Phthalic acid di(2-ethyl) hexyl ester degrading bacterium as well as culture method and application thereof

A technology of phthalate di- and culture method, which is applied in the field of dihexyl phthalate-degrading bacteria, can solve DEHP pollution and other problems, and achieve strong substrate specificity, rapid growth and remarkable effect

Inactive Publication Date: 2018-09-04
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
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  • Claims
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Problems solved by technology

[0005] The invention provides a bis(2-ethyl)hexyl phthalate degrading bacterium and its cultivation method and application, which provide tools for effectively solving the pollution problem of DEHP

Method used

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  • Phthalic acid di(2-ethyl) hexyl ester degrading bacterium as well as culture method and application thereof
  • Phthalic acid di(2-ethyl) hexyl ester degrading bacterium as well as culture method and application thereof
  • Phthalic acid di(2-ethyl) hexyl ester degrading bacterium as well as culture method and application thereof

Examples

Experimental program
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Embodiment 1

[0031] Embodiment 1: The bis(2-ethyl)hexyl phthalate degrading bacterium of this embodiment is Arthrobacter sp.DNEH-S1, which is preserved in the China Center for Type Culture Collection (CCTCC), and the preservation address is Wuhan University, Wuhan, The date of deposit is June 8, 2017, and the deposit number is CCTCC No: M 2017315.

[0032] In this example, Arthrobacter sp.DNEH-S1 is a Gram-positive bacterium, and the colony is milky white and shiny, with a wet surface and a bulge in the middle, and neat edges. The transmission electron microscope photo of Arthrobacter sp.DNEH-S1 is as follows figure 1 with figure 2 shown.

[0033] Physiological and biochemical identification of Arthrobacter sp.DNEH-S1 with reference to the eighth edition of "Berger's Bacteria Identification Manual" and "Common Bacterial System Identification Manual": the urease test, glycolysis test, methyl Red test, citrate utilization test, indole test are all positive, starch hydrolysis test, V-P te...

Embodiment 2

[0036] Example 2: Domestication and isolation of Arthrobacter sp.DNEH-S1

[0037] In this example, Arthrobacter sp.DNEH-S1 was isolated from the soil of an abandoned and polluted facility in Harbin. The separation method is carried out according to the following steps: 1. Take 500 g of the soil surface sample of 0-20 cm, put it in a sterilized waterproof paper bag, and store it in a refrigerator at 4°C. 2. Add a small amount of plastic debris to the retrieved soil, and place it in a constant temperature incubator at 30°C for enrichment and cultivation for 2 weeks. 3. Take 3 soil samples after enrichment culture, each weighing 5g, put them into a triangular flask containing 90mL sterilized water and 5-8 glass beads and seal them with a sealing film. Shake in a constant temperature shaking incubator for 30 minutes, then let stand for 30 minutes.

[0038] Take 10mL supernatant and add it into 100mL basic inorganic salt culture medium containing DEHP for cultivation. First, the...

Embodiment 3

[0040] Example 3: Molecular characterization of Arthrobacter sp.DNEH-S1

[0041] The molecular identification of Arthrobacter sp.DNEH-S1 obtained by isolation and screening was carried out according to the following steps: extract the total DNA of the strain, use bacterial 16S rDNA general primers 27f and 1522r, and use genomic DNA as a template for PCR amplification. Then use the gel recovery kit to recover and purify the PCR product; after that, carry out cloning and transformation, screen the colony of positive clones, and sequence them after expansion and cultivation. The electrophoresis results of Arthrobacter sp.DNEH-S1 genomic DNA are as follows image 3 . The PCR amplification result of Arthrobactersp.DNEH-S1 16S rDNA sequence is as follows Figure 4 .

[0042] The 16S rDNA sequence of Arthrobacter sp.DNEH-S1 was submitted to GenBank and the sequence number obtained was KC874838. The sequence result was compared with the 16S rDNA sequence in GenBank. The phylogeneti...

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Abstract

The invention discloses a phthalic acid di(2-ethyl) hexyl ester degrading bacterium as well as a culture method and application thereof and relates to a phthalic acid di(2-ethyl) hexyl ester degradingbacterium as well as a culture method and application thereof. By adopting the phthalic acid di(2-ethyl) hexyl ester degrading bacteriu as well as the culture method and application thereof, a tool for effectively solving pollution problems of DEHP (Di-2-Ethyl Hexyl Phthalate) is provided. The strain is Arthrobacter sp.DNEH-S1 and is preserved in the China Center for Type Culture Collection, thepreservation date is June 8, 2017, and the preservation number is CCTCC No.: M 2017315. The phthalic acid di(2-ethyl) hexyl ester degrading bacterium disclosed by the invention is capable of degradingDEHP in soil, and the toxic and harmful effects of EDHP upon soil microorganism activity can be effectively degraded.

Description

technical field [0001] The invention relates to a bis(2-ethyl)hexyl phthalate degrading bacterium and its cultivation method and application. Background technique [0002] Di(2-ethyl)hexyl phthalate (DEHP) is a high molecular weight phthalic acid esters (Phthalic acid esters, PAEs), and it has been widely used as a plasticizer In the manufacture of polyvinyl chloride. Because DEHP is biologically toxic and has "three-cause" toxicity to the human body, it has been listed as a priority pollutant by the US Environmental Protection Agency and the China National Environmental Monitoring Station. The restoration of DEHP-polluted environments has therefore become one of the hotspots of research at home and abroad. Among them, microbial remediation has the advantages of economy, safety, and high efficiency, and has great application potential. [0003] DEHP is the largest and most widely used plasticizer variety. As the output of China's plastic manufacturing industry continues t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20B09C1/10C12R1/06
CPCB09C1/10C12N1/20C12N1/205C12R2001/06
Inventor 张颖王蕾焦雅琪周长健陶月孟庆娟王一帆
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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