A kind of synthetic method of trichothecenes B group toxin
A toxin and a single technology, applied in the field of synthesis methods belonging to a trichothecene B group toxin, can solve the problems of long synthesis process route, cumbersome and complicated treatment, strong reagent toxicity, etc., and achieve simple sample processing method and high product quality The effect of high purity and simple method
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Embodiment 1
[0028] The preparation of embodiment 1 culture medium
[0029] Preparation of potato dextrose agar medium (or recovery medium PDA, or PDA medium for short): wash commercially available potatoes with distilled water, peel them, and cut them into 3 cm long and wide pieces with a knife. Take 200g of potato pieces, put them into a 1L beaker, add 400mL of distilled water and boil for 10min, then filter with gauze. Discard the potato pieces, keep the filtrate, add glucose 20g; agar 20g; heat to dissolve, add distilled water to 1000mL, sterilize under high pressure steam at 121°C for 20min, set aside, put the sterilized culture medium in a microwave oven before use Heat to dissolve, pour the plate on a conventional ultra-clean workbench, seal it with parafilm after cooling, and set aside.
[0030] Preparation of conidia suspension medium (CMC medium): weigh 7.5g sodium carboxymethylcellulose, 0.5g potassium dihydrogen phosphate, 0.5g yeast powder, 0.5g ammonium nitrate, 0.25g magnes...
Embodiment 2
[0032] The recovery of embodiment 2 strains and the preparation of conidia liquid
[0033] Strain recovery: Take out the dry powder bottle containing the strain Fusarium graminearum DSMNo.4528 (commercially purchased from DSMZ Company in Germany, and the Chinese import agent is Beijing Zhongyuan Heju Economic and Trade Co., Ltd.), place the sealed bottle mouth on an alcohol lamp and burn for 2 minutes. Use a pipette gun to draw a small amount of sterile distilled water and drop it on the burnt bottle mouth. The burnt bottle mouth will crack under cold water soaking, open the bottle mouth with sterile tweezers, and carefully remove the glass fragments. Take out the glass sleeve containing the dry powder, take out the powder block with sterile tweezers, put it into a 10mL sterilized culture bottle, add 2-3mL of sterilized distilled water, seal it with a parafilm, and let it stand overnight at room temperature. Carefully mix the bacterial suspension that has been rehydrated overn...
Embodiment 3
[0035] Example 3 Utilize the toxin-producing medium to cultivate toxin-producing
[0036] After cultivating the conidia liquid obtained in Example 2 in a desktop refrigerated constant temperature oscillator for 7 days, take the CMC conidia culture medium cultured on a shaker, and scrape off the conidia carefully with an inoculation loop in a conventional ultra-clean workbench , mixed into a suspension, take the sterilized pipette tip with a pipette holster, carefully draw 2mL of conidia suspension in the aseptic area of the alcohol lamp and add it to 90 sterilized Erlenmeyer flasks packed with rice After the inoculation is completed, place the inoculated Erlenmeyer flask in a sterilized mold incubator (the culture temperature is set at 28°C and the relative humidity is set at 50%), away from light; Take out the Erlenmeyer flask from the mold incubator and shake it evenly by hand every day to ensure sufficient oxygen and even distribution of toxins to promote the production o...
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