Burkholderia pyrrocinia and application thereof

A technology of Burkholderia pyrrole and Holderia, which is applied in the direction of bacteria, biochemical equipment and methods, microorganisms, etc., to achieve the effects of high degradation rate, simple operation, and short degradation cycle

Active Publication Date: 2016-12-21
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] 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 DBP

Method used

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  • Burkholderia pyrrocinia and application thereof
  • Burkholderia pyrrocinia and application thereof
  • Burkholderia pyrrocinia and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Identification of Example 1 bacterial strain B1213

[0035] 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.

[0036] (1) 16S rDNA sequence analysis:

[0037] 16s rDNA forward primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3', as shown in SEQ ID NO.1;

[0038] 16s rDNA reverse primer 1492R: 5'-ACG GTT ACC TTG TTA CGA CTT-3', as shown in SEQ ID NO.2;

[0039] 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 the re...

Embodiment 2

[0045] Example 2 Preparation of Burkholderia pyrrole B1213 Liquid Shake Flask Culture Solution

[0046] (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.

[0047] (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 solution. The seed medium used 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 rest of distilled water , pH7.0, sterilized at 121°C for 2...

Embodiment 3

[0049] Example 3 DBP standard curve formulation and content determination

[0050] (1) High-performance liquid chromatography (HPLC) to make a standard curve: take 600 μL of DBP and use methanol as a solvent to prepare a 1 mg / mL DBP solution. After diluting ten times, take 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, Dilute 1.0mL, 1.2mL, 1.4mL into 2mL centrifuge tubes, that is, the concentration gradient (μg / mL) is 10, 20, 30, 40, 50, 60, 70 respectively, filter with 0.45μm organic filter membrane and place in liquid Phase vials to be tested. The HPLC detection conditions in Example 3 are: the chromatographic column is Sepax Gp-C18 column (150mm*4.6mm, 5.0μm); the mobile phase is acetonitrile-water (83:17, V / V); the injection volume 10μL; flow rate 1.0mL / min; column temperature 25C°; UV detector wavelength 210nm. Take the peak area of ​​DBP as the abscissa and the concentration of DBP as the ordinate to make a standard curve; then obtain the peak area-concentration equation.

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Abstract

The invention relates to burkholderia pyrrocinia and application thereof, which belong to the field of environmental microorganism application. The burkholderia pyrrocinia B1213 provided by the invention has the preservation No. CGMCC No.12806, is preserved on July 21, 2016, belongs to a bacilliform gram-negative bacterium, and cannot form gemma. The burkholderia pyrrocinia B1213 provided by the invention can degrade dibutyl phthalate (DBP); the degradation rate of the DBP can reach up to 45.5 to 75.35 percent. A method for adopting the burkholderia pyrrocinia B1213 provided by the invention to degrade the DBP is characterized in that in the presence of a yeast extract, the burkholderia pyrrocinia B1213 is contacted with DBP. Compared with a method for adopting bacteria and fungus to degrade the DBP, the method for degrading the DBP provided by the invention is novel in strain source, high in degradation rate, short in degradation period, and simple to operate.

Description

technical field [0001] The invention relates to a strain of Burkholderia pyrrole and its application, belonging to the field of environmental microorganism application. Background technique [0002] Dibutyl phthalate (Dibutyl phthalate), abbreviated as DBP, is a kind of phthalic acid esters (Phhtalic Aicd Easters, PAEs). It is an important class of organic synthetic compounds of environmental hormones. It has light color, It is characterized by low volatility, low odor and low temperature resistance. It is the plasticizer with the largest output and the most consumption in recent years. It is widely used in rubber, plastics, spices and other industries. [0003] The connection between DBP and the carrier is unstable, and it is easy to diffuse into the environment. It can enter the human body and be enriched through various channels such as food, air, drinking water, and cosmetics. DBP has a toxic effect on aquatic plants, has a significant interference effect on animal estr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/01A62D101/28
CPCA62D3/02A62D2101/28C12N1/205C12R2001/01
Inventor 郦金龙李秀婷滕超熊科申卫家魏然
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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