Polypeptide derived from CIRBP, protein and preparation method and application thereof

Abstract

The invention discloses a CIRBP-derived polypeptide and protein as well as a preparation method and application thereof, the polypeptide and the assembled protein thereof have the characteristics of high activity and good bioavailability, are easily absorbed by a human body and effectively penetrate through a skin barrier, so that the core biological functions of accurately repairing DNA damage and maintaining genome integrity are fully exerted; the anti-aging composition shows a remarkable anti-aging effect and can be applied to the field of skin care products.

Description

Technical Field

[0001] This application belongs to the field of synthetic biology technology, and in particular relates to a polypeptide and protein derived from CIRBP, their preparation methods and applications. Background Technology

[0002] Cold-induced RNA-binding protein (CIRBP) is a protein whose expression is upregulated under stress conditions such as low temperature. Recent studies have found that it plays an important role in the DNA damage response. CIRBP can participate in and promote the "precise repair" of DNA damage in a mild and sustained manner, thereby helping to maintain the integrity of the genome.

[0003] However, in existing technologies, directly utilizing full-length CIRBP proteins in skincare products faces numerous limitations, such as low transdermal or absorption efficiency due to excessively large molecular weight, stability issues, and potential immunogenicity. Although the biological functions of CIRBP have been partially elucidated, there are currently no reports of designing and preparing small-molecule peptides from key functional regions derived from CIRBP, or further tandemly assembling multiple such functional peptides in a specific sequence to construct novel recombinant proteins with smaller molecular weights, and using them in the synthesis of anti-aging related products. Summary of the Invention

[0004] This application provides a polypeptide derived from CIRBP, its tandemly assembled protein, its preparation method and application. The polypeptide and its assembled protein have the characteristics of small molecular weight and strong activity, and are easily absorbed by the human body or penetrate the skin barrier, effectively exerting their anti-aging effects.

[0005] In a first aspect, this application provides a polypeptide comprising one or more amino acid sequences as shown in any one of SEQ ID No: 1-13; preferably, the polypeptide is derived from CIRBP protein and has a molecular weight of 0.5~0.7 kDa.

[0006] In a second aspect, this application provides a protein comprising the polypeptide described in the first aspect, wherein the molecular weight of the protein is 0.5 to 7.2 kDa; preferably, the protein comprises one or more amino acid sequences as shown in any one of SEQ ID No. 14-15.

[0007] Thirdly, this application provides a nucleic acid that encodes the polypeptide described in the first aspect or the protein described in the second aspect.

[0008] Fourthly, this application provides a vector comprising the nucleic acid described in the third aspect.

[0009] Fifthly, this application provides a host cell comprising the nucleic acid described in the third aspect or the vector described in the fourth aspect; preferably, the host cell comprises at least one of eukaryotic cells and prokaryotic cells.

[0010] In a sixth aspect, this application provides a method for preparing the polypeptide described in the first aspect, the method comprising introducing a nucleic acid or vector encoding the polypeptide into a host cell and then inducing its expression; preferably, the method further comprises an enzymatic digestion step.

[0011] In any embodiment of this application, the enzyme in the enzymatic hydrolysis step includes at least one of trypsin, subtilisin, thermophilic protease, papain, and alkaline protease, preferably trypsin.

[0012] In a seventh aspect, this application provides the use of the polypeptide described in the first aspect, the protein described in the second aspect, the nucleic acid described in the third aspect, the carrier described in the fourth aspect, or the host cell described in the fifth aspect in the preparation of anti-aging related products.

[0013] In any embodiment of this application, the product includes skincare products.

[0014] In any embodiment of this application, the skin care product includes at least one of the following: bacterial strain, fermentation broth, fermentation broth extract, lysate, fermentation broth precipitate, and culture obtained by said bacterial strain.

[0015] This application provides a CIRBP-derived polypeptide, its tandemly assembled protein, its preparation method, and its application. The polypeptide and its assembled protein are characterized by high activity and good bioavailability, are easily absorbed by the human body, and can effectively penetrate the skin barrier, thereby fully exerting their anti-aging effects. In particular, they can effectively upregulate the expression levels of type I and type III collagen in the skin, further enhancing their application value in the field of skin health and repair. Attached Figure Description

[0016] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments of this application will be briefly introduced below. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 A comparison diagram showing the advantages and disadvantages of the present invention compared to the prior art is provided.

