Pseudomonas and application of the same in biological removal of nitrogenous heterocyclic compounds

A technology of nitrogen heterocyclic compounds and pseudomonas, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of microbial growth and biological removal, and achieve the effect of broad application prospects

Inactive Publication Date: 2008-06-11
PEKING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Previous studies mainly focused on the degradation process and metabolic pathways of microorganisms under single-substrate conditions. Under dual-substrate conditions, especially when the contents

Method used

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  • Pseudomonas and application of the same in biological removal of nitrogenous heterocyclic compounds
  • Pseudomonas and application of the same in biological removal of nitrogenous heterocyclic compounds
  • Pseudomonas and application of the same in biological removal of nitrogenous heterocyclic compounds

Examples

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Embodiment 1

[0023] Example 1, Isolation, Purification and Identification of Pseudomonas sp. BC001

[0024] 1. Isolation and purification of Pseudomonas sp. BC001

[0025] Pseudomonas sp. BC001 is a Gram-negative bacterium obtained by sampling from the activated sludge in the secondary sedimentation tank of the sewage treatment system of Shougang Coking Plant, after domestication, isolation and purification. Specific steps are as follows:

[0026] Activated sludge samples were taken from the wastewater treatment system of Shougang Coking Plant, and intermittently aerated in a container (about 2 L) sealed with multiple layers of gauze. Regularly add 10mL pyridine (50-300mg / L), 10mL quinoline (50-300mg / L) and about 50mL coking wastewater raw water, and add 2mL phosphate buffer (each L contains 8.5gKH 2 PO 4 , 21.75g ​​K 2 HPO 4 , 33.4 g Na 2 HPO 4 ·7H 2 O, 1.7 g NH 4 Cl, pH 7.2) and 2mL trace element solution (1.5g FeCl per L 3 ·6H 2 O, 0.024g NiCl 2 ·6H 2 O, 0.19g CoCl 2 ·6H ...

Embodiment 2

[0038] Example 2, Pseudomonas sp. (Pseudomonas sp.) BC001 CGMCC No. 2223 adsorption and degradation detection of nitrogen-containing heterocyclic compounds

[0039] 1. Detection of Pseudomonas sp. BC001 for bioremoval of pyridine and quinoline

[0040] Under the conditions of pyridine single substrate, quinoline single substrate, pyridine-quinoline double substrate, pyridine-glucose double substrate, quinoline-glucose double substrate, implement Pseudomonas sp. BC001 CGMCC №.2223 on pyridine, Biological removal of quinolines. It was found that the bioremoval of pyridine by Pseudomonas sp. BC001 CGMCC №.2223 was mainly biosorption, while the bioremoval of quinoline included biosorption and biodegradation, adding an appropriate amount of carbon source could promote the degradation of quinoline. The specific implementation steps are as follows:

[0041] 1) Preparation of Pseudomonas sp. BC001 CGMCC №.2223 bacterial suspension

[0042] Inoculate Pseudomonas sp. BC001 CGMCC №.22...

Embodiment 3

[0067] Embodiment 3, the plasmid detection of pseudomonas (Pseudomonas sp.) BC001

[0068] The plasmid of Pseudomonas sp. BC001 was extracted by improved alkaline lysis method, and it was found that Pseudomonas sp. BC001 did not carry the plasmid.

[0069] Reagents used:

[0070] P1: Dissolve 6.06g Tris base and 3.72g Na 2 EDTA·2H 2 O in 800mL distilled water, adjust the pH to 8.0, dilute to 1 liter, and then add 100mg RNase A.

[0071] P2: Add 8.0g NaOH and 50mL SDS solution with a concentration of 20% (g / mL) in 950mL distilled water, mix well, and dilute to 1 liter.

[0072] P3: Add 294.5 g of potassium acetate to 500 mL of distilled water, adjust the pH to 5.5 with glacial acetic acid (about 110 mL), and dilute to 1 liter.

[0073] TE: Add 1.576g Tris·Cl per liter of distilled water to adjust the pH to 8.0; add 0.3722g EDTA·Na 2 .

[0074] The specific implementation is as follows:

[0075] 1. Use LB medium to cultivate Pseudomonas sp. BC001 to logarithmic growth pha...

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Abstract

The invention discloses a plant of pseudomonad and the application thereof in the biological removal of nitrogen heterocyclic compound. The pseudomonad adopts Pseudomonas sp. BC001 CGMCC NO.2223. The strain not only can utilize quinoline as a sole carbon source and a nitrogen source to perform metabolism, but also can well grow under the condition of highly concentrated pyridine and quinoline double stroma (pyridine is about 400 mg / L, quinoline is about 500 mg / L). The removal of the invention to the pyridine mainly passes through the biological absorption, the removal to the pyridine comprises two phases of biological absorption and biological degradation, and simultaneously, the additional carbon source with proper amount has the acceleration function to the pyridine degradation. Through the detection, the intermediate product of the pyridine degradation mainly adopts 2-hydroxyquinoline and 8-hydroxyl coumarin, the main end product adopts NH 4 <+>. BC001 bacteria have no resistance to penbritin, kanamycin sulfate, chloromycetin, cyclomycin and spectinomysin, and the used ecological security is ensured.

Description

technical field [0001] The invention relates to a strain of pseudomonas and its application in biological removal of nitrogen-containing heterocyclic compounds. Background technique [0002] At present, industrial production produces and discharges a large amount of aromatic compounds every day, and about two-thirds of them are heterocyclic compounds. Heterocyclic compounds are widely used in industrial solvents, dyes, explosives, medicine and pesticides and other fields. Pyridine and quinoline are two typical nitrogen-containing heterocyclic compounds, commonly found in coking wastewater, oil refining wastewater, pesticide wastewater, etc. They are extremely harmful to the environment and are difficult to biodegrade organic substances due to their toxic effect on most microorganisms. [0003] The use of bioaugmentation technology to treat heterocyclic refractory organics has become one of the hotspots of current research, and the bioaugmentation method of adding specific ...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/38C02F101/38
Inventor 温东辉柏耀辉孙庆华赵翠唐孝炎
Owner PEKING UNIV
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