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A method for analyzing the distribution of amino acid bonded folds in wool keratin

A wool keratin and amino acid technology, applied in the field of wool detection, can solve the problems of difficulty in applying wool keratin amino acid analysis, protein mishydrolysis/enzymolysis, inaccurate analysis results, etc., and achieves easy analysis, little damage to amino acids, and results. accurate effect

Active Publication Date: 2021-04-02
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there are few analyzes on the distribution of amino acids in wool keratin at present, and only scholars use acid, alkaline solution or various proteases to treat wool keratin to different degrees (by controlling the length of hydrolysis / enzymolysis time to achieve radial gradual layer erosion) to obtain residual keratin fibers, and then use infrared spectroscopy, X-ray diffraction and other techniques to analyze the radial distribution of amino acids in keratin
However, the above-mentioned dissolution method is easy to damage amino acids, and it is easy to mishydrolyze / enzymolyze the protein in the inner layer of keratin that is not expected to be hydrolyzed temporarily, resulting in inaccurate analysis results
In addition, the above method cannot be used for more detailed quantitative research, and it is difficult to apply it to the amino acid analysis of wool keratin

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] A method for analyzing the distribution of amino acid bond folds in wool keratin, comprising the following steps:

[0042] 1) Weigh wool as a sample, wash it repeatedly with deionized water to remove surface impurities, and dry it.

[0043] 2) Boil the dried sample in a mixed solution containing 0.07% dodecanol polyoxyethylene ether and 0.3% sodium sulfate at a bath ratio of 1:100 at 55°C for 30 minutes to remove grease and impurities, repeat once.

[0044] 3) After washing and removing grease, the samples were rinsed with deionized water and placed in an oven at 45°C to obtain dry wool.

[0045] 4) Divide the wool dried in step 3) into 12 groups of A1, A2, B1, B2, C1, C2, D, E, F1, F2, G, H, etc., among which group A1 is used as a control group, and no Any treatment; put A2 into a three-necked flask, at 50°C, in 0.75mol / L tris(2-carboxyethyl)phosphine with pH=5 at a liquor ratio of 1:20, pass nitrogen, and add 200r / The reaction was stirred at a speed of 1 min for 3...

Embodiment 2

[0059] A method for analyzing the distribution of amino acid bond folds in wool keratin, comprising the following steps:

[0060] 1) Weigh wool as a sample, wash it repeatedly with deionized water to remove surface impurities, and dry it.

[0061] 2) Boil the dried sample in a mixed solution containing 0.06% dodecanol polyoxyethylene ether and 0.2% sodium sulfate at a bath ratio of 1:95 at 50 °C for 25 minutes, wash off grease and impurities, and repeat once.

[0062] 3) After washing and removing grease, the samples were rinsed with deionized water and placed in an oven at 40°C to obtain dry wool.

[0063] 4) Divide the wool dried in step 3) into 12 groups of A1, A2, B1, B2, C1, C2, D, E, F1, F2, G, H, etc., among which group A1 is used as a control group, and no Any treatment; put A2 into a three-necked flask, at 45°C, in 0.7 mol / L tris(2-carboxyethyl) phosphine with pH=4 at a liquor ratio of 1:18, pass nitrogen through it, and at 180r / The reaction was stirred at a speed...

Embodiment 3

[0077] A method for analyzing the distribution of amino acid bond folds in wool keratin, comprising the following steps:

[0078] 1) Weigh wool as a sample, wash it repeatedly with deionized water to remove surface impurities, and dry it.

[0079] 2) Boil the dried sample in a mixed solution containing 0.08% dodecanol polyoxyethylene ether and 0.4% sodium sulfate at a bath ratio of 1:105 at 60 °C for 35 minutes, wash off grease and impurities, repeat once.

[0080] 3) After washing and removing grease, the samples were rinsed with deionized water and placed in a 50°C oven to obtain dry wool.

[0081] 4) Divide the wool dried in step 3) into 12 groups of A1, A2, B1, B2, C1, C2, D, E, F1, F2, G, H, etc., among which group A1 is used as a control group, and no Any treatment; put A2 into a three-necked flask, at 55°C, in 0.8 mol / L tris(2-carboxyethyl) phosphine with pH=6 at a liquor ratio of 1:22, pass nitrogen through it, and at 220r / The reaction was stirred at a speed of 1 m...

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PUM

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Abstract

The invention discloses a method for analyzing the distribution of bonded and folded amino acids in wool keratin, belonging to the technical field of wool detection. The method of the invention comprises the following steps: removing oil and impurities in wool by using a neutral wool scouring process; then dividing the wool into 12 equal parts; then carrying out layer-by-layer enzymatic hydrolysison the wool from exterior to interior with different proteases used in a permutation and combination manner, wherein tris(2-carboxyethyl)phosphine is used for specifically break disulfide bonds in the wool which is only subjected to enzymatic hydrolysis with an acidic protease; carrying out amino acid analysis on each group of wool with unbroken disulfide bonds at first so as to obtain the radialdistribution of amino acids; and comparing the groups of wool before and after the breakage of disulfide bonds so as to obtain the distribution of bonded and folded amino acids in wool keratin. The method of the invention employs permutated and combined multitime enzymatic hydrolysis manners for dissolving of keratin according to the characteristic that the distribution of amino acids in wool keratin has different levels, and is small in damage to amino acids, high in accuracy, good in specificity and friendly to environment.

Description

technical field [0001] The invention relates to the technical field of wool detection, in particular to a method for analyzing the bonding and folding distribution of amino acids in wool keratin. Background technique [0002] Wool is a slender solid cylinder with a curled shape, and the fiber structure is divided into three layers, namely the scale layer, the cortex layer and the medulla layer. The cortex is the main component of wool fibers and consists of many keratin proteins. The cortex is divided into two types: the positive cortex and the paracortical layer. The cells on the outside of the curled wave are called positive cortical cells, and those on the inside are called paracortical cells. [0003] The extraction of keratin by dissolving wool is an important means to solve the shortage of resources and waste of resources. However, the structure of wool keratin is very complex, mainly in the form of a-helix and β-sheet, of which the a-helix structure accounts for mo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12P21/06
CPCC12P21/06G01N33/68
Inventor 王秉王钟元尚亚廷周莲杨慧彭志勤
Owner ZHEJIANG SCI-TECH UNIV