Application of yingluo mycin in preparing drug resisting plant pathogenic fungi
A technology of enromycin and antimycin, which is applied in the field of application of enluomycin in the preparation of anti-phytopathogenic fungal drugs, can solve the problems that the anti-fungal activity of the same compound has not been seen, and achieve significant anti-phytopathogenic fungi. The effect of fungal action
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Embodiment 1
[0031] : the fermentation of englomycin producing bacteria
[0032] (yeast extract 0.4%, malt extract 1%, glucose 0.4%, agar 1.2%, pH7.2) streptomyces strain Streptomyces albidoflavus I07A-0182 grown on the slant was inoculated in the seed medium (glucose 0.5%, yeast extract 0.5%) ,, peptone 0.5%, beef extract 0.5%, corn steep liquor 0.4%, starch 2%, soybean cake powder 1%, CaCO 3 0.4%, CoCl 2 0.002%, pH 7.2) 50ml / 250ml shake flask, cultured on a rotary shaker at 28°C for 36hrs, then transferred to a 1L / 5L fermentation medium (the formula is the same as the seed medium) 1L / 5L shake flask with a 5% inoculation amount , cultivated on a rotary shaker at 28° C. for 72 hours, and harvested a total of 40 L of fermentation broth.
Embodiment 2
[0033] : the extraction of Yingluomycin
[0034] After centrifugation (4000r / min, 20min) of 40 liters of fermentation broth of Streptomyces albidoflavus I07A-0182, the supernatant was extracted with an equal volume of ethyl acetate, and the organic phase was washed with anhydrous Na 2 SO 4 Dehydration and concentration under reduced pressure gave about 20 g of an oily tan crude extract.
Embodiment 3
[0035] : the separation of Enromycin
[0036] The above crude extract was dry-loaded on a forward silica gel chromatography column (250g, 100-200 mesh, column volume 3x40cm), using cyclohexane-ethyl acetate 19:1, 4:1, 2:1, 1:1 500 ml each, graded elution, activity tracking showed that cyclohexane-ethyl acetate 2:1 part showed strong activity against rice blast fungus.
[0037] The cyclohexane-ethyl acetate 2:1 part was concentrated under reduced pressure to obtain about 1 g of the yellow crude product, which was applied to a reverse silica gel chromatography column (50 g, 40-60 μm, column volume 1×50 cm) by wet method, and methanol- Water carries out graded gradient elution (40% methanol 100ml, 60% methanol 300ml, 75% methanol 400ml, 80% methanol 300ml, 90% methanol 200ml), activity tracking shows that 75% methanol part has stronger activity against rice blast.
[0038] Merge the main active fractions, remove methanol by rotary evaporation, freeze-dry to obtain semi-pure prod...
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