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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 hydrocarbons have been isolated (Table 1). Therefore, it is of great innovation and application value to develop strains with a broad substrate spectrum

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

Embodiment 1

[0037] Example 1, Screening and Preservation of Pseudomonas aeruginosa CCTCC M 208114 Strain

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

[0039] The formula of the above-mentioned ion mixture is that every 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 above-mentioned vitamin mixed solution is, every liter of distilled water contains: 400mg calcium pantothenate (Calciumpantothenate); 200mg inositol (Inositol); 400mg nicotinic acid / nicotinic acid (Niacin); p-aminobenzoic acid); 0.5mg VB12 (Cyanocobalamin).

[0041] The formula...

Embodiment 2

[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 saturation, take 1.5 mL of culture, and centrifuge at 6000 rpm for 2 minutes; add 565 μl of TE buffer to the precipitate (TE buffer formula: 10 mmol Tris / L tris (Tris), 1mmol / L ethylenediaminetetraacetic acid (EDTA), adjust the pH to 8.0 with hydrochloric acid), resuspend it by blowing repeatedly with a pipette, add 30μl mass / volume ratio 10% sodium dodecylsulfonate (SDS) and 5μl 20mg / mL proteinase K, mix well, incubate at 37°C for 1 hour; add 100μl 5mol / L NaCl, mix well, then add μl CTAB / NaCl solution (CTAB / NaCl solution formula: dissolving 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 12000 rpm for 5...

Embodiment 3

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

[0049] (1) Strain selection: select Pseudomonas aeruginosa CCTCC M 208114;

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

[0051](3) Seed culture: the bacterial strain cultivated in step (2) was placed in 5mL LB liquid medium (see Example 1 for the formula) with an inoculation loop under aseptic conditions and cultured with shaking at 30°C After 12 hours, the culture was washed three times with sterile 0.85% saline to obtain a seed solution;

[0052] (4) Inoculate 10 mL of the inorganic salt medium (see Example 1 for the recipe) containing 20 g / L diesel oil with the seed solution obtained in step (3) according to the inoculum size of 5%, and shake and cultivate for 7 days at 30° C. The medium without inoculation was used as...

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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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