Pseudomonas sp. strain as well as cultivation method and application thereof

A Pseudomonas, enrichment medium technology, applied in the direction of chemical instruments and methods, microorganism-based methods, biochemical equipment and methods, etc., can solve the problems of bacteria that have not yet degraded phenol, and achieve high-efficiency phenol degradation performance , less cumulative effect

Active Publication Date: 2017-09-26
JIANGSU SUNTIME ENVIRONMENTAL REMEDIATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no reports of bacteria with both high-effi

Method used

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  • Pseudomonas sp. strain as well as cultivation method and application thereof
  • Pseudomonas sp. strain as well as cultivation method and application thereof
  • Pseudomonas sp. strain as well as cultivation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: Screening and performance determination of Pseudomonas CGMCC No.13433

[0031] The activated sludge is taken to a sewage treatment tank of a factory, and 5mL of mud-water mixture is taken from the activated sludge into 45mL of sterilized heterotrophic nitrification medium. The formula of heterotrophic nitrification medium is: (NH 4 ) 2 SO 4 0.945g / L, sodium citrate 6.536g / L, MgSO 4 ·7H 2 O 1g / L, NaCl 0.12g / L, MnSO 4 ·H 2 O 0.01g / L, FeSO 4 ·7H 2 O 0.02g / L, KH 2 PO 4 0.2g / L, Na 2 HPO 4 0.3g / L, pH 7.0-7.5. Then placed at 30°C, 200rpm air bath shaker enrichment culture for 12h. After gradient dilution of the enriched solution, spread evenly on the heterotrophic nitrification solid medium (agar 20g / L, the rest of the components are the same as the heterotrophic nitrification medium). After culturing in a constant temperature incubator at 30°C for 1 day, single clones with different shapes and sizes were picked, streaked and purified, and number...

Embodiment 2

[0035] Embodiment 2: Molecular biology identification of bacterial strain

[0036] The strains were identified by 16S rDNA sequence alignment. The total DNA of Pseudomonas CGMCC No.13433 was extracted, and the strain 16S rDNA was amplified by a pair of universal primers. The upstream primer was 8f (5'-AGAGTTTGATCCTGGCTCA-3'), and the downstream primer was 1492r (5'-GGTTACCTTGTTACGACTT-3'). PCR reaction system (50 μL): template DNA 1 μL, PCR Taqmix 25 μL, upstream and downstream primers 1.5 μL, DMSO 2 μL, add ddH 2 O to 50 μL of the reaction system. PCR program: 94°C for 5min, 30 cycles of 94°C for 30s, 55°C for 30s, 72°C for 1min and 30s, 72°C for 10min, 4°C for 5min. PCR products were purified and sequenced by Meiji Biomedical Technology Co., Ltd. The 16S rDNA sequence obtained by sequencing was submitted to NCBI, and the homology sequence comparison analysis was carried out through the software Blast and GenBank, and the phylogenetic tree of the strain was constructed us...

Embodiment 3

[0040] Embodiment 3: the resistance research of bacterial strain

[0041] Prepare LB plate with common resistance, LB medium formula: peptone 10g / L, yeast powder 5g / L, NaCl 10g / L, agar 20g / L. Antibiotics were selected for common resistance, including kan, ampicillin (amp), chloramphenicol (cm), streptomycin (sm) and tetracycline (tc), and antibiotics were added to LB medium at a ratio of 1:1000. Pick the strains and smear them on the resistant plates, and culture them in a 30°C incubator. After 1 day, the growth of the strain was observed. As shown in Table 1, Pseudomonas CGMCC No.13433 was resistant to amp, cm, sm and tc, but not to kan.

[0042] Table 1 The resistance result table of Pseudomonas CGMCC No.13433

[0043] can

[0044] In the table, "+" means positive; "-" means negative.

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Abstract

The invention discloses a pseudomonas sp. as well as a cultivation method and application thereof. The pseudomonas sp. is preserved in General Microorganism Center of China Committee for Culture Collection of Microorganisms, and the preservation No. is CGMCC No. 13433. The cultivation method comprises the following steps: inoculating the pseudomonas sp. preserved on a solid medium in an enrichment medium and performing shake cultivation at the constant temperature of 30 DEG C at 200rpm for 12h. The application of the pseudomonas sp. comprises application in biological denitrification of sewage and application in degradation of phenol. The pseudomonas sp. CGMCC No. 13433 has a very good application prospect in treatment of water with overproof ammonia nitrogen and water polluted by phenol in industrial and agricultural water and underground water.

Description

technical field [0001] The invention belongs to the technical field of environmental microorganisms, and relates to a pseudomonas strain, a culture method and application thereof. Background technique [0002] The rapid development of industry and agriculture has intensified the pollution discharge to the surrounding environment. The country's emphasis on environmental pollution discharge and governance is expected to change this situation. One of the main problems in the current water environment pollution is excessive nitrogen, which can cause eutrophication of water bodies and endanger aquatic organisms and human health. Among them, inorganic nitrogen (ammonia nitrogen, nitrate nitrogen and nitrite nitrogen) and organic nitrogen ( Such as protein, amino acid, etc.). Most of the polluted water is ammonia nitrogen exceeding the standard, which will cause eutrophication of the water body, cause algae to multiply or even explode; reduce the dissolved oxygen in the water body...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C02F3/02C12R1/38C02F101/34
CPCC02F3/02C02F3/34C02F2101/345C12N1/20C12N1/205C12R2001/38Y02W10/10
Inventor 唐晓声李海建
Owner JIANGSU SUNTIME ENVIRONMENTAL REMEDIATION
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