Deinococcus weizhi and use thereof

Abstract

The present invention provides a novel microbial strain resource, specifically relates to the identification of Deinococcus Weizhi as a new species, and provides specific Deinococcus weizhi VB142 and Deinococcus weizhie VB226. A fermentation broth, a fermentation product filtrate, a fermentation lysate, and extracts of the fermentation broth can be obtained by means of conventional fermentation, are applicable for the preparation of antioxidant products, and have wide application value and market prospects.

Description

Deinococcus weizhi and application thereof TECHNICAL FIELD

[0001] The present application relates to the isolation and identification of a new species of Deinococcus and its application. BACKGROUND

[0002] Strains of Deinococcus are a kind of bacteria with strong resistance to radiation. The strains do not form endospores, and the cells are spherical or rod-shaped and non-motile. In 1956, Anderson discovered the first strain of Deinococcus (Deinococcus radiodurans) R1 from meat cans treated by radiation sterilization. The strain has been widely studied in order to elucidate its mechanism of radiation resistance. At present, the genus has covered 20 valid published species. Due to the ability of Deinococcus strains to tolerate radiation, which is thousands of times higher than that of Escherichia coli, the application research of Deinococcus has attracted widespread attention.

[0003] Common radiation includes solar radiation, electromagnetic radiation, thermal radiation, etc. Among them, UVA and UVB contained in sunlight have the greatest damage to human skin, and UVB is the main cause of skin damage. In order to prevent radiation damage to human skin caused by external radiation, people often add various ultraviolet absorbers to various skin care products. Compared with ordinary ultraviolet absorbers, natural ultraviolet protection agents not only have the ability to protect against ultraviolet radiation, but also have good safety performance and are non-toxic and non-irritating.

[0004] In addition, the isolation and screening of natural microorganisms and the application of natural ingredients in cosmetics have become a hot spot in the research and development of the medical and cosmetic industries, and have high practical application value. SUMMARY

[0005] An object of the present application is to provide a new microbial strain resource.

[0006] The appended claims set forth particular aspects of the present application, as well as preferred embodiments.

[0007] The microbial strain resource provided in one aspect of the present application relates to a Deinococcus weizhi species. The Deinococcus weizhi described in the present application can also be referred to as Deinococcus sp.

[0008] Another aspect of the present application relates to a fermentation broth of Deinococcus weizhi, a fermentation product filtrate, a fermentation cytolysis product, or an extract of the fermentation broth.

[0009] Another aspect of the present application relates to the use of Deinococcus weizhi or its fermentation broth, fermentation product filtrate, fermentation lysate, extract of fermentation broth in the preparation of probiotic fermented cosmetic raw materials.

[0010] Further, more specifically, the strain or its fermentation broth, fermentation product filtrate, fermentation lysate, extract of fermentation broth is used as one or any combination of antioxidant, moisturizing, anti-inflammatory, anti-photoaging or anti-glycation active ingredients in cosmetic raw materials.

[0011] Another aspect of the present application relates to the use of Deinococcus weizhi or its fermentation broth, fermentation product filtrate, fermentation lysate, extract of fermentation broth in the preparation of antioxidant products.

[0012] Further, as a specific embodiment, the antioxidant product is selected from health products, pharmaceutical compositions or cosmetics.

[0013] Another aspect of the present application relates to a health product comprising the Deinococcus weizhi or its fermentation broth, fermentation product filtrate, fermentation lysate or extract of fermentation broth.

[0014] Another aspect of the present application relates to a pharmaceutical composition comprising the Deinococcus weizhi or its fermentation broth, fermentation product filtrate, fermentation lysate or extract of fermentation broth.

[0015] Another aspect of the present application relates to a cosmetic composition comprising one or any combination of the strain or its fermentation broth, fermentation product filtrate, fermentation lysate, extract of fermentation broth.

[0016] Further, more specifically, the cosmetic or skin care product is characterized by including cosmetic or skin care products in the form of cosmetic water, essence, cream, cream, milk, gel, mask, etc.

[0017] Further, as a specific embodiment, the Deinococcus weizhi is Deinococcus sp. VB142, deposited with the China Center for Type Culture Collection (CCTCC) of Wuhan University, with the accession number CCTCC M2024185 and the deposit date of January 23, 2024, and also deposited with the Korean Culture Center for Type Cultures (KCTC), with the accession number 15470BP and the deposit date of June 19, 2023.

[0018] Further, as a specific embodiment, the Deinococcus weizhi is Deinococcus sp. VB226, which is preserved in the China Center for Type Culture Collection (CCTCC) of Wuhan University, and the preservation number is CCTCC M2024186, and the preservation date is January 23, 2024.

[0019] As used herein, the term "sequence identity" refers to the extent of identity, over the entire length of the aligned sequences, between the amino acid residues or bases of the sequences after the two sequences are aligned to be as consistent as possible in a particular comparison region.

[0020] The present application provides a new microbial strain resource, specifically relates to Deinococcus weizhi identified as a new species, and provides specific Deinococcus sp. VB142 and Deinococcus sp. VB226, which are obtained by conventional fermentation to obtain fermentation broth, fermentation product filtrate, fermentation lysate, extract of fermentation broth, and can be used for preparing antioxidant products selected from health products, pharmaceutical compositions or cosmetics, and have wide application value and market prospect.

[0021] The second aspect of the present application provides a polypeptide, wherein the polypeptide sequence has at least 90% sequence identity with the amino acid sequence shown in SEQ ID NO. 3.

[0022] The polypeptide provided by the present application has high safety, strong cell repair ability, anti-inflammatory, scar lightening and moisturizing effects. The polypeptide of the present application is derived from Deinococcus weizhi. Through further research on genomics, the inventors analyzed the differences in the genomes of Deinococcus weizhi and other Deinococcus species, found a series of exopolypeptides specific to Deinococcus weizhi, and isolated some polypeptides with repair and anti-inflammatory effects. According to the amino acid sequence of the isolated polypeptide, those skilled in the art can more quickly obtain the polypeptide of the present application by chemical synthesis.

[0023] The term "peptide" used in the present specification can refer to a linear molecule formed by the interconnection of amino acid residues through peptide bonds. The peptide can be prepared according to the chemical synthesis method known in the art, especially according to the solid-phase synthesis technique.

[0024] According to the specific embodiments of the present application, the polypeptide provided by the present application can be directly chemically synthesized, or can be isolated from the secreted polypeptide of Deinococcus weizhi, or can be expressed and secreted in other host cells. The present application provides the amino acid sequence of the above-mentioned polypeptide, and those skilled in the art can obtain the polypeptide described in the present application by using the prior art. Any method known in the art for obtaining a polypeptide is within the scope of the present application. The synthesized polypeptide can be directly stored in the form of powder, which is convenient for storage and transportation.

