Bacillus subtilis capable of effectively degrading vomitoxin and application of bacillus subtilis
A technology of Bacillus subtilis and deoxynivalenol, which is applied in the field of microorganisms, can solve the problems of characteristic research on the anti-stress function of unseen deoxynivalenol strains and the small number of bacillus, and achieve broad market application prospects, specific effects, and detoxification activity high effect
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Embodiment 1
[0040] The screening of embodiment 1 bacterial strain
[0041] Specific steps are as follows:
[0042] 1) Screening of bacterial strains: screening of bacterial strains from animal intestinal contents, moldy feed, moldy food and natural environment.
[0043] Seed liquid preparation: Mix each intestinal content sample and the sample obtained from the environment with twice the volume of sterile water, place in a sterilized 18mm×180mm test tube, shake overnight; draw 0.1mL of the supernatant, respectively Mix with 0.8 mL of screening medium, shake and culture in a 15°C incubator for 12 hours to obtain a seed solution.
[0044] Wherein, the formula of screening medium is: NaH 2 PO 4 0.25g, NH 4 NO 3 1.0g, MgSO 4 .·7H 2 O0.25g, FeSO 4 0.001g, 17g agar, 2.0g glucose, add distilled water to 1000mL, pH7.0; sterilize by steam at 121℃ for 20min.
[0045] Preliminary screening of samples: add 100 μl of vomitoxin (100 μg / mL) to each seed solution, use non-inoculated screening me...
Embodiment 2
[0053] Embodiment 2 The cultivation method of Bacillus subtilis ANSB471 of the present invention
[0054] Take Bacillus subtilis (Bacillus subtilis) ANSB471 (preservation number CGMCC No.7344) 1.5mL (the concentration of live bacteria is 10 9 CFU / mL), inoculated in 80mL medium for shake flask fermentation, the fermentation temperature was 37°C, the pH value was 7.0, the rotation speed was 200r / min, and the fermentation time was 24h.
[0055] Among them, the shake flask fermentation medium is composed of the following components: tryptone 10g, yeast extract 2g, glucose 2g, beef extract 3g, sodium chloride 4g, disodium hydrogen phosphate 3g, magnesium sulfate heptahydrate 1.0g, distilled water 1000mL , pH 7.2.
[0056] After the fermentation, the fermentation broth was stored in a refrigerator at 4°C for later use.
Embodiment 3
[0057] Embodiment 3 The cultivation method of Bacillus subtilis ANSB471 of the present invention
[0058] Take Bacillus subtilis ANSB4711mL (viable bacteria concentration is 10 9 CFU / mL), inoculated in 50mL medium for shake flask fermentation culture, the fermentation temperature was 37°C, the pH value was 7.0, the rotational speed was 180r / min, and the fermentation time was 20h.
[0059] Among them, the shake flask fermentation medium is composed of the following components: tryptone 8g, yeast extract 1.5g, glucose 1.5g, beef extract 2g, sodium chloride 3g, disodium hydrogen phosphate 2.5g, magnesium sulfate heptahydrate 0.5g , Distilled water 800mL, pH value 7.0.
[0060] After the fermentation, the fermentation broth was stored in a refrigerator at 4°C for later use.
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