Variovorax paradoxus DEA-3 and immobilization and application thereof

A technology of immobilizing small balls and phages, which is applied in the field of environmental microorganisms, can solve problems such as entering the environment and water pollution, and achieve the effects of low production cost, good degradation effect, and convenient use

Inactive Publication Date: 2017-04-26
JIANGSU NANZI ENVIRONMENTAL PROTECTION SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since dimethylethanolamine is miscible with water, a large amount of dimethylethanolamine

Method used

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  • Variovorax paradoxus DEA-3 and immobilization and application thereof
  • Variovorax paradoxus DEA-3 and immobilization and application thereof
  • Variovorax paradoxus DEA-3 and immobilization and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0023] Example 1 Controversy on the isolation and identification of DEA-3

[0024] The controversial bacterium DEA-3 was obtained from the activated sludge pond of the sewage treatment station of the fine chemical plant.

[0025] The specific process of screening is:

[0026] 1) Take 5ml of activated sludge from the sewage treatment station of a fine chemical plant and inoculate it into an inorganic salt medium containing 200mg / LN,N-dimethylethanolamine. The formula is: 200mg / LN,N-dimethylethanolamine, 1.5 g / LK2HPO4, 0.5g / L KH2PO4, 0.2g / L MgSO4·7H2O, 1.0g / L NaCl, incubate at 30℃-37℃ for 7 days in a shaker incubator, remove the mixed solution 5m and inoculate to 400mg / LN, N -In the inorganic salt medium of dimethylethanolamine, the formula is: 400mg / LN,N-dimethylethanolamine, 1.5g / L K2HPO4, 0.5g / L KH2PO4, 0.2g / L MgSO4·7H2O, 1.0g / L NaCl, the enrichment solution is obtained after 7 days of cultivation;

[0027] 2) Take 1.0ml of N,N-dimethylethanolamine-enriched bacteria solution, add 9...

Example Embodiment

[0030] Example 2 Free controversy that DEA-3 can grow with N,N-dimethylethanolamine as the sole carbon source and carbon source

[0031] Pick a single colony from the plate with the controversial greedy bacterium DEA-3, inoculate it in LB liquid medium, culture for 12h (OD600≈1.0) at 30℃, 160rpm shaker, centrifuge at 8000-12000rpm, discard The supernatant was removed, washed three times with sterile water, and resuspended in an equal volume of sterile water as seed solution.

[0032] A 250mL Erlenmeyer flask was filled with an inorganic salt medium containing 200mg / L para-N,N-dimethylethanolamine. The formula was: 200mg / LN,N-dimethylethanolamine, 1.5g / L K2HPO4, 0.5g / L L KH2PO4, 0.2g / L MgSO4·7H2O, 1.0g / L NaCl, inoculate the PTA-2 seed solution at 1% (volume fraction) of the inoculum, cultivate in a shaker at 30℃, 160rpm, and take out 4mL every 2h The culture broth, the sample was centrifuged in a 8000rpm centrifuge, the supernatant was discarded, and the OD600 was determined by res...

Example Embodiment

[0033] Example 3 Debate on the method of preparing immobilized pellets from greedy phage DEA-3

[0034] 1) Use an inoculating loop to pick a small amount of bacteria on the slant medium and inoculate it in the LB liquid medium at 30°C and shake culture to the logarithmic phase at 160r / min;

[0035] 2) Centrifuge the above-mentioned cultured bacteria liquid at 10000r / min for 3min, pour out the supernatant, add the same volume of sterile water, shake well, centrifuge at 10000r / min for 3min, wash twice, use the same volume Suspended in sterile water to make bacterial liquid.

[0036] 3) Weigh 0.8g of sodium alginate and dissolve it in 20ml of distilled water, add 1000mg / L bamboo charcoal, and ultrasonically disperse;

[0037] 4) Drop the solution obtained in step 3 with a syringe into 3% CaCl at room temperature 2 The solution was cross-linked and calcified at 4°C for 6 hours to obtain calcium alginate gel immobilized pellets, which were washed twice with sterile water and stored at 4°C ...

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Abstract

The invention discloses variovorax paradoxus DEA-3 which is classified and named as variovorax paradoxus DEA-3, the preservation date is February 9, 2015, the full name of the preservation institution is China General Microbiological Culture Collection Center (CGMCC), and the preservation accession number is CGMCCNO.10543. The variovorax paradoxus DEA-3 can grow by use of N, N-dimethyl ethanolamine as the sole carbon source and nitrogen source, the 200mg /L N, N-dimethyl ethanolamine degradation rate of the variovorax paradoxus DEA-3 can reach 94.5%; the 200mg /L N, N-dimethyl ethanolamine degradation rate of an immobilization small ball prepared by the variovorax paradoxus DEA-3 can reach more than 90%, the immobilization small ball prepared by the variovorax paradoxus has a very good degradation effect, and in addition, the immobilization small ball prepared by the variovorax paradoxus is low in production cost, convenient to use, and suitable for promotion and use in the treatment of wastewater containing the N, N-dimethyl ethanolamine.

Description

technical field [0001] The present invention relates to the technical field of environmental microorganisms, in particular to An Argumentative Phagocytosis DEA-3 and its immobilization and application . Background technique [0002] N,N-Dimethylethanolamine is a widely used organic intermediate, it is a colorless or light yellow liquid, it can be used to synthesize cation exchange resin, it can be used as a stabilizer for paint, and an additive for fuel oil , Anti-scaling agent for boilers, with the development of polyurethane industry, it can also be used as an important auxiliary agent for polyurethane processing. Since dimethylethanolamine is miscible with water, a large amount of dimethylethanolamine will enter the waste water and enter the environment during use, causing water pollution. [0003] Microbial degradation has the advantages of low cost, high efficiency, no secondary pollution, and good ecological restoration. The wastewater of ethanolamine is of grea...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N11/14C12N11/10C02F3/34C12R1/01C02F101/38
CPCC02F3/34C02F2101/38C12N11/10C12N11/14C12N1/205C12R2001/01
Inventor 董晓梦袁鹏张永乐周乃亮李珊珊
Owner JIANGSU NANZI ENVIRONMENTAL PROTECTION SCI & TECH
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