Mussel byssus polypeptide with antibacterial property and application thereof

Abstract

The invention discloses a mussel byssus polypeptide with antibacterial performance and application thereof.The mussel byssus polypeptide has an obvious inhibiting effect on gram-positive bacteria and gram-negative bacteria, can inhibit formation of biological membranes of the gram-positive bacteria and the gram-negative bacteria, shows different antibacterial activities on multiple drug-resistant bacteria, and has the advantages that the mussel byssus polypeptide can be used for preparing a drug-resistant agent; and the hydrogel has low cytotoxicity and good biocompatibility, and shows good adhesion capability. The polypeptide is short in peptide chain and small in molecular weight, is a micromolecular polypeptide with great value, shows wide application potential in multiple fields, and has important value in the fields of medicine preparation, medical equipment and daily chemical products.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a mussel byssus polypeptide with antibacterial properties and application thereof. Background Art

[0002] Infectious diseases caused by pathogenic bacteria are one of the major diseases that threaten human life and health. The most common and effective method for preventing and treating bacterial infections is antibiotic therapy. However, many traditional antibiotics have weakened or even almost lost their antibacterial activity against many diseases caused by bacteria. In addition, methicillin-resistant Staphylococcus aureus (MRSA) is the main risk pathogen that induces the development of drug resistance. Therefore, new antimicrobial agents are urgently needed to combat multidrug-resistant bacteria. In addition, bacterial colonization and development of biofilms is another serious clinical problem. Since biofilms can survive in harsh environments, higher doses of antibiotics are required to completely eradicate pathogens. Therefore, alternative therapeutic strategies to treat bacterial infections by combating pathogens and their biofilms through a variety of different actions are urgently needed.

[0003] Mussels are an important species in marine aquaculture. Marine mussel adhesive proteins have high strength, high toughness and water resistance, as well as strong adhesion to the substrate, which is related to its special molecular structure, DOPA-mediated interchain crosslinking and interaction with the substrate. In addition, it has good biocompatibility and degradability, and is a class of bioadhesives with great advantages and potential. Type 5 mussel byssus protein is considered to be the adhesive protein molecule with the highest DOPA content in mussel attachment plaques, and plays a direct adhesion role in interfacial adhesion. Direct extraction of mussel byssus protein has limitations, and is a difficult, non-economical, and low-yield method of obtaining it. The preparation of natural mussel byssus protein Mfps by genetic engineering recombination is an effective green biomanufacturing approach to improve the yield and purity of the protein. At present, there are few studies on the core area of ​​type 5 mussel byssus protein.

[0004] Antimicrobial peptides (AMPs) have gradually attracted widespread attention due to their broad antimicrobial activity, unique mechanism of action and low drug resistance. However, the clinical application of natural AMPs is hampered by their high sensitivity to proteolytic degradation and nonspecific toxicity to mammalian cells, while existing antimicrobial preparations have poor inhibitory effects on bacterial biofilms. The high lysine content in Mfp-5 is reminiscent of the high occurrence of this residue in antimicrobial peptides, which confer their effects by disrupting the bacterial cell membrane. Based on relevant studies, it has been shown that lysine- and DOPA-rich peptides derived from Mfp-5 can indeed show antimicrobial activity, and based on this discovery, the most unique region in the type 5 mussel byssus was further searched, in order to serve as a replacement for similar antimicrobial peptides to solve the problems of existing antimicrobial peptides. Summary of the invention

[0005] The purpose of the present invention is to overcome the deficiencies of the prior art and provide a mussel byssus polypeptide with antibacterial properties and application thereof.

[0006] In order to achieve the above-mentioned purpose of the invention, the present invention adopts the following technical solutions: A mussel byssus polypeptide, the amino acid sequence of which is shown in SEQ ID NO.1.

[0007] The mussel byssus polypeptide is used in the preparation of antibacterial drugs; the antibacterial drugs refer to anti-gram-positive bacteria, gram-negative bacteria or multi-drug resistant bacteria; the gram-positive bacteria are Staphylococcus aureus; the gram-negative bacteria are Escherichia coli or Pseudomonas aeruginosa.

[0008] The multidrug-resistant bacteria are methicillin-resistant Staphylococcus aureus or multidrug-resistant Escherichia coli.

[0009] The mussel byssus protein has good biocompatibility and degradability, and has multiple applications in the field of medical devices. It can be used as a bridge between implantable metal stents and bioactive substances, and as a micro-nanoscale protective film, it can effectively protect the wound surface or skin mucosa. It can be used as a medical adhesive for the treatment of surgical incisions, burns, persistent ulcers and skin diseases, and has the effects of analgesia, antibacterial and promoting healing.

