Acinetobacter sp. and preparation method of acinetobacter sp. agent

A technology of Acinetobacter and inoculum, applied in the field of preparation of Acinetobacter and inoculum, can solve the problem of low ammonia nitrogen degradation efficiency, and achieve the effects of high degradation efficiency, strong environmental adaptability, and easy large-scale production

Active Publication Date: 2014-04-30
WUHAN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the problem of low ammonia-nitrogen degradation efficiency in p...

Method used

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  • Acinetobacter sp. and preparation method of acinetobacter sp. agent
  • Acinetobacter sp. and preparation method of acinetobacter sp. agent
  • Acinetobacter sp. and preparation method of acinetobacter sp. agent

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1, Isolation and purification of strains

[0018] (1) Source of the strain

[0019] The bacteria source was collected from the activated sludge of the wastewater biological treatment system of Wuhan Iron and Steel Coking Company.

[0020] (2) Isolation and purification of strains

[0021] The activated sludge collected is mixed, and 2 mL is inoculated in 100 mL of sterilized LB medium, the composition of the LB medium is: peptone is 1 wt%, sodium chloride is 1 wt%, yeast extract is 0.5 wt%; then cultured on a constant temperature shaker at 30°C and 150r / min for 15 h; then took 5 mL of the culture solution and transferred it to the sterilized nitrification medium, transferred 4 times, and gradually increased during the transfer process. The concentration of ammonium sulfate is as follows: 300mg / L, 600mg / L, 900mg / L, 1200mg / L. During the acclimatization period, phenol was added to the nitrification medium at the same time so that its final concentration was...

Embodiment 2

[0022] Embodiment 2, Identification of strains

[0023] (1) Morphological observation of the strain

[0024] Strain Y 3 After gradient dilution of the culture solution, spread it evenly on the top of the solid nitration medium, place it in a 30°C incubator for 22 hours, and observe the morphology. The results are as follows: figure 1 shown. Depend on figure 1 It can be seen that the bacterial colony is milky white, round, with a raised middle, smooth and moist surface, and neat edges. Pick a few colonies, after Gram staining, observe under the oil lens of an optical microscope, the results are as follows: figure 2 As shown, it can be clearly seen that the individual bacteria of the strain with the preservation number CCTCC NO: M2013445 are short rod-shaped, red in color, and are Gram-negative bacteria.

[0025] (2) 16S rDNA identification of the strain

[0026] Extract strain Y with DNA extraction kit 3 The genomic DNA was then cloned and sequenced for the 16S rDNA...

Embodiment 3

[0027] Embodiment 3, Acinetobacter ( Acinetobacter sp.) Y 3 Preparation of bacteria

[0028] Prepare 3L of nitrification medium, add it to a 5L fermenter, and add 1.5mL of Dow antifoaming agent, then place it in a high-pressure steam sterilizer, and sterilize it at 121°C for 25 minutes, and the culture to be sterilized After cooling the base, the Acinetobacter

[0029] ( Acinetobacter sp.) Y 3 Inoculate 3 vol% into the nitrifying medium, ferment at 30°C, 150 r / min and 0.5 (V / V min) aeration, stop fermentation after 24 hours, and add 25 vol% glycerol as a protective agent , for cryopreservation.

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Abstract

The invention discloses acinetobacter sp. and a preparation method of an acinetobacter sp. agent. According to the technical scheme, the provided acinetobacter sp. Y[3] is collected in the China Center for Type Culture Collection (CCTCC) on September 25, 2013, has a collection number of CCTCC NO:M2013445 and is heterotrophic nitrification bacteria; the acinetobacter sp. agent is prepared by inoculating the acinetobacter sp. Y[3] into a nitrification culture medium and fermenting for enlargement culture. The preparation method of the agent is simple and easy for large-scale production. The acinetobacter sp. can smoothly have a nitrification reaction in higher-concentration phenol-containing industrial wastewater and have simultaneous nitrification and denitrification reactions by taking phenol as a carbon source to convert ammonia nitrogen and organic nitrogen into nitrogen gas without nitrite nitrogen accumulation, achieves simultaneous efficient degradation of ammonia nitrogen and phenol in the wastewater, and has a good application prospect in the biological denitrification process of the phenol-containing industrial wastewater.

Description

technical field [0001] The invention belongs to the technical field of biological denitrification of environmental pollutants. In particular, it relates to a preparation method of Acinetobacter and bacterial agent thereof. Background technique [0002] In recent years, in order to improve the treatment capacity of wastewater biological systems, the so-called high-efficiency bacteria technology (bioaugmentation technology) has been developed rapidly by adding high-efficiency dominant bacteria to the system or engineering bacteria produced by genetic engineering. research hotspots in the field. [0003] In foreign countries, especially in developed countries, environmental microbial agents are widely concerned and valued by people, and it has become a new development trend. The nitrifying bacteria sold by Novozymes in Denmark and Bioform in the United States are composite bacterial agents, which contain a large number of autotrophic nitrifying bacteria and a certain amount o...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/16C02F101/38C02F101/34
Inventor 颜家保许龙龙杨洋余永登张浩武文丽陈佩陈冬冬
Owner WUHAN UNIV OF SCI & TECH
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