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Method for rapidly screening bacterial biofilm inhibitor from plant

A bacterial biofilm and inhibitor technology, applied in the field of natural products, can solve the problem of time-consuming for a single plant, and achieve the effect of speeding up the screening speed, improving the screening efficiency, and convenient screening

Inactive Publication Date: 2017-05-31
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the many potential biofilm inhibitory components in natural plants, the existing method uses the method of extracting and purifying one by one, enriching and then testing the activity. This mode makes the research of a single plant require a lot of time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Preparation of plant components

[0023] Firstly, 30% ethanol was used to extract the burdock leaves to obtain the crude burdock leaf extract, and then the burdock leaf extract was purified by macroporous resin column chromatography to obtain two groups of water eluted fraction and 34% ethanol eluted fraction. point.

[0024] (2) Determination of the inhibitory activity of plant components on bacterial biofilm by crystal violet staining

[0025] On the ultra-clean workbench, respectively add Pseudomonas aeruginosa bacteria solution (logarithmic phase bacteria) and 34% ethanol elution fraction of burdock leaf to 24-hole PVC plate, and make 4 parallels, and set LB at the same time The blank control and the negative control group without burdock leaf components were cultured for 24 hours. After the cultivation, suck off the culture, wash away the planktonic bacteria with sterile PBS, add 1 mL of anhydrous methanol to each well to fix the biofilm for 15 min, dry the c...

Embodiment 2

[0039] (1) Preparation of plant components

[0040] The dried product of Houttuynia cordata was crushed, extracted with 30% ethanol for 12 hours, the obtained mixed solution was first centrifuged, and the filter residue was removed by suction filtration, and the extract was rotatively evaporated at 50° C., and then concentrated to obtain a concentrated solution.

[0041] The Houttuynia cordata concentrate was added to the treated HPD100 macroporous resin for column chromatography, 3 times the column volume of pure water and 30% ethanol solution were used for column chromatography to elute, and the eluate was collected. The 30% ethanol eluate was taken, concentrated by rotary evaporation, and then dried in a vacuum drying oven. The obtained sample was the Houttuynia cordata column chromatography fraction eluted with 30% ethanol, which was set aside.

[0042] (2) Determination of the inhibitory activity of plant components on bacterial biofilm by crystal violet staining

[0043...

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PUM

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Abstract

The invention relates to a method for rapidly screening bacterial biofilm inhibitor from plant. The method comprises the following steps: preparing a plant extract product or a plant column chromatography component, employing a crystal violet staining method for researching the plant extract product or the plant column chromatography component to inhibit the activity of a harmful bacterial biofilm; then employing a chromatogram-mass spectrometry technology for analyzing the component change before and after the plant component is effected on bacteria, performing multi-data statistics analysis, by combing with active verification experiment result, screening the main effective component for inhibiting the bacterial biofilm in the plant. The method can rapidly screen the active ingredients for inhibiting the bacterial biofilm in the plant, and provides a novel convenient approach for high-flux rapid screening of anti-biofilm activity compound in the natural plant.

Description

technical field [0001] The invention belongs to the field of natural products and relates to a method for screening bacterial biofilm inhibitors from plants. Background technique [0002] Biofilm is a relatively unique group lifestyle adopted by bacteria when growing on a solid surface, and is a highly organized multicellular structure. Biofilm bacteria constitute a self-protective mode that differs in physiology and behavior from planktonic bacteria. In the food industry, harmful bacterial biofilms are a persistent source of contamination that can cause food spoilage and easily cause public health problems. Harmful bacterial biofilms are the primary cause of various bacterial-related diseases and a major threat to human health. However, biofilm bacteria are 10-1000 times more resistant to antibiotics and bactericidal disinfectants than planktonic non-biofilm bacteria, and conventional cleaning and disinfection therapies are ineffective in controlling biofilms. The resear...

Claims

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Application Information

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IPC IPC(8): C12Q1/18
Inventor 娄在祥李雅芹王洪新于福浩李成
Owner JIANGNAN UNIV