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Magnetic-graphene-oxide-immobilized CPO (chloroperoxidase), and preparation method and application thereof

An oxide stone, magnetic technology, applied in chemical instruments and methods, immobilized on or in inorganic carriers, biological water/sewage treatment, etc. problems, to achieve considerable economic benefits, improve stability and operational stability, reduce friction effects

Inactive Publication Date: 2016-03-30
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, chloroperoxidase in the free state has poor stability under extreme conditions, is easily inactivated, and is not easy to separate from the reaction environment for reuse, which restricts its large-scale industrial application.

Method used

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  • Magnetic-graphene-oxide-immobilized CPO (chloroperoxidase), and preparation method and application thereof
  • Magnetic-graphene-oxide-immobilized CPO (chloroperoxidase), and preparation method and application thereof
  • Magnetic-graphene-oxide-immobilized CPO (chloroperoxidase), and preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0040] 2. Preparation of graphene oxide (GO)

[0041] Add 1.0 g of graphite powder and 0.5 g of sodium nitrate to 23 mL of 98% concentrated sulfuric acid with a balance, place the system in an ice bath, and cool the temperature to 0°C. Add 3.0g of potassium permanganate into the mixed solution under vigorous stirring, the speed of addition should be limited, and the temperature of the mixed solution should be controlled below 20°C. Transfer the system from the ice bath, place it on a heating mantle, set the temperature at 35°C, and let it stand for 30 minutes, then slowly add 46 mL of warm water to the mixture, then set the temperature of the heating mantle at a constant temperature of 98°C, and keep 15min. Dilute the mixture to 140mL with warm water, add 2.5mL of 30% hydrogen peroxide, centrifuge, wash 3 times with 5% dilute hydrochloric acid, wash the supernatant with distilled water until the pH is neutral, freeze at -86.5°C for 12h, and then use a freeze dryer dry. In t...

Embodiment 1

[0059] 1. In a nitrogen environment, according to the molar ratio of Fe 3+ : Fe 2+ : volume fraction is 25% ammoniacal liquor=1.5mol: 1mol: 675.7mL, (NH 4 ) 2 Fe(SO 4 ) 2 ·6H 2 O and NH 4 Fe(SO 4 ) 2 12H 2 O was dissolved in ultrapure water to obtain a mixed solution, and 10 mL of ammonia water with a volume fraction of 25% was quickly added to the mixed solution, and slowly dropped into 0.01 g / mL of graphene oxide suspension, graphene oxide and Fe 2+ The mass molar ratio of the mixture was 50g:1mol; then stirred at 85°C for 45min, cooled to room temperature, and separated the synthesized MGO with a magnet. The synthesized MGO was washed 3 times with ultrapure water and 3 times with absolute ethanol, and dried in vacuum at 70°C for 12 hours. After drying, it was ground with a mortar for later use.

[0060] 2. Take 980uL of MGO suspension with a concentration of 1mg / mL and let it stand for 2min after centrifugation. After the solid is separated from the clear liquid, ta...

Embodiment 2

[0062] Maintain the pH=3.5 of the reaction solution by phosphate buffer solution, the CPO dosage and temperature are the same as those in Example 1, and set different reaction times as 5min, 10min, 15min, 20min, 25min, 30min, 35min, 40min, 45min, 50min , 60min, 150min, and 180min to investigate the effect of time on the immobilization amount of CPO loaded onto magnetic graphene oxide. As the reaction proceeds, before 20 minutes, the absorbance of CPO at its characteristic absorption peak at 398 nm gradually decreases, and after 20 minutes, the absorbance of CPO at its characteristic absorption peak at 398 nm gradually increases, so the comparison shows the best fixation time for 20min.

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Abstract

The invention relates to magnetic-graphene-oxide-immobilized CPO (chloroperoxidase), and a preparation method and application thereof. The preparation method comprises firstly getting magnetic graphene oxide, successively adding a phosphoric acid buffer and a CPO solution, and uniformly mixing, so as to obtain a reaction solution, wherein the mass mole ratio of magnetic graphene oxide to CPO is 0.98 mu g:(1.5-3.5)*10<-4> mu mol; oscillating the reaction solution for 5-180 min, then performing separation to obtain a solid A, and cleaning the solid A, so as to obtain magnetic-graphene-oxide-immobilized CPO. In the preparation method, magnetic graphene oxide is utilized as a carrier for immobilizing CPO, and the enzyme biocompatibility is improved. The structural stability of the enzyme molecule and the operation stability are improved. The obtained immobilized CPO can be easily separated out from a reaction environment for reuse, the reuse rate is high, the rate of CPO degrading anthraquinone dyes is high, and industrial application of the immobilized CPO is facilitated.

Description

【Technical field】 [0001] The invention belongs to the technical field of organic dye wastewater treatment, and in particular relates to CPO immobilized by magnetic graphene oxide and its preparation method and application. 【Background technique】 [0002] my country is a relatively developed country in the dye industry and textile printing and dyeing industry, with an annual dye output of 150,000 tons, ranking among the top in the world. With the development of the dye industry and the progress of printing and dyeing processing technology, the stability of the dye structure has been greatly improved, which has increased the difficulty of decolorization treatment. At present, the decolorization of printing and dyeing wastewater has become a major problem that needs to be solved urgently in wastewater treatment at home and abroad. [0003] In addition to biochemical treatment and adsorption, commonly used dye wastewater treatment methods also include physical chemical or chemic...

Claims

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Application Information

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IPC IPC(8): C12N11/14C02F3/34
CPCC02F3/342C12N11/14
Inventor 蒋育澄王凤娇胡满成李淑妮翟全国
Owner SHAANXI NORMAL UNIV
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