Hydramacin-1 antimicrobial peptide mutant and preparation method thereof

A mutant and antibacterial peptide technology, which is applied in the field of wild-type Hydramacin-1 gene mutants, recombinant proteins and their preparation fields of the large papillary hydra, can solve the problems that affect the further expression of antibacterial peptides, do not have the ability to process and modify proteins, and post-processing the expression products. trouble, etc.

Active Publication Date: 2013-02-13
冯鹏波
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

1) It does not have the gene expression regulation mechanism of the eukaryotic system and the post-translational processing and modification ability of proteins, and its products often form inactive inclusion bodies, which must be deformed and refolded before they can be used; 2) host cell toxicity
The antimicrobial peptide expressed by the host cell will feedback inhibi

Method used

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  • Hydramacin-1 antimicrobial peptide mutant and preparation method thereof
  • Hydramacin-1 antimicrobial peptide mutant and preparation method thereof
  • Hydramacin-1 antimicrobial peptide mutant and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment

[0036] Embodiment Hydramacin-1 antimicrobial peptide mutant preparation and biological activity assay

[0037] 1. Design and synthesis of antimicrobial peptide Hm-1 gene

[0038] According to the global public sequence database GenBank, the sequence number is DQ449931.1, the nucleotide sequence encoding the mature antimicrobial peptide Hydramacin-1 is used as a template, and the restriction endonuclease XhoI cleavage site is added to the front end of the antimicrobial peptide Hm-1, A restriction endonuclease XbaI restriction site was added to the back end, and two stop codons were continuously added to the front end of the restriction endonuclease XbaI restriction site. The codons for two amino acids (LE) and two basic amino acids (KR) introduced by XhoI belong to the α signal peptide of Pichia pastoris, and are also the intrinsic enzyme cleavage sites of Pichia pastoris. The nucleotide sequence was changed according to the codon preference of Pichia pastoris, and the designe...

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Abstract

The invention relates to a wild type Hydramacin-1 antimicrobial peptide mutant and a preparation method of the Hydramacin-1 antimicrobial peptide mutant. By searching for natural antimicrobial peptide Hydramacin-1 active center, on the basis of determining the active center, replacing the eighth amino acid threonine with aspartic acid or other amino acid, the result shows that the mutant T8D has more excellent antimicrobial property. The invention also discloses a method for expressing the mutant protein in yeast. The method is simple, and the expressed protein has higher antimicrobial activity, and can be applied in the fields such as bio-pharmaceuticals, breeding industry, food industry and feed industry, thereby having wide application prospect and economic significance.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a wild-type Hydramacin-1 gene mutant of Hydra magna, a recombinant protein encoded by it and a preparation method thereof. Background technique [0002] The discovery of penicillin made people no longer helpless against various diseases caused by pathogenic microorganism infection, and thus developed a large number of β-lactam antibiotics and other various antibiotics, which made a great contribution to the protection of human health. However, the beneficial effects of these antimicrobial drugs have also brought some adverse consequences. For example, streptomycin, used to treat tuberculosis, is ototoxic and causes vestibular dysfunction. Penicillin may cause allergic reactions in some allergic patients. Especially with the widespread and large-scale use of these traditional antibiotics, the problem of drug resistance of pathogenic bacteria has become an increasingly serious ...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21
Inventor 徐建华
Owner 冯鹏波
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