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Microbacterium bn-88, microbial agent and its application in degrading petroleum hydrocarbons

A microbial inoculum and microbacteria technology, applied in the field of microorganisms, can solve the problems of complex mechanism of action, difficult to ferment and expand culture, and unstable bacterial function effect.

Active Publication Date: 2021-04-27
北京本农环保科技集团有限公司 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing method for screening petroleum hydrocarbon-degrading strains, the screened mixed flora is a variety of degrading bacteria that work together to degrade petroleum hydrocarbons. Practical application promotion

Method used

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  • Microbacterium bn-88, microbial agent and its application in degrading petroleum hydrocarbons
  • Microbacterium bn-88, microbial agent and its application in degrading petroleum hydrocarbons
  • Microbacterium bn-88, microbial agent and its application in degrading petroleum hydrocarbons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045]Screening of the strain

[0046]Enrichment medium: Nutrient broth culture medium, protein 胨 10g, sodium chloride 5g, beef milk paste 3g, distilled water 1000ml, such as a solid medium, 2% of agar.

[0047]Degradation medium: NaCl 1.0g, (NH4)2· SO4 0.617G, KH2PO4 0.50g, k2HPO41.0G, MGSO40.50g, CACL2 0.1g, KCL 0.10g, FESO4· 7h2O 0.01g, the crude oil 1000mg, the distilled water is 1000ml, the pH is adjusted to 7.0-7.2, such as a solid medium, 2% of agar, 20 minutes of high pressure steam sterilization pot 121 ° C, spare.

[0048]Take 10G Dongying's median petroleum hydrocarbon pollution soil, add 90 mL of distilled water, stir well, use pipetting gun to take 10 ml of mixed liquid into 100 ml of enrichment medium, 35 ° C, 190 r / min constant temperature culture 24 h, 10% inoculated The amount is transferred to a new enrichment medium, and the 3 generations of continuous enrichment, the above-mentioned enrichment gradient dilution, coated into the oil-containing selective medium plate, 35 ...

Embodiment 2

[0049]Example 2 identification of strains

[0050]The strains were identified by cell morphology, colonies, physiological and biochemical properties, and 16S rRNA sequences.

[0051]According to Dong Xiuzhu, Cai Miao Ying's "Common Bacterial System Identification Manual" to determine the morphology and physiological and biochemical characteristics of strains.

[0052]The cloning and sequence analysis method of strain 16S rRNA gene is as follows:

[0053](1) Extraction of strain genomic DNA uses the bacterial genome DNA extraction kit of Beijing Fort-Biological Biotechnology Co., Ltd.;

[0054](2) Universal primers with 16S rRNA gene

[0055]27F: 5'-agagttgatcctggctcAg-3 '(SEQ ID NO.1),

[0056]1492R: 5'-ggttaccttgttacgactt-3 '(SEQ ID No.2),

[0057]PCR amplification was performed in genomic DNA as a template.

[0058]The amplification conditions were: 95 ° C predetermitability was 2min, then 95 ° C degenerate 30s, 58 ° C annealing 30s, 72 ° C extension 90s, a total of 35 cycles, and finally 72 ° C extends for...

Embodiment 3

[0065]Example 3 Growth curve of strain BN-88

[0066]Strawl Growth Curve Draw: Picking a Ring Bacillus Moss, inoculation into the nutrient broth culture medium, 30 ° C, 150 r / min, cultured for 20 hours, and prepared as a seed liquid. Submersible cultivation tubes, each tube dispensing the nutrient broth culture medium 5ml, each culture tube is used in a seed liquid, add 30 ° C, 150R / min culture, at 2 h, 4h, 6h, 8h. Sampling is sampled at 10H, 12H, 16H, 20H, 24H, 28H, 32H, 36h, 48h, and the concentration of the bacterial body is determined in a turbid method. The concentration of the OD600 average represents the concentration of the sampling time point culture fluid each time the OD600 average is measured. The absorbance values ​​of different culture times were determined, and the growth curve of strains was drawn.image 3 Indicated.

[0067]It can be seen from the results that growth reproduction of strains is divided into 4 periods: late slime, regular period, stable period, and decli...

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Abstract

The present invention relates to the field of microorganisms, in particular, provides a microbacterium Bn-88, a microbial agent and its application in degrading petroleum hydrocarbons. The present invention obtains the microbacterium Bn-88 with the preservation number of CGMCC NO.16607 through strain screening. The strain is easy to cultivate, grows and reproduces quickly, shows high efficiency and diversity when degrading petroleum hydrocarbons, and has good application prospects. In addition, Exobacterium Bn‑88 can reduce the pH value of a highly alkaline environment for the treatment of highly alkaline wastewater.

Description

Technical field[0001]The present invention relates to the field of microorganisms, and in particular, to a Microbiobacillus BN-88, a microbacterial agent, and its degraded petroleum hydrocarbon applications.Background technique[0002]The rapid development of the petroleum industry has brought many environmental issues and directly endangered human production and life. Some components of the petroleum hydrocarbons have a role in the human body, and the teratogenic changes can be used through the food chain in the animals and plants and the human body. They are listed as a focus pollutant. The environmental governance of petroleum hydrocarbon pollution has attracted high attention in many countries. At present, the common petroleum hydrocarbon pollution repair methods mainly include physical law, chemical method and biological law. However, physical law is generally complex and has high preparations in the previous period, high repair costs, and is not conducive to technology. The chem...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/34C10G31/00C12R1/01
CPCC02F3/34C10G31/00C12N1/20C12N1/205C12R2001/01
Inventor 韩建均梁雪杰王玉文赵川
Owner 北京本农环保科技集团有限公司
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