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Bacterial strain for degrading diethyl terephthalate and application thereof

A technology of diethyl phthalate and bacterial strains, which is applied to bacterial strains and application fields that can efficiently degrade diethyl terephthalate, and can solve problems such as carcinogenicity, imperfect treatment process of diethyl terephthalate, and environmental pollution

Active Publication Date: 2017-07-04
NANJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First of all, as a chemical raw material, diethyl terephthalate itself is an environmental hormone, which will cause serious pollution to the environment
Secondly, diethyl terephthalate will release terephthalic acid (TPA) under the action of microorganisms and hydrolytic enzymes, which is a very important chemical raw material and is widely used in synthetic resins, polyester fibers, plastic films and dyes and other industries, but it has inhibitory effect on the regeneration of microorganisms in water, and has mutagenic and carcinogenic effects on animals, and is recognized as a toxic pollutant.
At present, there are many studies on the biodegradation of terephthalic acid, but the treatment process of diethyl terephthalate is still far from perfect

Method used

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  • Bacterial strain for degrading diethyl terephthalate and application thereof
  • Bacterial strain for degrading diethyl terephthalate and application thereof
  • Bacterial strain for degrading diethyl terephthalate and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Isolation and screening of diethyl terephthalate degrading strain Delftia WL-3:

[0045] Take 5mL of sewage samples and place them in 100mL enrichment medium containing 500mg / L diethyl terephthalate, at 30°C, 180r min -1 Culture 7d. Measure the degradation situation of the diethyl terephthalate in the enrichment solution with an ultraviolet scanner, after confirming that the diethyl terephthalate is degraded, insert the solution containing 750mg / L diethyl terephthalate with 10% inoculum In the ethyl ester-rich medium, continue to enrich and measure the degradation, follow this method until the concentration of diethyl terephthalate increases to 1500mg / L, and pass for 3 times.

[0046] The diethyl terephthalate enrichment solution was serially diluted and spread on an inorganic salt plate with 1000 mg / L diethyl terephthalate as the sole carbon source, and cultured in a 30°C incubator for 3 to 4 days. Single colonies with different colony shapes were streaked on LB plat...

Embodiment 2

[0051] Identification and growth characteristics of strain Delftia WL-3 degrading diethyl terephthalate and its metabolites:

[0052] Identification of WL-3:

[0053] 16S rDNA identification of WL-3: Use primers 27F: 5`-AGAGTTTGATCCTGGCTCAG-3` and 1492R: 5`-TACCTTGTTACGACTT-3` to amplify the 16S rDNA of strain WL-3, and connect it to the clone by T / A cloning Vector pMD19T, construct the recombinant cloning vector pMD19T-16S, transform it into the cloning host strain Escherich coli DH5α to obtain the recombinant microorganism Escherich coli DH5α (pMD19T-16S), sequence the exogenous fragment of the recombinant microorganism obtained, and compare the NCBI database 16S rDNA sequence, the strain WL-3 was identified to the genus Delftia at the molecular level, and the nucleotide sequence of its 16S rDNA is shown in SEQ ID NO: 1 in the sequence table.

[0054] Growth characteristics of WL-3:

[0055] Delftia WL-3 grows slowly on LB plate, 30℃, 48h can form a milky white colony with...

Embodiment 3

[0058] Degradation characteristics of diethyl terephthalate and its metabolite-degrading bacteria Delftia WL-3:

[0059] Pick a single colony of strain WL-3 from the LB plate, inoculate them in 3 mL LB liquid medium at 30°C, shaker 180r min -1 Cultivate for 12h. Then the culture solution of the bacterial strain was transferred to 100 mL of fresh liquid LB medium, and the culture was continued for 12 h. 6000r·min -1 Centrifuge for 10min, collect the bacteria, wash twice with sterilized inorganic salt medium, and make OD 600nm =1.0 bacterial suspension, that is, seed liquid. In the inorganic salt medium with a final concentration of diethyl terephthalate of 1000mg / L, insert OD at 5% inoculum 600nm =1.0 Strain WL-3 seed liquid, at 30°C, 180r·min -1 Shake the culture, take a sample every 24h, measure the OD 600nm and concentrations of diethyl terephthalate.

[0060] Depend on Figure 4a ~ Figure 4d It can be seen that the bacterial strain WL-3 was inoculated in the medium ...

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Abstract

The invention discloses a bacterial strain for degrading diethyl terephthalate and application thereof. The bacterial strain is classified and named as Delftia sp., with the bacterial strain number being WL-3, the bacterial strain is preserved in China center for type culture collection, with the preservation number being CCTCC NO: M 2017136, and the preservation data being March 21, 2017. The invention further discloses application of the bacterial strain in degrading diethyl terephthalate. The bacterial strain WL-3 is aerobic microorganism, the optimal growth temperature is 30 DEG C, and the optimal growth pH is 7.0. The bacterial strain can completely degrade 1000mg / L diethyl terephthalate within 7 days and can use the same as a unique carbon source to grow to completely degrade metabolite of terephthalic acid. The bacterial strain disclosed by the invention has important application value on degrading the diethyl terephthalate, and the metabolite thereof, and biological treatment on industrial wastewater.

Description

technical field [0001] The invention belongs to the technical field of biological treatment, and relates to a bacterial strain for efficiently degrading diethyl terephthalate and its application. Background technique [0002] First of all, with the rapid development and wide application of the polyester plastic (PET) industry, a large amount of PET waste is produced every year. About 400kt of polyester waste is produced in my country every year. Although these polyester wastes are not very toxic and will not directly cause harm to the environment, due to the huge number of wastes and the difficulty of degrading them under natural conditions, they have caused great harm. White pollution. Due to the hydrophobicity and high crystallinity of PET, it is very difficult to directly screen for microorganisms or enzymes that can degrade efficiently from PET. [0003] Therefore, in this experiment, diethyl terephthalate (DET) was used as a structural mimic to domesticate and cultivat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/34
CPCC02F3/34C02F2101/34C12R2001/01C12N1/205
Inventor 董维亮刘嘉唯信丰学马江锋姜岷
Owner NANJING UNIV OF TECH
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