A Strain of Staphylococcus equine and Its Application in Degrading Heavy Metal Ions

A technology of heavy metal ions and staphylococci, which is applied in the detection and application of microorganisms to achieve the effects of low cost, wide application range and high processing efficiency

Active Publication Date: 2017-05-17
NANJING TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to screen out heavy metal Cr from activated sludge through the enrichment and screening of anti-heavy metal strains. 6+ Staphylococcus rumen, the strain is applied to the restoration of heavy metal pollution, improving the efficiency of heavy metal pollution treatment, reducing secondary pollution, reducing costs, and providing more options for microbial treatment of heavy metal pollution in complex environments

Method used

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  • A Strain of Staphylococcus equine and Its Application in Degrading Heavy Metal Ions
  • A Strain of Staphylococcus equine and Its Application in Degrading Heavy Metal Ions
  • A Strain of Staphylococcus equine and Its Application in Degrading Heavy Metal Ions

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1: anti-heavy metal ion Cr 6+ strain screening.

[0055] 1 Experimental materials

[0056] 1.1 Experimental strains

[0057] Source of soil sample: Activated sludge from Nanjing Chengbei Sewage Treatment Plant

[0058] 1.2 Experimental Instruments

[0059] Pipette gun (Eppendorf, Germany); desktop high-speed centrifuge (Eppendorf 5417R, Germany) vertical pressure steam sterilizer (Shanghai Shen'an Medical Instrument Factory LDZX-50KBS) desktop constant temperature oscillator (IS-RSD3) UV-visible spectrophotometer ( Analytical balance (SHIMADZU AUY120) from Shanghai Mapuda Instrument Co., Ltd. UV-1100)

[0060] 1.3 Experimental reagents

[0061] Potassium dichromate (K 2 C r 2 o 7 ) Chemical reagents are analytically pure and calculate the mass fraction of metal ions, which are respectively dissolved in deionized water to form a 10g / L storage solution.

[0062] 1.4 Medium

[0063] Basal medium: LB medium (g / L)—peptone 10g, sodium chloride 10g, yeast ...

Embodiment 2

[0074] Embodiment 2: diphenylcarbonyl spectrophotometric method measures Cr 6+ concentration.

[0075] 2.1 Drawing of standard curve

[0076] Take nine 50ml colorimetric tubes, add 0, 0.2, 0.5, 1.00, 2.00, 4.00, 6.00, 8.00 and 10.00ml of chromium standard solution in sequence, add 0.5ml each of 1+1 sulfuric acid and 1+1 phosphoric acid, add 2ml for color development solution, shake well, and dilute to the mark with water. After 5-10min, measure the absorbance at a wavelength of 540nm, and make a blank correction. Draw a standard curve with the absorbance as the ordinate and the corresponding hexavalent chromium content as the abscissa.

[0077] 2.2 Determination of hexavalent chromium in the bacterial liquid supernatant

[0078] Take 200 μl of the overnight cultured bacterial solution in a 50ml colorimetric tube, and the determination method is the same as that of the standard solution. After blank calibration with distilled water, the hexavalent chromium content is check...

Embodiment 3

[0083] Embodiment 3: strain identification.

[0084] 16sRNA sequence analysis method: PCR amplification uses bacterial 16sRNA universal primers:

[0085] 27F (5'-AGAGTTTGATCMTGGCTCAG-3')

[0086] 1492R (5'-GGTTACCTTGTTACGACTT-3')

[0087] PCR reaction system (25 μL): 5× amplification buffer (5 μL),

[0088] Genomic DNA (0.1 μL),

[0089] dNTP (2μL),

[0090] Primer F (0.5μL),

[0091] Primer R (0.5 μL),

[0092] Taq DNA polymerase (0.25 μL)

[0093] Deionized water (16.65 μL)

[0094] Reaction conditions: pre-denaturation at 98°C for 10 minutes, deformation at 98°C for 10 seconds, annealing at 58°C for 10 seconds, extension at 72°C for 90 seconds, a total of 30 cycles, and full extension at 72°C for 10 minutes. The PCR products were sequenced by Nanjing GenScript Co., Ltd.

[0095] The measured gene sequence is shown as SEQ ID No:1.

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Abstract

The invention discloses a strain of Staphylococcus equorum, which is classified and named as Staphylococcus equorum ss equorum (Staphylococcus equorum ss equorum). NO: M 2014486, the deposit date is October 15, 2014. The invention also discloses the application of the above-mentioned staphylococcus equineus in degrading heavy metal ions. The bacterial strain of the invention is applied to the treatment of heavy metal pollution, and has the advantages of no secondary pollution, high treatment efficiency, wide application range, low cost and the like.

Description

technical field [0001] The invention relates to a microorganism and its application in degrading heavy metal ions, in particular to a strain of Staphylococcus rumenum subsp. rumen and its application in heavy metal pollution, belonging to the technical field of microorganism detection and application. Background technique [0002] Soil is an important part of the earth's ecological environment and one of the important non-renewable resources on which human beings depend, but with the development of human economy and society. In particular, the industrial and mining industry has developed and utilized a large number of natural mineral resources, resulting in high-concentration heavy metal pollution in mining wasteland, which is one of the important environmental pollutants and has exacerbated the deterioration of the soil environment. Heavy metal pollution mainly comes from the mining and processing of industrial and mining industries, industrial wastewater, landfill, mining,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A62D3/02B09C1/10C12R1/44A62D101/43
CPCA62D3/02A62D2101/43B09C1/10C12N1/205C12R2001/44
Inventor 谢婧婧金月娟宋天顺张鸿鹍郭亭刘婷婷
Owner NANJING TECH UNIV
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