Pseudomonas aeruginosa and application thereof

A technology of Pseudomonas aeruginosa and bacterial agent, which is applied in the field of microbial technology and environmental biology, can solve the problems of limitation and low degradation substrate spectrum, and achieve the effect of high-efficiency emulsification ability

Inactive Publication Date: 2012-07-04
DAQING OILFIELD CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A lot of research has been done on several common toxic compounds in the world, and many strains that can degrade polycyclic aromatic hydroc

Method used

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  • Pseudomonas aeruginosa and application thereof
  • Pseudomonas aeruginosa and application thereof
  • Pseudomonas aeruginosa and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0037] Example 1. Screening and preservation of Pseudomonas aeruginosa CCTCC M 208114 strain

[0038] The formula of liquid basic inorganic salt culture medium is: KH 2 PO 4 0.5 g / l, K 2 HPO 4 4 g / l, NH 4 Cl 1 g / L, 1% by mass CaCl 2 2ml / L, 10% by mass MgCl 2 ·6H 2 O 2ml / L, FeCl with a mass percentage of 1% 3 200 microliters / liter, vitamin mixture 200 ml / liter, ionic mixture 5 ml / liter. Sterilize at 121℃ for 20 minutes.

[0039] The formula of the above ionic mixture is that each liter of distilled water contains: ZnCl 2 0.5g; FeCl 2 0.5g; MnCl 2 ·4H 2 O0.5g; Na 2 MoO 4 ·2H 2 O 0.1g; CuCl 2 ·2H 2 O 0.05g; Na 2 WO 4 ·2H 2 O 0.05g; HCl 120mmol / L.

[0040] The formula of the above-mentioned vitamin mixture is that each liter of distilled water contains: 400mg calcium pantothenate; 200mg inositol; 400mg niacin / niacin; 400mg VB6 (Pyridoxine hydrochloride); 200mg p-aminobenzoic acid ( p-aminobenzoic acid); 0.5 mg VB12 (Cyanocobalamin).

[0041] The formula of LB medium is: 1 liter of ...

Example Embodiment

[0046] Example 2. Extraction and sequencing of 16S rDNA gene of Pseudomonas aeruginosa CCTCC M 208114 strain

[0047] Cultivate 5 mL of Pseudomonas aeruginosa (Pseudomonas aeruginosa) CCTCC M 208114 bacterial culture to a saturated state, take 1.5 mL of the culture and centrifuge at 6000 rpm for 2 minutes; add 565 μl of TE buffer to the precipitate (TE buffer formula: 10 mmol / L tris (Tris), 1mmol / L ethylenediaminetetraacetic acid (EDTA), adjust the pH to 8.0 with hydrochloric acid), resuspend it by pipetting repeatedly with a pipette, add 30μl mass / volume ratio 10% sodium dodecyl sulfonate (SDS) and 5μl 20mg / mL proteinase K, mix well and incubate at 37°C for 1 hour; add 100μl 5mol / L NaCl, mix well, and then add μl CTAB / NaCl solution (CTAB / NaCl solution formula: dissolve cetyltriethylammonium bromide (CTAB) with a mass / volume ratio of 10% in 0.7mol / L NaCl), mix well, and incubate at 65°C 10 minutes; add an equal volume of chloroform / isoamyl alcohol, mix well, centrifuge at 12,0...

Example Embodiment

[0048] Example 3, Pseudomonas aeruginosa (Pseudomonas aeruginosa) CCTCC M 208114 growth system to the degradation of diesel

[0049] (1) Selection of strains: Pseudomonas aeruginosa (Pseudomonas aeruginosa) CCTCC M 208114;

[0050] (2) Slant culture: inoculate the strains in LB solid medium containing 1.6% agar with a mass / volume ratio, and statically culture for 24 hours at 30°C;

[0051] (3) Seed culture: The strains cultured in step (2) are cultured in 5 mL of LB liquid medium (see Example 1 for the recipe) in 5 mL of LB liquid medium (see Example 1 for the recipe) under aseptic conditions with an inoculum loop, and shake culture After 12 hours, the culture was washed three times with sterile 0.85% physiological saline to prepare seed liquid;

[0052] (4) Inoculate 10 mL of an inorganic salt medium containing 20 g / L diesel oil (see Example 1 for the recipe) with the seed solution prepared in step (3) according to the 5% inoculum amount and shake culture at 30°C for 7 days. Non-inf...

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Abstract

The invention discloses a pseudomonas aeruginosa CCTCCM208114 and application of the pseudomonas aeruginosa in petroleum pollutant degradation. The pseudomonas aeruginosa provided by the invention has the following characteristics that: 1) the pseudomonas aeruginosa has high-efficient petroleum hydrocarbon degradation capability; 2) the pseudomonas aeruginosa is capable of co-metabolizing fluorene in an inorganic salt culture medium in the presence of n-tetradecane; 3) a resting cell system of the pseudomonas aeruginosa has good polycyclic aromatic hydrocarbon degradation capability and can tolerate high polycyclic aromatic hydrocarbon concentration; 4) the pseudomonas aeruginosa has a wide substrate spectrum; and 5) the pseudomonas aeruginosa grows in an LB (Luria Bertani) culture medium or an inorganic salt culture medium (straight-chain paraffin or polycyclic aromatic hydrocarbon serves as the unique carbon source and energy source) and can generate biological surfactant. The pseudomonas aeruginosa can be widely applied to the field of bioremediation of petroleum compound pollution and is suitable for large-area popularization and application.

Description

technical field [0001] The invention belongs to the field of microbial technology and environmental biotechnology, in particular, it relates to a strain of Pseudomonas aeruginosa (Pseudomonas aeruginosa) DQ8 CCTCC M 208114 and its application in the degradation of petroleum pollutants. Background technique [0002] With the sharp increase in the global demand for energy, oil tanker leakage accidents frequently occur at sea, and oil wells and refineries also discharge a large amount of oily wastewater, which has caused huge environmental pollution. If we do not pay attention to crude oil pollution, it will endanger the environment and affect the living environment of human beings in the long run. How to deal with these crude oil pollution has become a major issue faced by many countries. [0003] According to the calculation of the national authoritative department, more than 600,000 tons of oil enter the environment in my country every year, causing serious pollution to the...

Claims

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Application Information

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IPC IPC(8): C12N1/20A62D3/02B01F17/00C12R1/385C09K23/00
Inventor 徐正顺牛彦良王凤兰伍晓林侯兆伟郭盟华金锐
Owner DAQING OILFIELD CO LTD
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