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Method for treating water pollutants with immobilized laccase based on protein mediator

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

Active Publication Date: 2018-09-21
SHANGHAI JIAOTONG 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 treating water pollutants with immobilized laccase based on protein mediator
  • Method for treating water pollutants with immobilized laccase based on protein mediator
  • Method for treating water pollutants with immobilized laccase based on protein mediator

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Preparation of immobilized laccase

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

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

[0034] Step 3: Drop the above liquid into 0.05mol / L CuSO with a syringe 4 solution, stand at room temperature for 0.5h until it is completely solidified.

[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, and the prepared immobilized laccase is light blue-green spherical with a diameter of about 2 mm and a weight of about 12.5 mg.

Embodiment 2

[0037] Activity assay of immobilized laccase

[0038] The o-toluidine solution was selected 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 1.00g o-toluidine, and draw 5mL of prepared hydrochloric acid into a glass mortar and grind it into a paste. Then put it into a 1000mL container and add 150mL of distilled water to dilute. At the same time, add 495mL of dilute hydrochloric acid and stir thoroughly with a glass rod, and finally dilute to 1000mL with distilled water.

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

[0041] Step 3: Use an ultraviolet spectrometer (Agilent 8453) to detect the content change of the product after the catalysis of o-toluidine, and detect once every 30 s on average. The relativ...

Embodiment 3

[0044] Treatment of benzidine hydrochloride with immobilized laccase

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

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

[0047] Step 3: Use an ultraviolet spectrometer (Agilent 8453) to detect the content change of the product after the catalysis of benzidine hydrochloride, and detect once every 30 s on average. The relative reaction rate of the 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. like figure 1 As shown in Table 1, by comparing the reaction rates of immobilized laccase and immobilized hemoglobin-laccase, the relative ...

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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 immobilized laccase for treating organic pollutants in water based on a protein mediator. Background technique [0002] The "three wastes" in the industry and the toxic chemicals produced during the decomposition of chemical pesticides are very polluting to the environment, among which phenol, chlorophenol and aniline are the main organic pollutants. Such pollutants accumulate in large quantities in the environment and cause great harm to human health. The 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 problems such as residual toxicity, high energy consumption, and secondary pollution. series of questions. With the increasing maturity of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C02F3/34C02F101/30
Inventor 赵一雷潘婷陈启华
Owner SHANGHAI JIAOTONG UNIV
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