Polypeptide, dna molecule encoding the polypeptide, vector, preparation method and use

Abstract

A polypeptide, a DNA molecule encoding the polypeptide, a vector, a preparation method and a use therefor are disclosed. The polypeptide comprises an amino acid sequence represented by formula (I) or formula (II): formula (I) comprises an amino acid sequence represented by SEQ ID NO: 1; formula (II) comprises an amino acid sequence obtained by subjecting the amino acid sequence represented by SEQ ID NO: 1 to modification, substituted, and deletion or addition of one or more amino acids. The polypeptide may be used in the preparation of drugs that treat or prevent diseases related to infections caused by bacteria, and can also be used in the preparation of drugs that promote tissue repair and wound healing.

1 . A polypeptide characterized in having any one of the amino acid sequences as set forth in (I) and (II):
 (I) the amino acid sequence as set forth in SEQ ID NO: 1;   (II) amino acid sequences obtained from the amino acid sequence as set forth in SEQ ID NO: 1 with modification, substitution, deletion or addition of one or more amino acids.   
     
     
         2 . The polypeptide of  claim 1 , characterized in that,
 the modification includes amidation, phosphorylation, methylation, acetylation, ubiquitination, glycosylation or carbonylation; the substitution is substitution of 1, 2, 3, 4 or 5 amino acids; the deletion is deletion of 1, 2, 3, 4 or 5 amino acids; and/or the addition is addition of 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids.   
     
     
         3 - 5 . (canceled) 
     
     
         6 . The polypeptide of  claim 1 , characterized in that, the polypeptide does not have the amino acid sequence as set forth in SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 53 or SEQ ID NO: 54. 
     
     
         7 . The polypeptide of  claim 1 , characterized in that, the polypeptide has an amino aide sequence as set forth in any one of SEQ ID NO: 2, SEQ ID NOs: 22 to 29, SEQ ID NO: 32, SEQ ID NOs: 36 to 45, or SEQ ID NO: 52. 
     
     
         8 - 12 . (canceled) 
     
     
         13 . A method for the improvement of condition in a subject comprising administering the polypeptide of  claim 1  to the subject, wherein the improvement of condition is selected from the group consisting of:
 the inhibition of bacteria; 
 the prevention and/or treatment of a bacterial infection disease and/or eczema; 
 the promotion of tissue repair and/or wound healing; and 
 the treatment of a burn injury, cold injury, crush injury, war injury, animal bite, nuclear radiation injury or combined injury. 
 
     
     
         14 . The method of  claim 13 , characterized in that, the bacterium is a gram-positive bacterium and/or gram-negative bacterium. 
     
     
         15 . The method of  claim 14 , characterized in that, the gram-positive bacterium belongs to  Staphylococcus, Streptococcus, Enterococcus, Mycobacterium , anaerobic bacterium or  Corynebacterium ; and/or the gram-negative bacterium belongs to  Neisseria, Enterobacteriaceae, Pseudomonas, Acinetobacter, Bordetella  or  Haemophilus.    
     
     
         16 . (canceled) 
     
     
         17 . The method of  claim 13 , characterized in that, the infection is a nonspecific infection and/or specific infection. 
     
     
         18 . The method of  claim 17 , characterized in that, the nonspecific infection is furuncle, carbuncle, erysipelas, acute lymphangitis, acute lymphadenitis, paronychia, felon, lateral pyogenic tenosynovitis of fingers, bursitis, palm deep space infection, pyemia or bacteremia; and/or the specific infection is tuberculosis, tetanus, gas gangrene, anthrax, pertussis, epidemic encephalomyelitis, gonorrhea, typhia, bacillary dysentery or diphtheria. 
     
     
         19 . (canceled) 
     
