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A method for analyzing the radial distribution of amino acids in mulberry silk fibroin

A silk fibroin, radial distribution technology, applied in the field of silk, can solve the problems of protein mishydrolysis/enzymolysis, inaccurate analysis results, easy damage to amino acids, etc., to achieve mild working conditions, improve degumming efficiency, and reduce damage.

Active Publication Date: 2018-08-24
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above dissolving method is easy to damage the amino acid, and it is easy to cause mishydrolysis / enzyme hydrolysis of the protein that is not expected to be hydrolyzed in the inner layer of the silk fiber temporarily, resulting in inaccurate analysis results
In addition, the above methods are not able to carry out more detailed quantitative research

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method for analyzing radial distribution of amino acids in mulberry silk fibroin protein, comprising the following steps:

[0034] 1) Take mulberry silk as a sample, wash it with deionized water, remove surface pollutants, and dry it.

[0035] 2) The dried sample was mixed with 0.5wt% Na at a bath ratio of 1:100 2 HPO 4 and 1wt%C 17 h 35 Boil in the mixed solution of COONa 30min, carry out degumming treatment, totally degumming twice;

[0036] 3) After degumming, the sample is rinsed with deionized water, and placed in an oven at 60° C. to obtain dried silk fibroin fibers.

[0037] 4) Take silk fibroin fibers with similar diameters and divide them into five groups A, B, C, D, and E with equal mass, wherein group A is used as a control group without any treatment.

[0038] 5) Put groups B, C, D, and E into the pre-prepared 1.5g / L trypsin solution at a bath ratio of 1:40, control the reaction temperature at 48°C, the pH of the solution at 8, and the reaction time f...

Embodiment 2

[0046] A method for analyzing radial distribution of amino acids in mulberry silk fibroin protein, comprising the following steps:

[0047] 1) Take mulberry silk as a sample, wash it with deionized water, remove surface pollutants, and dry it.

[0048] 2) The dried sample was mixed with 0.4wt% Na at a bath ratio of 1:95 2 HPO 4 and 0.8 wt% C 17 h 35 Boil in the mixed solution of COONa 35min, carry out degumming treatment, totally degumming twice;

[0049] 3) After degumming, the sample is rinsed with deionized water and put into an oven at 55° C. to obtain dried silk fibroin fibers.

[0050] 4) Take silk fibroin fibers with similar diameters and divide them into five groups A, B, C, D, and E with equal mass, wherein group A is used as a control group without any treatment.

[0051] 5) Put groups B, C, D, and E into the pre-prepared 1.5g / L trypsin solution at a bath ratio of 1:40, control the reaction temperature at 45°C, the pH of the solution at 7, and the reaction time ...

Embodiment 3

[0059] A method for analyzing radial distribution of amino acids in mulberry silk fibroin protein, comprising the following steps:

[0060] 1) Take mulberry silk as a sample, wash it with deionized water, remove surface pollutants, and dry it.

[0061] 2) The dried sample was mixed with 0.6wt% Na at a bath ratio of 1:105 2 HPO 4 and 1.2 wt% C 17 h 35 Boil in the mixed solution of COONa 25min, carry out degumming treatment, totally degumming twice;

[0062] 3) After degumming, the sample is rinsed with deionized water and placed in an oven at 65° C. to obtain dried silk fibroin fibers.

[0063] 4) Take silk fibroin fibers with similar diameters and divide them into five groups A, B, C, D, and E with equal mass, wherein group A is used as a control group without any treatment.

[0064] 5) Put groups B, C, D, and E into the pre-prepared 1.5g / L trypsin solution at a bath ratio of 1:40, control the reaction temperature at 50°C, the pH of the solution at 9, and the reaction tim...

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PUM

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Abstract

The invention relates to the field of silk and discloses a method for analyzing the radial distribution of amino acids in silk fibroin of mulberry silkworms. In the present invention, the mulberry silk is first degummed by alkaline degumming to obtain silk fiber, and the silk fiber is grouped, and then different proteases are used to enzymolyze the silk fiber from the outside to the inside in sequence, and finally the amino acid is used to analyze the silk fiber. The radial distribution of each amino acid in silk can be obtained by measuring the content of various amino acids in residual silk fibroin with different degrees of hydrolysis. According to the characteristics of the amino acid distribution in the radial direction of the mulberry silk fibroin fiber, the invention adopts multiple enzymatic hydrolysis methods to dissolve the silk fibroin, with high specificity, mild action conditions, little damage to amino acids, and more accurate results.

Description

technical field [0001] The invention relates to the field of silk, in particular to a method for analyzing the radial distribution of amino acids in mulberry silk fibroin. Background technique [0002] Silk is an animal protein fiber material, mainly composed of silk fibroin and sericin, of which silk fibroin accounts for about 70%, which is the main component of silk and determines the properties and uses of silk. Mulberry silk is one of the more common silks in life, and it has practical applications in various fields. [0003] The biochemical synthesis of silk fibroin matrix in silkworms is very complicated. The silk fibroin stock solution in different silk glands is different, and in the process of silk spinning, the silk fibroin stock solution is affected by the pressure in the silkworm body and the mouthparts, and it will show different flow patterns. Denaturation eventually leads to different microstructures such as the molecular structure and supramolecular structur...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/6806
Inventor 王秉陈茹茹李津梁军龙徐城锋
Owner ZHEJIANG SCI-TECH UNIV