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Human antibiotic peptide and use for derivative thereof

An antibacterial peptide, human-derived technology, applied in the field of human semen coagulation protein degradation small peptides, can solve the problem of uncontrolled killing of microorganisms and bacteria, and achieve the effects of inhibiting the growth of bacteria and fungi, convenient artificial synthesis and simple structure

Inactive Publication Date: 2008-04-30
KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology has been found effective for preventing or killing harmful microorganisms by creatively combining different types of molecules together through chemical reactions called synthetic methods. These techniques are simpler than traditional ways like chemotherapy but they still work well even if there's only one type of substance used on top.

Problems solved by technology

This patented technical issue addressed in this patents relates to finding novel compounds called antimicrobium peptids which can fight against various forms of harmful microbes like those causing tuberculosis disease. These molecules may be naturally occurring small proteins isolated during natural selection processes, making their identification challenges due to lack of immune response. However, these agents could potentially lead to developing resistance mechanisms towards drugs commonly used today without being able to stop fighting off even dangerous ones.

Method used

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  • Human antibiotic peptide and use for derivative thereof
  • Human antibiotic peptide and use for derivative thereof
  • Human antibiotic peptide and use for derivative thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Separation and purification of human antimicrobial peptides:

[0045] Normal people masturbate after 3-7 days of abstinence, take the semen in a dry and sterilized measuring cup, and after liquefaction at room temperature, the sperm microbiological dynamic and static computer-aided automatic analysis system (model: CASAS-QH-III, produced by Beijing Tsinghua Tongfang) confirms that the sperm indicators are normal The samples were centrifuged at 10,000 rpm for 10 minutes to separate the sperm and semen, and the supernatant was carefully collected and stored at -20°C for later use.

[0046] The first step: SP-Sepharose ion exchange: the normal human semen (n=40) obtained by the above method is loaded on the SP-Sepharose (Pharmacia product) cation exchange column equilibrated with 50mM Na2HPO4-NaH2PO4, PH=7.4 buffer in advance (130mm long, 20mm diameter). After loading the samples were linearly eluted with 0-0.5M NaCl. Collect 3ml per tube. Antibacterial activity test...

Embodiment 2

[0055] 1. Preparation, separation and purification of human antimicrobial peptides:

[0056] This embodiment is carried out by solid-phase chemical synthesis, using a fully automatic peptide synthesizer to synthesize their full sequences respectively. The names of the synthetic antimicrobial peptides and their full-sequence amino acid primary structures are as follows:

[0057] SgI-29 (SEQ ID NO: 1):

[0058] His Asn Lys Gln Glu Gly Arg Asp His Asp Lys Ser Lys Gly His Phe His Arg Val Val Ile His His Lys Gly Gly Lys Ala His

[0059] SgI-29R7K (SEQ ID NO: 2):

[0060] His Asn Lys Gln Glu Gly Lys Asp His Asp Lys Ser Lys Gly His Phe His Arg Val Val Ile His His Lys Gly Gly Lys Ala His

[0061] SgI-25 (SEQ ID NO: 3):

[0062] Lys Gln Glu Gly Arg Asp His Asp Lys Ser Lys Gly His Phe His Arg Val Val Ile His His Lys Gly Gly Lys.

[0063] Desalted and purified by HPLC reverse phase C18 column chromatography.

[0064] 2. Identification of human antimicrobial peptides:

[0065] The p...

Embodiment 3

[0067] 1. Expression of human antimicrobial peptide SgI-29 gene in Escherichia coli

[0068] According to the known gene sequence encoding mature human semen coagulation protein I from position 86 to position 112, it was cloned into the expression vector pMAL-p2X plasmid, and then transformed into Escherichia coli DH5α, which was induced to express MBP-SgI-29 by IPTG The fusion protein was placed in the periplasm of Escherichia coli, and part of the protein in the periplasm of the bacteria was treated with hypotonicity and then passed through the MBP affinity column to obtain the purified MBP-SgI-29 fusion protein, which was then cleaved by FXa to obtain the antimicrobial peptide SgI-29.

[0069] The high-fidelity pfu enzyme used in the examples, T4 DNA ligase was purchased from Promega, USA, restriction enzymes Xmn I, Hind III, affinity resin (Amylose resin) and expression vector pMAL-p2X were purchased from New England BioLabs, UK Company, tryptone and yeast extract were pur...

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Abstract

The invention relates to human antimicrobial peptide and the purpose of the derivant thereof. The human antimicrobial peptide has amino acid sequences chosen from a SEQ ID NO: 1 listing, and also comprises function equivalents obtained by substitution, cyclization, change from L-amino acid into D-amino acid, lacunae or addition of respective amino acid in amino acid sequence polypeptide. Compared with other source base antimicrobial polypeptide, the human antimicrobial peptide has the advantages that the structure is simple, the artificial synthesis is convenient, and the human antimicrobial peptide is applied to human body non-antigenicity. The human antimicrobial peptide and the derivant of the human antimicrobial peptide of the invention has the remarkable functions of inhibiting bacteria and funguses from growing, thus the human antimicrobial peptide can be used to prepare anti-microorganism infective preparation.

Description

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Claims

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Application Information

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Owner KUNMING INST OF ZOOLOGY CHINESE ACAD OF SCI
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