Application of Agromyces sp. MT‑E in Degradation of Various Phthalates
A technology of dibutyl phthalate and diisooctyl phthalate, which is applied in the field of soil fungus MT-E in degrading various phthalates, can solve the problem of degradable phthalates that have not yet been seen. Phthalates and other problems, to achieve the effect of enriching the germplasm resource bank, easy to cultivate, and strong adaptability
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Embodiment 1
[0029] Example 1 Isolation and identification of strains
[0030] Collect farmland soil that has been used in plastic greenhouses all the year round, weigh 5 g of fresh soil sample into a 150 mL Erlenmeyer flask containing 50 mL sterile water, incubate at 30°C, 150 rpm for 3 days, and add 5 mL of sludge suspension to 100 mL DBP and DEHP (50 mg / L respectively) in MSM medium. After culturing at 30°C and 150 rpm for 7 days, the inoculum was continuously enriched and transferred 10 times at a volume of 1 mL each time, and the content of PAEs in the medium was increased to 1000 mg / L accordingly. Then dilute the culture solution 10 times 3 ~10 5 Spread on the LB solid plate and incubate at 30°C for 1 to 3 days. After a single bacteria grows on the plate, a single colony is picked and streaked for purification several times, and a bacteria strain is isolated, numbered MT-E. Then the strain was inoculated on the LB solid plate and cultured upside down at 30°C for 7 days to observe the...
Embodiment 2
[0038] Example 2 Soil mold ( Agromyces sp.) Degradation effect of strain MT-E on DBP and DEHP
[0039] S1. Preparation of bacterial suspension: The purified MT-E strain was connected to 10 mL of LB liquid medium for overnight activation and culture to the logarithmic phase. The bacteria were collected by centrifugation at 5000 rpm for 10 min, and the bacteria were washed 3 times with PBS Resuspend afterwards, adjust OD 600 nm =0.8 as a bacterial suspension.
[0040] S2. Inoculate 1 mL of the above bacterial suspension into 100 mL MSM culture medium containing 200 mg / L DBP and DEHP (each containing 200 mg / L), use non-inoculation as a negative control, and adjust the pH to 7.5. Set of three repetitions. Incubate at 30°C and 150 rpm in a constant temperature shaker for 7 days. Take samples and extract samples on 0, 1, 3, 5, and 7 days respectively. Determine the degradation of DBP and DEHP in the samples by GC / MS, and use a spectrophotometer. Measure the bacterial growth OD at ea...
Embodiment 3
[0045] Example 3 Remediation effect of strain MT-E on soil contaminated by DBP and DEHP
[0046] S1. Preparation of test soil: The soil is the paddy soil of the farm of South China Agricultural University. After air-drying, it is passed through a 60-mesh sieve, and the pH is 6.7. DBP and DEHP are added to the soil so that the content in the soil reaches 100 mg / kg, and double Distilled water is adjusted to the field water holding capacity (about 30%) and aged for 15 days in the dark.
[0047] S2. Weigh the above-mentioned 100 mg kg -1 Put 200 g of DBP and DEHP soil in a triangular flask, add 50 mL of bacterial suspension to the soil and mix well. In addition, the treatment without inoculation was used as the control group. Adjust the soil to the field water holding capacity (about 30% of the water content), cultivate in a 30°C incubator protected from light, and sample and extract regularly at 0, 2, 4, 6, 8, and 10 days, and then use GC / MS Determine the residual amount of DBP and ...
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