A Rapid Screening Method for Antibacterially Active Filamentous Fungi

A technology of filamentous fungi and antibacterial activity, applied in the direction of fungi, etc., can solve the problems of long cycle, high cost, high labor intensity, etc., and achieve the effect of efficient screening, simple operation, and increased screening probability

Active Publication Date: 2017-06-27
HUAIHAI INST OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method has high cost, long period and high labor intensity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, a method for rapid screening of antibacterial active filamentous fungi,

[0018] (1) Sample collection: collect seawater and soil samples with sterile sampling bottles, send them to the laboratory quickly, and store them at 4°C;

[0019] (2) Medium preparation: Nutrient agar medium: 3g beef extract, 10g peptone, 5g NaCl, 1000ml distilled water, pH7.2; PDA medium: 200g potato, 20g glucose, 1000ml distilled water, natural pH; if preparing solid culture Add agar 20g / L to the base plate; for the isolation of filamentous fungi, after the PDA medium is sterilized and the temperature drops to 60°C, add filter-sterilized streptomycin and ampicillin to a final concentration of 100 mg / ml and 50 mg / ml, mixed well and poured onto the plate; if it is a marine sample, use seawater instead of distilled water during the preparation process in the above culture;

[0020] (3) Preparation of intercepting tips: take a 1ml pipette tip, and cut off 12mm from the wide mouth end...

Embodiment 2

[0026] Example 2, step (5) of a method for rapidly screening antibacterial activity filamentous fungi described in Example 1, by intercepting the tip Escherichia coli Co-cultivate with filamentous fungi to be screened, and put them on the indicator bacteria plate, measure the diameter of the inhibition zone after the indicator bacteria culture, and quickly screen the antibacterial active strains according to the presence or absence and size of the inhibition zone.

Embodiment 3

[0027] Embodiment 3, comparative experiment:

[0028]1. Screening of prior art methods. Seawater and sea mud were collected from Liandao, Lianyungang sea area, and soil and freshwater samples were collected from Huaguo Mountain. Filamentous fungi were isolated using PDA medium by dilution coating method. Seawater was used instead of distilled water in the filamentous fungus isolation medium in marine samples. The purified single colony was inoculated on a PDA slant and stored at 4°C. A total of 122 strains of filamentous fungi were isolated. Different strains were inoculated into PDA liquid medium, cultured at 30°C and 160rpm for 2-3 days, and the fermentation broth was filtered with double-layer filter paper, and the supernatant was passed through 0.22 µ After the microporous filter membrane is the sample to be tested, the antibacterial activity is determined by the tube-and-disk method. Add 200 to each Oxford mu L fermentation samples, using Staphylococcus aureus , ...

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PUM

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Abstract

The invention relates to a method for rapidly screening filamentous fungi with antibacterial activity. The method includes (1) sample collection; (2) medium preparation; (3) preparation of intercepting tips; (4) preparation of co-cultured strains; (5) isolation and co-cultivation of filamentous fungi; (6) antibacterial activity Strain screening. The method of the present invention rapidly screens antibacterially active filamentous fungi by means of co-cultivation. Compared with purebred culture for screening antibacterially active filamentous fungi, the screening probability of novel antibacterially active bacterial strains and antibacterially active substances can be increased, and the whole screening process is fast, The operation is simple, and the filamentous fungi with antibacterial activity can be quickly, intuitively and efficiently screened, which provides great convenience for the screening research of filamentous fungi with antibacterial activity.

Description

technical field [0001] The invention relates to a rapid separation method of microorganisms, more specifically a method for rapid screening of filamentous fungi with antibacterial activity. Background technique [0002] The long-term and extensive use of antibiotics has led to an increase in the types of drug-resistant pathogenic bacteria, and even the emergence of "super bacteria", and the emergence of new infectious diseases, making the research and development of new antibacterial substances imminent (Li Yuezhong, Chen Qi. Marine Microbial Resources and Studies on the production of bioactive metabolites, Advances in Bioengineering, 2000, 20 (5): 28-31). For a long time, actinomycetes have been considered to be the most important source of active secondary metabolites, but recent studies have shown that the potential of filamentous fungi to produce active secondary metabolites is also very large, so the screening of filamentous fungal active secondary metabolites Become a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14
Inventor 刘姝房耀维吕明生王淑军焦豫良
Owner HUAIHAI INST OF TECH
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