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Tolerant serratia grimesii for enriching heavy metals and application of serratia grimesii

A technology of Serratia griseri and heavy metals, which is applied in the direction of bacteria, chemical instruments and methods, and methods based on microorganisms, can solve the problems of application and resistance mechanism that need to be deepened, and research is not mature enough to achieve strong adaptability and growth Fast, versatile effects

Inactive Publication Date: 2015-04-29
HUBEI UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In recent years, the research on heavy metal bacteria has made great progress, but the research in this field is still immature, and the application and resistance mechanism need to be further studied

Method used

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  • Tolerant serratia grimesii for enriching heavy metals and application of serratia grimesii
  • Tolerant serratia grimesii for enriching heavy metals and application of serratia grimesii
  • Tolerant serratia grimesii for enriching heavy metals and application of serratia grimesii

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Embodiment 1: Screening of Serratia bacterium DJ16

[0027] Isolation of the original strain of Serratia bacterium DJ16: Take a sample from the soil of the sewage treatment plant, weigh 1.0g of the sample and add it to a test tube filled with 9mL of sterile water, shake it on a vortex for 10 minutes to fully disperse the soil particles, Then stand still for 5min, and adopt 10-fold gradient dilution method to make the supernatant into 10 -2 、10 -3、10 -4 、10 -5 The dilutions were coated on LB plates respectively (adding a certain amount of Cd 2+ , selection separation), and placed upside down in a 28°C incubator for 7 days. According to the different morphological characteristics of the colony, pick a single colony from the plate and inoculate it on the corresponding slant medium. If there are non-pure cultured strains, use an inoculation loop to pick a little lawn and streak on the plate for separation.

[0028] Mutagenesis of the original strain of Serratia bacter...

Embodiment 2

[0029] Embodiment 2: Serratia bacterium DJ16 (preservation number: CCTCC M 2014578), Biolog (GN2 / GP2) automatic identification analysis

[0030] Pick the bacterial lawn of the strain to be tested and inoculate it on a Biolog plate, and cultivate it to the logarithmic growth phase at 28°C. Dip the Biolog IF liquid with a sterilized cotton swab, scrape off the bacterial lawn on the plate, then dissolve it in the Biolog IF liquid, shake and mix well; use a Biolog turbidimeter to adjust the turbidity to the specified range (GP plate: 28% ± 3% , GN plate: 52% ± 3%); then use a pipette gun to add the bacterial suspension to the Biolog identification plate, 150 μL per hole; then take out the identification plate when it is incubated at 28°C for 16-24 hours, and use the Biolog MicroStation analyzer to test the identification plate Perform data reading and analysis.

[0031] Table 1: Biolog (GN2 / GP2) automatic identification results

[0032]

[0033]

[0034]

[0035]

[...

Embodiment 3

[0037] Embodiment 3: Bacteria DJ16 (preservation number: CCTCC M 2014578) 16S rRNA gene sequence analysis

[0038] (1) Extraction of chromosomal DNA (small amount):

[0039] 1. Take 1.5mL bacterial liquid in a 1.5mL Eppendorf tube and centrifuge at 12 000r / min for 5min.

[0040] 2. Discard the supernatant, resuspend the pellet in 900 μL phosphate buffered solution (PBS), and centrifuge at 12 000 r / min at 4°C for 5 min.

[0041] 3. Discard the supernatant, add 300TE and 200μL 10mg / mL lysozyme, blow and aspirate to mix, incubate at 37°C for 1h, and mix by inverting every 15min.

[0042] 4. Add 600TENS lysate (200mmol / L NaCl, 100mmol / L Tril-HCl pH8.0, 2.0% SDS, 50mmol / L EDTA, 0.5% Triton X-100) and 20mg / mL proteinase K 10μL to the precipitate, mix by inverting, Incubate at 55°C for 1 hour, and mix by inverting every 15 minutes.

[0043] Centrifuge at 12 000 r / min at 5.4°C for 5 min, and take the supernatant.

[0044] 6. Add an equal volume of P:C:I (25:24:1) to the supernatan...

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Abstract

The invention discloses tolerant serratia grimesii for enriching heavy metals and an application of the serratia grimesii. Bacterium is subjected to 16S rRNA gene sequence analysis to obtain a gene nucleotide sequence of the 16S rRNA as shown in SEQ ID NO:1 in a sequence table. By homology analysis, the strain is proven to belong to the serratia grimesii, named as DJ16 and preserved in China Typical Culture Collection Center on 19th Nov, 2014, and the preservation number is CCTCC NO: M 2014578. The serratia grimesii DJ16 disclosed by the invention can be used for enriching the heavy metals in soil and water.

Description

technical field [0001] The invention relates to the technical field of environmental microbial remediation, in particular to Serratia bacteria that tolerate and enrich heavy metals in soil and water and applications thereof. Background technique [0002] With the development of industry and the widespread use of fossil fuels, heavy metals are enriched in soil and water sources, and their diffusion is intensified through sewage irrigation and other ways, and they are enriched in the human body through the food chain, endangering health, such as Minamata caused by mercury pollution in Japan disease, bone pain disease caused by cadmium pollution. With the rapid development of my country's industry, the resulting heavy metal pollution is becoming more and more serious. According to the statistics of the Ministry of Environmental Protection, in 2009, my country's heavy metal pollution incidents caused 4035 people's blood lead to exceed the standard, and 182 people's cadmium excee...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34B09C1/10C12R1/425C02F101/20
CPCB09C1/10C02F3/34C02F2101/20C12N1/205C12R2001/425
Inventor 代俊张毅周梦舟陈雄
Owner HUBEI UNIV OF TECH
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