[0018] Figure 2 This is a comparison chart of the effects of Examples 1-2 and Comparative Examples 1-3 of this application on promoting type I collagen expression (where "a", "b", and "c" are used to indicate statistically significant differences between different groups).

[0019] Figure 3 This is a comparison chart of the effects of Examples 1-2 and Comparative Examples 1-3 of this application in promoting type III collagen expression (where "a", "b", and "c" are used to indicate statistically significant differences between different groups). Detailed Implementation

[0020] To make the purpose, technical solution, and beneficial technical effects of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the implementation details described in this specification are merely for illustrative purposes and are not intended to limit the scope of this application.

[0021] For simplicity, this application only explicitly discloses some numerical ranges. However, any lower limit can be combined with any upper limit to form a range not explicitly stated; and any lower limit can be combined with other lower limits to form a range not explicitly stated, just as any upper limit can be combined with any other upper limit to form a range not explicitly stated. Furthermore, although not explicitly stated, every point or individual value between the endpoints of the range is included within that range. Therefore, each point or individual value can be used as its own lower or upper limit and combined with any other point or individual value or with other lower or upper limits to form a range not explicitly stated.

[0022] It should be noted that the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Without further limitations, an element defined by the phrase "comprising..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes said element.

[0023] Unless otherwise stated, the values ​​of the parameters mentioned in this application can be measured using various measurement methods commonly used in the art (e.g., they can be tested according to the methods given in the embodiments of this application). Unless otherwise stated, the test temperature for all parameters mentioned in this application is 25°C and the test pressure is standard atmospheric pressure.

[0024] The foregoing description of this application is not intended to describe every disclosed implementation or method. Instead, the following description provides more specific examples of exemplary embodiments. Throughout the application, guidance is provided through a series of embodiments, which can be used in various combinations. The examples listed are merely representative and should not be construed as exhaustive.

[0025] In view of the aforementioned deficiencies in the prior art, this application provides a polypeptide derived from CIRBP, its tandemly assembled protein, and its preparation method and application.

[0026] The first aspect of this application provides a polypeptide comprising one or more amino acid sequences as shown in any one of SEQ ID No: 1-13; preferably, the polypeptide is derived from CIRBP protein and has a molecular weight of 0.5~0.7kDa.

[0027] Compared to the complete CIRBP protein, the peptides with the above-mentioned amino acid sequence have smaller molecular weights and superior structural characteristics, thereby significantly improving their bioavailability and transdermal absorption efficiency. This allows the peptides to enter cells and skin tissues more effectively, directly acting on DNA damage repair pathways and exerting their core anti-aging effects by enhancing genome stability. The peptides in this application are derived from smaller molecular weight peptides of the CIRBP protein, exhibiting stronger transdermal and anti-aging effects compared to larger molecular weight peptides derived from the CIRBP protein. The peptides in this application demonstrate stronger biological activity in key skin anti-aging indicators such as promoting the expression of type I and type III collagen, further demonstrating that they can effectively repair skin damage and maintain skin elasticity and firmness by improving skin structure, enhancing barrier function, and delaying the skin aging process.

[0028] The second aspect of the embodiments of this application provides a protein comprising the polypeptides described in the first aspect, which is a small molecular weight protein formed by tandemly connecting the polypeptides described in the first aspect in a specific order, and the molecular weight of the protein is 0.5~7.2kDa; preferably, the protein comprises one or more amino acid sequences as shown in any one of SEQ ID No. 14-15.

[0029] The aforementioned protein comprises one or more specific amino acid sequences assembled in tandem as shown in SEQ ID No. 14-15. It is constructed based on functional fragments of the polypeptide described in the first aspect, retaining the highly active core sequence of the polypeptide while significantly enhancing overall structural stability and solubility through sequence optimization. Furthermore, its smaller molecular weight improves its bioavailability and transdermal absorption. Compared to the natural full-length CIRBP protein, this protein is not only more easily absorbed by the human body and penetrates transdermally, but also exhibits significantly stronger activity in promoting precise DNA repair and maintaining genome stability, thus more effectively achieving the core effects of anti-aging, extending healthy lifespan, and preventing cancer.

[0030] A third aspect of the embodiments of this application provides a nucleic acid that encodes the polypeptide described in the first aspect or the protein described in the second aspect.

[0031] A fourth aspect of the embodiments of this application provides a carrier comprising the nucleic acid described in the third aspect.

[0032] The vector can express the expression under in vivo, in vitro, or ex vivo conditions. Preferably, the vector is a prokaryotic expression vector, a viral expression vector, or a eukaryotic expression vector. Examples include E. coli vectors, bacteriophages, etc.

