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Method for treatment of water pollutants by protein mediator based immobilized laccase

A technology for immobilizing laccase and water treatment, applied in water pollutants, chemical instruments and methods, biological water/sewage treatment, etc., can solve the problems of high cost and unstable use properties, and achieve low cost, easy operation, The effect of shortening the reaction time

Active Publication Date: 2016-08-10
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the practical application of this technology requires the use of small mediator molecules that are unstable, costly, and may produce toxic products.

Method used

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  • Method for treatment of water pollutants by protein mediator based immobilized laccase
  • Method for treatment of water pollutants by protein mediator based immobilized laccase
  • Method for treatment of water pollutants by protein mediator based immobilized laccase

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of immobilized laccase

[0031] Step 1: Weigh 2g of sodium alginate, add water to 100mL, and heat until completely dissolved.

[0032] Step 2: After cooling, add a certain amount of laccase and hemoglobin, and stir for 5 minutes to make it uniform.

[0033] The laccase is white rot fungus (T. versicolor) laccase, purchased from Sigma-Aldrich Company.

[0034] Step 3: Use a syringe to drop the above liquid into 0.05mol / L CuSO 4 In the solution, let stand at room temperature for 0.5h until it solidifies completely.

[0035] Step 4: Wash the prepared immobilized enzyme repeatedly with distilled water until no blue precipitate is produced after adding NaOH to the washing solution. The prepared immobilized laccase is light blue-green spheres with a diameter of about 2mm and a weight of about 12.5mg.

Embodiment 2

[0037] Activity determination of immobilized laccase

[0038] The o-tolidine solution was chosen as the substrate for the determination of immobilized laccase activity.

[0039] Step 1: Measure 150mL of concentrated hydrochloric acid and dilute to 500mL with distilled water. Weigh 1.00 g o-tolidine, and suck 5 mL of the prepared hydrochloric acid into a glass mortar and grind into a paste. Then put it into a 1000mL container and add 150mL distilled water to dilute, at the same time add 495mL dilute hydrochloric acid and fully stir with a glass rod, and finally dilute to 1000mL with distilled water.

[0040] Step 2: Take 0.2mL o-tolidine solution and add 1.8mL NaAc-HAc buffer (pH=4.6), add 72.5mg of immobilized laccase, continuously blow in air and stir, and react for 5min.

[0041] Step 3: Use an ultraviolet spectrometer (Agilent 8453) to detect the change in the content of the o-tolidine catalyzed product, and detect once every 30s. The relative reaction rate of the immobilized lac...

Embodiment 3

[0044] Immobilized laccase treatment of benzidine hydrochloride

[0045] Step 1: Weigh 0.026 g of benzidine hydrochloride and add it to 100 mL of water to prepare a solution with a concentration of 1 mM.

[0046] Step 2: Take 0.2 mL of benzidine hydrochloride solution and add 1.8 mL of NaAc-HAc buffer (pH=4.6), add 0.25 g of immobilized laccase, continuously blow in air and stir, and react for 5 min.

[0047] Step 3: Use an ultraviolet spectrometer (Agilent 8453) to detect the change in the product content after benzidine hydrochloride catalyzed by an average of every 30s. The relative reaction rate of immobilized laccase was obtained by using the spectral results.

[0048] The characteristic wavelengths of benzidine hydrochloride are 360nm, 617nm and 850nm, and 360nm is selected as the detection point. Such as figure 1 As shown in Table 1, by comparing the reaction rates of immobilized laccase and immobilized hemoglobin-laccase, the relative reaction rate of immobilized hemoglobin-l...

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Abstract

The invention relates to a preparation method of immobilized laccase for treatment of organic pollutants in water in the field of biological environment governance. Laccase and hemoglobin serving as a mediator are added into a sodium alginate solution, the substances are mixed, then the mixed solution is dripped into a CuSO4 solution to carry out reaction so as to obtain precipitate, i.e. immobilized laccase. According to the invention, hemoglobin is used as a protein mediator and is immobilized on a copper alginate carrier together with laccase to improve the activity of laccase, at the same time the level for treatment of organic pollutants in water is also enhanced. With a treatment concentration of 0.1mM for pollutants in a water sample, the method can be applied to treatment of water samples in polluted areas.

Description

Technical field [0001] The invention relates to a technology in the field of biological environment treatment, in particular to a method for preparing an immobilized laccase based on a protein mediator for treating organic pollutants in water. Background technique [0002] The "three wastes" in the industry and the toxic chemicals produced during the decomposition of chemical pesticides pollute the environment. Among them, phenol, chlorophenol and aniline are the main organic pollutants. Such pollutants accumulate in large amounts in the environment and cause great harm to human health. Common treatment methods mainly include adsorption method, ultrafiltration method, ozone method, extraction method, ultrasonic catalytic degradation method, chemical sedimentation method, activated sludge method, etc., but they all face toxicity, high energy consumption, secondary pollution, etc. Series of questions. With the increasing maturity of biotechnology, the use of biotechnology to trea...

Claims

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Application Information

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IPC IPC(8): C02F3/34C02F101/30
Inventor 赵一雷潘婷陈启华
Owner SHANGHAI JIAO TONG UNIV
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