A polycyclic aromatic hydrocarbon degrading bacterium and its screening method and application

A polycyclic aromatic hydrocarbon, degrading bacteria technology, applied in chemical instruments and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as reducing the service function and value of marine ecosystems, sticking to polluted beaches, and lack of oxygen.

Active Publication Date: 2021-06-18
THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Oil spill pollution accidents will also cause severe hypoxia in large areas of the sea, destroy marine productivity, cause a large number of fish, shrimp, shellfish and seabirds to die, and cause local "ocean desertification"; Cause deserted beaches, destroy mariculture and salt pan production, pollute or destroy coastal tourist areas, and reduce the service function and value of marine ecosystems

Method used

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  • A polycyclic aromatic hydrocarbon degrading bacterium and its screening method and application
  • A polycyclic aromatic hydrocarbon degrading bacterium and its screening method and application
  • A polycyclic aromatic hydrocarbon degrading bacterium and its screening method and application

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: strain morphological characteristics

[0027] Streak inoculate a single colony into HLB solid medium, place the plate upside down in a constant temperature incubator, and incubate at 28°C for 48 hours. The colony is round, yellow and opaque, with smooth and moist surface, regular edges, no halo, and a raised center. 2~3mm in diameter, see figure 1 .

Embodiment 2

[0028] Embodiment 2: Screening and identification of bacterial strains

[0029] (1) Pacific deep-sea sediments (sample number 45II-CC-S06-MC01, 4-6cm away from the surface layer of sediments, CC sea area of ​​the second voyage of Oceania 45 voyage, 153°23.1205′W, 12°58.1135′N), The sediment is yellow-brown, odorless, weakly viscous, and the surface layer is semi-fluid, with a slightly silty feeling when rubbed by hand. After the sediment samples were aseptically collected, they were cryopreserved and then transported to the laboratory for the next stage of research.

[0030] Take the ocean surface seawater and sterilize it with high-pressure steam at 121°C for 20 minutes, then add 1% volume of sterile NH 4 NO 3 solution (100g / L) and KH 2 PO 4 solution (10g / L, pH6.7) and 0.1% volume of FeSO sterilized by a 0.22μm filter 4 (0.4g / L), the pH value of the medium obtained at last is about 7.5. Before inoculation, 1% (V / V) polycyclic aromatic hydrocarbons (carbon source concent...

Embodiment 3

[0032] Example 3: Determination of bacterial strain polycyclic aromatic hydrocarbons and alkane degradation ability

[0033] Take the ocean surface seawater and sterilize it with high-pressure steam at 121°C for 20 minutes, then add 1% volume of sterile NH 4 NO 3 solution (100g / L) and KH 2 PO 4 solution (10g / L, pH6.7) and 0.1% volume of FeSO sterilized by a 0.22μm filter 4 (0.4g / L), the pH value of the medium obtained at last is about 7.5. Add 1% (V / V) alkanes and polycyclic aromatic hydrocarbons (carbon source concentration is 50mg / L) before inoculation

[0034] After cultivating for five days, open the bottle mouth, obtain the mass spectrogram of the sample by GC-MS (gas chromatography-mass spectrometry), measure the residual amount of phenanthrene, and obtain a degradation rate of 50% to 60%. The residual amount of pyrene was measured, and the degradation rate was 60%-70%. The degradation rate of 16-alkane is 10%-20%, the degradation rate of 12-alkane is 15%-25%, and ...

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Abstract

A polycyclic aromatic hydrocarbon degrading bacterium and its screening method and application relate to marine petroleum hydrocarbon degrading bacteria. In order to transform the heterologous alcanivorax (Alcanivorax xenomutans) 45II-AX, transform the heterologous alcanivorax into a Gram-negative strain of the genus Alcanivorax, with biological characteristics of non-fermentative, obligate aerobic, The shape of the bacteria is non-bacillus, the colony is round, yellow and opaque, the surface is smooth and moist, the edges are regular, no halo, the center is raised, the diameter is 2-3mm, the optimum growth conditions are: pH=7.0-8.5, temperature 25 ~28°C; can be used in the degradation of organic compounds. It was identified that the 16S rDNA gene sequence of a transformed heterologous alcanivorax was 99.93% similar to the type strain Alcanivorax xenomutans JC109(T).

Description

technical field [0001] The invention relates to marine petroleum hydrocarbon degrading bacteria, in particular to a strain of polycyclic aromatic hydrocarbon rock degrading bacteria and its screening method and application. Background technique [0002] Because crude oil contains unstable hydrocarbons, including polycyclic aromatic hydrocarbons, toluene, ethylbenzene and xylene, these harmful substances can enter the blood vessels of the human body through breathing, skin absorption and eyeballs, which may cause nausea, headache and dizziness Unwell feeling, even cancer, so this oil spill is a great threat to the health of nearby residents. Oil spill pollution accidents will also cause severe hypoxia in large areas of the sea, destroy marine productivity, cause a large number of fish, shrimp, shellfish and seabirds to die, and cause local "ocean desertification"; Cause deserted beaches, destroy mariculture and salt pan production, pollute or destroy coastal tourist areas, a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20B09C1/10C02F3/34C12R1/01C02F101/32
CPCB09C1/10C02F3/343C02F3/344C02F2101/32C12N1/20C12N1/205C12R2001/01
Inventor 邵宗泽王万鹏曾令宇
Owner THIRD INST OF OCEANOGRAPHY MINIST OF NATURAL RESOURCES
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