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Abamectin strain and screening method thereof

A screening method, the technology of abamectin, applied in the directions of microorganism-based methods, methods of using microorganisms, biochemical equipment and methods, etc., can solve the problem of cumbersome cultivation process, low titer of cultivation results, and reduction of tap water and plates for fermentation. It can reduce the discharge volume and reduce the production and operation cost.

Active Publication Date: 2019-06-11
内蒙古新威远生物化工有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the culture result titer of this invention is low, and the culture process is loaded down with trivial details, and technology needs to be further simplified
[0007] At present, there is no abamectin strain in my country, and its screening method is safe and simple, and can be applied to and utilize residual nutrients and organic acids in waste water after mutagenesis and domestication of plate and frame waste water without affecting the release of fermentation tanks. Potency; can also reduce the discharge of tap water for fermentation and plate and frame waste water

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  • Abamectin strain and screening method thereof

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preparation example Construction

[0058] 4.1 Sample preparation

[0059] 4.1.1 Defoaming: Put the fermented liquid in a 100ml centrifuge cup, centrifuge at 3500 rpm for 1 minute in a centrifuge, take it out and stir well with a glass rod.

[0060] 4.1.2 Preparation: Accurately absorb 1 ml of homogeneous fermentation broth after defoaming into a 10 ml volumetric flask, add 5 ml of methanol, ultrasonicate for 15-20 min, take it out and place it at room temperature, dissolve and dilute to the mark with methanol, and shake well. Stand still, take the supernatant and filter it with a 0.45um microporous membrane for subsequent use.

[0061] 5. Preparation of standard sample: Accurately weigh 0.04 g of avermectin standard product, accurate to 0.0002 g, dissolve it in a 50 ml volumetric flask, dissolve it with methanol and set the volume to the mark, and shake well. Transfer 1 ml to a 10 ml volumetric flask, dissolve with methanol and bring to the mark.

[0062] 6 Under the above conditions, after the instrument is ...

Embodiment 1

[0078] (1) Preparation of germinated spore suspension: take 1 cm 2 Place the slant in 20ml of sterile water to fully disperse the spores into the water to obtain a spore suspension. Put the triangular flask with the spore suspension on a shaker and culture it at 100r / min at 28°C for 1.5h to germinate spore suspension;

[0079] (2) Combined mutagenesis of the spore suspension of germination: the spore suspension of 10ml germination is added to a mass concentration of 0.3% lithium chloride solution, then placed under a UV lamp at 30 cm and irradiated for 20s, and the power of the UV lamp is 30W. and stir, then add 0.5ml of NTG with a concentration of 5mg / ml into the bacteria solution treated with ultraviolet light-lithium chloride, mix evenly and immediately place it in a water bath at 28°C for 30min, then add 9ml of normal saline and shake well to obtain mutagenic bacteria solution;

[0080] (3) Plate and slant culture: draw 0.5ml mutagenic bacteria solution, dilute 10 5 Aft...

Embodiment 2

[0090] (1) Preparation of germinated spore suspension: take 1 cm2 Place the slant in 20ml of sterile water to fully disperse the spores into the water to obtain a spore suspension. Put the triangular flask with the spore suspension on a shaker and culture it for 2 hours at 100r / min at a temperature of 28.5°C to obtain germinated spore suspension;

[0091] (2) Combined mutagenesis of germinated spore suspension: 10ml of germinated spore suspension was added to a mass concentration of 0.5% lithium chloride solution, then placed under a UV lamp at 35 cm and irradiated for 80s, and the power of the UV lamp was 30W. and stir, then add 0.5ml of NTG with a concentration of 5mg / ml into the bacteria solution treated with ultraviolet light-lithium chloride, mix well and immediately place it in a water bath at 28.5°C for 30min, then add 9ml of normal saline and shake well. Obtain the mutagenic bacteria solution;

[0092] (3) Plate and slant culture: draw 0.5ml of the bacterial solution ...

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Abstract

The invention discloses an abamectin strain and a screening method thereof. The strain has the Latin name Streptomyces avermitilis in classification nomenclature, is preserved in China General Microbiological Culture Collection Center; and the preservation number is CGMCC No.17157, and the preservation date is January 10, 2019. The screening method comprises the following steps: preparing spore suspension; carrying out compound mutation on the spore suspension in a lithium chloride solution, then carrying out ultraviolet lamp irradiation mutation, then adding NTG into a bacterium solution treated by ultraviolet light-lithium chloride, and terminating the reaction; carrying out plate and slant culture; carrying out shaking culture and screening; and finally obtaining the strain. The strainis applicable, utilizes residual nutrient substances and organic acids in wastewater, without influencing a tank discharge titer of a fermentation tank, and can reduce the discharge amount of tap water for fermentation and plate frame wastewater.

Description

technical field [0001] The invention belongs to the field of microbial fermentation, and in particular relates to an abamectin strain and a screening method thereof. Background technique [0002] Abamectin, also known as avermectin, is a group of high-efficiency, low-toxicity, broad-spectrum macrolide agricultural antibiotics with a sixteen-membered ring structure, which has extremely strong killing activity against internal parasites. Abamectin is produced by Streptomycesavermitilis, which is a strain of Streptomyces isolated from soil without fungal or bacterial activity. Its secondary metabolites are produced by biological liquid culture and consist of 8 components. Composition, respectively named Ala, Alb, A2a, A2b, B1a, B2a, B1b, B2b, the basic structure of these compounds is a 16-membered macrocyclic lactone. Among the eight components, component B1 has the most significant and effective biological activity, and has good application value and broad market prospects in...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/36C12N13/00C12N15/01C12R1/465
Inventor 高鹤永李军华郑智新刘文彬
Owner 内蒙古新威远生物化工有限公司
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