Functionalized dendritic mesoporous SiO2 immobilized-chloroperoxidase reactor with surface charge regulation and application thereof

A technology of chloroperoxidase and surface charge, applied in the direction of oxidoreductase, immobilized on or in the inorganic carrier, fixed on/in the organic carrier, etc., can solve the problem of weak electricity and easy leakage of enzyme molecules , weak electrostatic interaction and other problems, to achieve good tolerance, overcome the leakage of enzyme molecules, and improve the effect of immobilized capacity

Active Publication Date: 2019-08-16
SHAANXI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the weak electrical properties of silica itself, the electrostatic interaction with CPO is relatively weak, and the enzyme molecules are easy to leak during repeated use.

Method used

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  • Functionalized dendritic mesoporous SiO2 immobilized-chloroperoxidase reactor with surface charge regulation and application thereof
  • Functionalized dendritic mesoporous SiO2 immobilized-chloroperoxidase reactor with surface charge regulation and application thereof
  • Functionalized dendritic mesoporous SiO2 immobilized-chloroperoxidase reactor with surface charge regulation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Disperse 1 g of DSP nanoparticles into 30 mL of anhydrous toluene, add 0.5 mL of 3-aminopropyltriethoxysilane aqueous solution with a volume concentration of 97% dropwise, reflux at 110 ° C for 15 h, and centrifuge the resulting mixture. Washed three times with acetone and dried under vacuum at room temperature for 12 h to obtain amino-modified dendritic mesoporous SiO 2 (DSP-NH 2 ).

[0033] 100mg DSP-NH 2 Disperse evenly in 100mL of deionized water to form solution a; add 0.37g of N-hydroxysuccinimide sulfonic acid sodium salt and 0.20g of 1-ethyl-(3-dimethylaminopropyl) carbon to 20mL of deionized water Diimine hydrochloride, dissolve the solid, then add 60mL of an aqueous solution containing 0.113g of hyaluronic acid (HA) to the solution, mix well to form solution b; mix solution a and solution b evenly, and use triethylamine Adjust the mixed solution to pH 9, stir at 38 °C for 12 h, centrifuge, wash the product three times with deionized water, and dry it in vac...

Embodiment 2

[0036] Prepare DSP-NH according to the method of embodiment 1 2 . 100mg DSP-NH 2 Evenly dispersed in 2.5mL0.1mol L - 1 PBS buffer solution with a pH of 7.5 and 100 μL of glutaraldehyde aqueous solution with a volume fraction of 50%, let it stand at room temperature for 1 hour, and then centrifuged the mixed solution with 0.1mol L -1The PBS buffer solution with a pH of 7.5 was centrifuged and washed three times, and the supernatant was removed to obtain DSP-NH cross-linked glutaraldehyde 2 Dispersions. Add 50mg chitosan (CHIT) to 2.5mL 0.1mol·L -1 In aqueous HCl solution, the chitosan solid was completely dissolved; after that, the above-mentioned cross-linked glutaraldehyde DSP-NH 2 The dispersion liquid was mixed with the chitosan solution, stirred at room temperature for 12 hours, centrifuged, and the product was washed three times with PBS buffer solution with pH 8, and dried in vacuum at room temperature to obtain dendritic mesoporous SiO2 of cross-linked chitosan. ...

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Abstract

The invention discloses a functionalized dendritic mesoporous SiO2 immobilized-chloroperoxidase reactor with surface charge regulation and application thereof. The reactor herein is made by crosslinking surface amino-modified dendritic mesoporous SiO2 nanoparticles with hyaluronic acid or with protonated amino chitosan, and using the crosslinked material as a carrier to immobilize chloroperoxidasethrough electrostatic action and hydrogen bond action. Ducts of the carrier used in the reactor of the invention carry rich negative charge or positive charge, rich hydroxyl groups and other functional groups are contained, and therefore, the carrier can bind with enzyme molecules under greater force, and enzyme carrying capacity is increased. The reactor of the invention has good catalytic activity, thermal stability and reusability, has good tolerance in the organic solvents, such as DMF (dimethylformamide), methanol and acetonitrile, and can degrade the drugs, such as levofloxacin and rifaximin, in wastewater under high degrading speed and high degrading rate.

Description

technical field [0001] The invention belongs to the technical field of enzyme immobilization, and specifically relates to a dendritic mesoporous SiO with rich charges in the pores and rich functional groups such as hydroxyl groups 2 Immobilized chloroperoxidase reactor and its application. 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 appl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/14C12N11/10C12N9/08C02F3/34C02F101/34C02F101/36C02F101/38
CPCC12N11/14C12N11/10C12N9/0065C12Y111/0101C02F3/342C02F2101/34C02F2101/36C02F2101/38Y02A50/30
Inventor 蒋育澄王兰兰宋艺超胡满成李淑妮翟全国
Owner SHAANXI NORMAL UNIV
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