[0025] The third aspect of the present application provides an isolated polynucleotide encoding the above-mentioned polypeptide, wherein:

[0026] According to the specific embodiments of the present application, the polynucleotide has at least one of the nucleotide sequences of SEQ ID NO. 4 with at least 90% sequence identity.

[0027] It should be noted that, for the nucleic acid mentioned in the specification and claims of the present application, those skilled in the art should understand that it actually includes any one of the complementary double strands, or both. For the convenience, in the specification and claims, although only one strand is given in most cases, the other complementary strand is actually disclosed. In addition, the nucleic acid sequence in the present application includes DNA form or RNA form, and the disclosure of one means the disclosure of the other. Those skilled in the art can easily synthesize the polypeptide of the present application by using the nucleic acid sequence or the amino acid sequence provided by the present application.

[0028] The fourth aspect of the present application is to provide a variety of expression vectors comprising the polynucleotide as described above, wherein:

[0029] According to the specific embodiments of the present application, the expression vector comprises the polynucleotide of SEQ ID NO. 4 with at least 90% sequence identity.

[0030] According to the embodiments of the present application, the expression vector can comprise optional control sequences, which are operably linked to the nucleic acid molecule. The control sequences are one or more control sequences that can direct the expression of the nucleic acid molecule in a host. The expression vector provided in the embodiments of the present application can efficiently and massively express the polypeptide in a suitable host cell.

[0031] The fifth aspect of the present application is to provide a recombinant cell carrying the above-mentioned polynucleotide, the above-mentioned expression vector, or capable of expressing the above-mentioned polypeptide.

[0032] According to the specific embodiments of the present application, the recombinant cell is obtained by introducing the aforementioned expression vector into a host cell.

[0033] It should be noted that the recombinant cell of the present application is not particularly limited, and can be a prokaryotic cell, a eukaryotic cell, or a bacteriophage. The prokaryotic cell can be E. coli, B. subtilis, Streptococcus, or Giardia lamblia, etc. The eukaryotic cell includes Pichia pastoris, Saccharomyces cerevisiae, Schizosaccharomyces, Trichoderma, etc. fungi, grasshopper, etc. insect cells, tobacco, etc. plant cells, BHK cells, CHO cells, COS cells, myeloma cells, etc. mammalian cells. In some embodiments, the recombinant cell of the present application is preferably a mammalian cell, including BHK cells, CHO cells, NSO cells, or COS cells, and does not include animal reproductive cells, fertilized eggs, or embryonic stem cells.

[0034] The host cell is transformed and / or transfected with the expression vector.

[0035] The sixth aspect of the present application provides a composition, which is a drug containing at least one of the aforementioned polypeptide, polynucleotide, expression vector, recombinant cell, or composition.

[0036] The seventh aspect of the present application is to provide a topical drug, which contains at least one of the aforementioned polypeptide, polynucleotide, expression vector, recombinant cell, or composition.

[0037] According to a specific embodiment of the present application, the drug contains a polypeptide of SEQ ID NO. 3 or a combination thereof having at least 90% sequence identity as an active ingredient.

[0038] According to a specific embodiment of the present application, the topical drug includes ointment, pill, water, wine, powder, medicine line (medicine Ding), etc. dosage forms, which can be directly used, and the usage includes ointment sticking, coating, applying, mixing, fumigating, washing, soaking, bathing, eye drops, ear drops, nose drops, etc.

[0039] According to a specific embodiment of the present application, the topical drug dosage form includes solution type, suspension type, emulsion type; ointment, which can also be divided into ointment, cream, gel; and tincture, liniment, spirit, powder, oil, paste, plaster, film coating, aerosol, etc.

[0040] The eighth aspect of the present application is to provide a cosmetic, which contains at least one of the aforementioned polypeptide, polynucleotide, expression vector, recombinant cell, or composition.

[0041] According to a specific embodiment of the present application, the cosmetic contains a polypeptide of SEQ ID NO. 3 or a combination thereof having at least 90% sequence identity as an active ingredient.

[0042] Cosmetic Identification Management Regulations, cosmetics refer to products that are applied to human body (skin, hair, nails, lips, teeth, etc.) by smearing, spraying, alcohol or other similar methods, for the purpose of cleaning, maintaining, beautifying, modifying and changing appearance, or modifying body odor, and keeping good condition.

[0043] According to a specific embodiment of the present application, the cosmetics include cleaning products, skin care products, oral products, etc.

[0044] The ninth aspect of the present application provides use of the aforementioned polypeptide, polynucleotide, expression vector, recombinant cell, and composition in the preparation of cosmetics.

[0045] According to a specific embodiment of the present application, the cosmetics have at least one of the effects of anti-inflammation, scar lightening, and moisturizing.

[0046] The tenth aspect of the present application provides use of the aforementioned polypeptide, polynucleotide, expression vector, recombinant cell, and composition in the preparation of drugs.

[0047] According to a specific embodiment of the present application, the drugs have at least one of the effects of anti-inflammation, accelerating wound healing of body surface skin, and reducing scar formation.

[0048] The present application also provides use of the aforementioned polypeptide sequence (SEQ ID NO. 3) and nucleotide sequence (SEQ ID NO. 4) for identifying Deinococcus.

[0049] Further, the present application also provides a Deinococcus identification method, comprising:

[0050] (1) designing a primer set reverse complementary to the sequence of the polynucleotide of the second aspect;

[0051] (2) amplifying the sample to be detected using the primer set,

[0052] wherein the sample to be detected that can amplify the nucleic acid fragment is a sample containing Deinococcus.

[0053] Further, as a specific embodiment, the primer set has the nucleotide sequence as shown in SEQ ID No. 5 and SEQ ID No. 6. BRIEF DESCRIPTION OF DRAWINGS

[0054] Fig. 1 is a result of Deinococcus weizhi VB142 and VB226 and Deinococcus species ANI analysis alignment;

[0055] Figure 2 is the result of DPPH free radical scavenging rate of Deinococcus weizhi fermentation lysate VB142-1, VB226-1 and fermentation product filtrate VB142-2, VB226-2 in antioxidant detection;

[0056] Figure 3 is the result of hydroxyl radical scavenging rate of Deinococcus weizhi fermentation lysate VB142-1, VB226-1 and fermentation product filtrate VB142-2, VB226-2 in antioxidant detection.

