₂ ) for 24 hours.

5) Detection: After the cells were incubated for 24 hours, discard the supernatant, add MTT working solution (0.5mg/mL), incubate at 37°C in the dark for 4 hours, after the incubation, discard the supernatant, add 150 μL DMSO to each well, Read the OD value here.

6) Calculation of relative cell viability: calculated according to the formula,

Relative cell viability (%)=(sample well OD-zero adjustment well OD)/(solvent control well OD-zero adjustment well OD)×100%.

The results are shown in Figure 2 and Figure 3. At different concentrations, the morphology of keratinocytes was clear and no significant change was observed, and the cell viability did not fluctuate significantly. Therefore, it was concluded that at the above concentrations, the carbohydrate isomers +The fermentation broth in the recombinant XVII collagen fermentation system did not show obvious cytotoxicity to keratinocytes.

2. UVB irradiation keratinocyte IL-6 expression inhibition experiment

Table 4. Test groups

(1) Positive control group (dexamethasone) working solution configuration

Aspirate 2 μL of 10% mother solution and dissolve it in 2 mL of culture solution to prepare 0.01% dexamethasone.

(2) Operation steps

1) Cell inoculation: inoculate the cells in a 6-well plate at a seeding density of 2.8×10 ⁵ cells/well, and incubate overnight in an incubator (37° C., 5% CO ₂ ).

2) Dosing: Test groups according to Table 4. When the cell plating rate in the 6-well plate reaches 40% to 60%, dosing in groups, add 2 mL of sample to each well, and set 3 duplicate wells for each group. After administration, the 6-well plate was placed in an incubator (37° C., 5% CO ₂ ) for 24 hours.

3) UVB irradiation: After the cells were washed with PBS, according to the test group, 300mJ/cm ² of UVB irradiation was performed on the group with UVB irradiation.

4) Post-incubation: After the cells were washed with PBS, the 6-well plate was placed in an incubator (37° C., 5% CO ₂ ) and incubated for 24 hours.

5) Cell collection: After 24 hours of incubation, the cell culture supernatant was collected in an EP tube and stored in a -80°C refrigerator.

6) ELISA detection: detection and analysis were performed according to the operating instructions of the ELISA (enzyme-linked immunosorbent) detection kit.

(3) Statistical analysis of results

GraphPad Prism was used for graphing (Figure 4-Figure 6), and the results were expressed as Mean±SD. The comparison among groups was analyzed by t-test statistics. Statistical analyzes were two-tailed. P<0.05 was considered to have a significant difference, and P<0.01 was considered to have a very significant difference.

The results show that:

1) Compared with the BC group, the expression of IL-6 in the NC group increased significantly, indicating that the stimulation conditions of this test were effective;

2) Compared with the NC group, the expression of IL-6 in the PC group decreased significantly, indicating that the positive control of this test is effective;

3) Compared with the NC group, the IL-6 content of samples C3, C17, C19, B-1 and B-10 (also known as fermentation system C3, C17, C19, B-1 and B-10) all decreased significantly .

The difference in IL-6 content between NC and BC shows that the keratinocytes irradiated by UVB can normally secrete IL-6 inflammatory factor, and the difference in IL-6 content between PC and NC shows that the anti-inflammatory drug dexamethasone can effectively inhibit the occurrence of inflammation . The content of IL-6 in C3, C17, C19, B-1 and B-10 was significantly decreased compared with NC, which proved that C3, C17, C19, B-1 and B-10 had anti-inflammatory effects. Through data analysis, it was found that the relative concentrations of butyric acid produced by these five samples were all between 1.2 and 7.9, and all samples within this range also had a good inhibitory effect on IL-6, and the carbon and nitrogen sources contained in them were The formed composition can be used as the composition for achieving anti-inflammatory effects by regulating specific skin microorganisms according to the present invention.

From Examples 1 and 2, it can be proved that GC-MS is used to quickly identify the content of short-chain fatty acids in the fermentation broth, and the IL-6 inhibition test is used to evaluate the anti-inflammatory and soothing effects of the fermentation broth, so that the rapid screening and evaluation of raw materials can be achieved through specific strains Anti-inflammatory effect.

