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Phyllobacterium myrsinacearum

A fracturing technology of Mytobacterium bovis and oilfield, which is applied in the direction of bacteria, microorganisms, biochemical equipment and methods, etc., can solve the problems of difficult degradation of fracturing flowback fluid and secondary pollution, and achieve clear water quality and high degradation rate , the effect of good application prospects

Active Publication Date: 2014-06-18
AEGIS GASOLINEEUM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to provide a microbial composite bacterial agent for treating oil field fracturing flowback fluid, and the microbial composite bacterial agent for treating oil field fracturing flowback fluid should solve the problem of treating oilfield fracturing flowback fluid in the prior art. The method of liquid drainage is difficult to degrade and accompanied by technical problems of secondary pollution

Method used

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  • Phyllobacterium myrsinacearum
  • Phyllobacterium myrsinacearum
  • Phyllobacterium myrsinacearum

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: Separation and purification of Cladosporium mold and Pyrobacterium bovis

[0024] Taking the oilfield fracturing fluid flowback fluid as the sampling source, the fracturing fluid flowback fluid was diluted to 10 -1 ~10 -10 Dilute it, spread it on a solid medium containing 3% high-molecular organic compound guar gum, and cultivate it at a temperature of 30°C for 5 days.

[0025] A single colony was picked on the above plate and separated and purified by streaking on the plate to obtain two strains EB1 and EB2 with excellent traits.

Embodiment 2

[0026] The sequencing and identification of embodiment 2 bacterial strain EB1

[0027] 1. Genome extraction and electrophoresis detection

[0028] (1) Genome extraction process: Extract according to the method described in the instruction manual of the SK1375 Fungal Genome DNA Extraction Kit of Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0029] The extraction steps are described as follows:

[0030] 1. Take 50-100mg of fresh fungus or 20mg of dried fruiting bodies or mycelia, and grind them into powder in liquid nitrogen. Add 700 μl of FPCB Solution preheated at 65°C and 7 μl of β-mercaptoethanol in sequence, mix well, incubate at 65°C for 25 minutes, and mix occasionally.

[0031] 2. Centrifuge at 12,000rpm for 10min, transfer the supernatant to a clean 1.5ml centrifuge tube, add 700μl chloroform, mix well, and centrifuge at 12,000rpm for 5min.

[0032] 3. Pipette the supernatant into a clean 1.5ml centrifuge tube, add 40μl RNase A (20mg / ml), mix well, an...

Embodiment 3

[0075] The sequencing and identification of embodiment 3 bacterial strain EB2

[0076] 1. Genome extraction and electrophoresis detection

[0077] (1) Genome extraction process: Extract according to the method described in the instruction manual of the SK1201-UNIQ-10 Column Bacterial Genomic DNA Extraction Kit of Shanghai Sangon Bioengineering Technology Service Co., Ltd.

[0078] The extraction steps are described as follows:

[0079] Steps:

[0080] 1. Adjust the water bath to 56°C and 70°C before use, and place the eluent Elution buffer in a 60°C water bath;

[0081] 2. Take overnight cultured bacterial cells (up to 2*109 cells), add them to a centrifuge tube, centrifuge at 10,000 rpm for 30 seconds at room temperature, collect the bacteria, and suck off the supernatant as much as possible;

[0082] 3. Add 180ul solution digestion buffer, resuspend the bacteria, add 20ul proteinase K solution (10mg / ml), mix well, and completely lyse the cells at 56°C for 30 minutes;

[...

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Abstract

The invention provides a phyllobacterium myrsinacearum which has a preservation number of CGMCC (China General Microbiological Culture Collection Center) No: 5783. Cladosporium sp. which has a preservation number of CGMCC No: 5782 and the phyllobacterium myrsinacearum strain are respectively put into two culture media to carry out shake cultivation, then respectively inoculated to two reactors of the specific culture media according to the inoculation size of 5-15 percent by volume and continuously cultured for 8-48 hours to obtain bacteria liquid, and then the well-cultured bacteria liquid is compounded to obtain a compound microbial agent. Microorganisms are constantly suitable for the growth environment of oilfield wastewater through growth, propagation and domestication of the microorganisms in fracturing fluid flowback liquid, and finally, the purpose of degrading the fracturing fluid flowback liquid to the chemical oxygen demand of is achieved. The phyllobacterium myrsinacearum can be used for effectively degrading macromolecules and organic matters in the fracturing fluid flowback liquid, the degradation rate and the water clarity are high, and secondary pollution to the water body environment and the soil environment is reduced.

Description

[0001] This application is a divisional application with the application number "201210264645.7", the filing date of "July 27, 2012", and the title of the invention "a microbial composite bacterial agent for treating oilfield fracturing flowback fluid and its preparation method". Technical field: [0002] The invention relates to the field of biotechnology, in particular to microorganisms and their composite bacterial agents, in particular to a microbial composite bacterial agent for treating oil field fracturing flowback fluid and a preparation method thereof. Background technique: [0003] Generally, the main measure to increase oil and gas well stimulation in oil fields is to use fracturing fluid for oil and gas wells. However, in the process of fracturing, a large amount of oilfield fracturing flowback fluid will be produced, and the composition of fracturing fluid flowback fluid is relatively complex. Since the fracturing fluid contains a large amount of polymeric organi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 李松棠王婷婷陈宏宇张方元单煜巽
Owner AEGIS GASOLINEEUM TECH
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