[0057] Figure 4 shows the polypeptide-promoted HaCaT cell scratch repair activity in Example 8 of the present application;

[0058] Figure 5 shows the zebrafish tail area in the polypeptide moisturizing efficacy test in Example 10 of the present application;

[0059] Figure 6 shows the water-moisturizing rate (%) of the polypeptide in Example 10 of the present application;

[0060] Figure 7 shows the nucleic acid electrophoretogram of the polypeptide nucleotide (SEQ ID NO. 4) of Deinococcus weizhi R12, Deinococcus radiodurans DSM20539, Deinococcus weizhi VB142 and VB226;

[0061] Figure 8 shows the sequencing comparison result of the polypeptide nucleotide sequence of Deinococcus weizhi R12, Deinococcus weizhi VB142 and VB226. DETAILED DESCRIPTION

[0062] The present application is further described below in conjunction with examples, but the present application is not limited by the following examples.

[0063] The present application provides the use of the polypeptide sequence and nucleotide sequence as described above for identifying Deinococcus weizhi.

[0064] Sequence analysis: alignment is performed by the sequence alignment method ClustalW with default parameters. Preferably, the parameter settings used are: for pairwise alignment: Gap open penalty: 10; Gap Extension Penalty: 0.1, for multiple alignment, Gap open penalty is 10 and Gap Extension Penalty is 0.2. Protein weight matrix is set to Identity. Both residue-specific and hydrophobicity penalties are “on”, gap separation distance is 4 and end gap separation is “off”. Negative matrix is not used, and finally the Delay Divergent Cut-off is set to 30%.

[0065] The determination method of the present application determines the similarity between sequences based on sequence identity. In terms of bacterial identification, according to the widely accepted criteria, if the similarity of 16S rRNA sequences of two bacteria is lower than 97%, it usually indicates that they can belong to different species. On the contrary, if the similarity is higher than 97%, it indicates that these species are more closely related in taxonomy. This threshold is based on extensive studies on the comparison of bacterial 16S rRNA sequences, reflecting the small but significant genetic differences between species. The 16S rRNA currently often used cannot even indicate that it belongs to a species after being higher than the taxonomic threshold of the species, and usually further judgment is needed from the whole genome perspective through Average Nucleotide Identity (ANI) to determine the similarity of two genomes from the genome level, which is an average value based on the comparison of all orthologous protein-coding genes between two genomes, and is an indicator for comparing the relationship between two genomes at the nucleotide level. ANI has a higher degree of differentiation between closely related species, and an ANI value of 95% is considered as the standard for defining a species, i.e., greater than 95% is considered as a species. For the identification of polypeptide sequences, the determination criteria for similarity are different. In the present application, if the identity of two polypeptide sequences reaches or exceeds 80%, they are considered to be similar. This criterion reflects that even if there is a certain degree of sequence difference at the protein level, the similarity in structure and function can still be retained.

[0066] Sequence identity: indicates a quantitative measure of the degree of homology between two amino acid sequences or two nucleotide sequences of equal length. If the sequences to be compared are not of equal length, they must be aligned to give the most likely correspondence, allowing for gap or (optionally) truncation at the ends of the polypeptide or nucleotide sequences. The sequence identity can be calculated, where Ndif is the total number of non-identical residues in the two sequences after alignment, and Nref is the number of residues in one of the sequences. Thus, the DNA sequence AGTCAGTC and the sequence AATCAATC (Ndif = 2 and Nref = 8) have 75% sequence identity. The gap is calculated as the number of non-identical residues at a particular position, i.e., the DNA sequence AGTGTC and the DNA sequence AGTCAGTC (Ndif = 2 and Nrep = 8) have 75% sequence identity. For all embodiments of the present application involving amino acid sequences, the percentage of sequence identity between one or more sequences can also be based on alignment using clustalW software (http: / / www.ebi.ac.uk / clustalW / index.html) using default settings.

[0067] Example 1, Isolation and identification of VB142.

[0068] 1. Isolation of VB142

[0069] Strain VB142 with anti-UV properties was isolated from the facial skin of an adult female by the following method: 20% glycerol was used as a sampling solution, a sample was collected from the facial skin of an adult female and placed in the sampling solution, 100 uL was taken and spread on a solid culture medium, and incubated at 30°C for 72 h. An orange-red strain was selected and repeatedly purified to obtain VB142.

[0070] 2. Identification of VB142

[0071] (1) Morphological characteristics: VB142 was incubated at 30°C for 72 h on a TGY solid culture medium, the bacterial cells were spherical, the colonies were round, small, convex, smooth, opaque and orange-red.

[0072] (2) Physiological and biochemical characteristics: The following physiological and biochemical characteristics of VB142 were determined according to the method of the "Common Bacteria System Identification Manual".

[0073] VB142 is a gram-positive bacterium, and is positive for contactase. Its utilization of different carbon sources is shown in Table 1.

[0074] Table 1. Carbon source utilization results of VB142 Note: "+" indicates that the carbon source can be utilized, and "-" indicates that the carbon source cannot be utilized.

[0075] (3) 16S rRNA gene

[0076] The 16S rRNA gene sequence of VB142 is shown in SEQ ID NO: 1, and the similarity comparison result shows that the 16S rRNA gene sequence most similar to this strain is Deinococcus wulumuqiensis R12 T , with a sequence identity of 94.28%.

[0077] (4) G+C mol% value

[0078] The G+C mol% content of VB142 is 63.99%.

[0079] (5) Whole genome ANI value

[0080] According to whole genome sequence analysis, as shown in Figure 1, the ANI value of VB142 compared with Deinococcus wulumuqiensis R12 T is 85.63%.

[0081] According to morphological characteristics, physiological and biochemical characteristics, 16S rRNA gene and whole genome sequence analysis, VB142 is named as a new species of Deinococcus weizhi.

[0082] Example 2, isolation and identification of VB226.

[0083] 1. Isolation of VB226

[0084] Strain VB226 with anti-ultraviolet properties was isolated and screened from the facial skin of an adult woman. The isolation method is as follows: 20% glycerol was used as sampling liquid, samples were collected from the facial skin of an adult woman and placed in the sampling liquid, 100 uL was taken and inoculated on solid culture medium, and cultured at 30°C for 72h. An orange-red strain was selected and repeatedly purified to obtain VB226.

[0085] 2. Identification of VB226

[0086] (1) Morphological characteristics: VB226 was cultured on TGY solid medium at 30°C for 72h, and the bacterial cells were spherical, the colonies were round, small, convex, smooth, opaque and light orange-red.

[0087] (2) Physiological and biochemical characteristics: The following physiological and biochemical characteristics of VB226 were determined according to the method of "Common Bacteria System Identification Manual".

[0088] VB226 is a gram-positive bacterium, and is positive for contact enzyme. The utilization of different carbon sources is shown in Table 2.

[0089] Table 2. Carbon source utilization results of VB226 Note: "+" indicates that the carbon source can be utilized, and "-" indicates that the carbon source cannot be utilized.

