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Bacillus mojavensis and application thereof

A technology of Bacillus weiba and Bacillus, which is applied in the field of environmental microbial applications to achieve the effects of short degradation cycle, high degradation rate and simple operation

Active Publication Date: 2017-05-10
BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] Bacillus mohaiwei ( Bacillus mojavensis ) is mainly used in the fermentation of lipase, and there is currently no report on the use of Bacillus mohaiwei to degrade DBP

Method used

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  • Bacillus mojavensis and application thereof
  • Bacillus mojavensis and application thereof
  • Bacillus mojavensis and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Identification of Example 1 bacterial strain B1811

[0035] Strain B1811 was isolated from soil, and its morphology was as follows: figure 1 As shown, it is a rod-shaped Gram-negative bacteria that can 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 1541R: 5′-AAG GAG GTG ATC CAG CC-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 1461bp. The amplified 16S rDNA sequence was compared with the gene sequence of the related b...

Embodiment 2

[0045] Example 2 Preparation of Bacillus mohewei B1811 liquid shake flask culture solution

[0046] (1) Slant culture: Bacillus mohaiwei B1811 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 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 at 121°C...

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 bacillus mojavensis and application thereof and belongs to the field of environmental microorganism application. The bacillus mojavensis B1811 is preserved at the general microbiology center of China Committee for Culture Collection of Microorganisms(CGMCC), the preservation number is CGMCCNo.12805, the preservation date is on July 21, 2016. The bacillus mojavensis can be used for degrading dibutyl phthalate. The degradation rate of the dibutyl phthalate can be as high as 99.7%. A method for degrading the dibutyl phthalate by using the bacillus mojavensis B1811 is adopted. In the presence of a yeast extract, the bacillus mojavensis B1811 is in contact with the dibutyl phthalate. Compared with methods for degrading the dibutyl phthalate by using bacteria and fungi, the method for degrading the dibutyl phthalate has a new conception in strain source, has high dibutyl phthalate degradation rate and is short in degradation period and simple to operate.

Description

technical field [0001] The invention relates to a strain of bacillus mohaiwei and its application, which belongs to the field of environmental microorganism application. Background technique [0002] Dibutyl phthalate (Dibutyl phthalate), referred to as DBP, is a kind of phthalic acid esters (Phhtalic Aicd Easters, PAEs). It is an important class of environmental hormone organic synthetic compounds with 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 estrogen, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20A62D3/02C12R1/07A62D101/28
CPCA62D3/02A62D2101/28C12N1/205C12R2001/07
Inventor 郦金龙李秀婷朱运平滕超魏然申卫家
Owner BEIJING TECHNOLOGY AND BUSINESS UNIVERSITY
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