[0010] The mussel byssus polypeptide has rich dopa groups and lysine components, can promote cell adhesion and crawling, accelerate wound healing, and has anti-inflammatory and moisturizing effects, and can be used to develop anti-aging and moisturizing skin care products.

[0011] The mussel byssus polypeptide has broad-spectrum adhesion and anti-inflammatory properties, and can be used in sensitive skin care products to help relieve skin inflammation and enhance skin barrier function.

[0012] The interfacial adhesion and bonding ability of the mussel byssus polypeptide gives it a unique advantage in drug preparation, and its property of promoting cell adhesion and migration gives it an important application prospect in the preparation of drugs that promote cell adhesion or migration.

[0013] The present invention fully analyzes the sequence characteristics of mussel foot silk polypeptide by means of bioinformatics and genetic engineering, analyzes the structural characteristics by structural modeling, and on the basis of association analysis, discovers the key adhesion area for extracting mussel foot silk polypeptide. The excavated mussel foot silk polypeptide solves the problems of low direct extraction yield and poor purity by means of genetic engineering, retains its high bonding strength, and has a significant inhibitory effect on both Gram-positive bacteria and Gram-negative bacteria, can inhibit the formation of their biofilm, has a broad spectrum of antibacterial activity and medical value, low cytotoxicity and good biocompatibility. In order to solve the unoptimistic treatment conditions caused by the abuse of existing antibiotics and the formation of bacterial biofilms, the defects of strong cytotoxicity and low biocompatibility of natural antimicrobial peptides are solved at the same time.

[0014] 1. Exploration of mussel byssus polypeptides According to the core functional region of mussel mucin mentioned in Patent 1 (Guangzhou Jinan University Medical Biotechnology Research and Development Center Co., Ltd. A new type of self-assembled mussel mucin and its application: China, CN117986340B.2024.07.30), based on the comprehensive analysis of the sequence characteristics, structural characteristics and molecular evolution characteristics of type 5 mussel byssin, through multi-dimensional information mining, it was found that the conserved domain Domain3 of type 5 mussel byssin is its key structural region. This region was extracted from the full-length type 5 mussel byssin, and the key mussel mucin core functional region cMfp was screened.

[0015] 2. Construction of E. coli vector of mussel byssus polypeptide and expression in E. coli The mussel byssus polypeptide of the present invention has significant innovation, and its key feature is that it can achieve efficient biosynthesis and expression optimization through genetic engineering means. Specifically, the synthesis process of the polypeptide can be achieved by optimizing the codon preference of different expression host systems and combining with suitable recombinant expression vectors. In the present invention, the selectable expression system includes but is not limited to Escherichia coli, Saccharomyces cerevisiae, Pichia pastoris, insect Sf9 host cells and mammalian HEK293 cells. By cloning the target gene into a recombinant expression plasmid and transforming it into a host cell, the host cell is activated by an inducer to achieve overexpression of the target protein. In a preferred embodiment of the present invention, it is recommended to use pET20b (+) as a recombinant expression vector and Escherichia coli BL21 (DE3) as an expression host. This combination can significantly improve the expression efficiency of the target protein while ensuring its biological activity and stability, providing a solid foundation for subsequent industrial applications and research.

[0016] 3. Isolation and purification of mussel byssus polypeptide expression products The fermentation broth was homogenized by a high-pressure homogenizer, and the supernatant was taken by high-speed centrifugation at 12000rpm for 30min using BeckMan (JXN-26) from the United States and loaded on a HiprepTM 16 / 10 CM FF cation exchange column for separation. The loading flow rate was 1ml / min, and the mobile phase was a three-phase eluent, with phase A being 0.02M sodium chloride 20mM PB, phase B being 0.5M sodium chloride 20mMPB, and phase C being 2M sodium chloride 20mM PB. The elution flow rate was 2ml / min, and the purification of the expression product was detected by SDS-PAGE electrophoresis.

[0017] 4. Determination of Minimum Inhibitory Concentration of Mussel Byssus Peptide First, the gram-negative bacteria Escherichia coli and the gram-negative bacteria Staphylococcus aureus were inoculated into LB medium and TSA medium and cultured for 12 hours. After obtaining the bacterial solution, it was diluted to 10 6 CFU / ml concentration, inoculated into 96-well plates, added 50μl cell suspension and 50μl diluted protein solution to each well, incubated for 24h, detected OD600 absorbance value to confirm MIC, and took the suspension after culture and diluted it to a certain multiple to apply on the plate for solid plate counting method.

