Immobilized microspheres for degrading bacteria of PAEs (phthalic acid esters) and preparation method and application of such immobilized microspheres

A technology for degrading bacteria and microspheres, applied in the field of agricultural biology, can solve the problems of low DBP degradation efficiency and inability to use efficiently, and achieve the effect of simple operation

Active Publication Date: 2017-06-13
QINGDAO AGRI UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it is only aimed at soil DBP pollution and has a low degradation efficiency ...

Method used

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  • Immobilized microspheres for degrading bacteria of PAEs (phthalic acid esters) and preparation method and application of such immobilized microspheres
  • Immobilized microspheres for degrading bacteria of PAEs (phthalic acid esters) and preparation method and application of such immobilized microspheres
  • Immobilized microspheres for degrading bacteria of PAEs (phthalic acid esters) and preparation method and application of such immobilized microspheres

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Screening and Identification of Phthalate Degrading Bacteria

[0036] (1) Screening of phthalate-degrading bacteria

[0037] A certain amount of soil samples were collected in vegetable greenhouses in the suburbs of Qingdao, 10g of fresh soil samples were accurately weighed, added to a conical flask filled with 90mL of distilled water and 10mL of glass beads, placed in a shaker at 30°C, 175rpm and oscillated to mix evenly. Set aside.

[0038] Take 1mL from the above mixed solution according to the inoculum size of 1%, add it to 100mL inorganic salt culture solution with an initial concentration of 100mg / L phthalate (both DBP and DEHP are 50mg / L), and place it on a shaker Shaking culture was carried out at 30°C and 175rpm in the dark for 7 days. Gradually increase the concentration of phthalates (including 200mg / L, 500mg / L, 800mg / L, 1000mg / L, 1500mg / L and 2000mg / L) for each transfer, transfer once every seven days as an acclimation cycle, After a total of 6 ...

Embodiment 2

[0043] Example 2 Detection of Pseudomonas putida RXX-01's ability to degrade phthalates

[0044] The culture solution of the RXX-01 strain was inoculated in an inorganic salt medium (50 mL) with a DBP and DEHP concentration of 50 mg / L according to an inoculum amount of 1%, and was shaken at 25° C. and 150 r / min. After culturing for 3 days, the concentrations of DBP and DEHP in the culture medium were measured, and the degradation rates of DBP and DEHP were calculated. The results showed that the degradation rate of DBP was 90.8%, and the degradation rate of DEHP was 54.7%, which indicated that the RXX-01 strain had a high ability to degrade phthalates.

[0045] Degradation rate calculation formula: degradation rate (%)=((initial concentration of phthalates-residual concentration of phthalates) / initial concentration of phthalates)×100%.

Embodiment 3

[0046] Example 3 The preparation method of Pseudomonas putida RXX-01 immobilized microspheres

[0047] In order to determine the optimal ratio of immobilized materials in this embodiment, different bacterial liquid content, sodium alginate concentration, activated carbon content, and calcium chloride concentration were selected as factors to determine the concentration of immobilized microsphere terephthalate. The degradation rate was targeted, and the immobilization conditions were optimized.

[0048] A total of 4 factors (bacterial suspension content, sodium alginate concentration, activated carbon content, and calcium chloride concentration) were set in the single factor experiment, and 6 concentration levels were set for each factor. The percentages involved are all mass percentages. The bacterium suspension refers to the OD that the bacterium is made with physiological saline 600 = 1.0 suspension.

[0049] Under the conditions of sodium alginate concentration of 2%, ac...

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Abstract

The invention relates to immobilized microspheres for degrading bacteria of PAEs (phthalic acid esters) in farmland soil and a preparation method of such immobilized microspheres. The preparation method includes (1), preparing a bacterial suspension, namely culturing pseudomonas putida RXX-01 (preservation number: CGMCC No.13224) to a logarithmic phase, cleaning bacteria with normal saline, and mixing the bacteria into the bacterial suspension with the normal saline so as to enable OD600 of the bacterial suspension to equal 0.8-1.2; (2), preparing mixed bacterial liquid, namely preparing a sodium alginate solution with sterile water, adding activated carbon into the sodium alginate solution with mixing uniformly to obtain mixed liquid, and adding the bacterial suspension obtained in the step (1) into the mixed liquid with mixing uniformly so as to obtain the mixed bacterial liquid, wherein the mixed bacterial liquid comprises, by weight, 15-25% of bacterial suspension, 2-3% of sodium alginate and 2-4% of activated carbon; (3) preparing the immobilized microspheres, namely preparing a 20-30 g/L calcium chloride solution with sterile water, dropwise adding the mixed bacterial liquid into the calcium chloride solution, and crosslinking for 20-30 hours so as to obtain the immobilized microspheres, wherein a volume ratio of the mixed bacterial liquid to the calcium chloride solution is 1:1.5-1:2. The preparation method is simple, and the immobilized microspheres are high in efficiency of degrading the phthalic acid esters in the soil.

Description

technical field [0001] The invention belongs to the field of agricultural biology, and particularly relates to a degrading bacteria immobilized microsphere for removing phthalate in farmland soil, a preparation method and application thereof. Background technique [0002] Phthalic Acid Esters (PAEs) is a typical class of endocrine disruptors, Dimethyl Phthalate (DMP), Dibutyl Phthalate (DBP) and Di-(2-ethylhexyl Phthalate, DEHP) and other six phthalate compounds are listed as priority pollutants by the U.S. Environmental Protection Agency and my country Environmental Monitoring Center. As an important chemical product, phthalates are widely used as additives in the production process of plastic products. The rapid development of facility agriculture in my country has made plastic products widely used, and phthalates will be slowly released as the plastic ages And eventually enter the soil, causing it to become one of the most frequently detected organic pollutants in farmland ...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/10C12N11/04B09C1/10C12R1/40
CPCB09C1/10B09C2101/00C12N11/04C12N11/10C12N11/14
Inventor 周震峰饶潇潇王亚娟
Owner QINGDAO AGRI UNIV
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