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New antimicrobial peptides, their variants and uses

一种抗微生物肽、抗微生物的技术，应用在杀死微生物或抑制微生物生长领域，能够解决失去生存力、无活性、不适于功能研究等问题，达到易于合成、生产成本小的效果

Active Publication Date: 2018-03-27
CHAIN ANTIMICROBIALS OY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Methods such as denaturing gradient gel electrophoresis (DGGE), restriction fragment length polymorphism (RFLP) or cloning and direct sequencing are used to analyze the diversity of non-culturable fungal colonies in plants, but such methods are not suitable for functional studies
A common problem with endoparasites is that they produce metabolites only for a certain period of time in vitro and then become inactive, losing viability or the ability to produce bioactive secondary metabolites

Method used

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  • New antimicrobial peptides, their variants and uses
  • New antimicrobial peptides, their variants and uses
  • New antimicrobial peptides, their variants and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0138] Example 1: Isolation of Total Plant DNA

[0139] Fresh young leaves of Empetrum nigrum L. were surface sterilized with 70% ethanol (v / v) for 1 min and placed in sodium hypochlorite (3.5% v / v) for 5 min and used to isolate genomic DNA ( et al. 2001).

Embodiment 2

[0140] Example 2: Metagenomic library construction and screening for antibacterial activity

[0141] Plant DNA isolated from crowbar is dissolved in sterile water to 0.1 μg μl -1 concentration. DNA was then subjected to preparative pulsed field gel electrophoresis in the CHEF-DRII (Bio-Rad) system. The electrophoresis conditions (pulse interval and duration) were: respectively, N / S-60s and E / W–60s for 6h; N / S-90s and E / W–90s for 6h; N / S-99s and E / W - 99s for 6h; voltage of 6V / cm, fixed angle of 120° and use of 0.15×Tris-borate-EDTA (TBE) buffer. During electrophoresis, the temperature was maintained at 10 °C. After electrophoresis, bands from each side of the gel were excised and stained with ethidium bromide to visualize DNA. High molecular weight DNA was then excised from the remaining unstained portion of the gel and electroeluted at 100 V for 1 hour in a dialysis bag containing 0.5 x TBE. Amplification was performed with primers specific for fungi and bacteria as prev...

Embodiment 3

[0143] Example 3: Strains, plasmids and growth conditions

[0144] EPI-300 TM -T1 R Phage T1-resistant E. coli cultures were grown at 37°C on Luria-Bertani (LB) agar or in LB broth supplemented with appropriate antibiotics + 10 mM MgSO 4 grow in. The following antibiotic concentrations were used for E. coli strains: Chloramphenicol 12.5 μg ml -1 and ampicillin 100 μg ml -1 . Plasmid pCC1FOS carrying two origins of replication, a single-copy origin (ori2) and an inducible high-copy origin (oriV) TM (Epicentre, Madison, USA) were used to construct a metagenomic library from Crowberry endoparasites and for subcloning the genes conferring the antibacterial activity. The pET11-c vector was used to express the genes responsible for the antibacterial activity in the host strain E. coli BL21(DE3).

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PUM

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Abstract

The present invention relates to novel antimicrobial peptide and variants thereof. The invention further relates to method of killing or inhibiting growth of microbes and use of the peptide here disclosed as a medicament, feed additive, preservative or surfactant.

Description

field of invention [0001] The present invention relates to antimicrobial peptides or variants thereof, to methods of killing or inhibiting the growth of microorganisms, and to the use of the peptides described herein as pharmaceuticals, feed additives, preservatives or surfactants. Background technique [0002] Among pharmaceutical compounds, antibiotics have had a huge impact on human life expectancy. However, the ability of microorganisms to develop resistance is almost unlimited. Multidrug resistance has become a very common and dangerous feature of many human pathogens, and resistance is spreading globally. [0003] Endoparasites are promising producers of a wide range of secondary metabolites with potential applications in the biomedical, pharmaceutical, and medical industries. Methodologies typically used to study the functional diversity of endoparasites are based on an emphasis on the isolation of rapidly growing strains and thus do not represent the full biodivers...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C07K7/06
CPCA61K38/00A61P31/04A61P31/10C07K14/415C07K14/4723Y02A50/30A01N47/44A23V2002/00A61L29/08A61L29/16A61L2300/404A23K20/147A23L33/18
Inventor 特赫斯威·梅索维沙坎特高达安娜·玛丽亚·皮尔蒂莱
Owner CHAIN ANTIMICROBIALS OY
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