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Preparation method for nanogold-protein composite material with enzyme catalysis performance

A technology of catalytic performance and composite materials, which is applied in the field of preparation of nano-gold-protein composite materials, can solve the problems of colloidal gold particles, biomolecules falling off, etc., achieve good stability, mild conditions, avoid chemical reactions and purify the effect of the process

Inactive Publication Date: 2013-07-10
苏州汉能环保材料科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This adsorption is closely related to the pH value and salinity of the solution, changing these properties of the solution can easily lead to the aggregation of colloidal gold particles or the shedding of biomolecules

Method used

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  • Preparation method for nanogold-protein composite material with enzyme catalysis performance
  • Preparation method for nanogold-protein composite material with enzyme catalysis performance

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] The preparation of nano-gold-EcoRI endonuclease composite material with enzyme catalytic performance, its preparation route is as follows figure 1 shown.

[0038] 1. Preparation of nano-gold colloid:

[0039] (1) Heat 50 mL of chloroauric acid solution (1 mmol / L) to boiling.

[0040] (2) 5mL freshly prepared sodium citrate solution (sodium citrate concentration is 1wt%) joins in the chloroauric acid solution rapidly, waits for the solution color to turn red, continues to reflux solution and obtains nano-gold colloid for 20 minutes, the diameter of nano-gold about 10 nanometers.

[0041] 2. Preparation of gold nanoparticles protected by mercapto undecanoate (MUA):

[0042] (2-1) The nano-gold colloid obtained in step (1) is purged and degassed with nitrogen for 30 seconds.

[0043] (2-2) Take 450 μL of degassed nano-gold colloid and disperse it in 450 μL of phosphate buffer solution containing Tween 20, keep it stable for at least 20 minutes, then add 100 μL of merca...

Embodiment 2

[0054] The preparation of the nano-gold-Nt.AlwI nicking endonuclease composite material with enzymatic catalytic performance, except that the enzyme is Nt.AlwI endonuclease, other steps are the same as in Example 1.

[0055] The nano-gold colloid obtained in this example, the nano-gold protected by mercaptoundecanoate MUA, and the nano-gold-Nt.AlwI nicking endonuclease composite material were detected to obtain the ultraviolet-visible absorption spectrum, from which it can be seen that the modified Nt.AlwI The final nano-gold colloidal composite material has no obvious plasmon peak shift in the ultraviolet-visible absorption spectrum, indicating that the properties of colloidal gold are not affected.

[0056] The nano-gold-Nt.AlwI nicking endonuclease composite material in this example was identified by supercoiled pUC101DNA (dam- / dcm-), and its Nt.AlwI nicking endonuclease activity was not affected.

Embodiment 3

[0058] The preparation of the gold nanometer-EcoRV restriction endonuclease composite material with enzymatic catalysis, except that the enzyme is EcoRV endonuclease, other steps are the same as in Example 1.

[0059] The nano-gold colloid obtained in this example, the nano-gold protected by mercapto undecanoate MUA, and the nano-gold-EcoRV restriction endonuclease composite material were detected to obtain the ultraviolet-visible absorption spectrum, from which it can be known that the modified EcoRV nano-gold The colloidal composite has no obvious plasmon peak shift in the ultraviolet-visible absorption spectrum, indicating that the properties of the colloidal gold have not been affected.

[0060] The gold nanometer-EcoRV restriction endonuclease composite material of this example was identified by λDNA, and its EcoRV restriction endonuclease activity was not affected.

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Abstract

The invention provides a preparation method for a nanogold-protein composite material with an enzyme catalysis performance. The preparation method comprises the steps of: firstly dispersing nanogold sol into phosphoric acid buffer solution containing Tween 20 after degassing the nanogold sol by nitrogen purging, mixing the solution with mercaptoundecanoic acid, and standing and reacting at the normal temperature; carrying out centrifuging, supernatant discarding and gravity suspension on the mixed solution until supernatant does not contain the mercaptoundecanoic acid; centrifuging and collecting nanogold protected by MUA (mercaptoundecanoic acid), and dispersing the nanogold into the phosphoric acid buffer solution to be standby; mixing the nanogold protected by the MUA with EDAC (Carbodiimide Hydrochloride), NHS (N-Hydroxysuccinimide) and incision enzyme, adding the mixture into MES (Morpholineethanesulfonic acid) buffer solution, and standing and reacting at the normal temperature; and finally repeatedly carrying out the centrifuging, the supernatant discarding and the gravity suspension, and dispersing the obtained composite materials into the phosphoric acid buffer solution. According to the preparation method provided by the invention, the complex chemical modification to enzyme is not involved, and the activity of the enzyme is not influenced; the reaction is carried out at the room temperature, and the reaction conditions are mild; and the prepared functional nanogold-protein composite material is hardly influenced by solution media and has excellent stability.

Description

technical field [0001] The invention relates to a preparation method of a functional nano composite material, in particular to a preparation method of a nano gold-protein composite material with enzyme catalytic performance. Background technique [0002] Nanogold generally refers to chloroauric acid (HAuCl 4 ) under the action of a reducing agent to polymerize gold particles of a certain size. These particles become a stable colloidal state due to electrostatic interaction, so they are also called colloidal gold. In addition to the surface effect and quantum size effect of ordinary nanoparticles, gold nanoparticles also have good biocompatibility. Since biological macromolecules and functionalized gold nanoparticles are in the same nanometer dimension, when functionalized gold nanoparticles are added to the corresponding biological system, they have directional adsorption effects and enrichment effects. By chemical or physical adsorption, the two can prepare bio / nano-compo...

Claims

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Application Information

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IPC IPC(8): G01N33/68B82Y5/00
Inventor 徐玮陈平程晓丹李光朔陈明富
Owner 苏州汉能环保材料科技有限公司