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Method for degrading oil and phenanthrene through bacterial strain P51

A strain and petroleum technology, applied in the field of microorganisms, to achieve the effect of complete restoration of the marine environment and high degradability

Active Publication Date: 2018-05-18
DALIAN NATIONALITIES UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In order to solve the problem of marine oil pollution treatment in the prior art, the inventor screened oil-polluted sea areas to obtain microbial strains with oil-degrading properties, determined their classification status and evaluated their oil-degrading properties. At the same time, the research found that strain P51 was While degrading petroleum, it also has the effect of degrading phenanthrene, laying the foundation for degrading polycyclic aromatic hydrocarbon pollutants

Method used

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  • Method for degrading oil and phenanthrene through bacterial strain P51
  • Method for degrading oil and phenanthrene through bacterial strain P51
  • Method for degrading oil and phenanthrene through bacterial strain P51

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Collection and isolation of strain P51

[0034] The samples were collected from the seabed sediments in the oil-polluted sea area of ​​Xingang, Dalian. After collection, they were stored in ice boxes and quickly transported back to the laboratory for the enrichment and separation of oil-degrading microorganisms.

[0035] About 10 g of freshly collected sediment samples were weighed and added to a 250 mL Erlenmeyer flask containing 100 mL of enrichment medium, enriched at 15°C for 7 days, and then isolated by gradient dilution separation to obtain oil-degrading strains.

[0036] The culture medium consists of:

[0037] Inorganic salt medium: MgSO 4 ·7H 2 O 0.7g, NH 4 NO 3 1g, KCl 0.7g, KH 2 PO 4 2g, Na 2 HPO 4 3g, 1000mL of natural seawater, pH 7.5, add 10mL of trace element mixture after sterilization.

[0038] Trace element mixture: CaCl 2 2mg, FeCl 3 ·6H 2 O 50mg, CuSO 4 0.5mg, MnCl 2 4H 2 O0.5mg, ZnSO 4 ·7H 2 O 10mg, distilled water 1000mL.

[00...

Embodiment 2

[0042] Morphological observation and identification of strains:

[0043] Streak culture of strain P51 on enriched medium with petroleum as the only carbon source and LB medium at 15°C to single colony, and observe the colony morphology of strain P51. At the same time, the bacterial colonies were picked and fixed with 2.5% glutaraldehyde solution for 1-2 hours, and then the bacterial suspension was dropped on the silicon chip, and naturally aired to a slightly wet and semi-dry state, and then the silicon chip was rinsed with a phosphate buffer solution with a pH of 7.2 for 10 minutes. After rinsing three times, they were dehydrated with 30%, 50%, 70%, 85%, 95%, and 100% ethanol gradients respectively, and then fixed by dropping isoamyl acetate to the silicon wafer, and left overnight. On the second day, the prepared samples were coated with a film and then observed with a scanning electron microscope. like figure 1 As shown, the bacterial strain P51 is on the solid LB medium,...

Embodiment 3

[0046] Determination of Petroleum Degradation Performance of Strain P51

[0047] Evaluation of oil degradation performance of strain P51 The oil degradation rate of strain P51 was determined by ultraviolet spectrophotometry. Using the extracted petroleum ether mixture as a sample, the U-5100 ultraviolet spectrometer was used to analyze the change of petroleum content before and after strain degradation. The calculation formula of bacterial strain to the degradation rate of oil is: η=(n 0 -n 1 ) / n 0 ×100%, where η is the oil degradation rate, n 0 is the blank control, n 1 is the residual petroleum content in the extract of the culture medium inoculated with the strain.

[0048] Add 150 μL of petroleum to the enrichment medium as the only carbon source to analyze the petroleum degradation performance of the strain. The bacterial strain (bacterial suspension concentration is 10 8 cfu / ml) was inoculated into 100ml enriched medium according to the inoculation amount of 1%. A...

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Abstract

The invention relates to a method for degrading oil and phenanthrene through a bacterial strain P51. The preservation number of the bacterial strain is CGMCC No.14411, and the 16SrRNA sequence is shown in SEQ ID NO:1. The screened bacterial strain P51 has the function of efficiently degrading oil, and while the bacterial strain is used for degrading oil, the high degradability of phenanthrene in polycyclic aromatic hydrocarbon is achieved, so that the ocean pollution problem caused after oil pollution is solved; the marine environment is completely recovered, and the foundation is laid for developing microbial remediation oil pollution products.

Description

[0001] This application is a divisional application with the application number 2017110671186, the application date is October 30, 2017, and the invention title is "Petroleum and phenanthrene degrading strain P51 suitable for submarine low temperature environment". technical field [0002] The invention belongs to the technical field of microorganisms, and in particular relates to a bacterial strain P51 with the function of degrading petroleum and using the bacterial strain to control the marine environment. Background technique [0003] With the development of society and economy, human beings' requirements for oil continue to expand. Petroleum products such as gasoline, diesel, lubricating oil, paraffin, asphalt, etc. are gradually approaching human life, and various pollution incidents that follow are not uncommon. In November 2007, the Russian oil tanker "Volga Petroleum 139" carrying 4,700 tons of heavy oil encountered strong winds in the Kerch Strait, disintegrated and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C02F3/34C12N1/20C12N1/02C12R1/01C02F101/32
CPCC02F3/34C02F2101/327C12N1/02C12N1/20C12N1/205C12R2001/01
Inventor 刘秋李佳怡于基成
Owner DALIAN NATIONALITIES UNIVERSITY
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