[0033] As an example, the vector can be the pET28a vector or the pPIC9K vector.

[0034] A fifth aspect of the embodiments of this application provides a host cell comprising the nucleic acid described in the third aspect or the vector described in the fourth aspect; preferably, the host cell comprises at least one of eukaryotic cells and prokaryotic cells.

[0035] In some embodiments, the host cell may be a eukaryotic cell or a prokaryotic cell.

[0036] In some implementations, eukaryotic cells include animal and plant cells, such as T cells, yeast cells, HEK293 cells, or CHO cells, etc.

[0037] In some implementations, prokaryotic cells, such as Escherichia coli, are used.

[0038] As an example, the host cell could be Escherichia coli or Pichia pastoris.

[0039] This invention employs a microbial expression system for protein production. This system offers advantages such as mild culture conditions, low cost, and ease of scale-up, enabling efficient and stable expression of the target protein with significant product yield. Therefore, the preparation process of this invention possesses significant cost-effectiveness and scalable production potential, providing a solid technical foundation for the industrial application of related products in fields such as anti-aging and health management.

[0040] A sixth aspect of the embodiments of this application provides a method for preparing the polypeptide described in the first aspect. The preparation method includes introducing a nucleic acid or vector encoding the polypeptide into a host cell and then inducing its expression. Preferably, the preparation method further includes an enzymatic digestion step.

[0041] In some embodiments, the enzyme in the enzymatic hydrolysis step includes at least one of trypsin, subtilisin, thermophilic protease, papain, and alkaline protease, preferably trypsin.

[0042] The enzymatic hydrolysis step utilizes hydrolytic enzymes, particularly trypsin, which specifically cleave recombinant proteins, releasing highly bioactive target polypeptide fragments. This process not only ensures the structural integrity and functional effectiveness of the polypeptide product but also optimizes its molecular weight distribution, significantly enhancing its solubility, stability, and transdermal absorption efficiency. The polypeptides obtained through this enzymatic hydrolysis process are more easily absorbed by the body and utilized by the skin compared to undigested full-length CIRBP protein, exhibiting stronger DNA damage repair activity and thus more effectively fulfilling their core efficacy in anti-aging, extending healthy lifespan, and cancer prevention.

[0043] The seventh aspect of this application provides the use of the polypeptide described in the first aspect, the protein described in the second aspect, the nucleic acid described in the third aspect, the carrier described in the fourth aspect, or the host cell described in the fifth aspect in the preparation of anti-aging related products.

[0044] In some implementations, the product includes skincare products.

[0045] In some embodiments, the skin care product includes at least one of a bacterial strain, a fermentation broth, a fermentation broth extract, a cell lysate, a fermentation broth precipitate, and a culture prepared from said bacterial strain.

[0046] Example The following embodiments describe the disclosure of this application in more detail. These embodiments are for illustrative purposes only, as various modifications and variations will be apparent to those skilled in the art within the scope of the disclosure of this application. Unless otherwise stated, all parts, percentages, and ratios reported in the following embodiments are based on mass, and all reagents used in the embodiments are commercially available or synthesized by conventional methods and can be used directly without further processing, and the instruments used in the embodiments are commercially available.

[0047] Example 1 Example 1 provides a small molecular weight polypeptide derived from CIRBP and its preparation method, which includes the following steps: (1) Selection of polypeptide sequence: Select one or more active amino acid sequence fragments from CIRBP protein as shown in SEQ ID No. 1 to SEQ ID No. 13, with a molecular weight of 0.5~0.7 kDa.

[0048] (2) Construction of new assembled protein: The polypeptide sequence selected in step (1) is tandemly linked in a preset order to form a new assembled protein, the amino acid sequence of which is shown in SEQ ID No. 14, and the molecular weight of the protein is 7.2 kDa.

[0049] (3) Gene sequence design and optimization: The amino acid sequence shown in SEQ ID No. 14 was converted into the corresponding nucleotide sequence using an online tool (such as EMBOSS Backtranseq; https: / / www.ebi.ac.uk / jdispatcher / st / emboss_backtranseq). Then, the nucleotide sequence was optimized for the E. coli expression system using a codon optimization tool (such as GenSmart™ CodonOptimization; https: / / www.genscript.com.cn / gensmart-free-gene-codon-optimization.html) to obtain the optimized nucleotide sequence SEQ ID No. 16.

