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Endosulfan degradation bacterium JBW4

A technology for degrading bacteria and endosulfan, applied in the directions of bacteria, microorganisms, biochemical equipment and methods, etc., can solve problems such as toxicity, and achieve the effects of low cost, good application prospects and high efficiency

Active Publication Date: 2012-12-19
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The metabolites are sulfate esters, alcohols, ethers, hydroxy ethers and lactones, but only endosulfan sulfate still contains sulfur elements and has basically the same toxicity as endosulfan

Method used

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  • Endosulfan degradation bacterium JBW4
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  • Endosulfan degradation bacterium JBW4

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1: Screening and identification of endosulfan-degrading bacteria JBW4

[0038] Medium:

[0039] Inorganic salt basal medium: 5.8g K 2 HPO 4 , 4.5g KH 2 PO 4 , 2.0g (NH 4 ) 2 SO 4 , 0.16g MgSO 4 , 0.02g CaCl 2 , 0.002g Na 2 MoO 4 , 0.001g FeSO 4 , 0.001g MnCl 2 , 1L of deionized water, adjusted to pH 7.0, and sterilized at 121°C for 30min.

[0040]Medium containing a small amount of carbon source: Add 5.0 g of peptone to the above inorganic salt basic medium, adjust to pH 7.0, and sterilize at 121°C for 30 minutes.

[0041] Separation and purification medium: add endosulfan to the medium containing a small amount of carbon source, so that the concentration of endosulfan in the medium is 100 μg·mL -1 , 15.0 g of agar, adjusted to pH 7.0, and sterilized at 121 °C for 30 min.

[0042] LB medium: 10.0g peptone, 5.0g yeast extract, 10.0g NaCl dissolved in 1L deionized water, adjust the pH to 7.0, and sterilize at 121°C for 30min.

[0043] 1) Strain enr...

Embodiment 2

[0051] Example 2 Sequence Analysis of Endosulfan-degrading Bacteria JBW4 16S rDNA

[0052] 1. Extraction of total DNA of endosulfan-degrading bacteria JBW4

[0053] 1) Bacterial culture: Inoculate endosulfan-degrading bacteria JBW4 in LB medium, and shake and culture at 30°C for 16-18h;

[0054] 2) Bacteria collection: take 1.5mL culture solution in a 1.5mL centrifuge tube, 12000r min -1 Centrifuge for 30s, discard the supernatant, and collect the cells.

[0055] 3) Add 565 μL TE buffer to the 1.5 mL centrifuge tube after discarding the supernatant in step 2), mix well on the shaker, add 30 μL 10% SDS and 3 μL 20 mg·mL -1 Proteinase K, mixed upside down by hand, incubated in a water bath at 37°C for 1 hour; then added 100μL of 5mol·L -1 NaCl, after fully mixing, add 80 μL CTAB / NaCl solution, mix well and incubate in a water bath at 65°C for 10 minutes.

[0056] 5) Add 780 μL of chloroform / isoamyl alcohol (24:1), mix well, centrifuge at 10,000 rpm for 10 min, and transfer t...

Embodiment 3

[0068] Embodiment 3: the degradation characteristics of degrading bacteria JBW4

[0069] Extraction of endosulfan from the isolation and purification medium: inoculate the endosulfan-degrading bacteria JBW4 into the isolation and purification medium and cultivate for 5 days, add 5mL of n-hexane, fully oscillate and extract on the vortex mixer, let it stand for stratification, and take the organic phase.

[0070] Determination of endosulfan: gas chromatographic analysis: Shimadzu GC-14C gas chromatograph, capillary column (OV-1701) 0.53mm×30m, FID detector. Injection port temperature 260°C, column temperature 240°C, detector 260°C, N 2 Flow rate 25mL·min -1 , the injection volume is 2 μL.

[0071] The formula for calculating the degradation rate of endosulfan by bacterial suspension:

[0072] R = ρ ck - ρ ρ ck ...

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Abstract

The invention provides an endosulfan degradation bacterium JBW4 (Alcaligenes faecalis.JBW4), which was preserved in the China center for type culture collection (CCTCC) on May 24th, 2012 with the preservation serial number being CCTCC No:M2012181. The endosulfan degradation bacterium JBW4 is prepared to be bacteria suspending liquid, and the bacteria suspending liquid is applied to the degradation of endosulfan in the soil in a mode that the bacteria suspending liquid is directly added into the soil, so that endosulfan remained on objects of water bodies, soils and the like can be safely, effectively and quickly degraded, genetic toxicity and ecotoxicity of the endosulfan to the environment are reduced and the actions of restoring the soil polluted by the endosulfan and protecting the ecological environment are exerted. The bactericide of the strain is simple in preparation process, low in cost and high in efficiency, and has good application prospect; and moreover, the secondary pollution is avoided.

Description

technical field [0001] The invention relates to an endosulfan-degrading bacterium JBW4, in particular to an endosulfan-degrading bacterium—Alcaligenes faecalis. JBW4, which belongs to the technical field of biodegradation treatment. Background technique [0002] Endosulfan, (1,2,3,4,7,7-hexachlorobicyclo-2,2,1-heptene-2,3-bishydroxymethyl-5,6-sulfite), English name endosulfan , the molecular formula is C 9 h 6 Cl 6 o 3 S, is a cyclopentadiene organochlorine insecticide. Endosulfan technical is solid at normal temperature and pressure, usually a mixture of two isomers (α-endosulfan, β-endosulfan, the ratio of the two is 7:3), the chemical structure is as follows: [0003] [0004] Pure endosulfan is a white crystal with a relative density of 1.745g / cm 3 , with the smell of sulfur dioxide, the saturated vapor pressure at 25°C is 1.33×10 3 Pa, melting point is 70-100°C; insoluble in water, soluble in xylene, chloroform, acetone and other organic solvents; unstable in ...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/05
Inventor 朱鲁生王军王金花谢慧苏坤昌孔玲芬
Owner SHANDONG AGRICULTURAL UNIVERSITY
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