Enramycin high-yield bacterial strain and screening method thereof

A technology of enramycin and high-yielding strains, applied in the directions of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of long fermentation period, unfavorable large-scale production and promotion of enramycin, low yield and the like, To achieve the effect of simple method, reduced breeding cost and good reproducibility

Inactive Publication Date: 2019-03-29
GUANGDONG RONGDA BIOENG CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Enramycin is mainly produced by the fermentation of the antifungal Streptomyces fungicidicus. At present, this strain generally has the characteristics of long fermentation cycle and low yield, which is not conducive to the large-scale production and further promotion of enramycin

Method used

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  • Enramycin high-yield bacterial strain and screening method thereof
  • Enramycin high-yield bacterial strain and screening method thereof
  • Enramycin high-yield bacterial strain and screening method thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0041] The preparation method of the spore suspension includes but is not limited to: In the ultra-clean bench, the antifungal Streptomyces slant that has been cultivated for 8 days is washed with 10ml of 5% sterile glycerin water and placed in a triangular flask with glass beads, and shaken at 200rpm for 20min to make the spores Fully disperse and filter with cotton to obtain spore suspension. The spore suspension is used for mutagenesis. After the spores are prepared into single cells, they not only fully contact with the nutrient solution, but also make each single cell exposed to the mutagenic conditions, and also make the cells unaffected, which is convenient for the experiment. .

[0042] In a preferred embodiment, the screening is followed by microbial identification followed by high performance liquid chromatography.

[0043] Using the microbial identification method to screen the mutagenic strains first, and high-performance liquid chromatography to screen again, you...

Embodiment 1

[0058] The preparation of embodiment 1 spore suspension

[0059] The slant medium includes: tryptone 1g / L, skipjack extract 0.8g / L, yeast extract 0.8g / L, maltose 8g / L, agar 30g / L, pH 7.0±0.3. Cultured at 28°C for 8 days.

[0060] In an ultra-clean bench, wash the QH105 slant cultivated for 8 days with 10ml of 5% sterile glycerin water, transfer it to a conical flask with glass beads, vibrate at 200rpm for 20min to fully disperse the spores, filter with cotton to obtain a spore suspension, and set aside.

Embodiment 2

[0061] Embodiment 2 Atmospheric room temperature plasma mutagenesis

[0062] The plane medium includes: tryptone 1g / L, skipjack extract 0.8g / L, yeast extract 0.8g / L, maltose 8g / L, agar 30g / L, pH 7.0±0.3.

[0063] In the ultra-clean bench, take 10 μl of the QH105 spore suspension in Example 1 on the surface of the slide, and use sterile tweezers to move the slide coated with the bacterial solution to the ARTP instrument for mutagenesis treatment, with a power of 100W and a gas flow rate of 10L / min Irradiate for 0s, 10s, 20s, 30s, 40s, 50s, 60s respectively under the conditions, dilute the bacterial solution on the slide, take 100μl and spread it on the plate. Incubate at 28°C for 8 days and calculate the lethal rate. The results are shown in Table 1:

[0064] The lethality of table 1QH105 different processing time

[0065]

[0066] It can be seen from the table that atmospheric room temperature plasma mutagenesis has a strong lethality to bacterial strain QH105. When the ...

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Abstract

The invention relates to the field of fermenting strains, and particularly provides an enramycin high-yield bacterial strain and a screening method thereof. A breeding method of the enramycin high-yield bacterial strain comprises the steps of adopting a constant-pressure room temperature plasma method for inducing fungus-resisting streptomycete, wherein the constant-pressure room temperature plasma method comprises the steps of performing irradiation under the condition that the power is 90-110W and the gas flow is 9-11L/min for 35-45min, and finally, performing screening, so that the enramycin high-yield bacterial strain is obtained. According to the method, a constant-pressure room temperature plasma technology is used as the inducing method, and in accordance with streptomyces antifungus, design is performed, so that the test condition suitable for the streptomyces antifungus is preferably selected. Under the test condition, that the mutation rate of the streptomyces antifungus is high, and the orthomutation rate is high. The method is good in repeatability. The enramycin high-yield bacterial strain can be obtained, wherein the yield can be at least increased by three times. Theheredity stability is good. The method is simple and convenient, easy to operate, time-saving, and effortless, and the breeding cost is greatly reduced.

Description

technical field [0001] The invention relates to the field of fermentation strains, in particular to an enramycin high-yield strain and a screening method thereof. Background technique [0002] Enramycin (Enramycin), also known as Enlaimycin, Enramycin, Persistomycin, Enramycin, was first discovered as a polypeptide secreted by the actinomyces Streptomyces fungicidicus No.B5477 isolated from soil samples Specific antibiotics for animals. Enleimycin is an organic base composed of 17 amino acid molecules of 13 different types and fatty acid molecules. The amino acid molecules form a cyclic polypeptide structure, and the fatty acid is located at the end of the polypeptide structure. The main components of enramycin are enramycin A and B, which are used in the form of hydrochloride. Enramycin has a good antibacterial effect on Gram-positive bacteria, especially against harmful Clostridium difficile in the intestine. It has broad-spectrum, low toxicity, low residue, and is not e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12N13/00C12N1/20C12R1/465
CPCC12N13/00C12N15/01C12N1/205C12R2001/465
Inventor 张文叶彩云孟葵开唐谢芳蒋顺进郑雪媚
Owner GUANGDONG RONGDA BIOENG CO LTD
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