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Antiviral actinomyces HA10206 and application thereof

A technology of actinomycetes and antiviral activity, applied in the field of microorganisms, to achieve a strong inhibitory effect

Inactive Publication Date: 2012-04-04
INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are relatively few research reports on antiviral actinomycetes, and screening antiviral antibiotics from actinomycetes is still one of the current research hotspots

Method used

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  • Antiviral actinomyces HA10206 and application thereof
  • Antiviral actinomyces HA10206 and application thereof
  • Antiviral actinomyces HA10206 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Isolation of strain HA10206 CGMCC No.4861

[0025] After washing the collected sponge sample with sterile seawater for 3-5 times, cut the sample into small pieces with a sterile scalpel, and grind it in a sterile mortar for 5-10 minutes to make it into a uniform slurry. Make 10-fold serial dilutions on the samples, each take 100 μL 10 -1 、10 -2 、10 -3 、10 -4 The diluted bacterial solution was spread on Gao's No. 1 medium, and cultured upside down at 28°C. Colonies with different shapes and colors were picked in due course for streaking and purification. For the purified actinomycetes, according to the morphological characteristics of the colonies, the same strains were excluded, transferred to a slant for storage at 4°C, numbered, and reserved for strain screening.

Embodiment 2

[0026] Embodiment 2: the cultivation of bacterial strain HA10206 CGMCC No.4861

[0027] (1) Incline culture: culture for 6 days. Medium: soluble starch 10g, potassium nitrate 1g, dipotassium hydrogen phosphate 0.5g, magnesium sulfate 0.5g, ferrous sulfate 0.01g, prepared with old sea water, pH 7.2.

[0028] (2) Seed culture: Pick an activated single colony from the slope and insert it into the seed medium, and cultivate it at 28° C. and 180 r / min for 48 hours. Medium: glucose 1%, soybean powder 1%, yeast extract 1%, soluble starch 0.5%, dipotassium hydrogen phosphate 0.025%,

[0029] Prepared from natural aged seawater (fresh seawater after standing for a week), pH7.2.

[0030] (3) Fermentation culture: 10% of the inoculum was inserted into the seed culture solution, and cultured in shake flasks at 28° C. and 180 r / min for 6 days. Medium: 0.5% glucose, 1.5% soluble starch, 1% soybean powder, 0.5% yeast extract, 0.025% dipotassium hydrogen phosphate, prepared with 75% natura...

Embodiment 3

[0031] Example 3: Anti-H1N1 virus activity detection of bacterial strain HA10206 CGMCC No.4861

[0032] Fermentation was carried out according to the method of Example 2, and the fermentation product fermented for 6 days was collected, centrifuged at 10,000 r / min for 10 min, and the supernatant was filtered with a 0.22 μm sterile filter membrane for subsequent use.

[0033] The fermentation broth was tested for anti-H1N1 virus activity according to the following CPE+MTT combination method:

[0034]CPE+MTT combination method: culture MDCK canine kidney cells, after the cells grow into a single layer, inoculate 50 μL of H1N1 virus solution, 37 ° C, 5% CO 2 Incubate for 2 hours, discard the supernatant, add 50 μL of 20-fold diluted fermentation broth, incubate at 37°C, 5% CO2, and observe the test cytopathic effect (CPE) under an inverted microscope. The recording method of CPE is as follows: no CPE is "-"; 25% of cells with pathological changes is "+"; 25% to 50% of cells with ...

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Abstract

The invention discloses an actinomyces HA10206 strain for generating substances with antiviral activity, and the preservation number is CGMCC No.4861, and the invention belongs to the field of microbiological technology. The present invention has the advantages that: the actinomyces HA10206 strain is a new species of streptomyces, and the fermentation liquor of the product has obvious inhibition effects on H1N1 and H2N2, H3N2, H5N1 viruses of the same series, and certain inhibition effects on HIV and HBV, thereby the actinomyces HA10206 has good medicament development prospects.

Description

technical field [0001] The invention relates to an antiviral streptomyces (Streptomyces) actinomycete HA10206, which belongs to the technical field of microorganisms. Background technique [0002] Viral diseases are a serious threat to human health, and currently there is no specific and effective means of prevention and treatment for viral infections. Therefore, the study of specific antiviral drugs has very important practical significance. Influenza virus caused by influenza virus is an acute viral respiratory infectious disease with fast transmission speed, which is one of the main causes of human death, especially H1N1 influenza virus has not been effectively controlled all over the world, which seriously threatens human beings. life safety. It is still an urgent and important task to find new and effective antiviral drugs, especially those that are highly selective to viruses and have no toxic side effects on host cells. [0003] In recent years, the development of m...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/74A61P31/12A61P31/16A61P31/18A61P31/20C12R1/465
Inventor 黄惠琴朱军吴春燕鲍时翔孙前光刘敏方哲
Owner INST OF TROPICAL BIOSCI & BIOTECH CHINESE ACADEMY OF TROPICAL AGRI SCI
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