Strain capable of generating avermectin B component and utilization thereof
A technology of avermectin and strains, applied in the direction of microorganism-based methods, bacteria, microorganisms, etc., can solve the problems of complex process, high cost, low extraction rate, etc.
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Embodiment 1
[0024] Example 1. Medium preparation
[0025] No. 1 medium (strain medium):
[0026] A. Medium ratio (by weight percentage):
[0027] Glucose 1.5%, DL-asparagine 0.05%, beef extract 0.3%, potassium dihydrogen phosphate 0.05%, agar powder 2.5%, distilled water to make up to 100%, pH (before sterilization) 7.4.
[0028] B. Preparation
[0029] Weigh the glucose first, put it in a large beaker, add a little water to dissolve it (distilled water), then pick up the beef extract with a spoon and put it in a small 100ml beaker, weigh it with an electronic balance, add a small amount of distilled water to melt it, and pour it into the dextrose In a large beaker, stir well, then put DL-asparagine and potassium dihydrogen phosphate into a small beaker, weigh it, add a little distilled water, heat it on an electric stove with asbestos tiles, and pour into the above solution , Dilute to the total volume used, stir evenly with a glass rod and adjust pH=7.4 (before sterilization).
[0030] Weig...
Embodiment 2
[0043] Example 2: Mutagenesis and screening of strains producing abamectin B component
[0044] 1. Mutagenesis treatment
[0045] 1.1. UV mutagenesis treatment
[0046] Taking the existing avermectin-producing Streptomyces avermitilis GY115 as the starting strain, the avermectin produced by this strain contains 8 components (see Figure 1). First, fresh slant spores of GY115 are used to prepare a single spore suspension according to a method known to those skilled in the art, and 5 ml is sucked and placed in a sterile petri dish with a diameter of 60 mm. The plate was placed 20 cm under an ultraviolet lamp with a wavelength of 253.7 nm and a power of 30 W, and the lid was opened and shaken on a 7921 magnetic stirrer at a speed of 1370 revolutions / min for 30 seconds. When the time is up, transfer to the separation plate of No. 1 medium containing an appropriate amount of antibiotics (gentamicin 0.0004% w / v or chloramphenicol 0.0008% w / v or streptomycin 0.001% w / v), and place Incubat...
Embodiment 3
[0062] Example 3. Rejuvenation of strains
[0063] Select the type II single colony of X-28, which is mouse gray, large and full, and straw hat type. In the sterile room, use the inoculation loop to scrape the spores and place them in the aseptic operating procedure. In a test tube of sterile water, add glass beads, shake, and filter with absorbent cotton. Dilute 0.5ml bacterial suspension according to the multiple dilution method (add 4.5ml sterile water) and then dilute it 5 times. Use a 1ml pipette to drop 2 drops of the bacterial suspension on the plate containing No. 1 medium, spread it evenly with a glass spatula, and incubate at 28°C for 11-14 days. The isolated morphology is rat-grey, large and full, and the single colony of straw hat type was subjected to secondary titer verification.
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