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Method for effectively improving ore leaching capacity of axidithiobacillus ferrooxidans

A technology of Thiobacillus ferrooxidans and acidophilus, applied in the field of genetic engineering, to achieve the effect of increasing the content of metal elements and broad development and application prospects

Pending Publication Date: 2020-02-18
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the search found that the function of the density sensing system genes (afeR, orf3, afeI) in the oxidative leaching of ores, or the ability to overexpress the bioleaching of ores, and the overexpression of the density sensing system genes (afeR, orf3, afeI) Methods There are no reports on the strategy of improving the leaching ability of acidophilic Thiobacillus ferrooxidans

Method used

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  • Method for effectively improving ore leaching capacity of axidithiobacillus ferrooxidans
  • Method for effectively improving ore leaching capacity of axidithiobacillus ferrooxidans
  • Method for effectively improving ore leaching capacity of axidithiobacillus ferrooxidans

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Experimental program
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Effect test

Embodiment 1

[0030] Example 1 Construction of recombinant bacteria A.ferrooxidans (pJRD215-QS)

[0031] 1. Construction of recombinant plasmid pJRD215-QS

[0032] (1) Design primers, according to the genome sequence of A.ferrooxidans ATCC No.23270 published by NCBI (https: / / www.ncbi.nlm.nih.gov / ), design primers for amplifying the AfeI / R operon of the density sensing system ;

[0033] The sequence information is as follows:

[0034] QS sen: CCGCTCGAGG GGTACC CGGGAAAATGCGTCACGTC

[0035] (The underlined part is the KpnI restriction site)

[0036] QS ant: TGGACTAGTGC TCTAGA GAGCAGCCCTTCAAATGGT

[0037] (The underlined part is the XbaI restriction site)

[0038] (2) Genome and plasmid extraction: Genome and plasmid extraction kits (purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd.) were used to extract wild-type A. ferrooxidans genome and pJRD215 plasmid for future use.

[0039] (3) PCR amplification to obtain the target fragment

[0040] Using the A.ferrooxidans ge...

Embodiment 2

[0051] Embodiment 2 Comparative determination of ore surface erosion ability of recombinant bacteria A.ferrooxidans (pJRD215-QS) and contrast A.ferrooxidans (pJRD215)

[0052] The ore is pyrite, which is produced in Yunfu City, Guangdong Province, China. The ore is processed and broken into a size smaller than 0.5cm before use 3 particle. The initial inoculum of the recombinant strain and the control strain is OD 600 nm =0.5, add the same amount of pyrite, 30 ℃, static culture, eight days later, take the ore under the culture of the recombinant bacteria and the control bacteria, fix it, dry it at the critical point, dehydrate, spray gold, and observe it with a scanning electron microscope Erosion condition of ore surface.

[0053] ore surface such as image 3 , the surface of the ore under the culture of recombinant bacteria has vertical and horizontal ravines and unevenness. The overall situation of the ore surface at low magnification and the local area under 20,000 tim...

Embodiment 3

[0054] Example 3 Comparative determination of metal leaching ability of recombinant bacteria A.ferrooxidans (pJRD215-QS) and control A.ferrooxidans (pJRD215)

[0055] The initial inoculum of the recombinant strain and the control strain is OD 600 nm =0.5, add the same amount of processing and break it into a size less than 0.5cm 3 The pyrite was cultured statically at 30°C. After 15 days, the leaching solution was filtered to remove slag and bacteria, and the content of elements in the leaching solution was determined by ICP-MS (inductively coupled plasma mass spectrometer).

[0056] The content of each element in the leachate is as Figure 4 , the contents of copper, zinc, iron, arsenic and chromium in the leaching solution of the ore treated by the recombinant bacteria A.ferrooxidans (pJRD215-QS) were significantly higher than those of the ore treated by the control strain.

[0057] Based on the above results, it can be seen that the recombinant engineering bacteria A.fer...

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Abstract

The invention discloses a method for effectively improving ore leaching capacity of axidithiobacillus ferrooxidans. The method comprises the following steps: obtaining an AfeI / R operon gene fragment containing three genes of afeR, orf3 and afeI on the basis of a genome of wild-type A.ferrooxidans ATCC No.23270; connecting the gene fragment to a vector pJRD215 to obtain a recombinant plasmid pJRD215-QS; then transferring the pJRD215-QS into A.ferrooxidans in a mode of conjugational transfer to obtain a recombinant bacterium A.ferrooxidans (pJRD215-QS) which simultaneously increases copy numbersof three genes of afeI, orf3 and afeR of the AfeI / R operon; and mixing the recombinant engineering bacteria with pyrite to form an ore leaching system, and carrying out an ore leaching reaction, wherein a standing heap leaching mode is adopted in an ore leaching process, the grain size of ores is less than 0.5 cm<3>, and the ore leaching reaction is carried out at normal temperature and is simpleand convenient to operate. The method has great theoretical value and practical significance and wide development and application prospect for improving ore leaching efficiency of the biological metallurgy industry.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and in particular relates to a method for effectively improving the ore leaching ability of acidophilic Thiobacillus ferrooxidans, a dominant bacteria in ore leaching. Background technique [0002] Axidithiobacillus ferrooxidans (A.ferrooxidans for short) is an obligate chemoautotrophic, Gram-negative bacterium that can fix carbon dioxide, ferrous oxide, elemental sulfur and reducing inorganic sulfur compounds to obtain energy , is the first discovered microorganism with ore oxidizing ability, and it is also the dominant strain of ore leaching microorganisms. The unique physiological and metabolic mechanism of acidophilic Thiobacillus ferrooxidans makes this bacteria widely used and studied deeply in the biometallurgy industry. [0003] In recent years, the biometallurgy industry has developed rapidly, and the use of microorganisms to recover heavy metals has been widely used. Compared with tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/74C12N15/31C12N1/21C22B3/18C12R1/01
CPCC12N15/74C07K14/195C22B3/18Y02P10/20
Inventor 陈林旭高雪彦林建群林建强刘相梅庞昕
Owner SHANDONG UNIV
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