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Immobilized cell of mutagenized bacterium M1 of Rhodococcus ruber SD3 and application thereof in degradation of phenol pollutants

A technology of immobilizing cells and Rhodococcus rhodochrous is applied in the application field of degrading phenol pollutants, and can solve the problems such as brittleness of small balls and easy destruction by microorganisms.

Active Publication Date: 2013-06-19
JIANGXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, pellets made of calcium alginate are brittle and easily destroyed by microorganisms

Method used

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  • Immobilized cell of mutagenized bacterium M1 of Rhodococcus ruber SD3 and application thereof in degradation of phenol pollutants
  • Immobilized cell of mutagenized bacterium M1 of Rhodococcus ruber SD3 and application thereof in degradation of phenol pollutants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Embodiment 1: the phenol degradation performance of the mutagen M1 of Rhodococcus rubrum SD3

[0015] Pick up the mutagen M1 of Rhodococcus erythrococcus SD3 with an inoculation loop, revive it in LB medium, and wait for the strain to grow to logarithmic phase. Inoculate in phenol inorganic salt medium (NaCl 1 g, NH 4 Cl 1 g, MgSO 4 ·7H 2 O 3 g, K 2 HPO 4 1.5 g, KH 2 PO 4 1.5 g of phenol, 1.5 g of phenol, and phenol-free distilled water to 1000 mL), 35 °C, 200 r / min shaker shaking culture for 72 h. The degradation rate of phenol measured by 4-aminoantipyrine spectrophotometry was 99.77% ( figure 1 ).

Embodiment 2

[0016] Embodiment 2: the immobilization method of the mutagen M1 of Rhodococcus rubrum SD3

[0017] 1. Pick out the mutagen M1 of Rhodococcus rubella SD3 from the preserved slant, inoculate it in LB medium liquid, and shake it at 35°C and 200 rpm for 28 h. The activated bacterial solution was inoculated according to the inoculum size of 2%, and then cultured on a shaking table at 35°C and 200 rpm.

[0018] 2. Centrifuge the cultured bacteria liquid at 4000 rpm for 10 min, collect the bacteria in a 50 ml sterile centrifuge tube, add 50 ml of sterile water to shake and wash, then centrifuge at 4000 rpm for 10 min, repeat the above steps and wash once again.

[0019] 3. Take 10 g of wet bacteria and 10 ml of 1% sodium alginate (SA)-1% polyvinyl alcohol (PVA) solution and mix evenly at room temperature.

[0020] 4. Add the above mixed solution into a sterile syringe, and slowly drop it into an ice-cold 0.1 mol / L sterile saturated boric acid-calcium chloride solution.

[0021] 5....

Embodiment 3

[0023] Example 3: Reusability of the immobilized cells of the mutagen M1 of Rhodococcus erythrococcus SD3

[0024] 1. Add the immobilized cells to the phenol inorganic salt medium (NaCl 1 g, NH 4 Cl 1 g, MgSO 4 ·7H 2 O 3 g, K 2 HPO 4 1.5 g, KH 2 PO 4 0.5 g of phenol, 2.0 g of phenol, and phenol-free distilled water to a volume of 1000 mL), cultured on a shaker at 35°C and 200 rpm for 72 h.

[0025] 2. Take out the immobilized cells from the culture medium and wash them twice with sterile water. Add the washed immobilized cells to new phenol inorganic salt medium (NaCl 1 g, NH 4 Cl 1 g, MgSO 4 ·7H 2 O 3 g, K 2 HPO 4 1.5 g, KH 2 PO 4 0.5 g of phenol, 2.0 g of phenol, phenol-free distilled water to 1000 mL), 35 ° C, 200 rpm shaker for 72 h, repeated use five times. The degradation rates of phenol from the first to the fifth times were 99.99%, 99.99%, 98.98%, 99.78% and 98.53%, respectively.

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Abstract

The invention relates to an immobilized cell of a mutagenized bacterium M1 of Rhodococcus ruber SD3 and application thereof in degradation of phenol pollutants. For the mutagenized bacterium M1 prepared by mutagenizing Rhodococcus ruber SD3 with 0.3% lithium chloride and screening, the degradation rate on 1.5 g / L phenol within 72 hours is 99.77%. The immobilized cell having a diameter of 6 mm is prepared by embedding the mutagenized bacterium M1 with 1% sodium alginate and 1% polyvinyl alcohol. When the immobilized cell is repeatedly used 5 times, the degradation rate on 2 g / L phenol within 72 hours is more than 98%. Thus, the mutagenized bacterium M1 of Rhodococcus ruber SD3 and the immobilized cell thereof can efficiently degrade phenol pollutants, and can be used for biological treatment of phenol-containing industrial wastewater.

Description

technical field [0001] The invention belongs to the field of environmental biology technology, and in particular relates to the application of immobilized cells of the mutagen M1 of Rhodococcus rubella SD3 in degrading phenol pollutants. Background technique [0002] Phenol is a major pollutant in industrial wastewater. my country has included phenol in the blacklist of environmental priority pollutants, and has strict regulations on the discharge of phenol-containing wastewater: Under normal conditions, the concentration of volatile phenol in drinking water is 0.001 mg / L, and the concentration of phenol in water source water is 0.001 mg / L. The maximum allowable concentration is 0.002 mg / L. Biological method is currently a more advanced method and the most widely used treatment technology in the field of wastewater treatment, and it is also the main method for the harmless treatment of phenol-containing wastewater in my country. However, due to the toxicity and refracto...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/08C02F3/34C12R1/01C02F101/34
Inventor 彭仁杨贵娟杜芸芸王启明
Owner JIANGXI NORMAL UNIV
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