A nanometer enzyme reactor prepared based on a hydroxy selective immobilization manner, a preparing method thereof and applications of the reactor

A selective, nano-enzyme technology, applied in the field of biological enzyme treatment, can solve the problems of long reaction time (from tens of minutes to tens of hours, low treatment concentration, many treatment links, etc., and achieve obvious fading effect and high reuse rate The effect of high and broad application prospects

Inactive Publication Date: 2016-05-04
SHAANXI NORMAL UNIV
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Problems solved by technology

This dye and its intermediate degradation products have side effects such as high toxicity, high residue, carcinogenicity, teratogenicity, and mutagenicity, and enter the environment and easily cause environmental pollution
Due to the complex structure of such dyes, the current treatment methods are mainly photodegradation and microbial degradation methods, but both of these methods have low treatment concentrations (<50mg / L), or long reaction times (from tens of minutes to tens of hours) ) shortcomings, the traditional dye industrial treatment process has many processing links, inconvenient operation, long time, high cost, adsorption decolorization has the characteristics of only adsorbing dyes, but not destroying its structure
Advanced oxidation decolorization is considered a promising method, but it is expensive and will cause secondary pollution

Method used

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  • A nanometer enzyme reactor prepared based on a hydroxy selective immobilization manner, a preparing method thereof and applications of the reactor
  • A nanometer enzyme reactor prepared based on a hydroxy selective immobilization manner, a preparing method thereof and applications of the reactor
  • A nanometer enzyme reactor prepared based on a hydroxy selective immobilization manner, a preparing method thereof and applications of the reactor

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preparation example Construction

[0033] The preparation and application adopted scheme of nano-enzyme reactor of the present invention is made up of following steps:

[0034] 1. Graphene oxide (GO) prepared by Hummers method

[0035] To 75mL of concentrated sulfuric acid, add 1g graphite powder, 0.75gNaNO 3 and 3gKMnO 4 , mixed evenly, the mixture was stirred in an ice-water bath for 2 h, and stirred at room temperature for 5 days. Afterwards, 200 mL of 5% sulfuric acid was slowly added to the solution, the temperature was rapidly raised to 98° C., and stirring was continued at 98° C. for 2 h. Cool down to 60°C, pour 6mL of 30% hydrogen peroxide directly into the mixture, and stir for another 2h. The mixture was separated with a centrifuge, the supernatant was discarded, and the mixture was washed with 5% sulfuric acid and 0.5% H 2 o 2 Wash the solid matter with the mixed solution, centrifuge at high speed for 15 minutes, and repeat 5 times; in the same way, continue to wash with 5% hydrochloric acid for...

Embodiment 1

[0053] (1) Graphene oxide (GO) was prepared by the Hummers method;

[0054] (2) Add 0.1mmol to 130μL . L -1 EDAC and 100ug of NHSS were added to the MES buffer solution with a pH value of 6.0, and the ratio of the amount of the substance to n EDAC :n NHSS =5:1, then add 50μL of graphene oxide solution with a concentration of 1mg / mL, stir the mixture at room temperature for 30min, then centrifuge at 13000r / min for 20min to obtain a suspension, and then use 0.05mmol . L -1 The 2-morpholineethanesulfonic acid buffer was thoroughly washed to remove excess EDAC and NHSS in the suspension to obtain esterified graphene oxide.

[0055] (3) Disperse the esterified graphene oxide in 700 μL concanavalin (ConA) solution (0.1mg / mL, pH7.0) to obtain a mixed solution, in which the esterified graphene oxide and conA The mass ratio was 5:7; the mixture was incubated at room temperature for 1 h, then centrifuged at 4°C and 13000 r / min for 20 min, and then 0.05 mmol . L -1 Washing with ME...

Embodiment 2

[0060] Adopt the method for embodiment 1 to prepare nanozyme reactor, change the pH of reaction system in embodiment 1 step (4), other conditions are constant, investigate the immobilized capacity of pH value in the interval of 2~6, from image 3 It can be seen that when the pH value is 2.5, the CPO immobilization rate is the highest, indicating that the optimal pH value of GO biospecific binding method to immobilize CPO is 2.5.

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Abstract

The invention relates to a nanometer enzyme reactor prepared based on a hydroxy selective immobilization manner, a preparing method thereof and applications of the reactor. The method includes dispersing esterified graphene oxide into a concanavalin A solution, incubating at room temperature for 1-1.5 h, separating, cleaning to obtain graphene oxide-concanavalin A, dispersing the graphene oxide-concanavalin A into a chloroperoxidase solution, oscillating until the chloroperoxidase is immobilized to the graphene oxide-concanavalin A, separating and cleaning to obtain the nanometer enzyme reactor. The esterified graphene oxide the surface of which is liable to functionalize is selected as a solid-phase carrier, and the CPO (namely the chloroperoxidase) is immobilized to the esterified GO (namely graphene oxide) by adopting -OH on the surface of the CPO as immobilization sites, adopting concanavalin A as an agglutinin and based on specific recognition to hydroxy, thus preparing the nanometer enzyme reactor high in activity. The enzyme reactor is capable of simplifying industrial processing steps, shortening degradation time and increasing the efficiency when being applied for degradation.

Description

【Technical field】 [0001] The invention belongs to the technical field of biological enzyme treatment of organic sewage, and in particular relates to a nano-enzyme reactor prepared based on a hydroxyl selective immobilization method and its preparation method and application. 【Background technique】 [0002] Chloroperoxidase (CPO) is a heme peroxidase (42kDa) isolated from the marine fungus Caldariomyces fumago. CPO has a wide range of oxidation activities, and is currently considered to be the most widely used enzyme in the peroxidase family. However, CPO has poor stability in extreme reaction environments such as high temperature, strong acid, strong alkali, and organic solvents, and is not easy to remove In order to solve this problem, the enzyme is often immobilized on the surface of the carrier and in the micropores to retain its catalytic activity, and can be recycled and reused. Traditional immobilization methods generally use amino acid residues -NH 2 Or -COOH is the...

Claims

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Application Information

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IPC IPC(8): C12N11/14C12N11/02C02F3/34C02F101/32
CPCC12N11/14C02F3/34C02F2101/32C12N9/0065C12N11/02C12Y111/0101
Inventor 蒋育澄赵睿南胡满成李淑妮翟全国
Owner SHAANXI NORMAL UNIV
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