[0050] (4) Synthesis and vector construction: The nucleotide sequence shown in SEQ ID No. 16 was chemically synthesized by Qingke Bio Shanghai Branch and cloned into the pET28a expression vector to construct a recombinant plasmid.

[0051] (5) Construction of recombinant engineered bacteria: The above recombinant plasmid was transformed into Escherichia coli expression strain BL21(DE3) to obtain recombinant engineered bacteria.

[0052] (6) Culture of engineered bacteria: The recombinant engineered bacteria were inoculated into a 500 mL Erlenmeyer flask containing 150 mL LB medium and cultured with shaking at 37℃ and 220 rpm until the OD600 reached 0.6.

[0053] (7) Induction of expression: The culture temperature was lowered to 22℃, and IPTG with a final concentration of 1 mM was added to induce protein expression. The culture was continued for 10 h.

[0054] (8) Collection of bacterial cells: After induction, the bacterial cells were collected by centrifugation at 6000 rpm for 3 min.

[0055] (9) Cell disruption: The collected cells were resuspended in LB medium and disrupted by sonication.

[0056] (10) Preparation of crude extract: Centrifuge to remove cell debris and insoluble impurities, and collect the supernatant as crude protein extract.

[0057] (11) Enzymatic hydrolysis: The crude protein extract was hydrolyzed with trypsin (the molar ratio of trypsin to protein was 1:50) to release the target polypeptide fragment.

[0058] (12) Preliminary purification: Microfiltration and ultrafiltration were performed sequentially to further purify and concentrate the polypeptide product.

[0059] (13) Drying the finished product: The purified polypeptide solution is freeze-dried to obtain the polypeptide freeze-dried powder.

[0060] (14) Efficacy verification: The obtained freeze-dried powder was prepared into a 0.1% wt aqueous solution, and its skin care efficacy in promoting collagen production was evaluated through cell experiments.

[0061] Example 2 Example 2 provides a small molecular weight polypeptide derived from CIRBP and its preparation method, which differs from Example 1 only in that... The amino acid sequence of the newly assembled protein tandemly in a preset order is SEQ ID No. 15; the corresponding optimized nucleotide sequence is SEQ ID No. 17; the expression system uses Pichia pastoris and the expression vector is pPIC9K.

[0062] Other steps, including polypeptide sequence selection, gene synthesis, engineered bacteria construction, fermentation culture, protein extraction, enzymatic hydrolysis, purification, and efficacy verification, were all carried out in accordance with the methods described in Example 1, and the culture and induction conditions were adjusted accordingly based on the characteristics of the Pichia pastoris expression system.

[0063] Comparative Example 1 Comparative Example 1 directly expresses the complete CIRBP protein, whose amino acid sequence is shown in SEQ ID No. 18 and the corresponding nucleotide sequence is SEQ ID No. 19.

[0064] Compared with the newly assembled proteins in Examples 1 and 2, the protein sequence expressed in Comparative Example 1 is fundamentally different; the molecular weight (21.7 kDa) of the protein expressed in Comparative Example 1 is much larger than that in Examples 1 (7.2 kDa) and 2 (7.2 kDa); in addition, Comparative Example 1 did not involve the steps of screening for active peptide fragments, tandem recombination of sequences and enzymatic digestion to prepare small molecule peptides, and the final product was a full-length CIRBP protein macromolecule without structural optimization.

[0065] Comparative Example 2 Comparative Example 2 provides a high molecular weight polypeptide derived from CIRBP and its preparation method, which includes the following steps: (1) Selection of polypeptide sequence: Select one or more active amino acid sequence fragments as shown in SEQ ID No.20 to SEQ ID No.31 from CIRBP protein. The molecular weight of the polypeptide is 0.7~2.3 kDa.

[0066] (2) Construction of new assembled protein: The polypeptide sequences selected in step (1) are tandemly arranged in a preset order to form a new assembled protein. Its amino acid sequence is shown in SEQ ID No. 32. The molecular weight of the protein is 12.6 kDa. The number of polypeptides (12) in the new assembled protein of Comparative Example 2 is less than that in Example 1 (13) and Example 2 (13).

[0067] (3) Gene sequence design and optimization: The amino acid sequence shown in SEQ ID No. 32 was converted into the corresponding nucleotide sequence using an online tool (such as EMBOSS Backtranseq; https: / / www.ebi.ac.uk / jdispatcher / st / emboss_backtranseq). Then, the nucleotide sequence was optimized for the E. coli expression system using a codon optimization tool (such as GenSmart™ CodonOptimization; https: / / www.genscript.com.cn / gensmart-free-gene-codon-optimization.html) to obtain the optimized nucleotide sequence SEQ ID No. 33.

