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Cupriavidus sp. IDO capable of aerobic degrading indole and application thereof

A technology of copper greedy bacteria and indole, which is applied in the field of copper greedy bacteria, can solve the problem of limited research on the genus

Inactive Publication Date: 2014-10-15
DALIAN UNIV OF TECH
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The genus Coppercoronas is a bacterium that has received widespread attention in recent years and can be used to control environmental pollution. This genus can degrade most aromatic pollutants and can be applied to repair heavy metal pollution and other fields. However, there are not many studies on this genus in China at present.

Method used

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  • Cupriavidus sp. IDO capable of aerobic degrading indole and application thereof
  • Cupriavidus sp. IDO capable of aerobic degrading indole and application thereof
  • Cupriavidus sp. IDO capable of aerobic degrading indole and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: the acquisition of copper greedy bacterial strain

[0020] The soil samples from the seaside of Heishijiao, Dalian were diluted and plated, and the samples were spread in 1 / 10 LB (NaCl 1 g / L, peptone 1 g / L, yeast powder 0.5 g / L, pH=7, ind Indole 100 mg / L) plate, cultivated at 30°C, picked a single colony and used inorganic salt medium ((NH 4 ) 2 SO 4 2.0 g / L, KH 2 PO 4 2.0 g / L, Na 2 HPO 4 12H 2 O 3.28 g / L, FeCl 3 0.00025 g / L, indole 100 mg / L) culture, 30 ℃, pH 7.0, 150 rpm to obtain the bacterial liquid. The cell morphology of IDO is short rod-shaped, with a size of about 0.5 μm, such as figure 1 .

Embodiment 2

[0021] Embodiment 2: 16S rRNA molecular identification of bacterial strain

[0022] Pick a single colony grown on an inorganic salt solid medium plate and dissolve it in 10 μL of sterilized water, denature at 99°C and centrifuge to take the supernatant as a template for PCR amplification reaction. Use TaKaRa 16S rDNA Bacterial Identification PCR Kit with Forward primer / Reverse primer 2 as primers to amplify the target fragment. The total volume of the PCR reaction system is 50 μL: PCR Premix 25 μL, Forward Primer 0.5 μL, Reverse Primer 20.5 μL, template DNA 5 μL, sterile ultrapure water 19 μL. The PCR reaction conditions were: pre-denaturation at 94°C for 5 min, after 30 cycles, including denaturation at 94°C for 1 min, annealing at 55°C for 1 min, extension at 72°C for 1.5 min, after 30 cycles, extension at 72°C for 5 min, Take 5 μL of PCR products for 1% agarose gel electrophoresis, and UV detection after EB staining. Use TaKaRa Agarose Gel DNA Purification Kit Ver. 2.0 (T...

Embodiment 3

[0025] Embodiment 3: the broad-spectrum determination of the substrate of copper greedy bacteria IDO

[0026] Utilizing the bacterial solution in Example 1, using inorganic salt medium for cultivation, 5% inoculum, specifically investigated the growth of bacterial strain IDO with multiple compounds as the only carbon source, including benzoic acid, tryptophan, phenol, and catechol , salicylic acid, gentisic acid, indole, isatin, quinoline, pyridine, naphthalene, phenanthrene, biphenyl, etc., the concentration of each compound is 50 mg / L. Table 1 shows the effect of IDO of C. spp. in the medium with different compounds as the only carbon source.

[0027] Table 1 Investigation on the broad-spectrum substrates of IDO of C.

[0028] substance result substance result benzoic acid + 3-Methylindole - Tryptophan + 2-Methylindole - phenol + indole acetic acid - Catechol + quinoline - salicylic acid + pyridine - Gentisic...

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Abstract

The invention relates to a cupriavidus sp. IDO capable of aerobic degrading indole and the application thereof, and belongs to the field of microbial biotechnology. The bacterial strain is separated from soil samples at the seaside of Black Reef in Dalian City, Liaoning Province, China is preserved in the China General Microbiological Culture Collection Center, CGMCC on June 4th, 2014, and the preservation number of the bacterial strain is CGMCC No.9265. The bacterial strain IDO is a Gram-negative bacterium and is in a short-rod shape, and through the determination of a 16SrRNA gene sequence, the bacterial strain IDO and the cupriavidus have higher likeness, so that the bacterium is classified and named as Cupriavidus sp. IDO, and the GenBank accession number of the16SrRNA gene sequence of the bacterial strain is KJ875862. The bacterial strain uses various aromatic compounds as a unique carbon source for growth, has a higher degrading capacity to the indole, also has better pH and temperature tolerance, and shows the application prospects in the treatment of wastewater containing high-concentration indole.

Description

technical field [0001] The invention relates to a strain of copper greedy bacteria for aerobically degrading indole and application thereof, belonging to the field of microbial biotechnology. Background technique [0002] With the development of industry, the situation of water pollution in our country is becoming more and more severe. Coking wastewater, pharmaceutical wastewater and other industrial wastewater contain a large amount of nitrogen heterocyclic compounds, which are seriously harmful to the environment. Indole is one of the common nitrogen heterocyclic organic pollutants in industrial wastewater. It has high fluidity and water solubility, so it has potential hazards to the natural environment and human health. [0003] Compared with physical and chemical methods, biological methods have the advantages of less impact on people and the environment, mild conditions, and low cost, so they are widely used in pollutant treatment. At present, scholars at home and abr...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/02C02F3/34C12R1/01C02F101/38
CPCY02W10/10
Inventor 曲媛媛马桥张照婧沈娥王经伟沈文丽李端行李会杰刘紫嫣周集体
Owner DALIAN UNIV OF TECH
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