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Method for detecting keratin by using ThioGlo1 fluorescent probe

A fluorescent probe and keratin technology, applied in the field of keratin detection, can solve the problems of difficult extraction of keratin and difficulty in dissolving natural keratin, and achieve the effect of sensitive detection and simple method

Pending Publication Date: 2022-07-12
广东省科学院生物与医学工程研究所
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, because of the three-dimensional network structure formed in the keratin molecule, there are a large number of disulfide bonds, which makes it difficult for natural keratin to dissolve in general solutions, which brings difficulties to the extraction of keratin

Method used

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  • Method for detecting keratin by using ThioGlo1 fluorescent probe
  • Method for detecting keratin by using ThioGlo1 fluorescent probe
  • Method for detecting keratin by using ThioGlo1 fluorescent probe

Examples

Experimental program
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Effect test

Embodiment 1

[0029] 1. At room temperature, accurately weigh 1-5mg ThioGlo1 sample and prepare it into 1×10 with DMSO -5 mol / L solution for later use.

[0030] 2. Use phosphate buffer solution with pH 7.4 to prepare waste wool keratin into solutions of different concentrations (1-100×10 -5 mol / L).

[0031] 3. Add 20μL of dissolved 1×10 to the 3.0mL colorimetric tube. -5 mol / L ThioGlo1 solution, 2.5mL phosphate buffer solution, different concentrations (5, 10, 20, 30, 40, 50, 60, 80, 90 and 100×10 -5 mol / L) waste keratin solution 20 μL, image 3 ThioGlo1 in represents the control without added waste keratin. Dilute the phosphate buffer solution to 3.0 mL, shake well, and react at room temperature for 15 min.

[0032] 4. Use a FluoroLog-3 steady-state transient fluorescence spectrometer to detect the emission wavelength λ em = Fluorescence intensity around 510 nm, excitation wavelength λ ex =340nm.

[0033] The result is as image 3 As shown, in the figure, ThioGlo1-1 to ThioGlo1-10...

Embodiment 2

[0036] 1. At room temperature, accurately weigh 1-5mg ThioGlo1 sample and prepare it into 1×10 with DMSO -5 mol / L solution for later use.

[0037] 2. Use a phosphate buffer solution with a pH of 7.4 to prepare waste chicken feather keratin into solutions of different concentrations (1-100×10 -5 ).

[0038] 3. Add 20 μL of dissolved ThioGlo1 solution, 2.5 mL of phosphate buffer solution, and different concentrations (concentrations of 10, 20, 30, 50, 60, 80, 90 and 100 × 10) into a 3.0 mL colorimetric tube. 5 mol / L) 20 μL of discarded keratin solution. Dilute the phosphate buffer solution to 3.0 mL, shake well, and react at room temperature for 15 min.

[0039] 3. Use a FluoroLog-3 steady-state transient fluorescence spectrometer to detect the emission wavelength λ em = Fluorescence intensity around 510 nm, excitation wavelength λ ex =340nm.

[0040] The result is as Figure 4 As shown in the figure, ThioGlo1-12 to ThioGlo1-19 in the figure are the fluorescence emission ...

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Abstract

The invention discloses a method for detecting keratin by using a ThioGlo1 fluorescent probe, which is characterized by comprising the following steps of: mixing the ThioGlo1 fluorescent probe and keratin in a liquid environment for reaction, then detecting the fluorescence intensity when the emission wavelength lambda em is about 510nm and the excitation wavelength lambda ex is 340nm, and judging whether the keratin exists or the concentration of the keratin exists. The ThioGlo1 fluorescent probe is very sensitive to detection of the waste wool keratin and the waste chicken feather keratin, and the fluorescence intensity is in direct proportion to the concentration of the waste keratin, so that the existence of the waste keratin can be indicated or the concentration of the waste keratin can be quantitatively determined. The method can also be used for rapidly screening keratin degrading bacteria, and lays a foundation for high-throughput screening of the bacteria.

Description

Technical field: [0001] The invention belongs to the field of keratin detection, in particular to a method for detecting keratin by using a ThioGlo1 fluorescent probe. Background technique: [0002] Keratin is mainly found in wool, feathers, hair, nails, horns and hooves (up to 90% or more by mass), and is the most abundant and underexploited protein source. According to statistics, the global annual output of feathers exceeds 65 million tons, of which more than 2.5 million tons of wool. The keratin-rich by-products of the poultry processing industry and down product manufacturers in my country are usually disposed of by landfill or incineration, and only a small amount is simply converted into fertilizers and feeds. Due to the high sulfur content of keratin, the use of landfilling or burning methods will cause serious environmental pollution. Moreover, keratin is a polypeptide composed of 18 amino acids, which has good biocompatibility, degradability, regeneration, film-f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/64C09K11/06
CPCG01N21/6428G01N21/6486C09K11/06C09K2211/1029C09K2211/1088
Inventor 李清心王春凤吴金川王钦庆冯莉班雯婷
Owner 广东省科学院生物与医学工程研究所
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