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A strain of Agromyces sp. MT‑E that can simultaneously degrade multiple phthalates

A soil mold, MT-E technology, applied in the field of biological treatment of environmental pollutants, can solve problems such as degradable phthalates that have not yet been seen, and achieve the effects of enriching the germplasm resource pool, easy cultivation and strong adaptability

Active Publication Date: 2017-09-01
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Soil mold ( Agromyces sp.) is a genus widely present in the soil. It has been reported that some strains of this genus can efficiently degrade phenol, pesticide thiamethoxam and resistance to heavy metals, but it has not been found that it can degrade phthalo formate reports

Method used

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  • A strain of Agromyces sp. MT‑E that can simultaneously degrade multiple phthalates
  • A strain of Agromyces sp. MT‑E that can simultaneously degrade multiple phthalates
  • A strain of Agromyces sp. MT‑E that can simultaneously degrade multiple phthalates

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Experimental program
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Effect test

Embodiment 1

[0024] The isolation and identification of embodiment 1 bacterial strain

[0025] Collect farmland soil that has been used in plastic greenhouses all year round, weigh 5 g of fresh soil samples into a 150 mL Erlenmeyer flask containing 50 mL of sterile water, incubate at 30°C and 150 rpm for 3 days, then add 5 mL of sludge suspension to 100 mL of DBP and DEHP (50 mg / L, respectively) in MSM medium. After being cultured at 30°C and 150 rpm for 7 days, the inoculum was continuously enriched and transferred 10 times with an inoculum size of 1 mL each time, and the content of PAEs in the medium was correspondingly increased to 1000 mg / L. Then the 10 times acclimatized culture solution was diluted 10 3 ~10 5 Spread on LB solid plates and incubate upside down at 30°C for 1-3 days. After a single colony grows on the plate, pick a single colony and streak it several times for purification, and obtain a strain of bacteria, numbered MT-E. Then the strains were inoculated on LB solid ...

Embodiment 2

[0032] Embodiment 2 soil mold ( Agromyces sp.) Degradation effect of strain MT-E on DBP and DEHP

[0033] S1. Preparation of bacterial suspension: Inoculate the purified MT-E strain into 10 mL of LB liquid medium for overnight activation to the logarithmic phase, collect the bacterial cells by centrifugation at 5000 rpm for 10 min, and wash the bacteria 3 times with PBS After resuspension, adjust OD 600 nm =0.8 as bacterial suspension.

[0034] S2. Inoculate 1 mL of the above-mentioned bacterial suspension into 100 mL of MSM culture solution containing 200 mg / L of DBP and DEHP (each containing 200 mg / L), and use no inoculation as a negative control, and adjust the pH to 7.5. Set of three repetitions. Incubate in a constant temperature shaker at 30°C and 150 rpm for 7 days, take samples and extract samples at 0, 1, 3, 5, and 7 days respectively, and measure the degradation of DBP and DEHP in the samples by GC / MS. Determination of bacterial growth OD at each time point 600...

Embodiment 3

[0039] The remediation effect of embodiment 3 bacterial strain MT-E to DBP and DEHP polluted soil

[0040] S1. Preparation of test soil: The soil is paddy soil from the farm of South China Agricultural University. After air-drying, pass through a 60-mesh sieve with a pH of 6.7. Adjust the steamed water to the field water holding capacity (about 30%), and age in the dark for 15 days.

[0041] S2. Weigh the above-mentioned 100 mg kg -1 Put 200 g of DBP and DEHP soil in a Erlenmeyer flask, add 50 mL of bacterial suspension to the soil, and mix well. In addition, the treatment without bacteria was used as the control group. Adjust the soil to the field water holding capacity (about 30% water content), culture in a 30°C incubator in the dark, take samples and extract regularly at 0, 2, 4, 6, 8, and 10 days, and then conduct GC / MS Determination of DBP and DEHP residues in soil.

[0042] Sample extraction: Weigh 1 g of air-dried and ground soil sample into a 35 mL glass centrifuge...

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Abstract

The invention belongs to the technical field of biological treatment of environmental pollutants, and specifically discloses a strain of Agromyces sp. MT‑E capable of simultaneously degrading various phthalates. The strain was deposited in China Center for Type Culture Collection (CCTCC) on January 22, 2015, and the preservation number is CCTCC NO: M 2015054. The bacteria can use DBP and DEHP as the only carbon source for aerobic degradation, and the degradation rates can reach 97.06% and 86.25% in 7 days in the inorganic salt medium containing 200 mg / L DBP and DEHP as the only carbon source respectively. . After inoculating the bacteria in contaminated soil for 10 days, the degradation rates of DBP and DEHP in the soil were up to 77.29% and 55.51%, respectively. The efficient degradation rate indicated that the bacteria had a good application prospect in the remediation of PAEs-contaminated soil.

Description

technical field [0001] The invention relates to the technical field of biological treatment of environmental pollutants, in particular to a novel phthalate-degrading bacterium, and more specifically to a soil mold that can simultaneously degrade a variety of phthalates ( Agromyces sp.) MT-E. Background technique [0002] Phthalate, also known as phthalate, abbreviated as PAEs, is a general term for esters formed by phthalic acid. Among them, dibutyl phthalate (di- n -butylphthalate, DBP) and di-(2-ethylhexyl)phthalate, DEHP) are two of the most widely used plastic plasticizers, and their content in PVC plastics can reach 40~60% . In recent decades, the use of plastic greenhouses and agricultural film in my country has increased dramatically. With the extensive use of plastic films, more and more DBP and DEHP are released into farmland soil, and they have become the most commonly detected organic pollutants in farmland soil, with the detection level reaching mg / kg. Stud...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/14B09C1/10C12R1/645
CPCB09C1/105C12N1/145C12R2001/645
Inventor 莫测辉赵海明李彦文蔡全英李慧杜欢林静冯乃宪
Owner JINAN UNIVERSITY
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