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Colloidal gold probe for detecting absorption characteristics of laccase and preparation method thereof

A technology of adsorption characteristics and colloidal gold, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of limited application and difficult realization, and achieve convenient operation, promotion of enzymatic hydrolysis, and simple preparation process Effect

Inactive Publication Date: 2012-12-26
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

First of all, lignin peroxidase and manganese peroxidase are strict secondary metabolites produced under the condition of both carbon and nitrogen limitation, and the presence of carbon and nitrogen source nutrients will limit the secretion of enzymes by cells; secondly , because lignin peroxidase and manganese peroxide need a large amount of H2O2 auxiliary agent when degrading organic pollutants, which is difficult in reality thirdly, laccase has a redox potential of 780 mV, and can directly reduce molecular oxygen to water, even without H2O2 and other secondary metabolites

Method used

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  • Colloidal gold probe for detecting absorption characteristics of laccase and preparation method thereof
  • Colloidal gold probe for detecting absorption characteristics of laccase and preparation method thereof
  • Colloidal gold probe for detecting absorption characteristics of laccase and preparation method thereof

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Embodiment 1

[0027] A colloidal gold probe for detecting the adsorption property of laccase of the present invention comprises laccase, and colloidal gold particles are combined on the surface of the laccase. The particle size of the colloidal gold particles is 15 nm to 20 nm, and the Zeta potential is -30.5 mV to -35.5 mV. The particle size of the colloidal gold probe is 450 nm to 500 nm.

[0028] The colloidal gold probe used to detect the adsorption properties of laccase in this embodiment was prepared by the following steps:

[0029] (1) Preparation of colloidal gold solution:

[0030] Take HAuCl with a mass concentration of 0.01 % 4 100 mL of aqueous solution was heated to boiling on a constant temperature magnetic stirrer and kept for 2 min. Under the condition of stirring at a speed of 1000 r / min, 6 mL of trisodium citrate solution with a mass concentration of 10 g / L was quickly added; Turn the speed until the color of the solution turns to clear orange, stop heating, continue to...

Embodiment 2

[0040] A colloidal gold probe for detecting the adsorption property of laccase of the present invention comprises laccase, and colloidal gold particles are combined on the surface of the laccase. The particle size of the colloidal gold particles is 15 nm to 20 nm, and the Zeta potential is -30.5 mV to -35.5 mV. The particle size of the colloidal gold probe is 450 nm to 500 nm.

[0041] The colloidal gold probe used to detect the adsorption properties of laccase in this embodiment was prepared by the following steps:

[0042] (1) Prepare colloidal gold solution with the method of embodiment 1;

[0043] (2) Preparation of colloidal gold probes:

[0044] The pH value of the prepared colloidal gold solution was adjusted to 5.0 with 0.1 M acetic acid, and 0.9 mg / mL laccase solution was added dropwise and mixed evenly. Under stirring conditions, add 2 mL of 10 mg / mL PEG-20000 solution dropwise to 20 mL of the above mixed solution, and the volume ratio of the colloidal gold soluti...

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Abstract

The invention discloses a colloidal gold probe for detecting absorption characteristics of laccase and a preparation method thereof. The colloidal gold probe comprises the laccase, wherein the surface of the laccase is bonded with colloidal gold particles. The preparation method comprises the following steps: preparing a colloidal gold solution and adjusting the pH value of the solution to be 4.5-5; adding a laccase solution into the colloidal gold solution, and then adding a PEG (polyethylene glycol)-20000 solution while stirring, wherein the volume ratio of the colloidal gold solution to the laccase solution to the PEG-20000 solution is 5:(1.1-1.15):(0.6-0.65); and after the mixed solution fully reacts, performing primary centrifugation on the reaction solution so as to remove an aggregate, then performing secondary centrifugation on the reaction solution so as to remove supernate, and re-dissolving the obtained precipitate in a sodium citrate buffer solution to finally obtain the product. The preparation method has the advantages that the cost is low, the bonding property is stable, the laccase activity is high, the preparation process is simple, and the operation is convenient.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a colloidal gold probe and a preparation method thereof. Background technique [0002] Lignin is a common and refractory substance in crop straw and municipal solid waste. Improper treatment will cause great waste of resources and environmental pollution. White rot fungi are microorganisms with the strongest ability to degrade organic pollutants such as lignin. During the process of decomposing lignin, they will produce an enzyme system that oxidizes and decomposes lignin, mainly including lignin peroxidases (Lignin Peroxidases), Manganese Peroxidases and Laccase. They determine the way, speed and degree of lignin degradation, so the enzymatic hydrolysis of microorganisms becomes the key in the process of lignin degradation. The composition unit and spatial structure of lignin determine the complexity of its enzymatic hydrolysis process, and the adsorption of enzymes on the lignin ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/26
Inventor 赖萃曾光明黄丹莲冯冲凌赵美花危臻许飘黄超李宁杰陈亮
Owner HUNAN UNIV