[0090] (3) 16S rRNA gene

[0091] The 16S rRNA gene sequence of VB226 is shown in SEQ ID NO. 2, and the similarity comparison result shows that the 16S rRNA gene sequence most similar to this strain is Deinococcus wulumuqiensis R12 T ), with a sequence identity of 94.28%.

[0092] (4) G+C mol% value

[0093] The G+C mol% content of VB226 is 64%.

[0094] (5) Whole genome ANI value

[0095] Genome-wide sequence analysis, as shown in Figure 1, the ANI value of VB226 compared with Deinococcus wulumuqiensis R12 T ) is 85.47%.

[0096] According to morphological characteristics, physiological and biochemical characteristics, 16S rRNA gene and whole genome sequence analysis, VB226 is named as a new species of Deinococcus weizhi.

[0097] Example 3

[0098] By the method of comparative genomics, the genomic information of the two new species of Deinococcus weizhi in Example 2 and other species of Deinococcus (such as Deinococcus wulumuqiensis, Deinococcus radiodurans) disclosed in NCBI were analyzed, and the sequence differences in the genome were found. It was found that Deinococcus weizhi had some specific sequences compared with other Deinococcus, for example, the inventors of the application unexpectedly found that Deinococcus weizhi had a specific exopolypeptide, the polypeptide as described in the application, the amino acid sequence was SEQ ID NO. 3, and the nucleotide sequence was as shown in SEQ ID NO. 4.

[0099] Further, the specific exopolypeptide of Deinococcus weizhi is located at Chromosome 1 (2576018..2576080) in the genome of Deinococcus weizhi VB142 and at Chromosome 1 (66689..66772) in the genome of Deinococcus weizhi VB226.

[0100] Example 4

[0101] A method for preparing a fermentation product filtrate and a fermentation lysate of Deinococcus weizhi:

[0102] (1) Preparation of seed / fermentation medium

[0103] According to the mass percentage of each substance in the total mass of the medium, 5.0 g of peptone, 5.0 g of yeast extract and 1.0 g of glucose were dissolved in water, and the volume was adjusted to 1 L as seed / fermentation medium TGY. Sterilize and cool to room temperature for standby.

[0104] (2) Activation of bacterial strain

[0105] VB142 and VB226 were inoculated on TGY solid medium and cultured at 30°C.

[0106] (3) Obtain VB142 and VB226 seed liquid

[0107] The activated VB142 and VB226 were inoculated into the seed culture medium prepared in (1) and cultured at 30°C for one day to obtain VB142 and VB226 seed liquid.

[0108] (4) Preparation of VB142 and VB226 fermentation liquid

[0109] The VB142 and VB226 seed liquid prepared in (3) was inoculated into the fermentation culture medium prepared in (1) and cultured at 30°C for 2-3 days to obtain VB142 and VB226 fermentation liquid.

[0110] (5) Preparation of fermentation product filtrate

[0111] The fermentation liquid obtained in (4) was centrifuged at 8000 rpm / min for 10 min to obtain VB142 and VB226 fermentation product filtrate and bacterial cells.

[0112] (6) Preparation of fermentation lysate

[0113] The VB142 and VB226 bacterial cells obtained in (5) were respectively subjected to cell disruption and then dissolved in water to obtain VB142 and VB226 fermentation lysate.

[0114] Example 5

[0115] The VB142 and VB226 fermentation product filtrate and fermentation lysate obtained in Example 4 were respectively subjected to antioxidant activity detection, and the method was as follows:

[0116] (1) DPPH free radical scavenging ability detection method

[0117] a. Sample detection

[0118] 10 μL of the sample to be detected and 190 μL of detection working solution were added into a 96-well plate, vortexed, and allowed to stand at room temperature for 30 min. The absorbance at 515 nm was measured. One control tube was set for each determination tube.

[0119] b. Calculation formula

[0120] DPPH free radical scavenging rate D% = {[A 空白 -(A 样品 -A 对照 )] ÷ A 空白} × 100%

[0121] The results are shown in Figure 2. The VB142 and VB226 fermentation supernatant and lysate both had the ability to scavenge DPPH free radicals.

[0122] (2) Hydroxyl radical scavenging ability detection

[0123] a. Sample detection

[0124] Add the sample and the detection reagent into the EP tube respectively, vortex to mix, and place in a 37℃ water bath for reaction for 60 min. Centrifuge at 10000 rpm at room temperature for 10 min. Take 200 μL supernatant to measure the absorbance at 536 nm in a 96-well plate.

[0125] b. Calculation formula

[0126] Hydroxyl radical scavenging rate D% = (A 测定 -A 对照 ) ÷ (A 空白 -A 对照 ) × 100%

[0127] The results are shown in Figure 3. The fermentation supernatant and the cell lysate of VB142 and VB226 both have the ability to scavenge hydroxyl radicals.

[0128] Example 6

[0129] Reagent preparation: The cell lysate of Micrococcus variabilis VB142 and VB226 obtained in Example 4 is respectively prepared into a 1% stock solution with standard dilution water, and is used immediately after preparation; the sodium hyaluronate produced by the Health Industry Research Institute of Sinopharm Group is stored in a cool, dry and dark place, and the original solution is used as the stock solution.

[0130] Experimental animals: Zebrafish are bred in water for fish at 28℃ (water quality: 200 mg of instant sea salt is added to 1 L of reverse osmosis water, with an electrical conductivity of 450-550 μS / cm, a pH of 6.5-8.5, and a hardness of 50-100 mg / L CaCO3); they are bred and provided by the fish breeding center of the company, and the use license number is SYXK(Zhe)2022-0004. The wild type AB strain of zebrafish is bred by natural pair mating. Zebrafish at 6 hours post-fertilization (6hpf) are used for sample water replenishment and moisturizing efficacy evaluation.

[0131] Detection method: randomly select 2dpf melanin allele mutant albino zebrafish in 6-hole plate, 30 zebrafish are treated in each hole (experimental group). Respectively, 0.025% of the sample is given in water, and 0.05% of the positive control sodium hyaluronate is given in water. At the same time, a normal control group and a model control group are set up, and the volume of each hole is 3mL. Except for the normal control group, the rest of the experimental groups are given sodium chloride to establish a zebrafish dehydration model. After 22h of treatment at 28℃, 10 zebrafish are randomly selected from each experimental group and placed under a dissecting microscope for photography. NIS-Elements D 3.20 advanced image processing software is used for analysis and data collection, and the tail area (A) of zebrafish is analyzed. The statistical analysis results of this index are used to evaluate the water replenishing and moisturizing effect of the sample. The statistical processing results are expressed as Mean±SEM. The water replenishing and moisturizing effect calculation formula is as follows:

[0132] Water replenishing and moisturizing rate % = (S 样品 -S 模型对照 ) ÷ (S 正常对照 -S 模型对照 ) × 100%

[0133] Statistical analysis was performed using SPSS 26.0 software, and P<0.05 indicated that the difference was statistically significant. The specific results are shown in Table 3.

