Method for quickly screening anti-Aspergillus-niger active substances

An active substance, a technology of Aspergillus niger, which is applied in the field of biology and achieves the effects of simple method, reliable experimental results and obvious color change

Inactive Publication Date: 2013-01-02
THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the abundance of marine microbial resources, it is difficult to sc

Method used

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  • Method for quickly screening anti-Aspergillus-niger active substances
  • Method for quickly screening anti-Aspergillus-niger active substances

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: the preparation of marine microbial fermentation liquid extract sample;

[0027] Experimental materials: 87 samples of three batches of extracts, respectively from: Citricoccus alkalitolerans, preserved in China Marine Microorganism Culture Collection and Management Center, strain preservation number MCCC 1A00280; platform resource number: 1535C0001000000149; collected from Pacific Ocean; biohazard level of the bacteria: category four; non-fatal bacteria; Gram-positive aerobic bacteria, catalase positive, oxidase negative; colonies are yellow spherical or approximately spherical. Aspergillus sydowii, preserved in the China Marine Microorganism Culture Collection and Management Center, strain preservation number MCCC 3A00240; platform resource number: 1535C0001000011892; collected from the Western Pacific Ocean; the colony texture is velvety to flocculent, dense; the color is Gray-blue to dark blue, slightly lighter at the beginning, gradually darker after...

Embodiment 2

[0044] Example 2 : Screening of Active Samples

[0045] sample:

[0046] Aspergillus niger spore suspension: Pick Aspergillus niger spores cultured on a fresh PDA plate for 2-3 days and shake them in 0.5% Tween-20 saline; then count them under a microscope with a hemocytometer. Calculate the various concentrations of the Aspergillus niger spore suspension by counting under the microscope, and then use RPMI-1640 medium to dilute the Aspergillus niger spore suspension to 2.5×10 4 spores / ml.

[0047] Resazurin solution: Prepare 200ug / ml resazurin solution with sterile water

[0048] experiment procedure:

[0049] 1. Mix 176 microliters of 2.5×10 4 spores / ml Aspergillus niger spore suspension, 20 microliters of 200ug / ml resazurin solution and 4 microliters of fermentation broth extract (87 extract samples obtained in the examples, added separately) were added to the 96 microwell plate at the same time ; Set up two negative controls and two positive controls in a 96 microw...

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Abstract

The invention relates to a method for quickly screening anti-Aspergillus-niger active substances, which comprises the following steps: adding a sample to be screened, a resazurin and Aspergillus niger spore bacterial suspension, into a 96 microporous plate, and culturing in a 37 DEG C constant-temperature culture box for some time to obtain the result, wherein a negative control and a positive control are arranged, and the preferable culture time is 12-36 hours; and screening to obtain the anti-Aspergillus-niger active substances according to the result. When the color of the microporous plate is blue or purple, the fluorescent value measured by the microplate reader is lower than the 15000; and when the inhibition rate is higher than 40%, the detected sample is the anti-Aspergillus-niger active substances. The method is simple, quick and accurate.

Description

Technical field [0001] The present invention involves a biological method, which specializes in a method of rapidly screening active substances that anti -black mold. Background technique [0002] The natural product of microorganisms has always played an important role in the treatment of human diseases.In the past 20 years, with the widespread use of traditional antibiotics, the resistance of pathogenic bacteria has increased rapidly, and the new active compounds that have been found in traditional soil microorganisms have a significant decline.In the process of source, marine microorganisms have attracted widespread attention.The Lu Sheng -microorganisms of the main precursor sources of marine microorganisms and traditional pharmaceutical industries have a significant growth environment. They can adapt to the harsh living environment through novel metabolic methods, which may produce new secondary metabolites that are different from Lu Shengwei.And marine microorganisms are ri...

Claims

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Application Information

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IPC IPC(8): C12Q1/18C12R1/685
Inventor 陈建明胡彬王蔚陈卓李芊杨慧
Owner THIRD INST OF OCEANOGRAPHY STATE OCEANIC ADMINISTATION
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