Microbial compound bacteria agent for treating oilfield fracturing flowback fluid, and preparation method of microbial compound agent

A technology of microorganisms and liquid bacteria, applied in the biological field, can solve the problems of secondary pollution and difficult degradation of fracturing flowback fluid, and achieve the effect of clear water quality, high degradation rate and reduction of secondary pollution.

Active Publication Date: 2012-11-14
AEGIS GASOLINEEUM TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 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 fracturin

Method used

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  • Microbial compound bacteria agent for treating oilfield fracturing flowback fluid, and preparation method of microbial compound agent
  • Microbial compound bacteria agent for treating oilfield fracturing flowback fluid, and preparation method of microbial compound agent
  • Microbial compound bacteria agent for treating oilfield fracturing flowback fluid, and preparation method of microbial compound agent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: Separation and purification of Cladosporium mold and Pyrobacterium bovis

[0023] 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.

[0024] 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

[0025] Example 2 Sequencing and identification of strain EB1

[0026] 1. Genome extraction and electrophoresis detection

[0027] (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.

[0028] The extraction steps are described as follows:

[0029] 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 65°C preheated FPCB Solution and 7 μl β-mercaptoethanol in sequence, mix thoroughly, and incubate at 65°C for 25 minutes, mixing occasionally.

[0030] 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.

[0031] 3. Pipette the supernatant into a clean 1.5ml centrifuge tube, add 40μl RNase A (20mg / ml), mix well, and place at room t...

Embodiment 4

[0112] Example 4 A preparation method of a microbial composite bacterial agent for treating oil field fracturing flowback fluid

[0113] The strains with the preservation number CGMCC No: 5782 and the preservation number CGMCC No: 5783 were put into two mediums for shaker culture respectively. The medium was composed of water, and each liter of water contained 0.1g-5g of guanidine Glue, 0.05g~1g of KH 2 PO 4 , 0.05g ~ 2g of K 2 HPO 4 , 0.01g~0.1g of MgSO 4 , 0.01g ~ 0.5g of CaCl 2 , 0.01g ~ 0.2g of NaCl, the cultivation time is 3 ~ 10 days, the cultivation temperature is 5 ~ 30 ° C, and then the inoculum amount is 5% ~ 15% by volume in the two reactors equipped with the above medium Inoculate the above two strains respectively, culture continuously for 8-48 hours to obtain a liquid bacterial liquid, and then mix the liquid bacterial liquid containing Cladosporium and the liquid bacterial liquid containing Bacillus bovis according to the ratio of 1 to 5:1 by volume Compou...

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Abstract

The invention provides cladosporium sp. of CGMCC (China General Microbiological Culture Collection) No. 5782. The invention also provides phyllobacterium myrsinacearum of CGMCC No. 5783. The invention also provides a microbial compound bacterium agent containing the two strains for treating an oilfield fracturing flowback fluid, and a preparation method of the microbial compound agent. The two strains are respectively put in two specific culture media for shaking culture, and two strains are respectively inoculated into two reactors filled with the specific culture media according to the inoculation amount of 5% to 15%, and are continuously cultured for 8 to 48 hours to obtain a liquid bacterium; and then the cultured liquid bacterium is compounded according to the volume ratio of cladosporium sp. to phyllobacterium myrsinacearum of 1 to 5:1 to obtain the microbial compound bacterium agent. The invention can effectively solve the problem that macromolecular organic compounds in the oilfield fracturing flowback liquid is difficult to biodegrade, and reduces secondary pollution of the environment.

Description

Technical field: [0001] 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: [0002] 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 organic compounds, the COD value concentration of the fracturing fluid is high, and the treatment technology is difficult. [0003] At present, domestic treatment methods for oilfield fracturing flowback fluid mainly include the following points: [0004] 1) Incineration: It mainly deco...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12N1/20C02F3/34C12R1/645C12R1/01C02F103/10
Inventor 李松棠王婷婷陈宏宇张方元单煜巽
Owner AEGIS GASOLINEEUM TECH
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