Bacterial strain used for arsenic polluted soil remediation and application method thereof

A technology of soil remediation and application method, which is applied in the field of arsenic-contaminated soil microbial remediation to achieve the effect of increasing mobility and reducing the harm of arsenic

Active Publication Date: 2014-08-27
CENT SOUTH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Bacterial strain used for arsenic polluted soil remediation and application method thereof
  • Bacterial strain used for arsenic polluted soil remediation and application method thereof
  • Bacterial strain used for arsenic polluted soil remediation and application method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0028] Example 1: Isolation, purification and domestication of the strain of the invention

[0029] Collect arsenic-contaminated soil from the realgar mining area in Shimen County, Hunan Province. Take 5g of soil and culture in 50mL sterilized LB liquid medium for 2 days. Take 1mL of upper mud and dilute to 10 by gradient dilution method. -5 , 10 -6 , 10 -7 , Using a pipette from 10 -6 , 10 -7 0.1 mL of the diluted solution was coated with a solid medium containing 100 mg / L As(III) and incubated at 30°C for 3 to 4 days. The solid medium composition is: 15g / L LB medium, 2g / L sodium lactate, 2g / L potassium dihydrogen phosphate, 2g / L magnesium sulfate and 15g / L agar, pH=7-8. Use a sterile inoculating needle to pick up colonies of different morphologies, and streak them into 100mg / L As(III) solid medium and cultivate them at 30°C for 3 days. Separate and purify continuously for more than 3 times to obtain single colonies of different forms.

[0030] Use 2mL1% AgNO 3 The solution sprea...

Embodiment 2

[0031] Example 2: Biological identification and morphological characteristics of the strain of the present invention

[0032] The bacterial genomic DNA extraction kit was used to extract the genomic DNA of the strain, and then the 16S rRNA gene was amplified and sequenced using the 16S rRNA gene universal primer.

[0033] A phylogenetic tree was constructed using the 16S rRNA gene sequence. The strain was clustered with Brevibacterium yomogidense, and the similarity reached 99%, and the similarity with Brevibacterium sp.JC4316S rRNA gene reached 100%, named Brevibacterium sp.YZ-1. The phylogenetic analysis tree of this strain is as figure 1 Shown.

[0034] The bacterial body of this strain is rod-shaped and belongs to Brevibacterium. Scanning electron micrograph of this strain, see figure 2 .

Embodiment 3

[0035] Example 3: The oxidation efficiency and tolerance of the strain of the present invention to As(III)

[0036] Use an inoculating loop to pick up the purified colonies and culture them in 100 mL sterilized liquid medium with shaking for 24 hours. Then, 5 mL of the bacterial solution was inoculated into 50 mL sterile medium with As(III) concentrations of 50 mg / L and 100 mg / L, respectively, and cultured with shaking at 30°C for 72 hours. During this period, the residual As(III) content was measured at 24, 36 and 72h respectively. The As(III) solution with a concentration of 100mg / L was oxidized by more than 75%, and the As(III) solution with a concentration of 50mg / L was almost completely oxidized. Such as image 3 Shown.

[0037] Inoculate with the same amount of As(III) 200mg / L, 400mg / L, 600mg / L, 800mg / L, 1000mg / L, 1200mg / L, 1500mg / L liquid medium with shaking culture at 30℃ for 24h. Ultraviolet visible spectrophotometer measures the absorbance value OD600 at 600nm. The to...

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Abstract

The invention discloses a bacterial strain Brevibacterium sp.YZ-1 used for arsenic polluted soil remediation and an application method of the bacterial strain used for arsenic polluted soil remediation Brevibacterium sp.YZ-1. The collection number of the strain is CGMCC No.8329. The toleration for As (III) reaches above 1000 mg/L, and an As (III) solution with the concentricity of 100 mg/L can be oxidized by over 75%. Through the strain, water-soluble As (III) of arsenic polluted soil can be reduced by 82.6%, and the effective state As (III) can be reduced by 84.1%. The arsenic virulence in the environment is greatly reduced. The research shows that the strain has good application prospects in the aspect of environment arsenic pollution control.

Description

Technical field [0001] The invention belongs to the field of arsenic-contaminated soil microbial restoration, and specifically relates to a strain for arsenic-contaminated soil restoration and its application. Background technique [0002] The provinces in my country that are heavily polluted by arsenic include Xinjiang, Inner Mongolia, Qinghai, Hunan, Yunnan and Guangxi. Arsenic pollution is mainly caused by humans, such as the mining and smelting of metal mines, the processing and use of arsenic products, and the burning of coal. Arsenic is a highly toxic metal. It mainly exists in the inorganic form of arsenite (As(Ⅲ)) and arsenate (As(Ⅴ)) in the natural environment and is transformed into each other through oxidation and reduction of microorganisms. As(Ⅲ) is more mobile without electricity and is more harmful to organisms. As(Ⅲ) is about 100 times more toxic than As(Ⅴ). Oxidizing As(Ⅲ) to As(Ⅴ) is a detoxification process. [0003] my country is severely persecuted by arsenic...

Claims

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Application Information

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IPC IPC(8): C12N1/20B09C1/10C12R1/13
Inventor 杨志辉柴立元廖映平闵小波刘琳姚文斌
Owner CENT SOUTH UNIV
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