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Method for degrading octyl phenol by utilizing laccase

A technology of octylphenol and laccase, which is applied in the field of bioengineering and environmental pollutant treatment, can solve the problems of low degradation efficiency, long treatment time, secondary pollution, etc., achieve high removal efficiency, reduce reaction time, and reduce usage Effect

Active Publication Date: 2013-04-24
NANJING FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have problems such as low efficiency, secondary pollution, and high cost.
The biological method mainly uses the metabolism of microorganisms, so the treatment time is longer and the degradation efficiency is lower; the physical and chemical methods have different degrees of shortcomings and secondary pollution

Method used

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  • Method for degrading octyl phenol by utilizing laccase
  • Method for degrading octyl phenol by utilizing laccase
  • Method for degrading octyl phenol by utilizing laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] The preparation of embodiment 1 laccase

[0052] 1.1 Preparation of recombinant laccases Lcc1 and Lcc2 from Coriolus versicolor

[0053] 1.1.1 Cultivation of Coriolus versicolor

[0054] Versicolor versicolor (Coriolus versicolor, the microorganism can be screened from nature, can also be purchased from commercial channels (for example, can be purchased from the China Industrial Microbiology Culture Collection Management Center)) medium is PDA: glucose 20g / L, Potato juice 20%wt, inoculated with fresh Versicolor versicolor mycelium, cultured at 180rpm at 28°C for 3-4 days and collected by filtration.

[0055] 1.1.2 Extraction of total RNA from Coriolus versicolor

[0056] The collected mycelia of Coriolus versicolor (Coriolus versicolor) was washed once with PBS buffer solution, ground the mycelium under liquid nitrogen to powder, collected into a 2mL centrifuge tube, added 1mL Trizol and shaken vigorously, 4 Centrifuge at 12000g at 12000g for 10min at ℃, transfer the...

Embodiment 2

[0106] Example 2 Specificity of different recombinant laccase isoenzymes to octylphenol

[0107] Add five kinds of laccases to 2U / mL in the reaction system containing octylphenol 50mg / L; adjust the pH to 4.5 with citric acid buffer, control the temperature at 40°C; react in a water bath shaker at a speed of 100rpm After 24h, the residual amount of octylphenol was detected. Specific results such as figure 1 As shown, the degradation rates of octylphenol by the five laccases Lcc1, Lcc2, LacA, LacB, and LacC after 24 hours of reaction were 43.6%, 0%, 80.7%, 75.3%, and 61.1%, respectively. It indicated that laccase Lcc2 could not act on octylphenol, while laccase LacA had the best degradation effect on octylphenol. They are 1.9, 1.1, and 1.3 times of Lcc1, LacB, and LacC, respectively.

[0108] The detection method of octylphenol is as follows: first, add an equal volume of petroleum ether to the 10mL reaction system for liquid-liquid extraction, and extract each sample three t...

Embodiment 3

[0111] Example 3 Effect of laccase LacA on degrading octylphenol pollutants under different octylphenol concentrations

[0112] The concentration of octylphenol in the reaction system is 25mg / L, 50mg / L, 75mg / L, 100mg / L, 150mg / L, 200mg / L, add laccase LacA to a concentration of 2U / mL, and use citrate buffer Adjust the pH to 4.5, control the temperature at 40° C., and detect the residual amount of octylphenol after reacting in a water-bath shaker at a speed of 100 rpm for 24 hours. Test results such as figure 2 Shown: degradation rate decreases gradually along with the increase of octylphenol concentration; When the concentration of octylphenol was lower than 100mg / L, the average degradation rate of octylphenol showed a linear growth trend, and when the concentration continued to increase, octylphenol The upward trend of the average degradation rate of phenol is slow. Therefore, the substrate concentration should be controlled within a certain range (octylphenol concentration ...

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Abstract

The invention discloses a method for degrading octyl phenol by utilizing laccase. The method comprises the steps of: preparing a LacA enzyme solution; adjusting the concentration of the octyl phenol within 25-200 mg / L; adding the laccase having a concentration of 0.5-3 U / mL in the LacA enzyme solution; controlling the pH of a reaction system to 3.5-6; and degrading under the condition of keeping the temperature at 25-50 DEG C. As a bio-enzyme catalysis technology is adopted, the method for degrading the octyl phenol by utilizing the accase has outstanding characteristics of high removal efficiency, good convenience and safety, no pollution and the like. By comparing specifity of different isozymes to the octyl phenol, selecting the laccase having the strongest specifity to the octyl phenol and adding a medium substance, the method for degrading thhe octyl phenol by utilizing the laccase increases the reaction efficiency of the laccas, redudes reaction time, therefore reduces usage amount of the laccase, and lowers operating cost. The method for degrading the octyl phenol by utilizing the laccase can degrade octyl phenol efficiently and can obtain the degradation rate up to 97.2% after 12-hour reaction under a suitable catalysis condition when the concentration of the octyl phenol is 100 mg / L.

Description

technical field [0001] The invention relates to the fields of bioengineering and environmental pollutant treatment, in particular to a method for degrading octylphenol by using laccase. Background technique [0002] The use of octylphenol polyoxyethylene ether began in the 1940s, and its use increased rapidly in the 1980s. It is currently the second largest commercial nonionic surfactant in the world. Alkylphenol is the main intermediate metabolite of octylphenol ethoxylates biodegraded in the environment, although its final degradation product is CO 2 and H 2 0, but this process is very slow due to the existence of the benzene ring. Therefore, a large amount of octylphenol is enriched in the environment. On the one hand, octylphenol enters the body through water or bioaccumulation, through the biological chain, and on the other hand, enters the body through contaminated food. As early as 1938, researchers discovered that alkylphenols have estrogenic activity. Since the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A62D3/02C12N9/02C12N15/81A62D101/28C12R1/84C12R1/645
Inventor 赵林果柴常升裴建军
Owner NANJING FORESTRY UNIV
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