[0068] (4) Synthesis and vector construction: The nucleotide sequence shown in SEQ ID No. 33 was chemically synthesized by Qingke Bio Shanghai Branch and cloned into the pET28a expression vector to construct the recombinant plasmid.

[0069] (5) Construction of recombinant engineered bacteria: The above recombinant plasmid was transformed into Escherichia coli expression strain BL21(DE3) to obtain recombinant engineered bacteria.

[0070] (6) Culture of engineered bacteria: The recombinant engineered bacteria were inoculated into a 500 mL Erlenmeyer flask containing 150 mL LB medium and cultured with shaking at 37℃ and 220 rpm until the OD600 reached 0.6.

[0071] (7) Induction of expression: The culture temperature was lowered to 22℃, and IPTG with a final concentration of 1 mM was added to induce protein expression. The culture was continued for 10 h.

[0072] (8) Collection of bacterial cells: After induction, the bacterial cells were collected by centrifugation at 6000 rpm for 3 min.

[0073] (9) Cell disruption: The collected cells were resuspended in LB medium and disrupted by sonication.

[0074] (10) Preparation of crude extract: Centrifuge to remove cell debris and insoluble impurities, and collect the supernatant as crude protein extract.

[0075] (11) Enzymatic hydrolysis: The crude protein extract was hydrolyzed with trypsin (the molar ratio of trypsin to protein was 1:50) to release the target polypeptide fragment.

[0076] (12) Preliminary purification: Microfiltration and ultrafiltration were performed sequentially to further purify and concentrate the polypeptide product.

[0077] (13) Drying the finished product: The purified polypeptide solution is freeze-dried to obtain the polypeptide freeze-dried powder.

[0078] (14) Efficacy verification: The obtained freeze-dried powder was prepared into a 0.1% wt aqueous solution, and its skin care efficacy in promoting collagen production was evaluated through cell experiments.

[0079] Comparative Example 3 Comparative Example 3 provides a small molecular weight polypeptide derived from CIRBP and its preparation method. The molecular weight of the protein expressed in Comparative Example 3 is the same as that in Example 1 (7.2 kDa). The only difference between Comparative Example 3 and Example 1 is the amino acid sequence of the polypeptide and its newly assembled protein. Specifically: (1) Selection of polypeptide sequence: Select one or more active amino acid sequence fragments from the CIRBP protein as shown in SEQ ID No. 34 to SEQ ID No. 45.

[0080] (2) Construction of new assembled protein: The polypeptide sequences selected in step (1) are tandemly linked in a preset order to form a new assembled protein, the amino acid sequence of which is shown in SEQ ID No. 46.

[0081] (3) Gene sequence design and optimization: The amino acid sequence shown in SEQ ID No. 46 was converted into the corresponding nucleotide sequence using an online tool (such as EMBOSS Backtranseq; https: / / www.ebi.ac.uk / jdispatcher / st / emboss_backtranseq). Then, the nucleotide sequence was optimized for E. coli expression system using a codon optimization tool (such as GenSmart™ CodonOptimization; https: / / www.genscript.com.cn / gensmart-free-gene-codon-optimization.html) to obtain the optimized nucleotide sequence SEQ ID No. 47.

[0082] (4) Synthesis and vector construction: The nucleotide sequence shown in SEQ ID No. 47 was chemically synthesized by Qingke Bio Shanghai Branch and cloned into the pET28a expression vector to construct the recombinant plasmid.

[0083] Other steps, including the construction of recombinant engineered bacteria, culture of engineered bacteria, induction of expression, collection of bacterial cells, disruption of bacterial cells, preparation of crude extract, enzymatic hydrolysis, purification, drying, and efficacy verification, are all performed in accordance with the methods described in Example 1.

[0084] Performance testing: 1. Effects of CIRBP-derived peptides prepared in Examples 1-2 and Comparative Examples 1-3 on the expression levels of collagen I and collagen III genes. Normal human skin fibroblasts (HSF, purchased from ATCC: PCS-201-012) were selected and cultured at 37℃ and 5% C Under normal conditions, the culture was performed using DMEM high-glucose medium containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The experimental groups are as follows: The control group cells were given a culture medium containing DMSO at a final concentration of 0.1% wt (the same solvent concentration as the sample treatment group); The cells in the sample groups were respectively added with the sample treatment solutions corresponding to the Examples 1-2 and Comparative Examples 1-3, which had a final concentration of 0.1%wt.

