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Immobilized microspheres for remediation of petroleum contaminated soil, preparation method thereof and application thereof

A technology for soil remediation and oil pollution, applied in the restoration of contaminated soil, fixed on/in organic carriers, etc., can solve the problems of slow development of microbial preparation technology

Inactive Publication Date: 2011-09-28
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of the deficiencies of existing oil-contaminated soil microbial remediation technology and the slow development of corresponding microbial preparation technology, the purpose of the present invention is to provide an immobilized microsphere that can be used for oil-contaminated soil remediation and its preparation method

Method used

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  • Immobilized microspheres for remediation of petroleum contaminated soil, preparation method thereof and application thereof
  • Immobilized microspheres for remediation of petroleum contaminated soil, preparation method thereof and application thereof
  • Immobilized microspheres for remediation of petroleum contaminated soil, preparation method thereof and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0043] (1) Preparation of bacterial suspension:

[0044] Preparation of medium A:

[0045] Enrichment medium: used to inoculate petroleum hydrocarbon-degrading bacteria, prepare LB liquid medium according to conventional methods of microbiology, and use conventional methods for high-temperature sterilization. It can be sterilized at 121°C for 30 minutes, cooled to room temperature, and enriched Medium.

[0046] Proliferation medium: used to provide nutrients for petroleum hydrocarbon degrading bacteria, including glucose 20g / L, yeast extract 3g / L, peptone 2g / L, NH 4 NO 3 1g / L MgSO 4 ·7H 2 O 0.2g / L, KCl 0.2g / L, pH=7.8, use conventional methods for high-temperature sterilization, can be sterilized at 115°C for 20min, cool down for later use.

[0047] B enrichment culture:

[0048] Rinse the bacteria liquid from the slant culture medium of preserved petroleum hydrocarbon-degrading bacteria into common LB liquid culture medium, and obtain seed liquid after 12 hours of shake...

Embodiment 1

[0081] (1) Bacterial suspension preparation:

[0082] Preparation of A medium: (the medium of the following examples all adopts the medium prepared in Example 1, so the step (1) A of the following examples is omitted, and the description starts from step (1) B)

[0083] Enrichment medium: prepare a few milliliters of LB liquid medium according to conventional methods of microbiology: tryptone 10g / L, yeast extract 5g / L, sodium chloride 5g / L, pH=7.0.

[0084] Divide the above-mentioned enriched culture medium into several 1000ml Erlenmeyer flasks, the volume of each flask is 4 / 5 of the volume, put it into a high-temperature sterilization device, sterilize at 121°C for 30min, and cool to room temperature.

[0085] Proliferation medium (prepared according to conventional biological methods): glucose 20g / L, yeast extract 3g / L, peptone 2g / L, NH 4 NO 3 1g / L, MgSO 4 ·7H 2 O 0.2g / L, KCl 0.2g / L, pH=7.8. Sterilize at 115°C for 20min, cool for later use.

[0086] B enrichment cultur...

Embodiment 2

[0099] (1) Bacterial suspension preparation:

[0100] B enrichment culture:

[0101] In an aseptic operating bench, inoculate the Paracoccus sp.D17 strain in the enrichment medium at an inoculation amount of 5%; after completion, place it in a constant temperature culture shaker, adjust the shaker speed to 150rpm, and cultivate at 25°C for 48h; The bacterial liquid was collected and centrifuged at 4000 rpm for 10 minutes to obtain wet bacterial cells, which were stored at 4°C.

[0102] C bacteria suspension preparation:

[0103] 4g of wet bacteria was mixed evenly with 10ml of proliferation medium to prepare a bacterial suspension, which was stored at 4°C for later use.

[0104] (2) Preparation of sodium alginate-attapulgite gel:

[0105] Mix 3.5g of sodium alginate and 2.0g of attapulgite evenly, put in 90ml of deionized water, sterilize at 121°C for 30min, and then place it in a 60°C water bath to keep warm.

[0106] (3) Bacterial suspension dosing:

[0107] Take 90ml o...

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Abstract

The invention relates to immobilized microspheres for remediation of petroleum contaminated soil, a preparation method thereof and applications thereof. The immobilized microspheres are spheroidal, have internal apertures and have diameters of from 2.5 to 3.0 millimeters. The immobilized microspheres comprise sodium alginate as an embedding material and attapulgite as an adsorbent for adsorptions of petroleum hydrocarbon degrading bacteria and nutrient elements. The preparation method of the immobilized microspheres comprises embedding petroleum hydrocarbon degrading bacteria and nutrient elements through sodium alginate as an embedding material and attapulgite as an adsorbent, and carrying out a series of preparation steps of high density bacteria liquid suspension, immobilization and balling, and enrichment culture to obtain the immobilized microspheres. The immobilized microspheres prepared through the preparation method provided by the invention have favorable aperture conditions and adsorption performances, thereby having strong load capabilities. The immobilized microspheres provide good internal survival environments for petroleum hydrocarbon degrading bacteria, thereby having strong environmental impact resistance capabilities. The immobilized microspheres have the advantages of environmental protection, safety, good biocompatibility and wide foreground of application in bioremediation for petroleum contaminated soil.

Description

technical field [0001] The invention relates to an immobilized microsphere for remediation of petroleum-contaminated soil, a preparation method and application thereof, and belongs to the field of microbial preparations. Background technique [0002] Microbial remediation technology is recognized as the most viable soil cleaning technology. It has been widely and long-term developed in the remediation of oil-contaminated soil, and many breakthroughs and progress have been made. Generally speaking, the soil microbial remediation technology is to use the indigenous microorganisms in the soil or put the domesticated and efficient microorganisms into the soil to degrade petroleum pollutants and repair the polluted soil through the metabolic activities of the bacteria under suitable conditions. In order to overcome the problems of low effective concentration of free bacteria, weak competitiveness, and poor ability to resist toxicity, many eyes have turned to microbial agents. At...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/04A62D3/02A62D101/20B09C1/10
Inventor 王红旗王璇熊樱王倩花菲
Owner BEIJING NORMAL UNIVERSITY
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