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Application of Burkholderia pyrrole and method for degrading naphthalene

A technology of Burkholderia pyrrole, Holderella, applied in the directions of microorganism-based methods, applications, chemical instruments and methods, etc., to achieve the effects of simple operation, high degradation rate, and short degradation cycle

Active Publication Date: 2018-09-18
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Burkholderia pyrrole ( Burkholderia pyrrocinia ), its application is mainly in promoting plant growth and biological control of plant diseases; there is no report about using Burkholderia pyrrole to degrade NAP

Method used

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  • Application of Burkholderia pyrrole and method for degrading naphthalene
  • Application of Burkholderia pyrrole and method for degrading naphthalene
  • Application of Burkholderia pyrrole and method for degrading naphthalene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Identification of Example 1 bacterial strain B1213

[0032] Strain B1213 was isolated from soil, and its morphology was as follows figure 1 As shown, it is a rod-shaped Gram-negative bacteria that cannot form spores. Genomic DNA was extracted, and the genomic DNA was subjected to polymerase chain reaction (Polymerase Chain Reaction, PCR) using 16s rDNA and BCR1 primers to propagate specific DNA sequences, and the National Center for Biotechnology Information (National Center for Biotechnology Information, NCBI) database for strain comparison.

[0033] (1) 16S rDNA sequence analysis:

[0034] 16s rDNA forward primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3', as shown in SEQ ID NO.1;

[0035] 16s rDNA reverse primer 1492R: 5'-ACG GTT ACC TTG TTA CGA CTT-3', as shown in SEQ ID NO.2;

[0036] The amplified 16S rDNA sequence is shown as SEQ ID NO: 3, and the full length of the sequence is 1411bp. The amplified 16S rDNA sequence was compared with the gene sequence of relate...

Embodiment 2

[0042] Example 2 Preparation of Burkholderia pyrrole B1213 Liquid Shake Flask Culture Solution

[0043](1) Slant culture: Burkholderia pyrrole B1213 was inoculated on the slant medium and cultured at 40° C. for 48 hours to obtain the slant strain. The slant medium used contained the following ingredients per 1L: 2.0g glucose, 1.0g peptone, 0.5g yeast extract, 1.8g agar, the rest of distilled water, neutral pH, and sterilized at 115°C for 20 minutes.

[0044] (2) Take the slant strain and inoculate it into the sterilized seed culture medium, and incubate at constant temperature and shaking for 24 hours under the conditions of 175 rpm and 30°C to obtain the seed culture medium. The seed culture medium contains the following ingredients per 1L: 0.5g of ammonium sulfate, 4.0g of sodium chloride, 0.5g of potassium phosphate trihydrate, 0.4g of magnesium sulfate heptahydrate, 15g of agar powder, 5g of yeast extract, and the remainder of distilled water. Quantity, pH7.0, sterilized ...

Embodiment 3

[0046] Example 3 NAP standard curve formulation and content determination

[0047] (1) Prepare 400mg / L naphthalene mother liquor with n-hexane as solvent, dilute 20 times, 25 times, 30 times, 35 times, 40 times, 80 times, 100 times, 200 times respectively for later use, take 2.5mL of different dilution factors The naphthalene solution is placed in a 3mL quartz cuvette, and the absorbance value under the wavelength of 275nm is measured respectively with an ultraviolet spectrophotometer, and the ordinate is used as the absorbance value, and the concentration of naphthalene is used as the abscissa to draw a standard curve; the standard curve is as follows figure 2 Shown; and then obtain the absorbance value-concentration equation: y=0.0424x-0.0033, x is the concentration of naphthalene, and y is the absorbance value.

[0048] (2) Take 2.5mL of the fermentation broth obtained in Example 2 and place it in a 3mL quartz cuvette, measure the absorbance at 275nm wavelength with a UV s...

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Abstract

The invention relates to the application of Burkholderia pyrrole and a method for degrading naphthalene, belonging to the field of environmental microorganism application. The purpose of the Rubockholderia B1213 of the present invention is to degrade NAP; the degradation rate of NAP can be as high as 92.45%-96.89%; the Rubockholderia B1213 is preserved in the Chinese Microbiological Culture Preservation Management Committee General Microbiology Center; the deposit number is CGMCCNo.12806; the deposit time is July 21, 2016. The method for degrading NAP by using Burkholderia pyrrole B1213 in the present invention is to contact Burkholderia pyrrole B1213 with NAP under the condition that yeast extract exists. Compared with the method for degrading naphthalene by bacteria and fungi before the present invention, the method for degrading naphthalene of the present invention has novelty in the source of strains, and its degradation rate of naphthalene is higher, the degradation period is short, and the operation is simple.

Description

technical field [0001] The invention relates to the application of Burkholderia pyrrole and a method for degrading naphthalene, belonging to the field of environmental microorganism application. Background technique [0002] Naphthalene (NAP) is a representative class of important persistent organic pollutants (POPs) ubiquitous in the environment. A large number of studies have proved that NAP has the "three causes" of chronic toxicity, carcinogenicity, teratogenicity, and mutagenicity. Atmospheric subsidence, sewage irrigation, solid waste landfill leakage, oil field exploitation and extensive use of petroleum products have caused local soil NAP pollution around the world, and serious pollution has occurred in many places in my country. Due to its stable nature and difficult to degrade, NAP tends to accumulate continuously in the soil environment, seriously endangering the production and ecological functions of the soil, the quality of agricultural products and human healt...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C09K17/14B09C1/10C12R1/01
CPCB09C1/10C09K17/14C12N1/20C12N1/205C12R2001/01
Inventor 郦金龙李秀婷滕超朱运平申卫家
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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