Immobilized bio-enzyme microreactor based on starch-based nanomaterial and application thereof

A micro-reactor, nano-material technology, applied in the direction of immobilization on/in organic carriers, biological treatment devices, chemical instruments and methods, etc., can solve the problem of poor mechanical properties of natural starch hydrogels, limiting starch hydrogels Wide application, low stretchability and other problems, to achieve the effect of enhancing immobilization efficiency, improving stability and operating stability, and good tolerance

Active Publication Date: 2021-12-10
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, native starch hydrogels generally have disadvantages such as poor mechanical properties, high brittleness, and low stretchability, which limit the wide application of starch hydrogels in bioenzyme catalysis.

Method used

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  • Immobilized bio-enzyme microreactor based on starch-based nanomaterial and application thereof
  • Immobilized bio-enzyme microreactor based on starch-based nanomaterial and application thereof
  • Immobilized bio-enzyme microreactor based on starch-based nanomaterial and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Ultrasonically disperse 10 mg of graphene oxide (GO) in 10 mL of deionized water to obtain a GO suspension; stir and disperse 1 g of starch (Starch) in 10 mL of deionized water to obtain a starch suspension; add 2 mL of The GO suspension was mixed well so that the final concentration of GO in the mixed solution was 0.2 mg / mL, then the mixed solution was heated to 90 °C and stirred at a speed of 300 rpm for 3 h, and then ultrasonically treated in an ultrasonic bath at 60 °C for 2 h to obtain The homogeneous hydrogel was cooled to room temperature and freeze-dried to obtain a graphene oxide-modified starch hydrogel, denoted as GO@Starch.

[0025] Weigh 5mg GO@Starch, add 1400μL PBS buffer solution with pH=5 and 100μL 0.25mmol·L - 1 HRP solution was shaken at a constant temperature of 29°C for 10 h, and after standing for 6 h, the supernatant was removed, and washed 2 to 3 times with PBS buffer to remove unfixed HRP, and a starch-based nanomaterial-based immobilized HRP m...

Embodiment 2

[0053] Application of the HRP@GO@Starch prepared in Example 1 to degrade bisphenol A in wastewater

[0054] Add 10 mg of HRP@GO@Starch (with HRP@Starch as a comparative test at the same time), artificial wastewater, and different concentrations of bisphenol A into a 10mL centrifuge tube, and finally add 36 μL of 0.1mol L -1 h 2 o 2 An aqueous solution was used to initiate the reaction, keeping the total solution volume at 3 mL. React at room temperature for 25 min under magnetic stirring, and extract 3 times with ethyl acetate after the reaction. Finally, all the extract was evaporated and removed with a rotary evaporator, and then the sample was dissolved with chromatographically pure acetonitrile to obtain a crude sample. The crude sample is filtered through a 0.22 μm organic phase filter membrane and used for high-performance liquid chromatography (HPLC-15C) analysis and determination. 乙腈 :V 水 =90:10) as the mobile phase, the flow rate is 0.5mL min -1 , the detection ...

Embodiment 3

[0058] Application of HRP@GO@Starch prepared in Example 1 to degrade 2,4-dichlorophenol in wastewater

[0059] A series of 2,4-dichlorophenol standard solutions with different concentrations were accurately prepared using methanol as a solvent, filtered through a 0.22 μm organic phase filter, and the peak areas corresponding to different substrate concentrations were determined by high performance liquid chromatography. Concentration and its corresponding peak area were plotted, and the standard curve equation was fitted. Add 10 mg of HRP@GO@Starch (while using HRP@Starch as a comparative test), artificial wastewater, and 2,4-dichlorophenol at different concentrations in a 10mL centrifuge tube, and finally add 36 μL of 0.1mol L - 1 h 2 o 2 An aqueous solution was used to initiate the reaction, keeping the total solution volume at 3 mL. After reacting for 20 minutes in the dark, centrifuge, take out 1000 μL of the reaction solution into the container with a pipette gun, add...

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Abstract

The invention discloses an immobilized bio-enzyme microreactor based on a starch-based nanomaterial and application of the immobilized bio-enzyme microreactor. The bio-enzyme microreactor is obtained by conducting graphene oxide modification in the preparation process of starch hydrogel, taking the starch hydrogel modified with the graphene oxide as a carrier and immobilizing bio-enzyme through electrostatic interaction and hydrogen-bond interaction. The bio-enzyme microreactor takes the starch-based nanomaterial as a carrier, is low in production cost and has industrial application prospects; and the carrier has a compact network structure and a high specific surface area, is rich in hydrophilic groups such as hydroxyl and the like, retains high catalytic activity of free enzymes, improves the loading capacity of the enzymes, has good catalytic activity, thermal stability and reusability and shows good tolerance in organic solvents such as DMF, methanol and acetonitrile; and the immobilized horseradish peroxidase microreactor is used for degrading organic matters such as bisphenol A and 2,4-dichlorophenol in wastewater, and is high in degradation speed and high in degradation rate.

Description

technical field [0001] The invention belongs to the technical field of enzyme immobilization, and in particular relates to a microreactor for immobilizing biological enzymes with a graphene oxide-modified starch hydrogel as a carrier and an application thereof. Background technique [0002] Biological enzyme is a catalytic organic substance produced by living cells, most of which are proteins. It is a non-toxic and environmentally friendly biocatalyst. Biological enzymes have very high catalytic efficiency and high specificity. An enzyme can only catalyze one or one type of chemical reaction, and the reaction conditions are mild. However, free enzymes are easily inactivated in environments such as high temperature, strong acid and strong alkali, and organic solvents. The use of immobilized enzymes can effectively improve the tolerance of enzymes to temperature, acidity, alkali, and organic solvents. Moreover, immobilized enzymes have broad application prospects in sewage tr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C02F3/34C12N11/04C02F101/34
CPCC02F3/342C12N11/04C02F2101/345C02F2203/006Y02W10/10
Inventor 蒋育澄何濛胡满成李淑妮翟全国
Owner SHAANXI NORMAL UNIV
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