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Mutant strain of high-yield abamectin B component and screening method thereof

A technology of abamectin and mutant strains, which is applied in the fields of bioengineering and enzyme engineering, and can solve the problems of high risk, restricted use, and low yield of abamectin mutant strains.

Inactive Publication Date: 2015-04-08
CHANGSHU INSTITUTE OF TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nitrosoguanidine (NTG) is known as a "super mutagen", but it is also a carcinogen or highly toxic drug, and its operation is dangerous, which greatly limits its use
In addition, in several reported patents, the positive mutation rate of the strains was not seen, and the yield of the obtained mutant strains of abamectin was not high

Method used

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  • Mutant strain of high-yield abamectin B component and screening method thereof
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  • Mutant strain of high-yield abamectin B component and screening method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] ARTP mutagenesis and microwave mutagenesis took turns twice, the results of Streptomyces avermitilis mutagenesis:

[0044] The specific implementation is as follows:

[0045] First carry out ARTP mutagenesis, the specific steps and technical parameters are as follows:

[0046] (1) Pick the mature spores on the slope of the starting strain of Streptomyces avermitilis (No. AVE-110) into a test tube equipped with an appropriate amount of normal saline, oscillate and disperse the suspension, filter it with absorbent cotton and adjust the spore concentration to 10 6 -10 7 pieces / ml.

[0047] (2) Preparation of sample slides: under aseptic conditions, the spore liquid prepared above was dropped onto the sterile slides in an amount of 10 μl.

[0048] (3) Air-drying of the sample slides: under sterile conditions, place the liquid slides prepared in the above step (2) under cold wind (10-20 degrees) to air-dry.

[0049] (4) Setting parameters: power supply 100W, radio freque...

Embodiment 2

[0075] Microwave mutagenesis and ARTP mutagenesis took turns twice.

[0076] The specific implementation is as follows:

[0077] First carry out microwave mutagenesis, the specific steps and technical parameters are as follows:

[0078] A microwave mutagenesis:

[0079] (1) Pick the mature spores on the slope of the starting strain of Streptomyces avermitilis (No. AVE-110) into a test tube equipped with an appropriate amount of normal saline, oscillate and disperse the suspension, filter it with absorbent cotton and adjust the spore concentration to 10 6 -10 7 pieces / ml.

[0080] (2) Draw the obtained spore suspension on the aseptic operating table, inject it into a flat sterile plate with a flat bottom, the suspension volume of each plate is 10mL, adjust the power of the microwave oven to 700W, the pulse frequency is 2450MHz, and the processing time is 30s , irradiate the spore suspension, and then take out 0.1mL of the bacterial suspension from each plate and dilute it a...

Embodiment 3

[0102] Follow the sequence of ARTP mutagenesis - ARTP mutagenesis - microwave mutagenesis - microwave mutagenesis:

[0103] First carry out ARTP mutagenesis, the specific steps and technical parameters are as follows:

[0104] (1) Pick the mature spores on the slope of the starting strain of Streptomyces avermitilis (No. AVE-110) into a test tube equipped with an appropriate amount of normal saline, oscillate and disperse the suspension, filter it with absorbent cotton and adjust the spore concentration to 10 6 -10 7 pieces / ml.

[0105] (2) Preparation of sample slides: under aseptic conditions, the spore liquid prepared above was dropped onto the sterile slides in an amount of 10 μl.

[0106] (3) Air-drying of sample slides: under aseptic conditions, place the liquid slides prepared in (2) above under cold wind (10-20 degrees) to air-dry.

[0107] (4) Setting parameters: power supply 100W, radio frequency power 13W, working distance 2mm, plasma temperature 28°C, airflow 10...

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Abstract

The invention discloses a mutant strain of a high-yield abamectin B component, which is classified and named as streptomyces avermitilis. The strain is collected in China General Microbiological Culture Collection Center (CGMCC) on November 6, 2014, and the collection number is CGMCC No.9950. The invention also discloses a screening method of the mutant strain of the high-yield abamectin B component. The invention also discloses a production method of the abamectin B component. The invention is designed for solving the technical problem of providing a simple, efficient and safe mutation method of a high-yield strain of an abamectin B component, a strain of the abamectin B component is also obtained according to the invention, the positive mutation rate of a high-yield abamectin strain is over 40%, and the fermentation yield of the abamectin B component is over 8000mu g / ml.

Description

technical field [0001] The invention belongs to the technical fields of bioengineering and enzyme engineering, and in particular relates to a mutant strain with high production of abamectin B component and a screening method thereof. Background technique [0002] Avermectins (Avermectins) are a group of structurally similar sixteen-membered macrolide zidon fructose disaccharide derivatives produced by Streptomyces avermitilis, which were obtained from Shizuoka, Japan by Kitasato Research Institute in Japan in 1975. isolated from a soil sample in the county. Abamectin has broad-spectrum, high-efficiency activity against various nematodes and human parasites. Abamectin insecticidal and anti-parasitic mechanism is relatively unique, and has no cross-resistance with other anti-parasitic drugs, low toxicity, no residue, etc., so abamectin is the most effective insecticide found so far (including Killing mites) one. Abamectin has eight components (4 components A and 4 component...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N15/01C12N13/00C12R1/465
CPCC12N1/20C12N13/00C12N15/01C12P19/623C12N1/205C12R2001/465
Inventor 梁剑光陈中兵朱益波王立梅齐斌黄海涛
Owner CHANGSHU INSTITUTE OF TECHNOLOGY
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