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High-producing strain containing abamectin and screening method thereof

A technology of abamectin and high-yield strains, applied in the direction of microorganism-based methods, biochemical equipment and methods, bacteria, etc., can solve the problems of small screening amount, low fermentation capacity and low efficiency of strains, and achieve optimal culture medium and culture conditions, the effect of strain titer improvement

Active Publication Date: 2016-12-07
NORTH CHINA PHARMA GROUP AINO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is exactly to provide a kind of abamectin high-yielding bacterial strain, provide a kind of screening method of this bacterial strain at the same time, to solve abamectin production process bacterial classification fermentation ability is low and traditional bacterial strain screening method efficiency is low, screening amount is little The problem

Method used

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  • High-producing strain containing abamectin and screening method thereof
  • High-producing strain containing abamectin and screening method thereof
  • High-producing strain containing abamectin and screening method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Embodiment 1: the deep well plate fermentation of liquid culture medium of the present invention

[0023] 1. Preparation of liquid medium: dextrin 100g, yeast hydrolyzate 24.1g, malt extract 3.1g, beef extract 13.1g, cobalt chloride 0.002g, manganese sulfate 0.002g, sodium molybdate 0.002g, when preparing, Dextrin is gelatinized separately, then other raw materials are added, and finally drinking water is added to make the volume to 1000mL, and the pH value is 7.4-7.6. Distribute the prepared liquid medium into 24-well plates, cover the plates, place them in baskets, and sterilize at 121°C for 30 minutes for later use.

[0024] 2. Sow the spores of the strain AV-185S in a 24-well deep-well plate with 3 mL of liquid medium with a toothpick. After the inoculation is complete, cover the plate and fix the plate on the fixed plate of the shaker at a speed of 200 rpm. , and cultivated for 6 days at a temperature of 28° C. and a relative humidity of 70±5%.

[0025] 3. After ...

Embodiment 2

[0033] 1. Preparation of solid medium: Glucose 15g, beef extract 3g, asparagine 0.5g, potassium dihydrogen phosphate 0.5g, agar 19g, add drinking water to dissolve to 1000mL, pH value 7.2-7.5, sterilize at 121℃ 30min. When using, melt it first, then pour about 30mL into the double dish, and set it up for later use.

[0034] 2, the preparation of liquid medium: with embodiment 1.

[0035] 3. The isolation and verification culture of the starting bacterial strain: the starting bacterial strain used in the present invention is Streptomyces avermitilis (Streptomyces avermitilis) (number: 2015-45-38) from the laboratory of North China Pharmaceutical Group Ainuo Co., Ltd., and the mature spores of the bacterial strain are taken out Make a spore suspension and adjust the spore concentration to 10 6 -10 7 cells / mL, serially dilute the spore suspension to 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 、10 -6 、10 -7 , take 0.1mL of the dilutions of each concentration, spread on the solid dou...

Embodiment 3

[0043] Embodiment 3: Screening of high-yield strains

[0044] 1. Preparation of solid medium: glucose 15g, beef extract 3g, asparagine 0.5g, potassium dihydrogen phosphate 0.5g, agar 19g, add drinking water to dissolve to 1000mL, pH value 7.2-7.5, sterilize at 121℃ 30min. Prepare a 10wt% lithium chloride solution, filter and sterilize it with a sterile membrane before use. Mix the lithium chloride solution with the solid medium to prepare a solid medium with a final concentration of lithium chloride of 0.3 wt%. Melt it before using it, then pour about 30 mL into a double dish, and set it aside after solidification.

[0045] 2. The preparation of the liquid culture medium is the same as in Example 1.

[0046] 3. Take out the mature spores of the bacterial strain to make a spore suspension, and adjust the concentration of the spores to be 10 6 -10 7 individual / mL; place the spore suspension under a 15W ultraviolet lamp, and treat it in the dark at a distance of 30 cm for 25 ...

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Abstract

The invention discloses a high-producing strain containing abamectin, and also provides a screening method of the high-producing strain containing abamectin. The high-producing strain containing abamectin is (Streptomyces avermitilis) AV-185S, the preservation unit of the high-producing strain is the China general microbiological culture collection center, the preservation date of the high-producing strain is March 15, 2016. The screening method mainly comprises the steps of a, preparing a spore suspension; b, conducting ultraviolet-lithium chloride compound mutation treatment; c, preparing a streptomyces avermitilis liquid medium; d, screening the high-producing strain through liquid culture in a deep well plate; and e, detecting the strain fermentation potency. The average fermentation potency of the (Streptomyces avermitilis) AV-185S screened according to the screening method can be up to 6842.8 ug / mL.

Description

technical field [0001] The invention relates to a bacterial strain for producing antibiotics and a screening method thereof, in particular to an avermectin high-yielding bacterial strain and a screening method thereof. Background technique [0002] Avermectins (AVM), also known as avermectins, are a group of macrolide antibiotics fermented by Streptomyces avermitilis. Abamectin is widely used in the treatment of various domestic animal internal and external parasitic diseases because of its insecticidal, acaricidal and nematicidal effects. Abamectin includes 8 kinds of homologues with different structures, i.e. 4 main components ( Ala, A2a, B1a, and B2a) and 4 minor components (A1b, A2b, B1b, and B2b), among which, the B1 component has the highest activity, especially B1a has the strongest anthelmintic effect. [0003] The mechanism of action of abamectin is relatively special, and it is not easy to make pests develop resistance. It mainly acts on the gamma-aminobutyric aci...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N13/00C12N15/01C12R1/465
CPCC12N13/00C12N15/01C12N1/205C12R2001/465
Inventor 孙耀华刘书琴曲志西韩娜张勍韩瑞霞
Owner NORTH CHINA PHARMA GROUP AINO
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