[0134] Table 3 Water replenishing and moisturizing efficacy data *: compared with the normal control group, P<0.05; ***: compared with the normal control group, P<0.001

[0135] As shown in Table 3, in the zebrafish dehydration model, the cell lysates of VB142 and VB226 both have water replenishing and moisturizing effect, and their effect is better than that of sodium hyaluronate.

[0136] Example 7

[0137] Reagent preparation: the cell lysates of VB142 and VB226 of Micrococcus varians obtained in Example 4 are prepared into 1% stock solution with standard dilution water, and are prepared for use; dipotassium glycyrrhizinate is prepared into 1% stock solution with standard dilution water, and is prepared for use.

[0138] Experimental animals: Transgenic neutrophil green fluorescent MPX strain zebrafish were bred in fish water at 28°C (water quality: 200 mg of instant sea salt was added to 1 L of reverse osmosis water, the conductivity was 450-550 μS / cm; the pH was 6.5-8.5; the hardness was 50-100 mg / L CaCO3), which were bred and provided by the fish breeding center of the company, the experimental animal use license number was: SYXK (Zhe) 2022-0004, and the breeding management met the requirements of international AAALAC certification (certification number: 001458). Zebrafish were bred in a natural pair mating way. Zebrafish of 2 dpf were used for sample anti-inflammatory efficacy detection.

[0139] Test method: 2 dpf transgenic neutrophil green fluorescent MPX strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). 0.006% of the sample, 0.031% of the positive control glycyrrhizic acid disodium, and normal control and model control groups were given by water dissolution, and the volume of each well was 3 mL. Except for the normal control group, the rest of the experimental groups were given sodium dodecyl sulfate by water dissolution to establish a zebrafish inflammation model. After 18 h of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and placed under a dissecting microscope for photography. NIS-Elements D 3.20 advanced image processing software was used for analysis and data collection, and the number of neutrophils in zebrafish (N) was analyzed. The statistical analysis results of this index were used to evaluate the anti-inflammatory efficacy of the sample. The statistical processing results were expressed as Mean ± SEM. The anti-inflammatory efficacy calculation formula is as follows:

[0140] Anti-inflammatory efficacy % = (N 模型 -N 样品 ) ÷ N 模型 × 100%

[0141] Statistical analysis was performed using SPSS 26.0 software, P<0.05 indicated that the difference was statistically significant, and the anti-inflammatory efficacy data is shown in Table 4.

[0142] Table 4 Anti-inflammatory efficacy data ***: Compared with the model control group, P<0.001

[0143] As shown in Table 4, in the zebrafish inflammation model induced by sodium dodecyl sulfate, the cell lysates of VB142 and VB226 both had anti-inflammatory efficacy, and were better than glycyrrhizic acid disodium.

[0144] Example 8

[0145] Reagent preparation: The cell lysates of Micrococcus variabilis VB142 and VB226 obtained in Example 4 were prepared into 1% stock solutions with standard dilution water, and were prepared and used immediately; tea polyphenol was prepared into a 30% stock solution with standard dilution water, and was prepared and used immediately.

[0146] Experimental animals: Zebrafish were bred in fish water at 28°C (water quality: 200 mg of instant sea salt was added to each 1 L of reverse osmosis water, conductivity was 450-550 μS / cm; pH was 6.5-8.5; hardness was 50-100 mg / L CaCO3), and were bred and provided by the fish breeding center of the company. The experimental animal use license number was SYXK(Zhe)2022-0004, and the breeding management met the requirements of international AAALAC certification (certification number: 001458). Wild type AB strain zebrafish were bred by natural pair mating. Zebrafish aged 2 dpf were used for sample anti-photoaging efficacy evaluation.

[0147] Detection method: 2 dpf wild type AB strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples (concentration see Table 5-2) were given by water dissolution, and the positive control tea polyphenol was 0.001%, and a normal control group and a model control group were set up at the same time, and the volume of each well was 3 mL. Except for the normal control group, the rest of the experimental groups were irradiated with ultraviolet light to establish a zebrafish photoaging model. After 22 h of treatment at 28°C, 10 zebrafish were randomly selected from each experimental group and placed under a dissecting microscope for photography. NIS-Elements D 3.20 advanced image processing software was used for analysis and data collection, and the area (A) of the zebrafish tail fin was analyzed. The statistical analysis results of this index were used to evaluate the anti-photoaging efficacy of the sample. The statistical processing results were expressed as Mean ± SEM. The anti-photoaging efficacy calculation formula is as follows:

[0148] Anti-photoaging rate % = (A sample - A model control) ÷ A model control × 100%

[0149] Statistical analysis was performed using SPSS 26.0 software, and P<0.05 indicated that the difference was statistically significant. The anti-photoaging efficacy data is shown in Table 5.

[0150] Table 5 Anti-photoaging efficacy data ***: Compared with the model control group, P<0.001

[0151] As shown in Table 5, in the zebrafish aging model induced by ultraviolet light, the cell lysates of VB142 and VB226 had anti-photoaging efficacy.

[0152] Example 9

[0153] Reagent preparation: The cell lysates of Micrococcus variabilis VB142 and VB226 obtained in Example 4 were prepared into 1% stock solutions with standard dilution water, and were prepared and used immediately; aminoguanidine hydrochloride was prepared into a 2% stock solution with ultrapure water, and was stored at -20°C in the dark.

[0154] Experimental animals: Zebrafish were bred in fish water at 28℃ (water quality: 200mg of instant sea salt was added to 1L of reverse osmosis water, conductivity was 450-550μS / cm; pH was 6.5-8.5; hardness was 50-100mg / L CaCO3), and were bred and provided by the fish breeding center of the company, with experimental animal use license number SYXK(Zhe)2022-0004, and were bred and managed in accordance with the requirements of international AAALAC certification (certification number: 001458). Wild type AB strain zebrafish were bred by natural pair mating. Zebrafish aged 2dpf were used for sample anti-glycation efficacy evaluation.

