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A Crude Oil Degrading Bacteria Strain and Its Application

A crude oil and strain technology, applied in the field of microorganisms, can solve problems such as no significant results, and achieve the effects of good application potential, high degradation efficiency and simple cultivation.

Active Publication Date: 2020-09-04
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are no significant results in the investigation of crude oil degrading strains with good economic benefits in high-saline environments

Method used

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  • A Crude Oil Degrading Bacteria Strain and Its Application
  • A Crude Oil Degrading Bacteria Strain and Its Application
  • A Crude Oil Degrading Bacteria Strain and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1: halophilic bacteria Salinicola Separation, purification, screening and identification of sp.X4

[0042] Isolation and purification of strains:

[0043] (1) Weigh 1-2g of crude oil-contaminated soil samples near Qinghai Oilfield, add crude oil high-salt inorganic salt medium (the mass percentage of each component of the medium is: crude oil 0.2-0.3%, NaCl 5%, NH 4 Cl 0.4%, KH 2 PO 4 0.05%, K 2 HPO 4 0.05%, MgSO 4 ·6H 2 O 0.033%, yeast extract 0.05%, metal ion mother solution 0.1%, the rest is distilled water, and adjust the pH to 6.5±0.1), 30-40 ° C, 200 rpm for 3 days.

[0044] (2) After the medium is turbid, transfer 5mL of the culture supernatant into a fresh sterilized high-salt inorganic salt medium, and subculture continuously for 3-5 times. Draw 5mL of the culture solution after multiple cultures and add it into sterile water, and dilute it in a gradient of 10 -3 , 10 -4 , 10 -5 、10 -6 , 10 -7 、10 -8 , take 100 μL of the diluted solu...

Embodiment 2

[0056] Embodiment 2: halophilic bacteria Salinicola sp.X4 can degrade hydrocarbons in high saline environment.

[0057] In this implementation, a high-salt oil-containing inorganic salt medium was used to simulate the high-salt wastewater environment, and to explore the growth and utilization of crude oil as the only carbon source and the degradation of crude oil by the bacterial strain in the present invention.

[0058] Specific steps are as follows:

[0059] (1) the halophilic bacteria provided by the invention Salinicola sp.X4 is activated in high-salt LB medium (the mass percentage of each component of LB medium is: peptone 1%, yeast powder 0.5%, sodium chloride 5%, and the rest is distilled water), 30-40°C, 200rpm for 24h Afterwards, transfer to the gradient high-salt inorganic salt medium with 1% inoculum amount (the mass percentage of each component of the medium is: crude oil 0.2-0.3%, NH 4 Cl 0.4%, KH 2 PO 4 0.05%, K 2 HPO 4 0.05%, MgSO 4 ·6H 2 O 0.033%...

Embodiment 3

[0063] Embodiment 3: halophilic bacteria Salinicola Optimal growth environment for sp.X4

[0064] In this case, the optimal growth conditions of the strain were explored, and the specific steps are as follows:

[0065] (1) The optimum growth salinity:

[0066] The halophilic bacteria X4 provided by the present invention is activated in high-salt LB medium (the mass percentage of each component of LB medium is: peptone 1%, yeast powder 0.5%, sodium chloride 5%, and the rest is distilled water). The culture solution was transferred to LB medium with a salt concentration of 10-250 g / L at a 1% inoculation amount, and cultured at 37°C and 200 rpm for 24 hours. After the culture was completed, 1 mL of the culture solution was taken and centrifuged in a centrifuge at 12,000 rpm. After 5 minutes, remove the supernatant and resuspend the bacteria with sterile water, shake and mix well, and measure the OD of the bacteria with a UV spectrophotometer 600 .

[0067] (2) Optimum growt...

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Abstract

The present invention relates to a crude oil degrading strain and its application, and the strain is classified and named as moderate halophilic bacteria ( Salinicola sp.) X4, the deposit number is CCTCC NO: M2018773, belonging to Gram-negative bacteria. The strain is opaque, beige, and the cells of the strain are short rods. Compared with the existing crude oil degrading strains, the strains provided by the invention have high degradation efficiency, simple cultivation, are suitable for high-salt environments, and have good application potential in the treatment of high-salt oil-containing wastewater in the petroleum industry and marine crude oil leakage accidents.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a strain of crude oil degrading bacteria and its application. Background technique [0002] Petroleum, also known as the "black blood" of the earth, plays a pivotal role in the economic development of countries and regions. With the continuous development and utilization of petroleum resources, the parallel pollution problems are becoming more and more serious. In the process of petroleum exploration and development, environmental problems such as oily sewage, oily sludge, and oily soil often occur during drilling, oil production, transportation, storage, and utilization. Accompanied by different crude oil conditions, geological conditions or oil production conditions, extreme conditions such as high salt, high pH, ​​and high temperature often appear. If petroleum hydrocarbons cannot be handled well in these extreme environments, serious environmental problems...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/32
CPCC02F3/34C02F2101/32C12N1/205C12R2001/01
Inventor 李霜林佳辉张坤成郑欣然
Owner NANJING TECH UNIV
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