Abamectin producing bacterium and preparation method thereof

A kind of abamectin, the technique of producing bacteria, applied in the field of abamectin producing bacteria and its preparation, can solve the problems such as the production level needs to be further improved, and achieve the improvement of abamectin B components, reducing components, and subculture stable effect

Active Publication Date: 2011-08-17
HEBEI XINGBAI AGRI SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN200410009639.2 discloses a method for utilizing ultraviolet rays and nitrosoguanidine mutagenesis screening to obtain abamectin high-yield strains; CN20061...

Method used

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  • Abamectin producing bacterium and preparation method thereof
  • Abamectin producing bacterium and preparation method thereof
  • Abamectin producing bacterium and preparation method thereof

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Experimental program
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Effect test

Embodiment 1

[0079] Taking the existing Streptomyces avermitilis (MF800-6 for short) producing abamectin as the initial starting strain, according to image 3 Shown flow process and following method steps carry out:

[0080] a. Ultraviolet compound lithium chloride mutagenesis treatment:

[0081] (1) Preparation of spore suspension: Take 15 ml of sterilized normal saline, pour it into the slant of a freshly cultivated test tube (the culture medium is medium No. 1), and gently infuse the MF800-6 The spores were scraped off to make a spore suspension, counted with a hemocytometer, and the spore concentration was adjusted to 10 6 individual / ml. Take 10ml of spore suspension in a φ90 sterile petri dish, add a magnetic rod, place it on a magnetic stirrer in a UV mutagenesis box, and place the prepared spore suspension at a place 20 cm away from a UV lamp with a wavelength of 253.7nm and a power of 30W. For mutagenesis, open the lid of the dish during irradiation, stir with magnetic force, st...

Embodiment 2

[0105] Embodiment 2 Shake flask fermentation method re-screens high-titer bacterial strains

[0106] a. Preparation of strains in shake flasks

[0107] Prepare the seed shake flask medium according to the ratio and configuration process of the No. 3 medium, and take 1 cm on the slant of the cultured high-titer strain 3 The left and right bacteria blocks were inoculated into the seed shaker flask aseptically, sealed with a cotton plug, wrapped with 2 layers of gauze, placed on a 200-220r / min shaker, and incubated at 28°C±0.5°C for 40-50 hours.

[0108] b. Preparation and inoculation of fermentation shake flasks

[0109] According to the ratio and configuration process of the No. 4 medium, prepare the fermentation shake flask medium, transfer the above-prepared seeds aseptically into each fermentation shake flask with 5-8% seed liquid, seal it with a cotton pad, and put it in 200-220r / min shaker, culture at 28°C±0.5°C for 10-11 days, put in the bottle and measure B1a titer acc...

Embodiment 3

[0110] The seed expansion cultivation of embodiment 3 seed jars

[0111] Preparation of spore suspension

[0112] Take 1-4 bottles of FT26-9 strain slant test tubes, add 50-100 ml of sterile distilled water, gently scrape off the slant spores with a sterile inoculation needle on the ultra-clean workbench, plug the inoculation needle, and use 12 layers of gauze and kraft paper Wrap it well and put it in the refrigerator at 4°C for later use.

[0113] b. Prepare the seed medium according to the ratio of No. 5 medium, adjust the pH to 7.0-7.5 with sodium hydroxide before sterilization, steam sterilize at 121°C and 0.1MPa for 30 minutes, and wait until the temperature drops to 28 At -30°C, the spores were inserted into the seed tank by the differential pressure method, sealed and cultivated. Culture conditions: aeration ratio 1:0.6-1.2, temperature 28±0.5°C, stirring speed 150-300rpm. After 40-60 hours of cultivation, the hyphae under the microscope are thick and branched, stain...

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Abstract

The invention discloses abamectin producing bacterium and a preparation method thereof. The collection name of the bacterium is FT26-9 and the collection number of the bacterium is CGMCC No.4341. The bacterium was collected on November 12th, 2010. The preparation method comprises: performing ultraviolet and lithium chloride combined mutation induction treatment of a starting strain; screening an induced-mutation strain; performing mutation induction by nitrosoguanidine and screening; performing mutation induction by diethyl sulfate and screening; performing mutation induction by hydroxylamine and screening; performing mutation induction by 5-bromouracil and ultraviolet and screening; and finally obtaining the FT26-9 strain which can greatly improve the abamectin B component content in a fermentation product and reduce the abamectin A component content in the fermentation product. When the bacterium is used in industrial production of abamectin, the fermentation unit is improved by about 40 percent, and the product cost is reduced by about 35 percent.

Description

technical field [0001] The invention relates to an abamectin-producing bacterium and a preparation method thereof, in particular to a variant strain of the abamectin-producing bacterium and a preparation method thereof. technical background [0002] Avermectin (avermectin, AVM) is a class of macrolide antibiotics produced by liquid fermentation of streptomyces avermitilis. It is a new pollution-free biological agricultural and veterinary drug with high efficiency and low toxicity It is environmentally friendly and has good control effects on related pests of vegetables, fruit trees, cotton, rice and other crops. It is currently the most promising biological insecticide and acaricide in the world. At present, the abamectin produced by Streptomyces avermitilis fermentation includes 8 components with similar structures (AVMA1aAVMA1b AVMA2a AVMA2b AVMB1a AVMB1b AVMB2a AVMB2b). Among them, the biological activity of component B is better than that of component A, especially the ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N15/01C12N13/00C12R1/465
Inventor 王琳慧范令涛高建辉
Owner HEBEI XINGBAI AGRI SCI & TECH CO LTD
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