[0155] Detection method: 1.5mL centrifuge tubes of wild type AB strain zebrafish at 5dpf were randomly selected, and 10 zebrafish were treated in each tube (experimental group). The samples were water-solubilized to give 0.05%, and the positive control aminoguanidine hydrochloride was 0.02%, while the normal control group and the model control group were set up, and the volume of each tube was 150μL. Except for the normal control group, 0.4M glucose solution was water-solubilized to establish a zebrafish AGEs increase model in the remaining experimental groups. Three biological replicates were set in parallel. After 24h of 60℃ shaking table oscillation treatment, the supernatant was centrifuged, data was collected using a multifunctional enzyme marker, and the statistical analysis results of the index were used to evaluate the anti-glycation efficacy of the sample. The statistical processing results are expressed as Mean±SEM. The anti-photoaging efficacy calculation formula is as follows:

[0156] Anti-glycation rate % = (S 模型对照 -S 样品 ) ÷ S 模型对照 × 100%

[0157] Statistical analysis was performed using SPSS 26.0 software, P<0.05 indicated that the difference was statistically significant, and the anti-glycation efficacy is shown in Table 6.

[0158] Table 6 Anti-glycation efficacy data ***: Compared with the model control group, P<0.001

[0159] As shown in Table 6, in the glucose-induced zebrafish glycation model, the cytolysate of VB142 and VB226 had glycation efficacy.

[0160] Example 10

[0161] The micrococcus variabilis specific exosomal polypeptide provided by the embodiment 3 of the present application can be prepared by solid phase synthesis technology, which has the following steps:

[0162] a) Reactor treatment:

[0163] 2 / 3 volume of dichloromethane (DCM, Aldrich) was added to the reactor and soaked for 3h;

[0164] b) Resin swelling:

[0165] Weigh 1.0 g Wang resin (sigma-aldrich) into the reaction column, add 20 mL N,N- dimethylformamide (DMF, Aldrich), soak for 30 min, and swell the resin thoroughly;

[0166] c) First amino acid connection:

[0167] Configure the reaction solution containing 8 mmol Fmoc-amino acid (sigma-aldrich, the polypeptide is Fmoc-Ala-OH), 0.9 mmol 4-dimethylaminopyridine (DMAP, Aldrich), 4.5 mmol N,N'-diisopropylcarbodiimide (DIC, Aldrich), and nitrogen for 2.5 hours.

[0168] d) Washing:

[0169] After the reaction is complete, blow off the reaction solution, and then use 10 mL of DCM, isopropanol, and DMF to wash twice, respectively.

[0170] e) Blocking of residual reactive groups:

[0171] Dissolve 0.6 mL benzoyl chloride (Aldrich) and 0.45 mL pyridine (Aldrich) in 10 mL DCM, add to the resin, and react for 1 hour, then wash with 10 mL of DCM, isopropanol, and DMF twice, respectively.

[0172] f) Removal of Fmoc protecting group:

[0173] Add 15 mL of 20% piperidine in DMF solution to the reaction vessel, and react with nitrogen for 5 minutes. After drying, add 20 mL of 20% piperidine in DMF solution and react with nitrogen for 20 minutes. After drying, wash with 10 mL of DCM, isopropanol, and DMF twice, respectively. Take a small amount of resin for ninhydrin (sigma-aldrich) detection to detect the removal. If the resin is black or purple black, it is completely removed.

[0174] g) Amino acid condensation:

[0175] According to the polypeptide sequence, a second amino acid reaction solution was prepared to contain 2 mmol of Fmoc-amino acid (the polypeptide was Fmoc-Ser-OH), 0.8 g of a 10 mL DMF solution of benzotriazole tetramethyl tetrafluoroborate (TBTU, Aldrich), 1 mL of 0.135 g / mL p-hydroxybenzonitrile (HoBt, Aldrich), and 0.4 mL of N,N-diisopropyl ethylamine (DIEA, Aldrich), which were added to the reactor and reacted for 2 hours under nitrogen.

[0176] h) Repeating d) to g) according to the amino acid sequence until the synthesis of the last amino acid is completed.

[0177] i) Cleavage of the peptide:

[0178] A cleavage reagent containing trichloroacetic acid 82.5%, benzyl sulfide 5%, water 5%, phenol 5%, 2.5% ethanedithiol was added, and the reaction was carried out at room temperature for 4 h. After the reaction was completed, the reaction was dried under nitrogen to obtain the polypeptide powder.

[0179] Example 11

[0180] The safety of the polypeptide provided in Example 10 of the present application was investigated by a cell proliferation test. The solution used in this example was prepared as follows:

[0181] The complete DMEM medium was prepared as follows: 89% Gibco high-sugar DMEM medium (Gibco), 10% fetal bovine serum (Gibco), 1% 100 x penicillin-streptomycin double antibody (sigma-aldrich);

[0182] The polypeptide solution in the experimental group: the polypeptide was dissolved in the complete DMEM medium, the mother liquor concentration was 5 mg / mL, and it was stored at 4°C. Before use, it was diluted with complete DMEM medium.

[0183] The experimental process of this example was as follows: HaCat cells in the logarithmic phase of growth were taken and plated in a 96-well plate, 200 μL of 1 x 10 4 After 24 hours of cell adhesion, the culture medium was discarded and replaced with 100 μL of DMEM complete medium containing peptides 1-7, so that the concentrations of peptides 1-7 were 0, 0.05, 0.1, 0.25, 0.5, 1, 2.5, and 5 mg / mL, respectively, with 3 replicate wells for each concentration. After 24 hours, 100 μL of Cell Luminescent Cell Viability Assay kit detection reagent was added to each well, and incubated at 37°C for 10 minutes. The intensity of spontaneous light was detected on a microplate reader, and the number of reactive cells was determined.

[0184] The detection results are shown in Table 7:

[0185] Table 7: Relative cell amount of each group in cell proliferation test

[0186] The results show that the polypeptides have no obvious inhibitory or promoting effect on cell amount compared with the blank control after co-incubation with HaCat cells for 24 h at a concentration of 5 mg / mL or less, indicating that the peptides provided by the present application have no cytotoxicity and high safety.

[0187] Example 12

[0188] In order to observe the cell repair ability of the polypeptides provided in Example 10, a scratch repair function test was performed using HaCat cells. The solution used in this example was prepared as follows:

[0189] Complete DMEM medium: 89% high-sugar DMEM medium (Gibco), 10% fetal bovine serum (Gibco), 1% 100 x penicillin-streptomycin double antibody (sigma-aldrich);

[0190] Polypeptide solution of experimental group: the polypeptides prepared in Example 5 were dissolved in complete DMEM medium at a concentration of 2 mg / mL and stored at 4°C.

