Application of Bacillus mohevi and its method for degrading 2-ethylhexyl phthalate
A technology of phthalic acid and ethylhexyl ester, which is applied in the field of environmental microbial application to achieve the effects of high degradation rate, short degradation period and simple operation
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0033] Identification of Example 1 bacterial strain B1811
[0034] 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.
[0035] (1) 16S rDNA sequence analysis:
[0036] 16s rDNA forward primer 27F: 5'-AGA GTT TGA TCC TGG CTC AG-3', as shown in SEQ ID NO.1;
[0037] 16s rDNA reverse primer 1541R: 5′-AAG GAG GTG ATC CAG CC-3′, as shown in SEQ ID NO.2;
[0038] 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 related strai...
Embodiment 2
[0044] Example 2 Preparation of Bacillus Mohevi B1811 Liquid Shake Flask Culture Solution
[0045] (1) Slant culture: Bacillus Mohaiavi 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.
[0046] (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
[0048] Example 3 DEHP standard curve formulation and content determination
[0049] (1) High-performance liquid chromatography (HPLC) to make a standard curve: take 600 μL of DEHP and use methanol as a solvent to prepare a 1 mg / mL DEHP solution. After diluting ten times, take 0.2 mL, 0.4 mL, 0.6 mL, 0.8 mL, 1.0mL, 1.2mL, and 1.4mL were fixed to 2mL centrifuge tubes, that is, the concentration gradient (μg / mL) was 10, 20, 30, 40, 50, 60, and 70 respectively. 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. With the peak area of DEHP as the abscissa and the concentration of DEHP as the ordinate, a standard curve was made; then the peak area-concentration equation was obtained.
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More