[0018] 5. Detection of different antibacterial activities of mussel byssus polypeptides against drug-resistant and non-resistant bacteria Take the bacterial suspension of Staphylococcus aureus, methicillin-resistant Staphylococcus aureus, Escherichia coli and multidrug-resistant Escherichia coli activated overnight, centrifuge and discard the supernatant, resuspend with PBS and dilute to about 10 6 CFU / mL, take 300μL of the diluted bacterial solution and 300μL of protein solution of different concentrations and mix them in a 2mL EP tube, culture at 37℃ 220rpm, take pictures at 2h and 6h to observe the turbidity changes and take samples for plating.

[0019] 6. Detection of the ability of mussel byssus polypeptide to inhibit biofilm The bacterial suspension of Staphylococcus aureus and Escherichia coli activated overnight was diluted to about 10 6CFU / mL. 500 μL of bacterial suspension and 500 μl of protein dilution were inoculated into 24-well microtiter plates. After 24 and 48 h of culture, the supernatant culture medium was removed and the underlying biofilm was washed three times with phosphate-buffered saline (PBS). The biofilm was then fixed in methanol for 15 min and stained with 0.1% (w / v) CV for 10 min. The wells were washed with sterile water and dried at 37°C for 30 min. Images were captured using an optical microscope. The crystal violet was then dissolved with 500 μl of 33% glacial acetic acid at 37°C with shaking (80 r / min) for 30 min. OD was measured using a microplate reader. 595 nm absorbance value.

[0020] 6. Cytotoxicity test of mussel byssus polypeptide The cytotoxicity of mussel byssus polypeptide was determined by MTT cell proliferation. The solutions with different protein concentrations were air-dried to modify the bottom of 96-well plates by coating method. After L929 cells were revived and activated, they were inoculated into 96-well plates, with 1x10 4个 cell / well and in a 5% CO 2 Incubate in a 37 °C incubator for 24 hours (for blank control, use PBS instead of protein solution). At the end of the incubation, remove the medium and add MTT solution with a final concentration of 0.5 mg / mL to each well and continue incubation for 4 hours. At the end of the incubation, carefully remove the medium and add 200 μL of formazan solution to each well and shake slowly using a decolorizing shaker to ensure complete dissolution. After 10 minutes, measure the absorbance of the solute at 570 nm using an enzyme labeling instrument.

[0021] 7. In vitro adhesion ability test of mussel byssus polypeptide 30 ng of protein solution was dripped onto the plastic culture dish, and gun tips of different sizes were bonded to the dish. The mixture was dried in a humid environment at 25°C for 12 h, and the bonding condition was observed.

[0022] Compared with the prior art, the present invention has the following beneficial effects: The mussel foot silk polypeptide of the present invention has a significant inhibitory effect on both Gram-positive bacteria and Gram-negative bacteria, can inhibit the formation of their biofilm, and at the same time exhibits different antibacterial activities against multi-drug resistant bacteria, has low cytotoxicity and good biocompatibility, and exhibits good adhesion and bonding ability. It has a short peptide chain and a small molecular weight, is a very valuable small molecule polypeptide, and has broad application prospects.

[0023] 1. As a new type of antibiotic used for bacterial treatment or prevention, it provides new ideas and treatment options for clinical multidrug-resistant bacterial infections.

[0024] 2. Solve the defects of strong cytotoxicity and low biocompatibility of natural antimicrobial peptides.

[0025] 3. As the most unique region in the type 5 mussel byssus, it is expected to be used as an adhesion domain in protein design. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] In order to more clearly illustrate the embodiments of the present invention, the drawings related to the embodiments are briefly introduced below.

[0027] Figure 1 This is the mass spectrometry detection image of mussel byssus polypeptide; Figure 2 This is the predicted map of alph-helical wheel of mussel byssus polypeptide; Figure 3 This is the plasmid map of the mussel byssus polypeptide expression vector; Figure 4 Electrophoresis diagram for screening bacteria expressing mussel byssus polypeptide; Figure 5 This is a diagram showing the purification effect of mussel byssus polypeptide by cation column exchange chromatography; Figure 6 This is a diagram showing the antibacterial effect of mussel byssus polypeptide on non-drug-resistant bacteria; Figure 7 This is a diagram showing the antibacterial effect of mussel byssus polypeptide on drug-resistant bacteria; Figure 8 Quantitative graph of crystal violet for the inhibition of biofilm formation by mussel byssus polypeptide; Fig. 9 This is a diagram showing the results of the cytotoxicity test of mussel byssus polypeptide; Fig.10 This is a diagram showing the in vitro adhesion ability of mussel byssus polypeptide. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with specific examples. It should be understood that the examples are only used to illustrate the present invention and are not used to limit the scope of the present invention.