[0191] The experimental process of this example is as follows: a scratch test wound healing 2-hole insert (ibidi) was pasted in the center of the 12-hole plate, the cell suspension concentration was adjusted to 3 x 10 5 After the addition of the cell suspension, the hole plate was avoided to be shaken, and was cultured in an environment of 37°C and 5% carbon dioxide for at least 24 hours. The cell density after 24 hours was checked under a microscope, and after obtaining the fused cell layer in the two-hole insert, the culture insert was gently taken out with a sterile forceps, and the cell layer was washed with PBS to remove cell debris and unattached cells. Complete DMEM medium containing small peptides at a concentration of 2 mg / mL was added to each hole of the 12-hole plate. After the end of sample addition, three photos were taken at each of the three positions in the central field of the scratch in each hole, and automatic image analysis was performed using Image J software to calculate the average value of the cell confluence area. Three photos were taken at the same positions after 12 hours, and automatic image analysis was performed using Image J software to calculate the average value of the cell confluence area. The detection results are shown in Figure 4 and Table 8:

[0192] Table 8: 12h healing rate of cell scratch test *: P<0.05; **: P<0.01 vs Control

[0193] The results show that the healing rate of the culture medium added with the polypeptide is obviously higher than that of the blank culture medium without the polypeptide, indicating that the polypeptide provided by the application has strong cell repair ability.

[0194] Example 13

[0195] In order to observe the anti-inflammatory effect of the polypeptide provided in Example 10 of the application, an anti-inflammatory activity inhibition test was performed using HaCat cells. The solution used in this example is prepared as follows:

[0196] Complete DMEM culture medium: 89% high-sugar DMEM culture medium (Gibco), 10% fetal bovine serum (Gibco), 1% 100x penicillin-streptomycin double antibody (sigma-aldrich);

[0197] Polypeptide solution of experimental group: the polypeptide is dissolved in complete DMEM culture medium with a concentration of 2 mg / mL and stored at 4°C;

[0198] Inflammation inducer: fresh E. coli DH5a is washed twice with PBS, resuspended with PBS, the bacterial solution is adjusted to OD = 1.0, boiled at 95°C for 10 min, and stored at -80°C.

[0199] Positive control: palmitoyl tetrapeptide-7 (Guangzhou Baiyu Biological) is prepared into 2 mg / mL with complete DMEM culture medium and stored at 4°C.

[0200] The experimental process of this example is as follows: HaCat cells are inoculated in a twelve-hole plate, 1x10 5 After the cells adhere, the culture medium is discarded, and fresh culture medium containing peptides 1-7 is added, and 50 μL of inflammation inducer is added to each small peptide three times. The positive control is culture medium with only inflammation inducer, and the blank control is added with 50 μL of PBS, and the culture is performed for 24 h. The supernatant is collected, and the IL-8 level in the supernatant is detected by human IL-8 detection kit (Dakewei Biotechnology Co., Ltd.).

[0201] The calculation process is as follows:

[0202] Inhibition rate = [1-(C 实验组 -C 空白组 ) / (C 阳性组 -C 空白组 )]x100%

[0203] The detection results are shown in Table 9 as follows:

[0204] Table 9: IL-8 level and inhibition rate of supernatant

[0205] As can be seen from the results of the above table, the polypeptides have certain inhibitory effect on IL-8 expression induced by E. coli inflammation inducer.

[0206] Example 14

[0207] The polypeptide provided in Example 10 of the present application was used to observe the moisturizing effect by zebrafish experiment. The solution used in this example was prepared as follows:

[0208] Polypeptide solution in experimental group: all small peptides were prepared into 1% with standard dilution water, and were used immediately after preparation.

[0209] Positive control: sodium hyaluronate fruit-flavored drink (Jiangsu Yijia Garden Health Technology Co., Ltd.), stored in the shade. The original solution was directly taken and used immediately.

[0210] The experimental process of this example is as follows: 2dpf melanin allele mutant Albino strain zebrafish were randomly selected in a 6-well plate, and 30 zebrafish were treated in each well (experimental group). The samples (concentration of 0.05%) were given respectively, and the positive control sodium hyaluronate fruit-flavored drink was given at a concentration of 15.6 μL / mL, and a normal control group and a model control group were set up at the same time, and the volume of each well was 3 mL. Except for the normal control group, sodium chloride was given to the rest of the experimental groups to establish a zebrafish dehydration model. After 22 h of treatment at 28℃, 10 zebrafish were randomly selected from each experimental group and placed under a dissecting microscope for photography, and NIS-Elements D 3.20 advanced image processing software was used for analysis and data collection, and the tail area (S) of zebrafish was analyzed, and the statistical analysis results of the index were used to evaluate the water replenishing and moisturizing effect of the sample.

[0211] The results are expressed as Mean ± SEM. The water replenishing and moisturizing effect calculation formula is as follows:

[0212] Water replenishing and moisturizing rate % = (S 样品 -S 模型对照 ) ÷ (S 正常对照 -S 模型对照 ) × 100%

[0213] Statistical analysis was performed using SPSS 26.0 software, and P<0.05 indicated that the difference was statistically significant.

[0214] The detection results are shown in Figures 5, 6 and Table 10:

[0215] Table 10: Zebrafish tail dehydration test for small peptide water replenishing and moisturizing effect Compared with the model control group, *: P<0.05, **: P<0.01, ***: P<0.001

[0216] As can be seen from the results of the above table, the polypeptides can all protect the reduction of zebrafish tail area caused by dehydration, and have water replenishing and moisturizing effect.

[0217] Example 15

[0218] This embodiment discloses a method for identifying *Micrococcus microcarpa*, specifically, an analysis method using specific primers and specific sequences, including:

[0219] 1. Genomic DNA extraction

[0220] The specimens in this embodiment were *Deinococcus wulumuqiensis* R12 isolated from Hangzhou Microbiotechnology Co., Ltd. T China General Microbiological Culture Collection Center), D. radiodurans (DSM 20539) T DNA template extraction was performed using a kit (product number: D3146) provided by Guangzhou Meiji Biotechnology Co., Ltd. (China Marine Microbial Culture Collection Center).

[0221] 2. Polypeptide nucleotide sequence analysis

[0222] The isolated *Morphococcus* was amplified by PCR using a primer set, wherein SEQ ID NO.5 and SEQ ID NO.6 are the upstream and downstream primers for the nucleotide sequence of the polypeptide, and the reaction system is shown in Table 11.

[0223] Table 11: Reaction System Used

[0224] This experiment used NEB products. High-fidelity PCR premix (catalog number #M0531) is used for PCR amplification. This premix combines high-fidelity polymerase and optimized reaction buffer to ensure extremely high amplification efficiency and accuracy in PCR reactions.

[0225] When preparing for the PCR reaction, first calculate the required amount of DNA template using the following formula: Added volume = Total DNA volume / Concentration. The total DNA volume should be controlled within the range of 50 ng to 250 ng to ensure the specificity and efficiency of the PCR reaction. By accurately calculating the amount of DNA template added, a uniform distribution of DNA in the reaction system can be ensured, thereby improving the reproducibility and accuracy of amplification. The amplification procedure is shown in Table 12.

