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A kind of preparation method of glutathione thiotransferase protected copper nanocluster and its application in the detection of chlortetracycline

A glutathione and copper nano-cluster technology, applied in copper sulfide, nanotechnology, nano-optics and other directions, can solve the problems of bacterial resistance, environmental pollution, poor stability, etc., and achieve unique photophysical properties and fast methods. Simple, Inexpensive Effects

Active Publication Date: 2022-03-18
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chlortetracycline is a broad-spectrum tetracycline antibiotic widely used in animal husbandry because of its high efficiency and low price. It is mainly used to inhibit the infection of Gram-positive bacteria and negative bacteria. In animal feed; however, improper dosage has also produced many side effects, such as causing environmental pollution, bacterial resistance, etc.
The traditional detection method for aureomycin has limitations such as poor stability, low sensitivity, and long time-consuming, so it is urgent to develop a fast and efficient detection method
Glutathione sulfur transferase protects cells from toxic chemicals, but so far there is no direct way to confirm its interaction with aureomycin, so it is crucial to develop new technologies such as fluorescent sensing important

Method used

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  • A kind of preparation method of glutathione thiotransferase protected copper nanocluster and its application in the detection of chlortetracycline
  • A kind of preparation method of glutathione thiotransferase protected copper nanocluster and its application in the detection of chlortetracycline
  • A kind of preparation method of glutathione thiotransferase protected copper nanocluster and its application in the detection of chlortetracycline

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Embodiment 1

[0035] Example 1: 100 μL concentration is 20 mg ml -1 Glutathione S-transferase was added to 5 x 10 -3 mol L -1 copper sulfate solution, stirring continuously at room temperature. Use 1.0mol L -1Sodium hydroxide solution adjusted the pH of the mixed solution to 12. After 10 minutes, 1 mg of dithiothreitol was added to the above solution, and the pH of the solution was adjusted to 7, and the mixed solution was stirred at room temperature for 2 hours. Using a dialysis bag with a molecular cut-off of 25kDa, dialyze the initial product in phosphate buffer for 24 hours to remove excess reactants and restore the protein to its natural state as much as possible to obtain glutathione sulfur transferase- copper nanoclusters.

Embodiment 2

[0036] Example 2: 100 μL concentration is 20 mg ml -1 Glutathione S-transferase was added to 5 x 10 -3 mol L -1 copper sulfate solution, stirring continuously at room temperature. Use 1.0mol L -1 Sodium hydroxide solution adjusted the pH of the mixed solution to 12. After 10 minutes, 1 mg of dithiothreitol was added to the above solution, and the pH of the solution was adjusted to 7, and the mixed solution was stirred at room temperature for 4 hours. Using a dialysis bag with a molecular cut-off of 25kDa, dialyze the initial product in phosphate buffer for 24 hours to remove excess reactants and restore the protein to its natural state as much as possible to obtain glutathione sulfur transferase- copper nanoclusters.

Embodiment 3

[0037] Example 3: 100 μL concentration is 20 mg ml -1 Glutathione S-transferase was added to 5 x 10 -3 mol L -1 copper sulfate solution, stirring continuously at room temperature. Use 1.0mol L -1 The sodium hydroxide solution adjusted the pH of the mixed solution to 12. After 10 minutes, 1 mg of dithiothreitol was added to the above solution, and the pH of the solution was adjusted to 7, and the mixed solution was stirred at room temperature for 6 hours. Using a dialysis bag with a molecular cut-off of 25kDa, dialyze the initial product in phosphate buffer for 24 hours to remove excess reactants and restore the protein to its natural state as much as possible to obtain glutathione sulfur transferase- copper nanoclusters.

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Abstract

The present invention relates to the technical field of synthesis of copper nanoclusters, in particular to a method for preparing copper nanoclusters protected by glutathione sulfur transferase and its application in the detection of aureomycin. The specific steps are as follows: The enzyme is used as a template, and glutathione sulfur transferase-copper nanocluster stock solution is prepared under specific pH conditions, and further purified by dialysis. Under the excitation wavelength of 325nm, the copper nanocluster exhibits blue fluorescence and has an emission peak at 400nm; taking quinine sulfate as a standard reference, the relative quantum yield of glutathione sulfur transferase-copper nanocluster is 1.36%; Copper nanoclusters can be used as an effective and environmentally friendly fluorescent probe for the detection of aureomycin (CTC) in solution. The fluorescent copper nanocluster of the present invention has unique photophysical properties, is cheap, is almost non-toxic and has excellent biocompatibility. The detection method of aureomycin is fast and simple, and the detection sensitivity is good. It is an ideal application in the fields of biology and medicine. Fluorescent nanomaterials.

Description

technical field [0001] The invention relates to the technical field of synthesis of copper nanoclusters, in particular to a preparation method of glutathione sulfur transferase-protected copper nanoclusters and its application in the detection of aureomycin. Background technique [0002] Glutathione sulfur transferase is a kind of multifunctional enzyme that exists in all organisms and can be combined with cytoplasm or cell membrane. important role. It protects cells from toxic chemicals such as drugs, endogenous peroxide radicals, and toxic metabolites by catalyzing the reduction of glutathione to generate a variety of electrophilic substances, many endogenous compounds such as prostate The metabolism of hormones and steroids is also related to the glutathione binding reaction. Therefore, glutathione sulfur transferase is an important biological enzyme that defends against reducing and toxic electrophilic substances produced during normal metabolic processes. [0003] Met...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C09K11/02C09K11/58B82Y20/00B82Y30/00B82Y40/00C01G3/12G01N21/64
CPCC09K11/02C09K11/58B82Y20/00B82Y30/00B82Y40/00C01G3/12G01N21/643
Inventor 付丁伊焦思凡丁姝姝曹蕾张建
Owner NANTONG UNIVERSITY
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