Bromoxynil degrading bacterium and application thereof

A technology for degrading bacteria and bromoxynil, which is applied in the field of environmental microorganisms to achieve good removal effects, protect the ecological environment, and reduce the cost of wastewater treatment

Active Publication Date: 2014-01-08
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Biodegradation is the main way to remove bromoxynil in nature. At present, there is no research report on bromoxynil-degrading bacteria in China. Therefore, microbial strains that efficiently degra

Method used

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  • Bromoxynil degrading bacterium and application thereof
  • Bromoxynil degrading bacterium and application thereof
  • Bromoxynil degrading bacterium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Isolation and identification of bromoxynil-degrading bacteria BX03

[0027] Take 5.0ml of activated sludge obtained from the wastewater treatment tank of a pesticide factory production workshop producing bromoxynil and add bromoxynil concentration to 100ml of inorganic salt liquid culture of 100mg / L. The formula is: 1.50g per liter K 2 HPO 4 , 0.50g KH 2 PO 4 , 0.20g MgSO 4 ·7H 2 O, 1.00g NaCl, 1.00g (NH 4 ) 2 SO 4 , pH 7.0, shake culture at 160r / min, 30°C, transfer to fresh inorganic salt medium every 5 days according to 5% inoculum, and transfer 5 times continuously.

[0028] Take the above bromoxynil-enriched bacterial solution and perform gradient dilution. The dilution is spread on the inorganic salt solid medium containing 100mg / L bromoxynil, and the medium formula is: every liter contains 1.5g K 2 HPO 4 , 0.5g KH 2 PO 4 , 0.2g MgSO 4 ·7H 2 O, 1.0g NaCl, 18g agar, pH=7.0, culture at 30°C for several days. A single colony was picked from i...

Embodiment 2

[0029] Embodiment 2 The method that bromoxynil degrading bacteria BX03 produces bacterial agent

[0030] Activate the original seed of the bromoxynil-degrading bacteria BX03 (CGMCC NO.7438) of the present invention on a petri dish and in an incubator, culture at 30°C for 30 hours, and measure the degradation performance, and inoculate it on the inclined surface of the test tube after the mature colonies spare. The test tube was inoculated in a 1000ml shake flask containing 200ml LB medium, LB medium formula (g / L): yeast extract 5.00, peptone 10.00, NaCl 10.00, pH 7.0, constant temperature shaking culture to logarithmic phase, ready for inoculation seed jar. The seed tank is 500 liters, the feeding volume is 400 liters, the medium formula is: glucose 0.8%, (NH 4 ) 2 SO 4 1%, K 2 HPO 4 0.2%, MgSO 4 0.05%, NaCl0.01%, CaCO 3 0.3%, yeast extract 0.02% (the above "%" means g / 100ml), pH value 7.2-7.5. After the feeding is completed, 121°C high-pressure damp heat sterilizatio...

Embodiment 3

[0031] Example 3 Cell Immobilization Experiment Containing Burkholderia BX-03 Bacterial Agent

[0032]Take the BX-03 (CGMCC NO.7438) bacterial culture solution cultured in LB medium for 8 hours, collect the bacteria by centrifugation, wash twice with sterile water, and make the bacteria concentration 20g L ‐1 bacterial suspension. Mix 1.5mL bacterial suspension with 20mL 2% sodium alginate solution, add 3% CaCl drop by drop 2 Immobilized gel beads of uniform size were formed in the solution, washed twice with sterile water, placed in a 4°C refrigerator for cross-linking and solidified for 4 hours, filtered and washed with sterile water, and set aside.

[0033] The first group: when the concentration of bromoxynil is 100mg·L ‐1 In 100ml of inorganic salt culture medium, add the bacterial suspension cultured by the above-mentioned LB according to the inoculum amount of 1.5%, at 30°C, 160r·min ‐1 Cultivate on a shaking table, take samples every 4 hours until 24 hours, measure ...

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Abstract

The invention discloses a bromoxynil degrading bacterium and application thereof, and belongs to the field of high biotechnologies. A bacterial strain of the bromoxynil degrading bacterium is a Gram staining reaction negative bacterium BX03, and belongs to Burkholderia bacterial genus (Burkholderia sp.) is identified as Burkholderia sp.. The bromoxynil degrading bacterium is mainly biologically characterized in that the bromoxynil degrading bacterium is Gram-negative, is in the shape of a short spar and is free of spores, and flagella grow at the ends of the bromoxynil degrading bacterium. Bromoxynil can be used as the only carbon source and the only nitrogen source to grow the bacterium, and the bacterium can be mineralized. The bromoxynil degrading bacterium and the application have the advantages that 95% of 100mg/L of the bromoxynil can be degraded by the bacterial strain within 24 hours under a flask shaking condition in a laboratory, and accordingly the problem of difficulty in biologically degrading bromoxynil during waste water treatment can be solved.

Description

technical field [0001] The invention relates to the technical field of environmental microorganisms, in particular to a bromoxynil-degrading bacterium and its application. Background technique [0002] The extensive use of herbicides has had a huge impact on the environment. First of all, the use of chemical herbicides not only effectively controls weeds, but also reduces the number of microorganisms in the soil and the activity of some enzymes in the soil to a certain extent, thereby affecting soil quality and fertilizer supply capacity, and the greater the amount of application, the greater the impact. Big. Second, herbicides can pollute groundwater, thereby affecting human health. [0003] Bromoxynil is widely used abroad as a contact herbicide for selective post-emergence stem and leaf treatment in the form of its octanoate, sodium salt, and potassium salt. It is mainly absorbed by the leaves, and rapidly causes tissue necrosis by inhibiting the various processes of p...

Claims

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Application Information

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IPC IPC(8): C12N1/20C02F3/34C12R1/01C02F101/38
Inventor 蔡天明陈立伟徐静刘时旸胡江
Owner NANJING AGRICULTURAL UNIVERSITY
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