     
         20 . The method of  claim 13 , characterized in that, the infection is an infection caused by MRSA, MRCNS, erythromycin-resistant  Streptococcus pyogenes , ESBL-producing  Escherichia coli , imipenem-resistant  Pseudomonas aeruginosa ; or
 the infection is a local infection, systemic infection or toxic disease caused by  Staphylococcus aureus ; or   the infection is a urinary system infection, septicemia or postoperative infection, caused by coagulase-negative  staphylococci ; or   the infection is a pyogenic inflammation, rheumatic fever and acute glomerulonephritis, scarlatina, bacterial pneumonia, saprodontia, subacute bacterial endocarditis or newborn infection, caused by  Streptococcus ; or   the infection is lobar pneumonia, trachitis, otitis media, meningitis, pleurisy, endocarditis or septicemia, caused by  Streptococcus pneumonia ; or   the infection is puerperal septicopyemia in pregnant women, neonatal meningitis, postpartum infection, bacteremia, endocarditis, skin and soft tissue infection or osteomyelitis, caused by  Streptococcus agalactiae ; or   the infection is a cardiovascular system infection or urinary tract infection, caused by  Enterococcus ; or   the infection is a urinary tract infection, pyogenic abdominal infection, septicemia, endocarditis or diarrhea fever, caused by  Enterococcus faecium ; or   the infection is endocarditis, cholecystitis, meningitis, urinary tract infection or wound infection, caused by  Enterococcus faecalis ; or   the infection is epidemic encephalomyelitis caused by  Neisseria meningitides ; or   the infection is gonorrhea caused by  Neisseria gonorrhoeae ; or   the infection is a urinary system infection, parenteral pyogenic infection, intestinal infection or hemorrhagic colitis, caused by  Escherichia ; or   the infection is bacillary dysentery caused by  Shigella ; or   the infection is typhia and paratyphoid caused by  Salmonella ; or   the infection is pneumonia, bronchitis, urinary tract infection, wound infection, meningitis or peritonitis, caused by  Klebsiella pneumonia ; or   the infection is antibiotic-associated hemorrhagic colitis caused by  Klebsiella oxytoca ; or   the infection is pneumonia or meningitis caused by  Citrobacter , or   the infection is skin soft tissue infection, urinary tract infection, respiratory tract infection, abdominal infection, central nervous system infection, eye infection, wound infection, endocarditis or septicemia, caused by  Enterobacter cloacae ; or   the infection is pneumonia, urinary tract infection, bacteremia or postoperative infection, caused by  Serratia ; or   the infection is tetanus caused by  Clostridium tetani ; or   the infection is gas gangrene or food poisoning caused by  Clostridium perfringens ; or   the infection is abdominal infection, female reproductive tract and pelvic infection or bacteremia, caused by asporous anaerobic bacterium; or   the infection is diphtheria caused by  Corynebacterium diphtheria ; or   the infection is acne, comedo caused by  Corynebacterium acnes ; or   the infection is a wound infection, burn tissue infection, lung infection, urinary tract infection, otitis media, keratitis, endocarditis or septicemia, caused by  Pseudomonas aeruginosa ; or   the infection is a respiratory tract infection, bacteremia, urinary tract infection, secondary meningitis, surgical site infection or ventilator-associated pneumonia, caused by  Acinetobacter baumannii ; or   the infection is pertussis caused by  Bordetella pertussis ; or   the infection is bacteremia in infants and children, acute bacterial meningitis, cellulitis, osteomyelitis or joint infection, caused by  Haemophilus  such as  Haemophilus influenza ; or   the infection is tuberculosis caused by  Mycobacterium tuberculosis ; or   the infection is chronic refractory infection wound surface.   
     
     
         21 - 22 . (canceled) 
     
     
         23 . A DNA molecule encoding the polypeptide of  claim 1 , characterized in having any one of the nucleotide sequences as set forth in I and II:
 I. the nucleotide sequence as set forth in SEQ ID NO: 3;   II. nucleotide sequences obtained from the nucleotide sequence as set forth in SEQ ID NO: 3 with modification, substitution, deletion or addition of one or more bases.   
     
     
         24 . (canceled) 
     
     
         25 . A method for preparing a polypeptide, characterized in comprising the steps of:
 obtaining a DNA molecule encoding the amino acid sequence as defined in (I) or (II); wherein   (I): the amino acid sequence as set forth in SEQ ID NO: 1; and   (II): amino acid sequences obtained from the amino acid sequence as set forth in SEQ ID NO: 1 with modification, substitution, deletion or addition of one or more amino acids;   fusing the DNA molecule with an expression vector to construct a recombinant expression vector;   transforming the recombinant expression vector into a host cell to obtain a transformant; and   inducing the transformant to express a protein, and then performing separation and purification, to obtain the polypeptide.   
     
     
         26 . The preparation method of  claim 25 , characterized in that, the DNA molecule comprises any one of the nucleotide sequences as set forth in I and II:
 I. the nucleotide sequence as set forth in SEQ ID NO: 3;   II. nucleotide sequences obtained from the nucleotide sequence as set forth in SEQ ID NO: 3 with modification, substitution, deletion or addition of one or more bases.   
     
     
         27 . The preparation method of  claim 26 , characterized in that, the host cell is a prokaryotic host cell or eukaryotic host cell. 
     
     
         28 . The preparation method of  claim 26 , characterized in that, the prokaryotic host cell is  Escherichia coli.    
     
     
         29 . A pharmaceutical formulation for the treatment of a wound surface infection and/or promotion of tissue repair and healing, characterized in that, the pharmaceutical formulation consists of the polypeptide of  claim 1  and a pharmaceutically acceptable carrier. 
     
     
         30 . The pharmaceutical formulation of  claim 29 , characterized in that, the polypeptide accounts for 0.01%˜1.5% by weight of the pharmaceutical formulation; and/or the carrier comprises hydroxypropylmethylcellulose. 
     
     
         31 - 32 . (canceled) 
     
     
         33 . The pharmaceutical formulation of  claim 29 , characterized in that, the pharmaceutical formulation is a gel, powder for injection, aerosol, spray, liniment, film, patch, cream, ointment, adhesive plaster, liquid, decoction, granule, tablet, pill, sustained-release agent, controlled-release agent, powder, paste, liniment, lotion, smeared film, penetration ions, eye drop, nasal drop, gargle, sublingual tablet, insufflating agent, suppository, aerosol, inhalant, fumicant, oral solution, oral tablet, injection, syrup, electuary, vinum, pulvis, granule, pill, tablet, capsule, enema or suppository. 
     
     
         34 . A method of manufacturing a feed, health product, food and cosmetic additive, comprising using the polypeptide of  claim 1 .