[0029] Example 1: Mining of mussel byssus polypeptide cMfp A total of 405 relevant protein information were retrieved from the NCBI database. Combined with the blastp comparison and manual verification, a total of 4 target entries with complete nucleic acid sequences and protein sequences were screened out. Mytilusgalloprovincialis, Mytilusedulis, Mytiluscoruscus, Mytiluscalifornianus. The amino acid composition of the four mussel foot silk proteins was analyzed, which was mainly composed of glycine (Gly), lysine (Lys), tyrosine (Try), and serine (Ser). On the basis of obtaining basic sequence information, the characteristics and distribution of specific sequence sites were further analyzed by multiple sequence alignment. The threshold was set to a Max score of more than 100 to ensure that the obtained sequences had high similarity and possible homology. The results showed that the four types of type 5 mussel foot silk proteins were clearly divided into three more conservative regions. MEME mined and predicted the protein conservative motifs to identify possible functional domains or binding site protein motifs. The mining results are consistent with the results of multiple sequence alignment. At the same time, from the visualized motif diagram, it can be intuitively found that the domain3 region is the region with the highest tyrosine and lysine enrichment in type 5 mussel byssus protein. The ProtParam tool is used to analyze the physicochemical properties of the sequence, including protein molecular weight, theoretical isoelectric point, amino acid composition, instability coefficient and fat solubility index. The fasta file of the protein sequence is uploaded to the SignalP 6.0 platform to predict whether there is a potential signal peptide cleavage site and its position in a given amino acid sequence. The AbinitioRelax application is used for Robetta de novo prediction to predict the initial structure of the protein. At the same time, the Rosetta all-atom force field is used for all-atom refinement to obtain a more accurate protein structure model. The structural models of four types of mussel byssin proteins were constructed from scratch using the Alphfold2 algorithm. PSI-PRED was used as a secondary structure predictor, and I-TASSER was used for homology modeling to predict the tertiary structure of the protein. The primary sequence characteristics and three-dimensional structural characteristics were associated with the distribution of molecular evolution sites, and the key mussel mucin core functional region cMfp was screened. The mass spectrometry detection of the obtained protein was as follows Figure 1 The results of the alph-helical wheel diagram indicate that the lysine and tyrosine in the cMfp region have the potential to form an alternating helical ring. Further spatial structure simulations found that a helical conformation can indeed be formed, as shown in Figure 2 shown.

[0030] Example 2: Construction and expression screening of mussel byssus polypeptide cMfp expression vector The codon preference of the core polypeptide of the type 5 mussel byssus protein was optimized by the codon preference of Escherichia coli. The core polypeptide fragment of the type 5 mussel byssus protein was fully synthesized by a gene synthesis company. The obtained cMfp gene fragment was inserted between NdeI and EcoRⅠ of the pET20b(+) vector to form the pET20b(+)-cMfp vector. The vector plasmid map is shown in Figure 3 shown.

[0031] According to the instructions of the TIANgel Midi Purification Kit provided by Takara, the obtained pET20b(+) - cMfp vector ligation product was introduced into TOP10 competent cells. Subsequently, the transformed cells were placed on a resistance plate containing ampicillin (Amp+) and cultured at 37°C. Single clone colonies were picked from the plate, and positive clones were identified by PCR. The positive colonies were inoculated into 5mL of ordinary LB medium containing 100μg / mL ampicillin and cultured overnight at 37°C. After the culture, the plasmid was extracted and sent for sequencing to confirm the accuracy of the sequence.

