Screening method for petroleum degrading bacteria, method for preparing petroleum degrading bacteria inoculant from screened bacteria, and application of inoculant

A technology of petroleum degrading bacteria and screening method, which is applied in the field of screening methods and the preparation of petroleum degrading bacteria inoculants by the bacteria screened therewith, can solve the problems of less selection, high strength, low oil and the like

Active Publication Date: 2015-03-04
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with liquid fermentation, solid-state fermentation has the following disadvantages: limited to microorganisms resistant to low water activity, less selection; slow fermentation rate and long cycle; complex natural raw material components, sometimes changing, affecting yield quality; difficult to detect and control process parameters; The operation consumes a lot of labor and is intensive
In this experiment, 1, 2, and 3 g of petroleum were added to 100ml of culture medium. In the absence of a solubilizer, the crude oil was almost insoluble. The concentration of petroleum in the medium was very low, and the pet

Method used

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  • Screening method for petroleum degrading bacteria, method for preparing petroleum degrading bacteria inoculant from screened bacteria, and application of inoculant
  • Screening method for petroleum degrading bacteria, method for preparing petroleum degrading bacteria inoculant from screened bacteria, and application of inoculant
  • Screening method for petroleum degrading bacteria, method for preparing petroleum degrading bacteria inoculant from screened bacteria, and application of inoculant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Screening of Efficient Petroleum Degrading Bacteria Strains

[0065] 1. Collection of oil-degrading indigenous bacteria sources

[0066] The oil-contaminated soil near the oil storage tank of the Shengli Oilfield refinery in Shandong Province was used as the source of indigenous oil-degrading bacteria. Sampling method: Use a small shovel to take soil samples with a thickness of 10cm, mix them evenly after sampling at multiple points, take 3kg soil samples and bring them back to the laboratory for later use.

[0067] 2. Preparation of Petroleum Polysorbate-80 Enhanced Solution

[0068] After the surfactant reaches the critical micelle concentration (CMC) in the aqueous solution, the solubility of some water-insoluble or slightly soluble substances in the micellar solution can be significantly increased to form a transparent colloidal solution. This effect is called solubilization. (solubilization). Polysorbate-80 (polyoxyethylene sorbitan fatty acid ester) is a water-...

Embodiment 2

[0117] Liquid fermentation production of petroleum degrading bacterial agent

[0118] 1. Primary seed cultivation

[0119] Under sterile conditions, the mixed strain 2 of petroleum-degrading bacteria purified in Example 1 was inoculated in the slant culture medium of the test tube. Culture medium composition Per 1000ml sterile water: petroleum polysorbate-80 enrichment solution containing petroleum 0.125g equivalent (wherein petroleum mass concentration is 2%, polysorbate-80 mass concentration is 4%, remainder is water.), beef Cream 3g, peptone 10g, NaCl 5g, agar powder 20g, pH value is 7, sterilized at 121°C for 20 minutes. The culture conditions are: 30°C, cultured for 24 hours, and used as first-class seeds.

[0120] 2. Secondary seed cultivation

[0121] Under sterile conditions, the two-ring primary seeds were inserted into a 500ml Erlenmeyer flask filled with 200ml medium for shaking flask culture. Shake flask medium composition per 1000ml sterile water: petroleum po...

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Abstract

The invention relates to a screening method for petroleum degrading bacteria. An enrichment medium, a screening medium and a domestication medium, which are used in the method, are media employing petroleum as a unique carbon source, and specifically, petroleum polysorbate-80 solubilizing fluid is specifically added into the media. The method has the beneficial effects that a nonionic surfactant polysorbate-80 is used as a solubilizing agent, so that the dissolution effects of petroleum in water can be improved, and the problem of difficulty in the dissolution of petroleum in the media is solved; petroleum is actually the unique carbon source, and the petroleum polysorbate-80 solubilizing fluid is used as the carbon source of the media in processes of primary screening, domestication, purification and the like of the petroleum degrading bacteria; the screening of the petroleum degrading bacteria comprises the processes of primary screening, domestication, purification and the like, a strain is gradually domesticated to be adapted to growth and reproduction under the condition of employing petroleum as the unique carbon source in the domestication medium of which the petroleum concentration gradient gradually rises, and tolerance to petroleum and the degradation efficiency are improved.

Description

technical field [0001] The invention relates to the technical field of microbial remediation of petroleum-contaminated soil, in particular to a screening method for petroleum-degrading bacteria, a method for preparing a petroleum-degrading bacterial agent from the screened bacteria, and an application of the bacterial agent in oil-contaminated soil remediation. Background technique [0002] Petroleum is a complex mixture containing a variety of hydrocarbons (normal alkanes, branched alkanes, aromatic hydrocarbons, alicyclic hydrocarbons) and a small amount of other organic compounds (sulfides, nitrides, naphthenic acids, etc.), and is the most important chemical substance for human beings. One of energy. In the process of oil extraction and transportation, oil leakage is often accompanied, causing environmental pollution. Especially polycyclic aromatic hydrocarbons (Polycyclic Aromatic Hydrocarbon-bon, PAHs) have become a huge threat to human health due to their poor bioava...

Claims

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Application Information

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IPC IPC(8): C12N1/00B09C1/10
CPCB09C1/10C12N1/00C12N1/36
Inventor 郑庆柱田侠崔德杰崔春月
Owner QINGDAO AGRI UNIV
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