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Method for optimizing activity of nano-enzyme particles

A nanoparticle and nanozyme technology, applied in the field of nanobiology and nanomedicine research, nanomaterials, can solve problems such as not universal

Active Publication Date: 2018-05-25
INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although nanozyme particles have high catalytic activity, their catalytic efficiency and substrate selectivity still need to be further optimized in practical applications. 2
The current optimization methods for nanozyme particles mainly focus on the control of the nanoscale of nanoparticles, including size, structure, morphology, doping, and surface charge modification, etc. These methods have certain limitations in improving the activity of nanozyme particles. , not universal

Method used

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  • Method for optimizing activity of nano-enzyme particles
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  • Method for optimizing activity of nano-enzyme particles

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Embodiment 1: Amino acid modification Fe 3 o 4 Preparation and Characterization of Nanozyme Particles

[0038] In this example, amino acid modified Fc 3 o 4 Nanozyme particles were synthesized using a hydrothermal method. Specifically as follows: 3g FeCl 3 Add 80mL of ethylene glycol and mix well, after fully dissolved, add 6g of anhydrous sodium acetate, 0.1g of alanine or histidine, mix thoroughly for 30 minutes, then put the sample into the reaction kettle, and react at 200°C for 14 hours . After cooling, magnetically separate, wash with ethanol, dry at 60°C, and store in separate packages. Fe synthesized by this method 3 o 4 The surface of nanozyme particles is modified with alanine or histidine, which are respectively denoted as Ala-Fe 3 o 4 , His-Fe 3 o 4 . where unmodified Fe 3 o 4 The nanozyme particles are named as Naked-Fe 3 o 4 .

[0039] amino acid modified Fe 3 o 4 Nanozyme particles and unmodified Fe 3 o 4Nanozyme Particles Nanopartic...

Embodiment 2

[0041] Example 2 Histidine modification on Fc 3 o 4 Effect of Nanozyme Particle Activity

[0042] 0.2 μg Fe with different amino acid modifications 3 o 4 Nanozyme particles and bare Fe 3 o 4 Nanozyme particles were added to 100 μL of reaction solution (0.2M sodium acetate, pH 4.5). Then add H at a final concentration of 0.1M 2 o 2 And 0.8mM 3,3,5,5-tetramethylbenzidine (TMB), microplate reader iMark TM Microplate Reader (Bio-Rad, USA) measured absorbance at 652 nm.

[0043] The result is as figure 2 As shown in A, histidine modification can significantly increase Fe 3 o 4 Peroxidase activity of nanozyme particles.

Embodiment 3

[0044] Example 3 Histidine modification on Fe 3 o 4 Nanozyme particle to substrate H 2 o 2 The affinity and reaction kinetics of the

[0045] Fe 3 o 4 Nanozyme particles and HRP on the substrate H 2 o 2 The reaction kinetics is carried out according to the following scheme:

[0046] Differently modified Fe 3 o 4 Add 0.2 μg or 0.1ng HRP of nanozyme particles to 100 μL reaction solution (0.2M sodium acetate, pH 4.5). Add 0.8mM TMB and gradient changes of H 2 o 2 , and then use iMark TM Microplate Reader (Bio-Rad, USA) detects the whole reaction system, draws the reaction curve, according to the Mie reaction equation ν=V max ×[S] / (K M +[S]) to calculate the Mie reaction constant. where V max is the maximum reaction rate; [S] is the molar concentration of enzyme added; K M is the Michaelis reaction constant; K cat Worth calculated according to equation K cat =V max / [S] Calculated.

[0047] The Mie reaction equation curve drawn is as follows figure 2 As sho...

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Abstract

The invention provides a method for optimizing activity of nano-enzyme particles. Specifically, a small molecule compound containing an imidazole ring is utilized for modification to improve the affinity of the nano-enzyme particles to a substrate H2O2, and the activity of nano-enzyme particles is optimized. By means of the method, the affinity of the nano-enzyme particles to the substrate H2O2 isimproved by 12 times, and the catalytic efficiency of the nano-enzyme particles is improved by 21 times. The activity of nano-enzyme particles is optimized by simulating the active center of a natural enzyme, more novel methods can be provided for optimization of the activity of the nano-enzyme particles, and the actual application of nano-enzyme particles in the bionanotechnology field can be promoted.

Description

technical field [0001] The invention belongs to the research fields of nanomaterials, nanobiology and nanomedicine. More specifically, the present invention provides a novel method for optimizing nanozyme particles. Background technique [0002] Since the middle of the 20th century, in order to make up for some defects in the practical application of natural enzymes, artificially synthesized enzymes or mimic enzymes have attracted great interest of researchers. However, because of the low activity and low selectivity of artificial synthetic materials, the application range of artificial mimetic enzymes is greatly limited. There are still many challenges to construct an ideal artificial mimetic enzyme. Recently, the inventors found that inorganic Fe 3 o 4 Nanoparticles etc. have inherent enzymatic activity 1 , and this nanomaterial with intrinsic enzyme activity is called nanozyme particle 2 . As a typical representative of nanozyme particles, Fe 3 o 4 The report of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J31/28
CPCB01J31/28
Inventor 阎锡蕴范克龙高利增段德民
Owner INSITUTE OF BIOPHYSICS CHINESE ACADEMY OF SCIENCES
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