[0032] The pET20b(+)-cMfp plasmid was transformed into E. coli BL21(DE3)pLysS competent cells. Ten monoclonal colonies were randomly selected and positive clones were screened by PCR. The positive colonies were inoculated into a 20 mL test tube, and 5 mL of LB medium containing 100 μg / mL ampicillin was added and cultured at 37°C overnight. Subsequently, 50 μL of the overnight culture was inoculated into 5 mL of LB medium containing 50 μg / mL ampicillin at an inoculum size of 1:100 and cultured at 37°C for about 3 hours until the OD600 value reached 0.6-0.8. When the OD600 value reached the target range, 200 μL of the bacterial solution was taken and centrifuged at 12000 rpm for 1 minute to collect the bacteria before induction of expression. Then, 1 mM IPTG was added to the remaining medium and cultured at 37°C for another 3 hours to induce protein expression. After induction, take 200 μL of bacterial solution again, centrifuge at 10000 rpm for 1 minute, and collect the induced expression bacteria. Add 80 μL of 5% SDS solution to the bacteria before and after induced expression, boil at 100℃ for 10 minutes until the solution is clear. Then add 20 μL of 5xloading buffer and boil at 100℃ for 10 minutes again to complete sample preparation. Finally, the samples before and after induced expression were identified by SDS-PAGE electrophoresis. The results are as follows: Figure 4 shown.

[0033] Example 3: Purification of mussel byssus polypeptide cMfp First, accurately weigh 100 g of the wet weight of BL21(DE3) / pET20b(+)-cMfp bacteria. Then, use 1000 ml of 20 mM phosphate buffer (PB) and 0.15 M sodium chloride solution (pH 7.4) as resuspension buffer to fully resuspend the bacteria. After resuspension, place the bacterial suspension in a low-temperature environment of 4°C and use a high-pressure homogenizer for three times of crushing, with the pressure set to 800 Pascals each time. After the crushing is completed, the sample is centrifuged at 12,000 rpm for 30 minutes to separate cell fragments and unbroken cells. Finally, collect the supernatant for subsequent experimental operations. According to the characteristics of cMfp protein, a cation exchange column was selected. Before loading the sample, the column was pretreated by washing the ethanol with 3-5 column volumes of distilled water, and the column was balanced with 5 column volumes of binding buffer. The recommended flow rate is 1 ml / min. After equilibration, the sample was loaded and balanced with binding buffer for at least 5 column volumes until the absorption peak reached a stable baseline. Unbound proteins and non-specific proteins were washed away with washing buffer (20mM PB 0.5MNacl) for at least 3-5 column volumes. Finally, elution buffer (20mMPB 2M Nacl) was used for elution, usually 5-10 column volumes, and the absorption peak was collected from the rise to the fall to the stable baseline, and the purified new self-assembled mussel mucin cMfp was obtained. The purification was detected by SDS-PAGE electrophoresis. The results are shown in the figure. Figure 5 The amino acid sequence of the obtained mussel byssus polypeptide is shown in SEQ ID NO.1.

[0034] Example 4: Determination of the minimum inhibitory concentration of mussel byssus polypeptide cMfp against Gram-positive and Gram-negative bacteria The minimum inhibitory concentration (MIC) of the peptide was determined by the microbroth dilution method. Gram-negative bacteria Escherichia coli and Gram-positive bacteria Staphylococcus aureus were inoculated into LB medium and TSA medium and cultured at 37°C for 12 hours. The bacterial solution was diluted to 10 6 CFU / ml concentration, the peptide was prepared into a 2mg / ml storage solution for standby use, a series of gradient peptide solutions were prepared in sequence using the two-fold dilution method, 50μl of cell suspension and 50μl of diluted protein solution were inoculated in a 96-well plate, and 100μl of culture medium was added to each well, and a positive control (containing bacterial solution but not peptide) and a negative control (containing neither bacterial solution nor peptide) were set up respectively. After 12-16h of incubation, the OD was detected by an ELISA instrument 600 The absorbance value confirms the MIC, and the suspension after culture is diluted to a certain multiple and plated for solid plate counting. The results are as follows Figure 6 shown.

[0035] Example 5: Detection of antibacterial activity of mussel byssus polypeptide cMfp against drug-resistant and non-resistant bacteria Gram-negative resistant bacteria - MDR-E. coli (multi-drug resistant Escherichia coli) and non-resistant Escherichia coli and Gram-positive resistant bacteria - MRSA (methicillin-resistant Staphylococcus aureus) and non-resistant Staphylococcus aureus were inoculated into TSB medium and cultured at 37°C for 12 hours. 200 μL of the overnight activated MRSA and MDR-E. coli bacterial suspension was centrifuged at 8000 rpm for 1 min, the medium was discarded, and the suspension was resuspended with 200 μL PBS and diluted to about 10 6 CFU / mL, take 200μL of diluted bacterial solution and 200μL of protein solution of different concentrations and mix them in 2mL EP tube, culture at 37℃ 220rpm, take pictures at 2h and 6h to observe the turbidity changes and take samples to plate. The results are as follows Figure 7 shown.