[0226] Table 12: Amplification Procedure

[0227] 3. PCR product processing and sequencing

[0228] Gel electrophoresis detection:

[0229] After the PCR reaction is completed, the purity of the PCR product is first checked by gel electrophoresis. Gel electrophoresis is a standard molecular biology technique used to separate and identify nucleic acid fragments. Through this step, it can be ensured that the PCR product has no non-specific amplification or contamination. The results are shown in Figure 7, the PCR product is set to 305 bp, the VB142 and VB226 products show less than 500 bp, and the Urumqi Deinococcus PCR product is greater than 1 kb, which is obviously different, and there is no PCR product for Deinococcus radiodurans.

[0230] Further, the inventors also purchased Deinococcus radiodurans DRR1, ATCC27603, 17438 from CCTCC and repeated the above steps, and found that there was no PCR product for Deinococcus radiodurans.

[0231] Purity and concentration determination:

[0232] The concentration of the PCR product was accurately measured using a Nanodrop spectrophotometer. The Nanodrop technique can provide detailed information about the concentration and purity of the sample, ensuring that the quality of the sample meets the sequencing requirements.

[0233] Sample submission:

[0234] The PCR product that passed the purity and concentration detection was sent to Sheng Wu Bioengineering (Shanghai) Co., Ltd. (hereinafter referred to as Sheng Wu Bio) for first-generation sequencing analysis. The polypeptide nucleotide sequence alignment is shown in Figure 8, the red box area is the polypeptide nucleotide sequence region, and the sequencing results of Deinococcus minor VB142 and VB226 are completely consistent, and Deinococcus urumqiensis R12 lacks a sequence. The above examples only express several embodiments of the present application, and the description is more specific and detailed, but it cannot be understood as limiting the scope of the patent. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, a number of modifications and controls can be made, which are within the scope of the present application. Therefore, the protection scope of the present application patent should be subject to the appended claims.

Claims

1. Deinococcus weizhi comprising Deinococcus sp. having at least 97% 16S rRNA sequence identity to SEQ ID NO. 1 or SEQ ID NO. 2 and average nucleotide identity (ANI) within 95%.

2. The Deinococcus weizhi of claim 1, which is Deinococcus sp. VB142 deposited with the China Center for Type Culture Collection (CCTCC) on January 23, 2024, under the accession number CCTCC M 2024185, or deposited with the Korean Collection for Type Cultures (KCTC) on June 19, 2023, under the accession number 15470BP.

3. The Deinococcus weizhi of claim 1, which is Deinococcus sp. VB226 deposited with the China Center for Type Culture Collection (CCTCC) on January 23, 2024, under the accession number CCTCC M 2024186.

4. A fermentation broth, a fermentation product filtrate, a fermentation lysate product or an extract of the fermentation broth of the Deinococcus weizhi of any one of claims 1 to 3.

5. Use of the Deinococcus weizhi of any one of claims 1 to 3 or a fermentation broth, a fermentation product filtrate, a fermentation lysate product, an extract of the fermentation broth thereof in the preparation of an antioxidant, moisturizing, anti-inflammatory, anti-photoaging or anti-glycation product.

6. Use according to claim 5, wherein: The antioxidant product is selected from a health product, a pharmaceutical composition or a cosmetic product.

7. A health product comprising the Deinococcus weizhi of any one of claims 1 to 3 or the fermentation broth, the fermentation product filtrate, the fermentation lysate product or the extract of the fermentation broth of claim 4.

8. A pharmaceutical composition comprising the Deinococcus weizhi of any one of claims 1 to 3 or the fermentation broth, the fermentation product filtrate, the fermentation lysate product or the extract of the fermentation broth of claim 4.

9. A cosmetic composition comprising the Deinococcus weizhi of any one of claims 1 to 3 or the fermentation broth, the fermentation product filtrate, the fermentation lysate product or the extract of the fermentation broth of claim 4.

10. The cosmetic or dermatological preparation according to claim 9, wherein The cosmetic or skin care product includes a dosage form of cosmetic water, essence, paste, cream, milk, gel, mask, etc.

11. An isolated polypeptide, comprising, The amino acid sequence has at least 90% sequence identity to SEQ ID No.

3.

12. An isolated polynucleotide, comprising: The polynucleotide encodes the polypeptide of claim 11; the nucleotide sequence has at least 90% sequence identity to any one of SEQ ID No.

4.

13. An expression vector, characterized in that, The polynucleotide of claim 12.

14. A recombinant cell, wherein, The polynucleotide of claim 12, the expression vector of claim 13, or the polypeptide of claim 11.

15. The recombinant cell of claim 14, wherein, The recombinant cell is obtained by introducing the expression vector of claim 13 into a host cell.

16. A composition characterized in that, The polypeptide of claim 11, the polynucleotide of claim 12, the expression vector of claim 13, the recombinant cell of claim 14 or 15.

17. An external medicament, characterized in that, The pharmaceutical comprises at least one of the polypeptide of claim 11, the polynucleotide of claim 12, the expression vector of claim 13, the recombinant cell of claim 14 or 15, and the composition of claim 16.

18. An external cosmetic use characterized by, The cosmetic comprises at least one of the polypeptide of claim 11, the polynucleotide of claim 12, the expression vector of claim 13, the recombinant cell of claim 14 or 15, and the composition of claim 16.

19. Use of the polypeptide of claim 11, the polynucleotide of claim 12, the expression vector of claim 13, the recombinant cell of claim 14 or 15, and the composition of claim 16 in the preparation of a cosmetic; Optionally, the cosmetic has at least one of the effects of anti-inflammation, scar lightening, and moisturizing.

20. Use of the polypeptide of claim 11, the polynucleotide of claim 12, the expression vector of claim 13, the recombinant cell of claim 14 or 15, and the composition of claim 16 in the preparation of a pharmaceutical; Optionally, the pharmaceutical has at least one of the effects of anti-inflammation, accelerating the healing of a wound on the skin, and reducing scar formation.

21. Use of the polynucleotide of claim 12 in identifying M. pachydermatis.

22. A method of identifying Micrococcus, characterized by: The method comprises (1) designing a primer set reverse complementary to the sequence of the polynucleotide of claim 12; (2) amplifying a sample to be tested using the primer set, wherein the sample to be tested that can be amplified to obtain a nucleic acid fragment is a sample containing M. pachydermatis.

23. The method of claim 22, wherein: The primer set has the nucleotide sequences as shown in SEQ ID No. 5 and SEQ ID No. 6.

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