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Bacterial preparation for degrading petroleum and restoring petroleum polluted soil ecology and preparation method thereof

A technology of oil pollution and bacterial preparations, applied in the restoration of contaminated soil, biochemical equipment and methods, and methods based on microorganisms, which can solve the problems of unseen high density

Inactive Publication Date: 2009-12-16
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been some studies at home and abroad on the restoration of oil-contaminated soil ecology. However, there have been no reports of high-density, high-activity bacteria preparations that can be used in situ to degrade oil and restore oil-contaminated soil ecology.

Method used

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  • Bacterial preparation for degrading petroleum and restoring petroleum polluted soil ecology and preparation method thereof
  • Bacterial preparation for degrading petroleum and restoring petroleum polluted soil ecology and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Follow these steps:

[0023] 1) Activate the slant planes of the four strains stored at 4°C and culture them in shake flasks respectively. Cultured in soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.2, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 24h.

[0024] 2) Secondary liquid expansion culture (6L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 24h. .

[0025] 3) Tertiary liquid expansion culture (60 L), the medium is the same as above; the inoculum size is 10%, and cultured at 28° C. for 24 hours.

[0026] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on it. Sealed in a polyethylene bag and stored at room temperature.

Embodiment 2

[0028] Follow these steps:

[0029] 1) Activate the slant planes of the four strains stored at 4°C and culture them in shake flasks respectively. Expand the culture with soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.0, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 12h.

[0030] 2) Secondary liquid expansion culture (10L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 12h. .

[0031] 3) Tertiary liquid expansion culture (100 L), the medium is the same as above, the inoculum size is 10%, and cultured at 28° C. for 12 hours.

[0032] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on the humic acid. Sealed in a polyethylene bag and stored at room temperature.

Embodiment 3

[0034] Follow these steps:

[0035] 1) Activate the slant planes of the three strains stored at 4°C, and culture them in shake flasks respectively. Expand the culture with soluble starch medium, the specific formula is as follows: soluble starch 20g, KNO 3 1g, NaCl 0.5g, K 2 HPO 4 0.5g, MgSO 4 .7H 2 O 0.5g, FeSO 4 0.01g, distilled water 1000mL, pH7.2, 0.1Mpa, autoclave at 121℃ for 30min. Incubate at 28°C for 12~24h.

[0036] 2) Secondary liquid expansion culture (20L), replace the soluble starch in the above medium with cornmeal, add 0.1% humic acid (mass fraction), inoculum size 5%, culture at 28°C for 24h. .

[0037] 3) Tertiary liquid expansion culture (200L), the medium is the same as above, the inoculum size is 10%, and cultured at 28°C for 12~24h.

[0038] 4) Using humic acid as a carrier to fix and adsorb the bacterial solution on the humic acid. Sealed in a polyethylene bag and stored at room temperature.

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Abstract

The invention relates to a bacterial preparation for degrading petroleum and restoring petroleum polluted soil ecology and a preparation method thereof. The bacterial preparation for degrading the petroleum and restoring the petroleum polluted soil ecology is a composite colony consisting of bacillus megaterium, pseudomonas fluorescens, streptococcus faecalis and candida tropicalis through optimized combination, wherein the matching ratio of the components is 1-2:1:1:0.5-1. The bacterial preparation has the advantage that the bacterial preparation can quickly form a micro-ecological advantage colony after the bacterial preparation is applied to the soil. If 2 to 5 percent of the bacterial preparation is added relative to the weight of surface soil (a cultivation layer of about 15 to 20 centimeters), the oil removal rate in three months is more than or equal to 75 percent (dichloromethane reflux extraction and gravimetric method), and the bacterial preparation has high effective colony number and strong adaptability and has remarkable effect of restoring the petroleum polluted soil ecological environment.

Description

technical field [0001] In the field of biological technology, the present invention relates to a bacterial preparation for degrading petroleum and restoring petroleum-contaminated soil ecology and a preparation method thereof. Background technique [0002] The pollution of the surrounding environment will inevitably be caused in the oil extraction, transportation and use, especially the pollution of the soil ecosystem is becoming more and more serious. There have been some studies at home and abroad on the restoration of oil-contaminated soil ecology. However, there have been no reports of high-density, high-activity bacterial preparations that can be used on a large scale in situ to degrade oil and restore oil-contaminated soil ecology. Contents of the invention [0003] The technical problem to be solved by the present invention is to provide a high-density, high-activity microbial preparation for degrading petroleum and restoring the ecology of petroleum-contaminated so...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/16B09C1/10C12R1/125C12R1/39C12R1/46C12R1/74
Inventor 张清敏齐建超郭婷乔俊陈威
Owner NANKAI UNIV
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