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Escherichia coli lipid A binding element PCK as well as preparation method and application thereof

A technology of medicinal salts and compounds, applied in the field of Escherichia coli lipid A binding motif PCK and its preparation and application, which can solve the problems of unreported research on cytotoxicity

Active Publication Date: 2021-11-26
FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Search data found that there is no report on the synthesis and cytotoxicity of the lipid A binding motif derived from E. coli PbgA protein

Method used

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  • Escherichia coli lipid A binding element PCK as well as preparation method and application thereof
  • Escherichia coli lipid A binding element PCK as well as preparation method and application thereof
  • Escherichia coli lipid A binding element PCK as well as preparation method and application thereof

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Embodiment 1

[0069] 1. Preparation of Escherichia coli lipid A binding motif PCK

[0070] The lipid A-binding motif PCK resin was synthesized by Fmoc solid-phase synthesis method, and the lipid A-binding motif PCK was prepared by trifluoroacetic acid (trifluoroacetic acid, TFA) cleavage, freeze-drying, HPLC purification and mass spectrometry identification.

[0071] The amino acid sequence of PCK is:

[0072] Ser Tyr Pro Met Tyr Ala Arg Arg Phe Leu Phe Lys Trp Gly Leu Leu Arg-NH 2 (as shown in Sequence Listing Sequence 1).

[0073] The specific steps for synthesizing Escherichia coli lipidA-binding motif PCK by Fmoc solid-phase synthesis are as follows: the preparation of lipidA-binding motif PCK is carried out from the C-terminal to the N-terminal direction. First, the first amino acid at the C-terminal, namely Fmoc-Arg-OH, is activated, Coupling to CTC resin, and then deprotecting the protecting group Fmoc to obtain X-Arg+CTC resin; then activate and couple the second amino acid at the...

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Abstract

The invention discloses a polypeptide specifically bound with escherichia coli lipid A and application of the polypeptide in inhibiting gram-negative bacteria. The amino acid sequence of the polypeptide is SEQ ID No.1 in a sequence table. The polypeptide can specifically inhibit gram-negative bacteria by being combined with lipid A, the effect of resisting gram-negative bacteria infection is achieved, the polypeptide has low hemolysis or no hemolysis, and the hemolysis rate is 0.4% when the concentration is 128 micrograms per milliliter. Aiming at escherichia coli, the FIC index value of the polypeptide and berberine combined drug is 0.34, and the polypeptide and berberine combined drug has a synergistic antibacterial effect and is free of hemolysis and low in cytotoxicity during combined drug use. The polypeptide or the pharmaceutical salt and the derivative thereof can be used as a novel bacteriostatic preparation, especially can be used in combination with berberine, and can be applied to development of bacteriostatic drugs.

Description

technical field [0001] The invention relates to an Escherichia coli lipid A binding motif PCK, a preparation method and application thereof. Background technique [0002] Livestock and poultry diarrhea diseases caused by Gram-negative pathogenic bacteria have seriously affected the health of the body and brought great harm to the livestock and poultry breeding industry. Among them, pathogenic Escherichia coli is one of the main pathogenic bacteria in human and animal intestines, which can cause diarrheal diseases. The long-term irrational use of antibiotics has led to the emergence of high resistance, multi-resistance, pan-drug resistance and the formation of superinfection in bacteria. So far, there is no satisfactory and effective treatment drug, and it is urgent to develop new green alternative antibiotic products. [0003] The E. coli cell membrane is composed of an inner membrane (i.e., the cytoplasmic membrane) and an outer membrane. The phospholipid bilayer forms the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/245A61K38/16A61K45/06A61K31/4375A61P31/04
CPCC07K14/245A61K38/164A61K45/06A61K31/4375A61P31/04A61K38/00Y02A50/30Y02P20/55
Inventor 王秀敏戴小枫李秀梅刘魏魏
Owner FEED RESEARCH INSTITUTE CHINESE ACADEMY OF AGRICULTURAL SCIENCES
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