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High-density ferment method for petroleum hydrocarbon degradation bacterium

A technology of petroleum hydrocarbon degrading bacteria and high-density fermentation, applied in biochemical equipment and methods, restoration of contaminated soil, microorganisms, etc., can solve the problems of low cost, achieve easy operation, increase the amount of bacteria, and reduce bioremediation cost effect

Active Publication Date: 2008-02-27
CHINA PETROLEUM & CHEM CORP +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The method is low in cost, simple in operation, and can produce a large number of petroleum hydrocarbon-degrading bacteria in a short time, which solves the problem of bacterial strains in the treatment of large-scale oil-polluted soil and the treatment of sudden oil pollution accidents

Method used

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  • High-density ferment method for petroleum hydrocarbon degradation bacterium

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preparation example Construction

[0041]The preparation steps of the fermented seed liquid are as follows: pick 2-3 activated colonies from the plate medium, place them in a 300mL shake flask filled with 50mL primary seed medium, and place them at a temperature of 32-37°C. Cultivate for 8-10 hours in a shaker with a rotational speed of 200rpm to obtain a culture solution; then divide the above seed solution into four 300mL shake flasks filled with 50mL of secondary seed medium, and cultivate for 10-12h under the same conditions. That is, the fermented seed liquid is obtained.

[0042] Preparation and inoculation before the fermentation: Dissolve the batch fermentation medium except glucose in 3.5L deionized water, pour it into the fermenter, calibrate the pH meter, sterilize, and correct the dissolved oxygen DO to be 0 during the sterilization process %; under the protection of the flame circle, the petroleum hydrocarbon degrading bacteria (Enterobacter cloacae, Enterobactere cloacae, screened and preserved fr...

example 1

[0052] Example 1: A high-density fermentation method of petroleum hydrocarbon degrading bacteria

[0053] This highly efficient petroleum hydrocarbon degrading bacteria was screened and preserved from the contaminated soil of Zhongyuan Oilfield in our laboratory.

[0054] ①Seed cultivation

[0055] Pick 2-3 activated colonies from the plate medium and place them in a 300mL shake flask filled with 50mL primary seed medium, and culture them in a shaking table with a temperature of 34°C and a rotation speed of 200rpm for 8-10h. Obtain the primary seed culture solution; divide the primary seed culture solution equally into four 300mL shake flasks filled with 50mL secondary seed medium, and cultivate under the same conditions for 10-12h. That is, the fermented seed liquid is obtained.

[0056] ② Preparation before planting

[0057] Dissolve the batch fermentation medium except glucose in 3.5L deionized water, pour it into the cleaned fermenter, calibrate the pH meter, sterilize,...

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Abstract

The invention discloses a high-density fermenting method of petroleum hydrocarbon degraded bacteria in the biological fermenting technical domain, which is characterized by the following: activating slope bacteria; enlarging the cultured seed liquid; preparing before inoculating; inoculating; supplementing the ferment culture medium batch by batch; adjusting the even growing speed of bacteria; reducing cost; simplifying the operation; producing large amount of petroleum hydrocarbon degraded bacteria within short period; solving the problem of soil pollution and the accident of petroleum pollution.

Description

technical field [0001] The invention belongs to the technical field of biological fermentation, in particular to a high-density fermentation method of petroleum hydrocarbon degrading bacteria. Background technique [0002] Bioremediation technology is the main remediation technology for soil contaminated by petroleum and related products. Due to the characteristics of complex composition, refractory biodegradation and low bioavailability of petroleum pollutants, it is difficult for indigenous microorganisms to degrade petroleum hydrocarbons in soil effectively, rapidly and thoroughly. The bio-addition method can improve the efficiency of bioremediation and effectively remove petroleum hydrocarbon pollutants by adding microorganisms with high-efficiency degradation of petroleum hydrocarbons. However, for the treatment of large-scale oil-contaminated soil or the treatment of sudden oil pollution accidents, there will be a large demand for microorganisms with high-efficiency d...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/00B09C1/10
Inventor 花秀夫刘铮张坤徐圆圆张敏莲
Owner CHINA PETROLEUM & CHEM CORP
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