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Reductive chromium ion bacillus thuringiensis YB-03 and cultivation method and application of reductive chromium ion bacillus thuringiensis YB-03

A technology of Bacillus aureus, YB-03, applied in reducing chromium ion Bacillus thuringiensis YB-03 and its cultivation method and application field

Active Publication Date: 2012-09-19
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In addition, genes for reducing hexavalent chromium have also been found in Vibrio harveyi. The proteases expressed by these genes can reduce hexavalent chromium under certain conditions, but domestic scholars have rarely reported on hexavalent chromium reducing genes.

Method used

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  • Reductive chromium ion bacillus thuringiensis YB-03 and cultivation method and application of reductive chromium ion bacillus thuringiensis YB-03
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  • Reductive chromium ion bacillus thuringiensis YB-03 and cultivation method and application of reductive chromium ion bacillus thuringiensis YB-03

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation and screening of hexavalent chromium-reducing bacteria

[0025] Take 10g of activated sludge (collection time: September 2010; collection location: Zhengzhou Wulongkou Sewage Treatment Plant; collection person: Wei Fei) in 90ml of sterile water, and dilute to 10 -1 ~10 -6 Six gradients. Take 1ml of the solution from each gradient and spread it on 6+ LB solid medium, cultured at 37°C for 24h.

[0026] Reductive identification of chromium-resistant bacteria: select colonies from the culture medium containing 100mg / L Cr 6+ In LB liquid medium, cultured at 37°C, 150r / min for 24h, took out an appropriate amount of solution and centrifuged to get the supernatant, and measured the remaining Cr in the solution 6+ content.

[0027] A total of two strains with different morphology were obtained in this screening and culture, which were named YB-01 and YB-03 respectively. The reducing effects of the two strains on hexavalent chromium are shown in Table 1....

Embodiment 2

[0031] Identification of embodiment 2 YB-03 bacteria

[0032] (1) Physiological and biochemical identification: Physiological and biochemical identification was carried out according to the "Bergey's Bacterial Identification Manual", and the results are shown in Table 2.

[0033] Table 2: Physiological and biochemical results of YB-03 bacteria

[0034]

[0035] (2) 16srDNA identification: The total bacterial DNA was extracted by alkaline lysis method, and PCR was performed with 16srDNA universal primers.

[0036] Universal primer: 16s-F: 5'-AGAGTTTGATCCTGGCTCAG-3' (see SEQ ID NO.2)

[0037] 16s-R: 5'-AAGGAGGTGATCCAGCCGCA-3' (see SEQ ID NO.3)

[0038] PCR system: 2×Mix: 25ul; upper primer: 1ul; lower primer: 1ul; template: 1ul; ddH 2 O: 22ul

[0039] The PCR program was as follows: first, pre-denaturation at 94°C for 5 min, and then cycled: DNA denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 1 minute, 30 cycles, and finally ex...

Embodiment 3

[0042] Embodiment 3 YB-03 bacterial resistance test

[0043] (1) will K 2 Cr 2 o 7 , AgNO 3 , HgCl 2 , Cu(SO 4 ) 2 , Zn(SO 4 ) 2 , CoCl 2 , MnCl 2 Configure mother liquors of various required concentrations, filter and sterilize with a sterile 0.45um filter, add to LB to make the medium contain metal ions of different concentrations, insert the reducing bacteria into the medium, 37°C, Shake at 150r / min for 24 hours, observe whether the bacteria grow, and record the highest concentration of reducing bacteria against metal ions.

[0044] As can be seen from Table 3, YB-03 bacteria except for Cr 6+ Ions are resistant, and also have varying degrees of resistance to other heavy metal ions.

[0045] Table 3: Resistance of YB-03 bacteria to metal ions

[0046]

[0047] (2) Add ampicillin, kanamycin, cephalosporin, streptomycin, tetracycline and hygromycin to the medium, and observe the resistance of the reducing bacteria.

[0048] It can be seen from Table 4 that YB-0...

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Abstract

The invention relates to reductive chromium ion bacillus thuringiensis YB-03 and a cultivation method and application of the reductive chromium ion bacillus thuringiensis YB-03. According to the invention, the separated reductive chromium ion bacillus thuringiensis YB-03 strain with CGMCC (China General Microbiological Culture Collection Center) NO.5653 not only can reduce Cr<6+>, but also can generate resistance to Ag<+>, Hg<2+>, Cu<2+>, Zn<2+>, Co<2+>, Mn<2+> at different degrees, is insensitive to penbritin and streptomycin and has important application value in treatment of chromium pollution.

Description

technical field [0001] The invention relates to a chromium-resistant bacterium, in particular to a chromium-ion-reducing Bacillus thuringiensis YB-03 and its cultivation method and application. Background technique [0002] Chromium and its compounds are widely used in industry, resulting in a large amount of chromium-containing wastewater and waste residues, causing serious pollution to the environment and endangering public health. Hexavalent chromium is extremely toxic and is one of the main heavy metals causing environmental pollution. Therefore, the control of chromium pollution has aroused widespread concern. Traditional physical and chemical treatment methods include pH adjustment, flocculation precipitation, ion exchange and activated carbon adsorption, etc., but these methods need to consume other energy or use a large amount of chemical reagents, the treatment cost is relatively expensive, and there is a possibility of secondary pollution. Therefore, the biologic...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/07C02F101/22
Inventor 杨丽荣薛保国魏斐全鑫马毅辉梁慎王恒亮武超
Owner HENAN ACAD OF AGRI SCI
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