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Tetrahydrofuran-degrading Pseudomonas Oleovorans DT4 and applications thereof

A technology of Pseudomonas oleagines and tetrahydrofuran, which can be applied to bacteria, microorganism-based methods, water/sludge/sewage treatment, etc., and can solve the problems of long microbial generation time and low tetrahydrofuran biodegradation efficiency.

Active Publication Date: 2010-09-29
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims at the shortcomings of low biodegradation efficiency of tetrahydrofuran and long generation time of microorganisms, and provides a bacterial strain capable of efficiently degrading tetrahydrofuran and its application

Method used

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  • Tetrahydrofuran-degrading Pseudomonas Oleovorans DT4 and applications thereof
  • Tetrahydrofuran-degrading Pseudomonas Oleovorans DT4 and applications thereof
  • Tetrahydrofuran-degrading Pseudomonas Oleovorans DT4 and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1: Isolation and identification of P.OleovoransDT4

[0023] (1) Sample collection and domestication

[0024] The activated sludge from the sewage treatment tank of a chemical plant in Shaoxing was collected on site, and THF was used as the only carbon source and energy source for domestication and enrichment. Several months later, the enriched activated sludge was inoculated into a 250mL sealed saline bottle containing 50mL of BSM medium, and THF was used as the only carbon source and energy source to continue the cultivation and enrichment. The experiment requires a constant temperature (30±1°C) and is kept under aerobic conditions.

[0025] BSM was formulated as follows: 4.5g Na 2 HPO 4 12H 2 O, 1.0g KH 2 PO 4 , 1.8g (NH 4 ) 2 SO 4 , 0.2g MgSO 4 ·7H 2 O, 0.03g CaCl 2 2H 2 O, 1mL trace element mother solution, make up to 1000mL with water.

[0026] Concentration composition of trace element mother liquor: FeSO 4 ·7H 2 O 1.0g / L, CuSO 4 ·5H 2 O ...

Embodiment 2

[0032] Example 2: Characteristics of THF degradation by P.Oleovorans DT4

[0033] Use THF as the sole carbon source of P.Oleovorans DT4, inoculate the bacterium from the seed solution to 50mL BSM medium, so that the initial bacterial concentration (by dry weight) is 3.2mg / L; add THF to make the initial THF concentration 5mM . Put it into a shaker with a temperature of 30°C and a rotation speed of 160r / min for cultivation, and take samples every 2 hours, see the results figure 1 . With the prolongation of time, the bacterial concentration increased gradually, and reached 275.6 mg / L (by dry weight) at 14 hours. This example shows that the degrading bacteria P.Oleovorans DT4 can use THF as the only carbon source and energy source for growth and reproduction, and has the ability to efficiently degrade THF.

[0034] Use 1mol / L NaOH or HCl solution to adjust the BSM medium to different pH values ​​(5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5), under the condition...

Embodiment 3

[0037] Embodiment 3: Degradation of benzene, toluene, ethylbenzene and xylene by P.Oleovorans DT4

[0038] Using benzene and toluene as the sole carbon source of P.Oleovorans DT4 respectively, inoculate 50mL of BSM medium with the seed liquid acclimated and cultivated in toluene, so that the initial cell concentration is 3.2mg / L, and the initial substrate (benzene or toluene ) at a concentration of 1 mM, and the degradation experiment was carried out in a shaker at 30° C. and 160 r / min. As a result, benzene and toluene were degraded completely in 18 and 24 hours respectively, as Figure 4 shown.

[0039] Add 2mM THF and 2mM ethylbenzene to 50mL BSM medium, insert P.Oleovorans DT4 bacterial solution to make the initial concentration reach 3.2mg / L (based on the dry weight of bacterial cells), and place on a shaker at 30°C and 160r / min In the degradation experiment. As a result, ethylbenzene began to be degraded when THF was completely degraded. Using the same method, P.Oleov...

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Abstract

The invention discloses a Pseudomonas Oleovorans DT4 capable of degrading tetrahydrofuran effectively, which is preserved in China Center for Type Culture Collection, Address: Wuhan University, Wuhan, China, 430072, Preservation Date: 07.15.2009, Preservation Number: CCTCC NO: M 209151. The tetrahydrofuran-degrading strain according to the invention, which is aerobic non-fermented Gram negative bacteria, can effectively degrade the substrate by taking tetrahydrofuran as carbon source during energy growth; the strain can degrade benzene and methylbenzene, and degrade ethylbenzene and dimethylbenzene in a manner of cometabolism; and the invention lays a foundation for application in purifying THF wastewater and waste gas containing projects by biological method.

Description

(1) Technical field [0001] The invention relates to a strain of Pseudomonas Oleovorans DT4 capable of degrading tetrahydrofuran and its application. (2) Background technology [0002] Tetrahydrofuran (THF) is the most widely used cyclic ether compound in industry. As a solvent, it can dissolve all compounds except polyethylene, polypropylene and fluororesin. It is known as "universal solvent"; THF is an anti-infective drug and hormone It is an important raw material for organic synthesis of drugs, and it can also be used to synthesize polytetrahydrofuran to produce spandex elastic fibers and polyurethane materials. In 2006, the total consumption of THF in the world was about 480,000 tons; in my country, it was about 106,000 tons, and the demand in 2010 is expected to reach 215,000 tons. However, THF is a cytochrome enzyme P450 inhibitor, which can invade the body through the respiratory tract, digestive tract, and skin, and has irritating effects on the skin and mucous memb...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C02F3/34C02F3/02A62D3/02B01D53/84C12R1/38C02F101/34C02F101/36A62D101/28A62D101/20
CPCY02A50/20Y02W10/10
Inventor 陈建孟陈东之周玉央金小君
Owner ZHEJIANG UNIV OF TECH
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