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Pseudomonas putida P.PAO-1 with dephosphorization function as well as acclimatization and culture method

A technology of domestication and cultivation of Pseudomonas putida, applied in the direction of using microorganisms, methods based on microorganisms, biochemical equipment and methods, etc., can solve problems such as limiting the application of phosphorus accumulating bacteria and affecting the phosphorus removal ability of phosphorus-removing bacteria strains

Inactive Publication Date: 2015-07-01
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the long proliferating process of bacteria needs to be accompanied by a high concentration of carbon source and a long time of microorganism / wastewater contact time, which is difficult to achieve in actual wastewater treatment projects. Application of Phosphorus Removal in Wastewater with Low Organic Concentration
However, if the traditional culture mode is changed, the enrichment-acclimation two-stage culture mode is adopted to cultivate the strains, that is, the first stage adopts enrichment culture, and the bacteria amount is amplified under the condition of high carbon source and nutritional balance, and the second stage adopts the condition of nutritional imbalance. Domestication and cultivation of the bacteria will induce the pre-synthesis and accumulation of PHB in the bacteria [5] , and a large amount of PHB pre-reserve in the bacteria will have a greater impact on the phosphorus removal ability of potential phosphorus removal strains

Method used

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  • Pseudomonas putida P.PAO-1 with dephosphorization function as well as acclimatization and culture method
  • Pseudomonas putida P.PAO-1 with dephosphorization function as well as acclimatization and culture method
  • Pseudomonas putida P.PAO-1 with dephosphorization function as well as acclimatization and culture method

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Embodiment 1

[0031] Pseudomonas putida strain P.PAO-1 forms a large amount of PHB primary accumulation in vivo and its ability to remove phosphorus under aerobic conditions:

[0032] Pseudomonas putida strain P.PAO-1 (preserved on August 26, 2013 in the General Microbiology Center of China Committee for Microorganism Culture Collection (CGMCC for short), the preservation number is CGMCC No.8073, the same below) was inoculated in 5ml sterilized enrichment medium (each L contains 3g beef extract, 10g peptone, 5g NaCl, 0.02g KH 2 PO 4 , Trace elements: every L contains 5g EDTA, 5g FeSO 4 ·7H 2 O, 1.6gCuSO 4 ·5H 2 O, 5g MnCl 2 4H 2 O, 50mg (NH 4 ) 6 Mo 7 o 24 4H 2 O, 50mgH 3 BO 3 , 10mg KI and 50mg CoCl 2 ·6H 2 (2, the same below) pre-cultured, and then inoculated into ten 100mL sterilized enrichment medium (250ml sterilized Erlenmeyer flask), respectively, at 30°C, 120r min -1 Under the conditions of shaking culture for 24 hours, washed with normal saline for 3 times, centrifu...

Embodiment 2

[0038] Effect of adding Pseudomonas putida strain P.PAO-1 on the phosphorus removal ability of activated sludge after domestication culture

[0039] Under the same culture conditions, equal amounts of domesticated Pseudomonas putida strain P.PAO-1, ​​activated sludge, and activated sludge + domesticated Pseudomonas putida strain P. PAO-1 (the addition ratio is 1:1) is used for anaerobic / aerobic alternate culture, in which anaerobic 5h, aerobic 3h, respectively take 5ml of bacterial liquid at the end of anaerobic culture and during aerobic culture, centrifuge, and measure the bacteria Phosphorus release, phosphorus uptake and COD in the centrifugate.

[0040] The activated sludge collected from the return sludge well of the sewage plant was used for the addition experiment of Pseudomonas putida strain P.PAO-1. Since the sewage plant adopts the anoxic / anaerobic / aerobic (A / A / O) activated sludge process, the sludge contains a large number of active phosphorus-accumulating bacteri...

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Abstract

The invention relates to pseudomonas putida P.PAO-1 with a dephosphorization function as well as an acclimatization and culture method. The collection number of the pseudomonas putida P.PAO-1 is CGMCC No. 8073, and the nucleotide sequence is shown in SEQ ID No. 1. The culture method comprises the following steps: inoculating the strain of the pseudomonas putida P.PAO-1 in 5-10ml of a sterilization and enrichment culture medium and pre-culturing, then inoculating the strain in 100-200ml of a sterilization and enrichment culture medium, carrying out shake culture for 12-24h under a condition of 30 DEG C, cleaning, centrifuging, and re-suspending to form bacterium solution; then carrying out anaerobic and aerobiotic alternating culture and acclimatization to obtain an acclimatized bacterium. The pseudomonas putida P.PAO-1 disclosed by the invention is cultured in a manner of enrichment-acclimatization two-phase culture, the initial accumulation of polyhydroxybutyric acid is formed in the body of the P.PAO-1 strain, and the P.PAO-1 strain is applied to a low-concentration carbon-containing organic wastewater treatment to realize the purpose of efficient dephosphorization.

Description

technical field [0001] The invention belongs to the field of phosphorus-removing functional bacteria and a domestication and cultivation method thereof, and in particular relates to a Pseudomonas putida P.PAO-1 with phosphorus removal function and a domestication and cultivation method thereof. Background technique [0002] In recent years, the eutrophication of water bodies in my country has become increasingly serious; studies have shown that excessive discharge of phosphorus in sewage from a large number of urban sewage treatment plants is an important cause of eutrophication in water bodies [1] . In the biological treatment process for nitrogen and phosphorus removal adopted by sewage treatment plants in my country at this stage, the low biological phosphorus removal ability of activated sludge is an important problem to be solved urgently. [0003] Phosphorus accumulating bacteria (PAOs) are the main contributors to the biological phosphorus removal method. They can de...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/36C12R1/40
CPCC12N1/36C12N1/205C12R2001/40
Inventor 田晴朱艳彬谢学辉杨波李方秦华星张成王康伟黄娜庄林杰孟丹杰王琦
Owner DONGHUA UNIV
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