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Preparing method and application of mimic enzyme with double catalysis functions based on hemin mediation gold mineralization path

A functional simulation, heme technology, applied in chemical instruments and methods, physical/chemical process catalysts, organic compound/hydride/coordination complex catalysts, etc. Problems such as low catalytic activity

Active Publication Date: 2017-04-26
QUFU NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the direct application of heme in various catalytic fields still faces great challenges. For example, heme itself has low catalytic activity and is insoluble in water. It is easy to aggregate into dimers with low catalytic activity in aqueous solution. The structure is easily destroyed in oxidizing media and the catalytic activity is weakened

Method used

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  • Preparing method and application of mimic enzyme with double catalysis functions based on hemin mediation gold mineralization path
  • Preparing method and application of mimic enzyme with double catalysis functions based on hemin mediation gold mineralization path
  • Preparing method and application of mimic enzyme with double catalysis functions based on hemin mediation gold mineralization path

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Embodiment 1

[0073] A method for preparing a heme-mediated gold mineralization pathway-based dual-catalysis-function mimetic enzyme, the steps comprising:

[0074] (1) Take hemin, add water and stir at room temperature, prepare Hemin solution with a concentration of 0.1 mg / mL, store at 4°C; take chloroauric acid and add water, prepare HAuCl with a concentration of 10mM 4Solution; get NaOH and add water, be prepared as the NaOH solution of concentration 1.0M;

[0075] (2) 1.0 mL HAuCl prepared in step (1) 4 The solution was mixed with 1.0 mL Hemin solution in equal volume. After 5 min, 20 μL NaOH solution was added under vigorous stirring, and then stirred in a water bath, reacted at 37 ° C for 8 h, centrifuged at 8000 r / s for 20 min, and the supernatant was dialyzed by 14 KDa The bag was dialyzed with deionized water for 12 hours to obtain a dual-catalytic function mimic enzyme (Hemin-AuNCs complex) based on the heme-mediated gold mineralization pathway, which was stored at 4°C.

Embodiment 2

[0077] A method for preparing a heme-mediated gold mineralization pathway-based dual-catalysis-function mimetic enzyme, the steps comprising:

[0078] (1) Take hemin, stir with water at room temperature, prepare Hemin solution with a concentration of 0.4mg / mL, and store at 4°C; take chloroauric acid and add water to prepare HAuCl with a concentration of 10mM 4 solution; take NaOH and add water to prepare a NaOH solution with a concentration of 1.0 M;

[0079] (2) 1.0 mL HAuCl prepared in step (1) 4 The solution was mixed with 1.0 mL Hemin solution in equal volume. After 5 min, 30 μL NaOH solution was added under vigorous stirring, and then stirred in a water bath, reacted at 80°C for 2 h, centrifuged at 8000 r / s for 20 min, and the supernatant was dialyzed by 14 KDa The bag was dialyzed with deionized water for 12 hours to obtain a dual-catalytic function mimic enzyme (Hemin-AuNCs complex) based on the heme-mediated gold mineralization pathway, which was stored at 4°C.

Embodiment 3

[0081] A method for preparing a heme-mediated gold mineralization pathway-based dual-catalysis-function mimetic enzyme, the steps comprising:

[0082] (1) Take hemin, stir with water at room temperature, prepare Hemin solution with a concentration of 0.6mg / mL, and store at 4°C; take chloroauric acid and add water to prepare HAuCl with a concentration of 10mM 4 solution; take NaOH and add water to prepare a NaOH solution with a concentration of 1.0 M;

[0083] (2) 1.0 mL HAuCl prepared in step (1) 4 The solution was mixed with 1.0 mL Hemin solution in equal volume. After 5 min, 50 μL NaOH solution was added under vigorous stirring, and then stirred in a water bath, reacted at 0°C for 13 h, centrifuged at 8000 r / s for 20 min, and the supernatant was dialyzed by 14 KDa The bag was dialyzed with deionized water for 12 hours to obtain a dual-catalytic function mimic enzyme (Hemin-AuNCs complex) based on the heme-mediated gold mineralization pathway, which was stored at 4°C.

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Abstract

The invention provides a preparing method and application of mimic enzyme with double catalysis functions based on a hemin mediation gold mineralization path. The one-pot type in-situ synthesis method is adopted, the hemin and chloroauric acid are mixed under the alkaline condition, then gold in-situ biological mineralization is achieved with the hemin as a reducing agent and a stabilizing agent, and a Hemin-AuNCs compound with Hemin peroxidase catalysis and gold catalysis activity is prepared. The function of a nano wire of the Hemin-AuNCs compound is brought into play, the electronic transmission capability of the Hemin catalysis reaction is promoted, meanwhile, the adsorption capability for a substrate is enhanced, and the catalysis activity of the Hemin-AuNCs compound is obviously higher than that of common Hemin by four or more times. Complex photoelectric instrument usage is not involved in the preparing method, and the beneficial effects that the catalysis activity is high, operation is simple, response is fast and cost is low are achieved. The cancer biomarker alpha fetoprotein immunoassay serves as the example, the Hemin-AuNCs compound is applied to marking of an AFP antibody, then, the enzyme linked immunosorbent assay is adopted, and high-sensitivity detection on the cancer biomarker alpha fetoprotein in blood is achieved through the enzymatic catalysis and gold catalysis silver deposition signal amplification path.

Description

technical field [0001] The invention belongs to the technical field of enzyme catalysis and sensing detection, and relates to a preparation method and application of a simulated enzyme with dual catalytic functions based on heme-mediated gold mineralization pathway. Background technique [0002] Enzyme-linked immunosorbent assay (ELISA) and gold-catalyzed silver deposition staining are two common and highly sensitive detection techniques that have been widely used in the detection of disease markers such as proteins and small biomolecules. These two types of detection technologies involve enzyme labeling and gold labeling of antibodies, respectively. For example, horseradish peroxidase (HRP) is commonly used to label antibodies, but there are limitations such as high price, easy inactivation, and harsh storage conditions. Therefore, choosing an enzyme-like catalyst with low price and high catalytic activity to replace HRP has become a research hotspot in the field of ELISA d...

Claims

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Application Information

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IPC IPC(8): B01J31/22G01N33/574G01N33/68
Inventor 王桦张立燕姜耀刘凤娟刘敏
Owner QUFU NORMAL UNIV
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