Fluorescent platinum nano-cluster based on papain as template and preparation method and application of fluorescent platinum nano-cluster

A papain and platinum nanotechnology, applied in the field of fluorescent probes, achieves the effects of stable fluorescent signal, simple detection and easy realization

Active Publication Date: 2020-02-18
ANHUI NORMAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

my country's Guangdong, Guangxi, Hainan, Yunnan, Fujian, Sichuan, Guizhou and ...

Method used

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  • Fluorescent platinum nano-cluster based on papain as template and preparation method and application of fluorescent platinum nano-cluster
  • Fluorescent platinum nano-cluster based on papain as template and preparation method and application of fluorescent platinum nano-cluster
  • Fluorescent platinum nano-cluster based on papain as template and preparation method and application of fluorescent platinum nano-cluster

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Preparation of papain-coated platinum nanoclusters

[0040] Specific steps are as follows:

[0041] (1) Get 500mg of papain and add 10mL of pure water to prepare a 50mg / mL papain solution;

[0042] (2) H 2 PtCl 6 Add water to make the concentration of 25mM H 2 PtCl 6 solution;

[0043] (3) Add 160 μL of H 2 PtCl 6 The solution was added to 5.6mL of papain solution, so that papain and H in the mixed solution 2 PtCl 6 The molar ratio is 3.5:1, and the vortex is fully mixed for 5 minutes;

[0044] (4) Add 1M NaOH to the solution obtained in step (3) to adjust the pH value to 12, and mix again with a vortex for 5 minutes;

[0045] (5) The prepared sample was placed in a dark water bath at 37°C for 36 hours to obtain a polymer of platinum nanoclusters. After centrifugation at 5,000rpm / min for 10min, the supernatant was taken and stored in a 4°C refrigerator in the dark for future use.

[0046] The transmission electron microscope figure (TED picture) of the platin...

Embodiment 2

[0051] Example 2: Preparation of papain-coated platinum nanoclusters

[0052] Specific steps are as follows:

[0053] (1) Get papain and add papain solution in pure water to be prepared into 50mg / mL;

[0054] (2) H 2 PtCl 6 Add water to make the concentration of 25mM H 2 PtCl 6 solution;

[0055] (3) H 2 PtCl 6 The solution is added to the papain solution, so that the papain and H 2 PtCl 6 The molar ratio of the two is 2.5:1, and the vortex is fully mixed for 10 minutes;

[0056] (4) Add 1M NaOH to the solution obtained in step (3) to adjust the pH value to 8, and mix again with a vortex for 10 min;

[0057] (5) The prepared sample was placed in a water bath at 25° C. for 14 hours in the dark to obtain a polymer of platinum nanoclusters. After centrifugation at 5,000 rpm / min for 10 minutes, the supernatant was taken and stored in a refrigerator at 4° C. in the dark for future use.

Embodiment 3

[0058] Example 3: Preparation of papain-coated platinum nanoclusters

[0059] Specific steps are as follows:

[0060] (1) Get papain and add papain solution in pure water to be prepared into 50mg / mL;

[0061] (2) H 2 PtCl 6 Add water to make the concentration of 25mM H 2 PtCl 6 solution;

[0062] (3) H 2 PtCl 6 The solution is added to the papain solution, so that the papain and H 2 PtCl 6 The molar ratio of the two is 4:1, and the vortex is fully mixed for 6 minutes;

[0063] (4) Add 1M NaOH to the solution obtained in step (3) to adjust the pH value to 9, and mix again with a vortex for 8 minutes;

[0064] (5) The prepared sample was placed in a dark water bath at 65°C for 48 hours to obtain a polymer of platinum nanoclusters. After centrifugation at 5,000 rpm / min for 10 minutes, the supernatant was taken and stored in a 4°C refrigerator in the dark for future use.

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Abstract

The invention provides a platinum nano-cluster based on papain as a template and a preparation method and application of the platinum nano-cluster. The preparation method specifically comprises the following steps: mixing a papain solution with an aqueous solution of a high-valence platinum complex, adding a NaOH solution to adjust the pH value to 8-13, and completely reacting in a dark water bath. The platinum nano-cluster prepared by the method is sensitive to lysozyme response, a detection system is simple, convenient and controllable to operate and short in consumed time, and the platinumnano-cluster can be applied to detection of lysozyme content in urine. The prepared papain platinum nano-cluster greatly enhances the activity of papain, so that the synthesis substance can be used for tracking the application effect of papain and detecting the content of lysozyme in a sample on the basis of the fluorescence characteristic besides the original application of the papain in the aspects of medicines, foods, feeds, textiles and the like.

Description

technical field [0001] The invention belongs to the technical field of fluorescent probes, and in particular relates to a platinum nano-cluster using papain as a template, a preparation method and application thereof. Background technique [0002] Papain is a sulfhydryl protease mainly derived from papaya green fruit milk. It has the characteristics of hydrolyzing amide bonds and ester bonds, and has a strong ability to decompose proteins. At the same time, papain has good thermal stability and high enzyme activity. It has a wide range of applications in medicine, food, textile, leather and other fields. Papain consists of 212 amino acids, three amino acids Cys25, His159 and Asp158 constitute its catalytic triad, and the other six cysteine ​​residues form three pairs of disulfide bonds, and none of them are on the active site. my country's Guangdong, Guangxi, Hainan, Yunnan, Fujian, Sichuan, Guizhou and Taiwan provinces are rich in papaya resources, but they have not been f...

Claims

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Application Information

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IPC IPC(8): C09K11/02C09K11/87C12Q1/34C12Q1/37G01N21/64B82Y30/00
CPCC09K11/87C09K11/02G01N21/6428C12Q1/34C12Q1/37B82Y30/00G01N2021/6432Y02A50/30
Inventor 王宝娟常欣陈杰吴爽高鹏吴雨芬王源秀
Owner ANHUI NORMAL UNIV
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