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A strain of Bacillus licheniformis resistant to high temperature degradation of crude oil to produce emulsifier and its application

A technology of Bacillus licheniformis and a biological emulsifier, which is applied in the fields of energy biotechnology and environmental biology, can solve the problem of not many bacterial species, and achieves improved colloid content, high emulsifying ability and temperature resistance, and improved crude oil recovery. Effect

Active Publication Date: 2019-09-03
CHINA UNIV OF PETROLEUM (BEIJING)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

For example, CN104593298A discloses a novel strain of thermophilic and salt-tolerant degradation of crude oil to produce emulsifiers and its application. Although the bacteria can grow at high temperature and produce bio-emulsifiers, the mass fraction of the optimum salt for the bacteria is 3.0% and Bioemulsifier cannot be produced at high temperature of 80°C to efficiently degrade crude oil
CN105441351A discloses a thermophilic surfactant-producing oil-degrading bacterium and its application. Although the bacterium also belongs to Bacillus licheniformis and can grow and produce bio-emulsifier at high temperature, the bacterium can only reach 70000 mg / L of salinity. salinity and the resulting bioemulsifier is only effective at emulsifying diesel at a temperature of 55°C
Among some petroleum microorganisms that have been isolated so far, there are not many strains that can degrade crude oil under anaerobic conditions, are thermophilic and salt-tolerant, and can produce biological emulsifiers at the same time.

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  • A strain of Bacillus licheniformis resistant to high temperature degradation of crude oil to produce emulsifier and its application
  • A strain of Bacillus licheniformis resistant to high temperature degradation of crude oil to produce emulsifier and its application
  • A strain of Bacillus licheniformis resistant to high temperature degradation of crude oil to produce emulsifier and its application

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Effect test

Embodiment 1

[0046] Screening, isolation, identification and growth curve determination of the high temperature resistant oil production bacteria T6189 of the present invention

[0047] The first step: Screening of high temperature resistant oil production bacteria T6189

[0048] The screening method of high temperature-resistant oil production bacteria T6189: according to the crude oil inorganic salt medium (its composition is: NaNO 3 10g / L, Na 2 MO 4 0.08g / L, FeSO 4 0.12g / L, MgSO 4 0.2g / L, CaCl 2 0.12g / L, KH 2 PO 4 1.0g / L, (NH 4 ) 2 HPO 4 1.0g / L, crude oil (derived from Xinjiang T6189 well production fluid) 2% (V / V), sucrose 0.2‰ (W / V) (the amount of crude oil and sucrose added is based on 100% of the inorganic salt medium); pH value Prepare medium for 7.0-7.2) formula, wherein add 0.2‰ sucrose in the medium to quickly activate the bacteria so that the screening process can be carried out quickly.

[0049] The experimental steps are as follows:

[0050] (1) Pack the pre...

Embodiment 2

[0071] Effects of Different pH Values, Temperatures and Salinity on the Growth of Thermostable Oil Production Bacteria T6189

[0072] (1) Effect of pH value on the growth of the bacterium: ① prepare LB liquid culture medium, pack in 150mL Erlenmeyer flasks, and the volume of each bottle is 100mL; use 1mol / L hydrochloric acid and 1mol / L sodium hydroxide to adjust each The pH values ​​in the bottle are 5, 6, 7, 8 and 9 respectively; ② Sterilize in a high-temperature and high-pressure sterilizer, and insert the seed liquid (that is, the bacterial liquid after LB medium activates the T6189 strain) according to the inoculum amount of 2wt% , and then cultivated at 55°C and 150rpm for 48 hours; ③ After the cultivation, take 4mL of the culture solution in a centrifuge tube, centrifuge to wash the precipitate and suspend the bacteria with an equal volume of distilled water, measure its absorbance under the condition of 600nm in a UV spectrophotometer and observe its Bacterial concentra...

Embodiment 3

[0077] Metabolic characteristics of oil production bacteria T6189 with high temperature tolerance to crude oil

[0078] Step 1: Microbial metabolism and extraction separation of crude oil

[0079] The crude oil used in the present invention is respectively from the oil samples of Well 4001, Well 4024, Well 4027, Well 4032 and Well 4045 in Henan Oilfield, wherein the oil samples of Well 4027 are liquid at normal temperature, and the oil samples of other oil wells are all solid at normal temperature . The specific experimental steps are as follows:

[0080] (1) Microbial metabolism of crude oil: 1. Prepare the medium according to the inorganic salt medium, add 0.2wt% sucrose in the medium as a rapid microbial activator, and then add 0.5wt% of different crude oils to each 100mL culture medium Sealed in a 150mL Erlenmeyer flask, and sterilized at 121°C for 15min; ②Take out each Erlenmeyer flask containing different crude oils and cool them in an ultra-clean workbench, and inocul...

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Abstract

The invention provides a strain of bacillus licheniformis capable of producing emulsifiers resistant to high temperature degradation of crude oil and applications thereof. Specifically, the present invention relates to Bacillus licheniformis with a deposit number of CGMCC No. 13054 and a Bacillus licheniformis preparation containing the bacterium. The present invention also relates to the application of the bacillus licheniformis or its preparation in fermenting and cultivating to produce biological emulsifiers, in degrading crude oil and flooding oil, and in emulsifying oil products. The Bacillus licheniformis of the present invention can grow with crude oil as carbon source and energy source, and synthesize biological emulsifier; the strain and its metabolites can effectively degrade the content of saturated components and aromatic components in crude oil; the biological emulsifier produced by the bacteria has Excellent high temperature resistance and strong emulsifying ability; the strain and the bioemulsifier produced by it can be used in microbial oil recovery to improve oil recovery.

Description

technical field [0001] The invention belongs to the fields of energy biotechnology and environmental biotechnology. Specifically, the invention relates to a strain of Bacillus licheniformis and its application. The agent can be used in microbial oil recovery, has the advantages of high temperature resistance and salt resistance, and can effectively degrade crude oil and improve oil recovery. Background technique [0002] Biotechnology has been more and more used in the field of oil extraction, microbial oil recovery technology is one of them. Microbial Enhanced Oil Recovery (MEOR) technology can further enhance oil recovery and prolong oilfield development life, and has broad application prospects. Compared with other oil recovery technologies, this technology has the advantages of low cost, wide application range, stable biological activity, long effective action time, and environmental friendliness. Microbial oil recovery technology is to improve the oil recovery by chan...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P1/04C09K8/582C12R1/10
CPCC09K8/582C12N1/20C12P1/04C12N1/205C12R2001/10
Inventor 孙珊珊张忠智朱攀峰崔凯董浩罗一菁张志勇侯吉瑞刘同敬王菁王欣怡
Owner CHINA UNIV OF PETROLEUM (BEIJING)
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