[0085] After 24 hours of treatment under appropriate conditions, total RNA was extracted from cells in each group, and cDNA was obtained through reverse transcription. GAPDH was then used as an internal control gene, and amplification was performed using real-time quantitative PCR (qRT-PCR) with specific primers targeting collagen I and collagen III genes. Their relative expression levels were calculated. The experimental results are as follows: Figure 2-3 As shown, compared with comparative examples 1-3, the expression levels of collagen I and collagen III genes in examples 1-2 were significantly increased, indicating that the peptides provided in this application have significant activity in promoting collagen synthesis and can effectively repair skin damage and maintain skin elasticity and firmness by improving skin structure, enhancing barrier function and delaying skin aging.

[0086] 2. Effects of CIRBP-derived peptides prepared in Examples 1-2 and Comparative Examples 1-3 on skin hydration, skin elasticity, fine line depth, and average area of ​​fine lines around the eyes. The lyophilized powders prepared in Examples 1-2 and Comparative Examples 1-3 were prepared as 0.1% (w / w) aqueous solutions. Water was used as a control group. 0.1 ml of each aqueous solution was applied to the left side of the test subject's face, and the control group's aqueous solution was applied to the right side of the face. This was done twice daily, morning and evening, for 30 consecutive days, with 10 test subjects in each group. After 30 days, the skin's moisture content, skin elasticity, fine line depth, and average area of ​​fine lines around the eyes were measured and averaged. The measurement results are shown in Table 2 below, where the measurement results for Comparative Example 4 are the average values ​​obtained from 40 test subjects.

[0087] Table 1 As can be seen from Table 1, applying the aqueous solution of the polypeptide composition provided in Examples 1 and 2 has a more significant and superior effect on average skin moisture content, average skin elasticity, average depth of fine lines, and average area of ​​fine lines around the eyes compared to applying the aqueous solution of the composition provided in Comparative Examples 1, 2, and 3.

[0088] The above description is merely a specific implementation of this application. Those skilled in the art will clearly understand that, for the sake of convenience and brevity, the specific working process described above can be referred to the corresponding process in the foregoing method embodiments, and will not be repeated here. It should be understood that the protection scope of this application is not limited thereto. Any person skilled in the art can easily conceive of various equivalent modifications or substitutions within the technical scope disclosed in this application, and these modifications or substitutions should all be covered within the protection scope of this application.

Claims

1. A polypeptide, characterized in that, The polypeptide comprises one or more amino acid sequences as shown in SEQ ID No: 1-13; Preferably, the polypeptide is derived from CIRBP protein, and the polypeptide has a molecular weight of 0.5~0.7 kDa.

2. A protein, characterized in that, The protein comprises the polypeptide of claim 1, and the molecular weight of the protein is 0.5~7.2 kDa; Preferably, the protein comprises an amino acid sequence as shown in any one or more of SEQ ID No. 14-15.

3. A nucleic acid, characterized in that, The nucleic acid encodes the polypeptide of claim 1 or the protein of claim 2.

4. A carrier, characterized in that, The vector comprises the nucleic acid as described in claim 3.

5. A host cell, characterized in that, The host cell comprises the nucleic acid of claim 3 or the vector of claim 4; Preferably, the host cell includes at least one of eukaryotic cells and prokaryotic cells.

6. A method for preparing the polypeptide according to claim 1, characterized in that, The preparation method includes introducing a nucleic acid or vector encoding a polypeptide into a host cell and then inducing its expression; Preferably, the preparation method further includes an enzymatic hydrolysis step.

7. The preparation method according to claim 6, characterized in that, The enzymes used in the enzymatic hydrolysis step include at least one of trypsin, subtilisin, thermophilic protease, papain, and alkaline protease, preferably trypsin.

8. The use of a polypeptide of claim 1, a protein of claim 2, a nucleic acid of claim 3, a carrier of claim 4, or a host cell of claim 5 in the preparation of anti-aging related products.

9. The application according to claim 8, characterized in that, The products include skincare products.

10. The application according to claim 9, characterized in that, The skincare product includes at least one of the following: bacterial strain, fermentation broth, fermentation broth extract, cell lysate, fermentation broth precipitate, and culture obtained from the bacterial strain.

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