Embodiment 3, rapid screening has the evaluation method of the raw material of skin anti-inflammation effect

The method includes the following steps:

S1. Obtain the draft genome data of the target strain through NGS sequencing or public database retrieval;

S2, using prokka software to predict genes from the draft genome data of the target strain;

S3. By collecting the nucleic acid sequences of key genes in the short-chain fatty acid synthesis pathway, use the clustalo software to perform multiple sequence alignments, output the sequence alignment in STOCKHOLM format, and then use the hmmer software to construct the hidden mark of the key genes in the short-chain fatty acid synthesis pathway Each key gene corresponds to a Hidden Markov Model, so there are n key genes in the short-chain fatty acid synthesis pathway corresponding to n Hidden Markov models of key genes in the short-chain fatty acid synthesis pathway, n> 0; use the hidden Markov model of key genes on the short-chain fatty acid synthesis pathway to determine whether each gene predicted in step S2 is a key gene on the short-chain fatty acid synthesis pathway: if the target strain has short-chain fatty acids If the key gene on the synthetic pathway is selected, the possibility of the target bacterial strain producing short-chain fatty acids is large; Fatty acid synthesis pathway, so as to delineate the selection range of raw materials and construct the target strain library;

S4. Ferment and co-culture the target strains selected from the target strain library and the raw materials to be screened within the defined range according to the set ratio and conditions to obtain a fermentation co-culture solution;

S5. Determining the content of short-chain fatty acids in the fermentation co-culture solution by using a targeted GC-MS analysis and determination method;

S6. It is used to irradiate keratinocytes with UVB to trigger the expression of IL-6 in the keratinocytes, and then add the fermentation co-culture solution to detect the decrease of IL-6 expression to obtain the IL-6 inhibitory effect of the fermentation co-culture solution, The purpose of evaluating the skin anti-inflammatory effect of the raw material to be screened is realized; the evaluation standard is that the better the IL-6 inhibitory effect is, the better the skin anti-inflammatory effect of the raw material to be screened is.

Wherein, the short-chain fatty acid is butyric acid. The synthetic pathway of butyrate includes 4, respectively the first pathway, the second pathway, the third pathway and the fourth pathway (see Figure 7), and these 4 synthetic pathways involve 3 common genes; the 3 common genes The hidden Markov models of the corresponding key genes are bcd.hmm, etfA.hmm, etfB.hmm; the hidden Markov models of the key genes corresponding to the first pathway are thl.hmm, bhbd.hmm, cro. hmm, but.hmm, buk.hmm; the hidden Markov models of key genes corresponding to the second pathway include gctA.hmm, gctB.hmm, hgCoAdA.hmm, hgCoAdB.hmm, hgCoAdC.hmm, gcdA.hmm, gcdB.hmm; the hidden Markov model of the key gene corresponding to the third pathway has 4hbt.hmm, abfD.hmm, abfH.hmm; the hidden Markov model of the key gene corresponding to the third pathway has kamA. hmm, kamD.hmm, kamE.hmm, kdd.hmm, kce.hmm, kal.hmm, atoA.hmm, AtoD.hmm;

bcd.hmm, etfA.hmm, etfB.hmm, thl.hmm, bhbd.hmm, cro.hmm, but.hmm, buk.hmm, gctA.hmm, gctB.hmm, hgCoAdA.hmm, hgCoAdB.hmm listed above , hgCoAdC.hmm, gcdA.hmm, gcdB.hmm, 4hbt.hmm, abfD.hmm, abfH.hmm, kamA.hmm, kamD.hmm, kamE.hmm, kdd.hmm, kce.hmm, kal.hmm, atoA .hmm and AtoD.hmm are hidden Markov models of key genes in the short-chain fatty acid synthesis pathway.

Using the method described above, different strains of Staphylococcus epidermidis (different ATCC codes) were identified for the key genes of the butyrate synthesis pathway, and the results are shown in Table 5. It can be seen from Table 5 that the butyrate synthesis pathways of these five strains of Staphylococcus epidermidis mainly focus on the first pathway (orange pathway) and the fourth pathway (gray pathway).

Table 5. Judgment results of key genes in the butyrate synthesis pathway of selected strains of Staphylococcus epidermidis

Selected strains shared gene first access Second access third pathway fourth pathway ATCC 12228 + + - - + ATCC 14990 + + - - + ATCC 35984 + + - - + ATCC 700926 + + - - + ATCC 51625 + + - - +

Note: + means yes, - means no.

The target strain is selected from Staphylococcus epidermidis; further, the Staphylococcus epidermidis is Staphylococcus epidermidis ATCC 12228, Staphylococcus epidermidis ATCC 14990, Staphylococcus epidermidis ATCC 35984, Staphylococcus epidermidis ATCC700926 and/or Staphylococcus epidermidis ATCC 51625. The experimental data obtained with Staphylococcus epidermidis ATCC 12228 are shown in Examples 1 and 2.