[0036] Example 6: Detection of the ability of mussel byssus polypeptide cMfp to inhibit biofilm formation Gram-negative bacteria Escherichia coli and Gram-positive bacteria Staphylococcus aureus were inoculated into LB medium and TSA medium and cultured at 37°C for 12 hours. The overnight activated Escherichia coli and Staphylococcus aureus bacterial suspensions were diluted to about 10 6 CFU / mL. 500μL of bacterial suspension and 500μl of protein dilution were inoculated into 24-well microtiter plates. After 24 and 48 hours of culture, the supernatant culture medium was removed and the underlying biofilm was washed 3 times with phosphate-buffered saline (PBS). The biofilm was then fixed in methanol for 15 minutes and stained with 0.1% (w / v) CV for 10 minutes. The wells were washed with sterile water and dried at 37°C for 30 minutes. Images were captured using an optical microscope. The crystal violet was then dissolved with 500μl of 33% glacial acetic acid at 37°C with shaking (80 r / min) for 30 minutes. The OD was measured using a microplate reader. 595 Absorbance value, the result is Figure 8 shown.

[0037] Example 7: Detection of cytotoxicity of mussel byssus polypeptide cMfp The purified and filtered cMfp protein solution was diluted with PBS to concentrations of 0.8, 0.4, 0.2, and 0.05 mg / ml. Take 10 μL of protein solution of different concentrations and add it to the wells of the 96-well plate to coat the bottom of the plate. After the protein solution is dried to form a protein film layer, wash it three times with PBS and dry it. The wells without any treatment serve as the control group. After the bottom of the plate is modified, the 96-well plate is placed under ultraviolet light for 1 hour. Subsequently, the well-grown L929 cells are taken, digested with trypsin, and 1×10 4The density of cells was inoculated into a protein-modified 96-well plate and an unmodified 96-well plate, respectively. The cells were cultured in a cell culture incubator at 37°C for 24 hours. After culturing to the specified time point, 10 μL of MTT solution was added to each well and cultured for another 4 hours. Carefully aspirate all the supernatant in the well, and then add 100 μL of formazan solubilization solution to each well. After the formazan is completely dissolved, use an enzyme reader to measure the absorbance at 570 nm and draw an absorbance curve. The experimental results are shown in Figure 2. Fig. 9 shown.

[0038] Example 8: In vitro adhesion ability test of mussel byssus polypeptide cMfp Take an appropriate amount of sample solution and spot it on the surface of the culture dish using a micropipette. The spot volume is 10μL, which is equivalent to about 30ng of sample. Then, place different specifications of gun tip consumables (including 1mL, 200μL and 10μL) on top of the sample solution to ensure that the gun tip is in full contact with the sample. After the solution is naturally air-dried, carefully turn the culture dish cover upside down and observe whether the gun tip is firmly adhered to the culture dish cover to evaluate its bonding effect. The specific observation results are as follows: Fig.10 As shown in the figure, the three specifications of 1mL, 200μL and 10μL pipette tips all showed good adhesion properties and could be firmly attached to the culture dish cover, indicating that these pipette tips have high stability and reliability in trace sample processing.

Claims

1. A mussel byssus polypeptide, the amino acid sequence of which is shown in SEQ ID NO.

1.

2. Use of the mussel byssus polypeptide according to claim 1 in the preparation of antibacterial drugs.

3. The use according to claim 2, characterized in that The antibacterial agent refers to anti-Gram-positive bacteria, Gram-negative bacteria or multi-drug resistant bacteria.

4. The use according to claim 3, characterized in that The Gram-positive bacteria are Staphylococcus aureus; the Gram-negative bacteria are Escherichia coli or Pseudomonas aeruginosa; and the multidrug-resistant bacteria are methicillin-resistant Staphylococcus aureus or multidrug-resistant Escherichia coli.

5. Use of the mussel byssus polypeptide according to claim 1 in preparing a bioadhesive for protecting wound surfaces or skin mucosa.

6. Use of the mussel byssus polypeptide according to claim 1 in the preparation of products for sensitive skin care, anti-inflammatory, moisturizing or anti-aging.

7. Use of the mussel byssus polypeptide according to claim 1 in the preparation of a drug for promoting cell adhesion or migration.

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