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Composite inoculant for degrading petroleum, preparation method and applications thereof

A technology of compound bacterial agent and oil, applied in the field of oil pollution control

Active Publication Date: 2016-07-06
YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, most of the studies of domestic and foreign scholars focus on the screening of oil-degrading strains, oil-degradation mechanism, and soil remediation technology. These studies have gradually confirmed the feasibility of microbial remediation of oil-contaminated soil, but few studies have involved the production of bacterial agents.

Method used

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  • Composite inoculant for degrading petroleum, preparation method and applications thereof
  • Composite inoculant for degrading petroleum, preparation method and applications thereof
  • Composite inoculant for degrading petroleum, preparation method and applications thereof

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preparation example Construction

[0040] The preparation method of the above-mentioned composite bacteria agent for degrading petroleum of one embodiment comprises the following steps:

[0041] S10, insert the seed solution of Acinetobacter venezius into the LB culture medium of pH4.5-pH5.5 with the inoculum size of 8%-12%, feed sterile air, place at 28°C-32°C, ventilation volume 0.20m 3 / h-0.30m 3 Fermenting and culturing for 14h-18h under the conditions of stirring speed of 140rpm-160rpm per hour and stirring speed of 140rpm-160rpm, to obtain the Acinetobacter venicei fermentation culture liquid.

[0042] The preparation method of the seed solution of Acinetobacter venezius is as follows:

[0043] S110. Introduce the oil-contaminated soil collected from Dongying into sterilized LB culture solution, culture on a shaking table at 28°C-32°C for 22h-26h, and obtain Acinetobacter venezia after separation and purification.

[0044] In S110, the method of separation and purification is as follows:

[0045] Use ...

Embodiment 1

[0080] (1) Isolation of Acinetobacter venezia and Dietzella: Put the oil-contaminated soil collected from Dongying into the sterilized LB culture medium, cultivate it on a shaker at 30°C for 24 hours, and obtain Acinetobacter venezia and Dietzella sp.

[0081] (2) Test tube culture medium: Sterilized liquid LB medium was used to inoculate Acinetobacter veniceus into a test tube culture medium at pH 4.5, and after inoculation, it was placed on a shaker at 30° C. for 24 h of activation. Using sterilized liquid LB medium, Dietzella was inoculated into a test tube medium with pH 7.5, and after inoculation, it was placed on a shaker at 30°C for activation and culture for 24 hours.

[0082] (3) Expansion medium: use sterilized liquid LB medium, transfer the Acinetobacter venezius in the test tube culture medium to the Erlenmeyer flask medium at pH 4.5, place it in a shaker at 30°C, and culture it with shaking 24h, as seed solution. Using sterilized liquid LB medium, the Dietzella ...

Embodiment 2

[0088] (1) Isolation of Acinetobacter venezia and Dietzella: Introduce oil-contaminated soil into sterilized LB culture medium, culture on a shaking table at 28°C for 26 hours, and obtain Acinetobacter venezia after separation and purification and Dietzella.

[0089] (2) Test tube culture medium: Sterilized liquid LB medium was used to inoculate Acinetobacter veniceus into a test tube culture medium at pH 4.5, and after inoculation, it was placed in a shaker at 28°C for 26 hours of activation. Using sterilized liquid LB medium, Dietzella was inoculated into a test tube medium with pH 7.0, and placed in a shaker at 28°C for activating culture for 26 hours after inoculation.

[0090] (3) Expansion medium: use sterilized liquid LB medium, transfer the Acinetobacter venezius in the test tube culture medium to the Erlenmeyer flask medium at pH 4.5, place it in a shaker at 28°C, and shake it. 26h, as seed solution. Using sterilized liquid LB medium, transfer the Dietzella in the t...

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Abstract

A composite inoculant for degrading petroleum is characterized by comprising acinetobacter venetianus LCL-1 and dietzia sp. CN-3, wherein the ratio of live acinetobacter venetianus LCL-1 to live dietzia sp. CN-3 is 1-50:10-200. The strain of acinetobacter venetianus LCL-1 is preserved in China Center for Type Culture Collection (CCTCC), the preservation number is M 2015538; and the strain of dietzia sp. CN-3 is preserved in China Center for Type Culture Collection (CCTCC), and the preservation number is M 2015537. The acinetobacter and dietzia are separated from soil polluted by petroleum, both have an efficient petroleum degrading performance, and can well cooperate with each other. Through the relationships such as commensalism, mutualism, and the like, the shortage of low degradation rate of single bacteria is overcome, and the petroleum degrading performance is largely improved. Moreover, the invention also provides a preparation method of the composite inoculant and an application of the composite inoculant in petroleum degradation.

Description

technical field [0001] The invention relates to the technical field of petroleum pollution control, in particular to a composite microbial agent for degrading petroleum, a preparation method and application thereof. Background technique [0002] Petroleum is the "blood" of modern industry, but with the increasing amount of oil extraction, a large amount of oil enters the environment during extraction, transportation, storage and accidental leakage, seriously polluting soil, groundwater, rivers and oceans. Especially in recent decades, petroleum has become one of the main pollutants in the environment such as soil and water. Among the remediation methods of oil pollution, microbial remediation is a low-investment, high-efficiency, easy-to-apply, and great-development-potential method for a clean environment because of its good treatment effect, low cost, strong adaptability, and no secondary pollution. Emerging Technologies. [0003] A lot of related research has been carri...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/01
CPCB09C1/10C12N1/20
Inventor 胡晓珂梁成林
Owner YANTAI INST OF COASTAL ZONE RES CHINESE ACAD OF SCI
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