Helicobacter pylori bacteriophage endolysins and preparation method thereof

A Helicobacter pylori, phage lysing enzyme technology, applied in the biological field, can solve the problem of weak, cracking Helicobacter pylori, etc.

Active Publication Date: 2020-07-07
江苏万邦医药科技有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There are currently no literature or patents on research on H. pylori phage lyase; H. pylori is a Gram-negative bacterium, and its lyase may not be able to lyse H. pylori from the outside in or be weak

Method used

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  • Helicobacter pylori bacteriophage endolysins and preparation method thereof
  • Helicobacter pylori bacteriophage endolysins and preparation method thereof
  • Helicobacter pylori bacteriophage endolysins and preparation method thereof

Examples

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Effect test

Embodiment 1

[0057] Preparation of Helicobacter pylori phage lyase containing polycation nonapeptide

[0058] 1. Construction of Helicobacter pylori phage lyase expression vector and engineering bacteria containing polycationic nonapeptide

[0059] 1. Construction of the nucleotide sequence of Helicobacter pylori phage lyase comprising polycation nonapeptide: the polypeptide and nucleotide sequences of polycation nonapeptide and Helicobacter pylori phage lyase are as follows respectively (Seq ID No.7 and Seq ID No. 8, Seq ID No. 11 and Seq ID No. 13). According to the codon preference of Escherichia coli, the nucleotides comprising the natural Helicobacter pylori phage lyase are optimized. This optimization method is a technique known to those skilled in the art. For example, see the Journal of Inner Mongolia University: Escherichia coli and yeast codon usage Compare, 2006, vOI.37, NO.1:34-39; the final nucleotide sequence is as follows (Seq ID No.12). Add polycation nonapeptide sequence...

Embodiment 2

[0083] Preparation of Helicobacter pylori Phage Lyase Containing Small Hydrophobic Peptides

[0084] 1. Construction of Helicobacter pylori phage lyase expression vector and engineering bacteria containing hydrophobic small peptides

[0085] 1. Construct the nucleotide sequence of the Helicobacter pylori phage lyase containing the hydrophobic small peptide: the polypeptide and nucleotide sequences of the hydrophobic small peptide and the Helicobacter pylori phage lyase are as follows (Seq ID No.9 and Seq ID No. 10, Seq ID No. 11 and Seq ID No. 13). According to the codon preference of Escherichia coli, the nucleotide containing the natural Helicobacter pylori phage lyase was optimized, and the final nucleotide sequence was obtained as follows (Seq ID No.12). Add hydrophobic small peptide sequence (gene sequence is Seq ID No.9) and enterokinase cleavage site VDDDDK sequence (gene sequence is Seq ID No.5) to the 5' end of Helicobacter pylori phage lyase to obtain hydrophobic sm...

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PUM

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Abstract

The invention belongs to the technical field of biology, and in particular relates to helicobacter pylori bacteriophage endolysins and a preparation method thereof. A natural helicobacter pylori bacteriophage endolysin protein sequence is screened out in the invention; furthermore, easily permeable polycationic 9 peptide and hydrophobic small molecule peptide are respectively added into the N endof the protein sequence; fermentation-induced expression of an improved helicobacter pylori bacteriophage endolysin gene is carried out through an escherichia coli expression system; the expression quantity is high; a soluble expression method without deformation and renaturation is further provided; furthermore, a purification step is simple and convenient; the fact that the lysis ability of theimproved helicobacter pylori bacteriophage endolysins on a standard helicobacter pylori strain ATCC700392 is high is found through an in-vitro experiment; and material and theoretical basis can be provided for research on treatment of helicobacter pylori infection.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a helicobacter pylori phage lyase and a preparation method thereof. Background technique [0002] Helicobacter pylori (HP) infection is the main pathogenic factor of chronic active gastritis, peptic ulcer, gastric mucosa-associated lymphoid tissue (MALT) lymphoma and gastric cancer. The following regimens are currently used for the treatment of Helicobacter pylori infection: 1) dual therapy: secret preparations are used in combination with antibiotics. 2) Triple or quadruple therapy: secret medicine + metronidazole + tetracycline + omeprazole; omeprazole + amoxicillin + clarithromycin; secret medicine + tinidazole + clarithromycin. Compliance with these treatments is very poor, and multiple strains of HP have developed drug resistance, and even multi-drug resistant bacteria and superbugs have emerged. Therefore, it is urgent to find a new antibacterial agent to deal with the threat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/80C12N15/60C12N15/70C12N1/21
CPCC12N9/80C12N15/70C12Y305/01028Y02A50/30
Inventor 梅丽赵珊珊文可馨苗森赵梅
Owner 江苏万邦医药科技有限公司
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