Solid microbe agent for degrading petroleum pollution, and petroleum products, and preparation method

A microbial agent and solid technology, which is applied in the field of bioremediation of oil-contaminated soil and emergency treatment of oil product leakage accidents, can solve the problems of soil secondary pollution and limited applicability, and achieve rich nutrition and easy operation in the preparation process , loose texture effect

Active Publication Date: 2007-10-10
山东百科利生态科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, fungi tend to produce toxic intermediate metabolites (such as trans-diol) in the process of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1: the preparation of mixed bacterial fermentation broth

[0028] The bacteria used were Bacillus subtillus and Sphingobacterium multivolum, respectively. Bacterial fermentation broth is obtained by culturing on inclined planes, activating culture in shake flasks, and fermentation in shake flasks respectively. The specific operation steps are as follows:

[0029] (1) Slant culture medium: Prepare beef extract peptone solid medium by conventional methods. The specific steps are: 3g beef extract, 5g peptone, 5g NaCl, 1000ml distilled water, pH7.2~7.4, 1.5%~2% agar, stir well , at 1.05kg / cm 2, Sterilize at 121.3°C for 20-30 minutes. Make slopes after sterilization, and inoculate the preserved strains onto the slopes of test tubes on ultra-clean workbench respectively, culture the strains at 25-30°C for 16-32 hours, and wait for the bacteria lawn to cover the slopes for later use;

[0030] (2) Preparation of seed liquid: the liquid medium is still beef extract...

Embodiment 2

[0032] Embodiment 2: the preparation of mixed bacterial fermentation broth

[0033] As described in Example 1, the difference is that in step (3), the ratio of the seed liquid of the two strains of bacteria is changed, and the mixing volume ratio of Bacillus subtilis and Sphingobacterium multivorum is 2.0: 1, and high yield is obtained under this condition. Active mixed fermentation broth, the effective number of viable bacteria of Bacillus subtilis is 10 6 ~10 10 individual / ml, the effective number of viable bacteria of Sphingobacterium multivorum is 10 6 ~10 12 pieces / ml.

Embodiment 3

[0034] Embodiment 3: Two strains of bacteria are tested for the degradation ability of crude oil

[0035] Pick a ring of bacterial lawn from the two strains of activated bacteria respectively, activate it in beef extract peptone liquid medium for 16-32 hours, make a certain concentration of bacterial suspension, and then use 2-10% The inoculum amount was transferred to crude oil medium, placed on a shaker at 20-30°C for 5-12 days, and the rotation speed was 150-250rpm. The culture medium without bacteria was used as blank control. At the end of the cultivation, add dichloromethane (DCM) to the culture medium, centrifuge to break the oil-in-water emulsion, filter and dehydrate the organic phase with granular anhydrous sodium sulfate, place it in a pre-constant weight beaker, blow it off with nitrogen at room temperature until constant Weight, the residual oil content in the culture medium was determined by gravimetric method. Adopt the following formula to calculate the biode...

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PUM

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Abstract

This invention discloses a solid microbial agent for degrading petroleum pollutants and petroleum products. The solid microbial agent is prepared by: mixing primary seed solutions of Bacillus subtilis and Sphingobacterium multivorum at a volume ratio of (0.5-20):1, fermenting to obtain mixed bacterial fermentation solution, and mixing the mixed bacterial fermentation solution and turfy soil at a weight ratio of (0.1-1):1. The active living bacteria number of Bacillus subtilis is 106-1010/mL, and that of Sphingobacterium multivorum is 107-1012/mL. The solid microbial agent has such advantages as high activity in soil, rapid growth and no secondary pollution to environment, and can be used for biological bioremediation of petroleum polluted soil and petroleum product leakage fields.

Description

technical field [0001] The invention relates to a solid microbial bacterial agent for degrading petroleum pollutants and petroleum products, and belongs to the technical field of bioremediation for emergency treatment of petroleum-contaminated soil and petroleum product leakage accidents. Background technique [0002] Bioremediation refers to the addition of some substances (such as nutrients, O 2 , microorganisms) or biotechnology that enhances the ability of these substances to accelerate the natural biodegradation process. As early as 1976, this bioremediation method was successfully used to control the oil spill caused by the "Queen Mary" at the Cape of Good Hope. In 1989, the bioremediation performed by Exxon valdez in the treatment of the oil tanker spill accident in the Gulf of Alaska was widely recognized by the society, and the bioremediation technology has developed rapidly since then. [0003] Bioremediation is a low-consumption, high-efficiency and environmenta...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/125
Inventor 王加宁李超敏黄国强杨合同孔学张新建李鑫钢廉景燕
Owner 山东百科利生态科技有限公司
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