The above content is only a specific implementation manner of the present application, and the protection scope of the present application is not limited thereto. Those skilled in the art may make changes or substitutions within the technical scope disclosed in this application, and these changes or substitutions shall all be within the protection scope of this application.

Claims (9)

1. A composition for achieving an anti-inflammatory effect by modulating specific skin microorganisms, comprising a carbon source and a nitrogen source; the carbon source is selected from one or more of saccharide isomer, sphingomonas fermentation product extract, glycerol, sodium hyaluronate and trehalose; the nitrogen source is one or more selected from recombinant type I collagen, recombinant type III collagen, recombinant type XVII collagen, arginine, cocoyl glutamic acid TEA salt and ceramide.

2. The composition of claim 1, wherein the carbon source is a carbohydrate; the nitrogen source is a protein.

3. The composition of claim 2, wherein the carbohydrate is a carbohydrate of human origin; the protein is a protein of human origin.

4. The composition according to claim 1, wherein the composition is used as a constituent of a fermentation medium for microbial fermentation such that the butyric acid content produced by the microorganism is 1.2 to 7.9 times the butyric acid content produced under the same conditions without adding the composition.

5. The composition of claim 1, wherein the butyric acid is produced by a microorganism as a constituent of a fermentation medium; the concentration of the carbon source in the fermentation medium is 0.1% -3%; the concentration of the nitrogen source in the fermentation medium is 0.1% -3%.

6. The composition according to claim 4, wherein the composition is a raw material having a skin anti-inflammatory effect obtained by screening by an evaluation method for rapidly screening raw materials having a skin anti-inflammatory effect;

the evaluation method for rapidly screening the raw materials with the skin anti-inflammatory effect comprises the following steps:

s1, obtaining target strain genome sketch data through NGS sequencing or public database retrieval;

s2, predicting genes from genome sketch data of the target strain through gene prediction software;

s3, collecting nucleic acid sequences of key genes on a short chain fatty acid synthesis path, performing multi-sequence comparison by using multi-sequence comparison software, outputting STOCKHOLM format of sequence comparison, and constructing a hidden Markov model of the key genes on the short chain fatty acid synthesis path by using hmmer software; each key gene corresponds to a hidden Markov model, so that the existence of n key genes on a short-chain fatty acid synthesis path corresponds to the hidden Markov models of the key genes on n short-chain fatty acid synthesis paths, and n is more than 0; determining whether each gene predicted in the step S2 is a key gene on the short-chain fatty acid synthesis pathway by using a hidden Markov model of the key gene on the short-chain fatty acid synthesis pathway: if the target strain has a key gene on a short chain fatty acid synthesis pathway, the target strain has high possibility of producing short chain fatty acids; otherwise, the small is; determining a short-chain fatty acid synthesis path of the target strain according to the key gene types on the short-chain fatty acid synthesis path of the target strain, thereby defining a selection range of raw materials and constructing a target strain library;

s4, fermenting and co-culturing the target strain selected from the target strain library and the raw materials to be screened in a defined range according to a set ratio and conditions to obtain a fermentation co-culture solution;

s5, determining the content of short-chain fatty acid in the fermentation co-culture solution by adopting a targeted GC-MS analysis and determination method;

s6, the method is used for facilitating UVB irradiation on the keratinocytes, triggering IL-6 expression of the keratinocytes, and then adding the fermentation co-culture solution to detect the IL-6 expression reduction condition to obtain the IL-6 inhibition effect of the fermentation co-culture solution, so that the purpose of evaluating the skin anti-inflammatory effect of the raw materials to be screened is achieved; the evaluation criterion is that the better the IL-6 inhibition effect is, the better the skin anti-inflammatory effect of the raw materials to be screened is.

7. Use of a composition according to any one of claims 1 to 6 for the preparation of a skin anti-inflammatory formulation.

8. A method for achieving an anti-inflammatory effect by modulating specific skin microorganisms, characterized by providing the microorganisms on the skin with a fermentation medium for producing short chain fatty acids to achieve the anti-inflammatory effect; the fermentation medium comprising the composition of any one of claims 1-6.

9. The method of achieving an anti-inflammatory effect by modulating a specific skin microorganism according to claim 8, wherein the short chain fatty acid is acetic acid, propionic acid, butyric acid, isobutyric acid, valeric acid, isovaleric acid and/or caproic acid; the regulation and control of the specific skin microbial flora is realized by changing the composition of the fermentation medium.

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