Bacillus velezensis Shui-6, production method and application
A bacillus and production method technology, applied in the field of microbiology and biodegradation, can solve problems such as liver toxicity, tumor occurrence, genotoxicity, etc., achieve efficient degradation, good application prospects, and efficient inhibition of fungal growth
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Embodiment 1
[0078] Screening, identification of embodiment 1 Veles bacillus Shui-6 (Bacillus velezensis)
[0079] Taking natural soil as the source of bacteria, take 5g soil sample and put it into a 50mL centrifuge tube, add 45mL deionized water, shake it on the shaker for 30min, let it stand for 30s, and make 10 -1 g / mL; Take 100μL 10 -1 Add 900 μL deionized water to the diluent and put it into a 1mL EP tube, vortex for 3-5s to make 10 -2 g / mL, take 100 μL 10 -2 Add 900 μL deionized water to the diluent, put it into a 1mLEP tube, vortex for 3-5s, and make 10 -3 g / mL, and so on to 10 -6g / mL; use NA plate, take 300 μL of the diluted solution to smear the plate, and incubate at 37°C for 24 hours; pick a single colony (with different shapes, sizes, and viscosities) from the NA plate by the four-section line method, and put the single Bacterial colonies were added to LB liquid medium, and cultured at 37° C. and 180 rpm (revolutions per minute) for 48 hours to obtain a bacterial liquid for...
Embodiment 2
[0082] Embodiment 2 Veles bacillus Shui-6 is to fungus bacteriostasis test
[0083] Use plate confrontation culture method for screening, take 10 μL of 1×10 7 CFU / mL of the fungal spore suspension to be tested is dropped on the center of the PDA plate, and at an equidistant distance of 3 cm around, take 10 μL of Bacillus veleisi Shui-6 (OD 600 =0.8) The suspension was dropped around, and 10 μL of LB liquid medium was dropped on one of them as a blank control group. At the alcohol lamp of the ultra-clean workbench, the liquid was dried on the PDA medium, and then put into a constant temperature incubator Cultivate at 28°C, invert for about 5 days, and observe the growth of mycelia every other day until the mycelia cover the plate. Each experiment was repeated 3 times. The diameter of the fungus to be tested was measured with a vernier caliper using the cross method, and the average value was taken as the measurement result. The antibacterial rate was calculated according to ...
Embodiment 3
[0089] Example 3 Degradation of ochratoxin A by Bacillus Velez Shui-6
[0090] 1. Inoculate Bacillus Velez Shui-6 in LB liquid medium to the initial OD 600 =0.05, cultured with shaking at 200 rpm (radius of rotation: 20 mm) for 24 hours at 37°C, and centrifuged at 10 000 r / min for 10 minutes to collect the supernatant.
[0091] 2. Dissolve 1 mg of OTA standard (MZ standard, product number M44001) in 10 mL of chromatographically pure methanol to obtain an OTA standard solution with a concentration of 100 ppm.
[0092] 3. Prepare the experimental group solution:
[0093] Take 5 mL of the bacterial solution collected in step 1, put it in a 10 mL centrifuge tube, add 5 μL of the OTA solution obtained in step 2 to the centrifuge tube, mix well, let stand at 37 ° C for 72 h, then centrifuge at 10 000 g for 10 min and collect the supernatant to obtain Experimental group solution.
[0094] 4. Prepare the control group solution:
[0095] According to the method in step 3, take 5 mL...
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