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Preparation method of gossypin enzymatic hydrolysis product coupled protein used for historical relic detection

A technology of cotton cellulose and enzymolysis products, applied in the field of cultural relics detection, can solve problems such as animal immunity, and achieve the effect of speeding up the speed of enzymolysis, shortening the process flow, and improving the degree of enzymolysis

Active Publication Date: 2017-10-20
ZHEJIANG SCI-TECH UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the main component of cotton fiber is cellulose, which cannot be directly used as a complete antigen for animal immunization, so it is difficult to use ELISA, Western Blotting and other methods to identify cotton fabrics in ancient textile relics

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1) Take 10 g of cotton fiber, grind it for 3 minutes, put it in a beaker, add 45 mL of benzene and 20 mL of ethanol, and soak for 6 h to degrease the fiber. After the treatment, the mixed solution is evaporated under reduced pressure to recover benzene and Ethanol, the degreased cotton fibers were rinsed 5 times with deionized water, and dried in an oven at 50 °C.

[0026] 2) Take 5 g of dried cotton fiber powder, add 200 mL of 4% acetic acid, heat in a water bath at 75 °C for 12 h, then filter with suction, take the precipitate, wash it with deionized water for 5 times, and then add 200 mL of 7 % sodium hydroxide, heated in a water bath at 75 °C for 12 h, then suction filtered, and the precipitate was washed 5 times with deionized water.

[0027] 3) Take the final precipitate obtained in step 2, add 8% hydrogen peroxide with a mass ratio of solid to liquid of 1:30, heat it in a water bath at 45 °C for 5 h, then filter with suction, rinse the precipitate with deionized ...

Embodiment 2

[0034] 1) Take 10 g of cotton fiber, grind it for 4 minutes, put it in a beaker, add 50 mL of benzene and 25 mL of ethanol, and soak for 8 hours to degrease the fiber. After the treatment, the mixed solution is evaporated under reduced pressure to recover benzene and Ethanol, the degreased cotton fibers were rinsed 5 times with deionized water, and dried in an oven at 50 °C.

[0035] 2) Take 5 g of dried cotton fiber powder, add 200 mL of 5% acetic acid, heat in a water bath at 75 °C for 12 h, then filter with suction, take the precipitate, wash it with deionized water for 5 times, and then add 200 mL of 8 % sodium hydroxide, heated in a water bath at 75 °C for 12 h, then suction filtered, and the precipitate was washed 5 times with deionized water.

[0036] 3) Take the final precipitate obtained in step 2, add 10% hydrogen peroxide with a mass ratio of solid to liquid of 1:30, heat it in a water bath at 45 °C for 5 h, then filter with suction, rinse the precipitate with deion...

Embodiment 3

[0043] 1) Take 10 g of cotton fiber, grind it for 5 minutes, put it in a beaker, add 55 mL of benzene and 30 mL of ethanol, and soak for 10 h to degrease the fiber. After the treatment, the mixed solution is evaporated under reduced pressure to recover benzene and Ethanol, the degreased cotton fibers were rinsed 5 times with deionized water, and dried in an oven at 50 °C.

[0044] 2) Take 5 g of dried cotton fiber powder, add 200 mL of 6% acetic acid, heat it in a water bath at 75 °C for 12 h, then filter with suction, take the precipitate, wash it with deionized water for 5 times, and then add 200 mL of 9 % sodium hydroxide, heated in a water bath at 75 °C for 12 h, then suction filtered, and the precipitate was washed 5 times with deionized water.

[0045] 3) Take the final precipitate obtained in step 2, add 12% hydrogen peroxide with a mass ratio of solid to liquid of 1:30, heat it in a water bath at 45 °C for 5 h, then filter with suction, rinse the precipitate with deion...

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Abstract

The invention relates to the field of historical relic detection, and discloses a preparation method of a gossypin enzymatic hydrolysis product coupled protein used for historical relic detection. The method comprises the following steps: degreasing gossypin, processing the degreased gossypin with acetic acid and sodium hydroxide to remove lignin and hemicellulose, carrying out enzymatic hydrolysis on the obtained cellulose by using cellulase and beta-glucosidase, preparing a deionized water-ammonium sulfate-polyethylene glycol-400 extract liquid, respectively freeze-drying an upper layer liquid and a lower layer liquid which are obtained after extraction to respectively obtain a recovered enzyme and glucose powder, dissolving the glucose powder and bovine serum albumin in deionized water and a potassium bromide solution, standing the obtained solution in a constant temperature and constant humidity box, and freeze-drying the solution to obtain a glucose-bovine serum albumin conjugate. Coupled macromolecules obtained through using the method are stable, and can be used to inoculate animals as a complete antigen to obtain a corresponding specific antibody, so feasibility is provided for the identification of cotton fabrics in ancient textile historical relics.

Description

technical field [0001] The invention relates to the field of cultural relic detection, in particular to a method for preparing a cotton cellulose enzymatic hydrolyzate coupled protein for cultural relic detection. Background technique [0002] In recent years, a large number of textile cultural relics have been unearthed in my country, and the identification of their types is of great significance to the study of our country's history and culture. There have been several accurate and sensitive methods for the identification of silk fabrics, among which enzyme-linked immunosorbent assay and Western Blotting are the representatives. Basic knowledge. Since silk itself contains a large amount of protein, silk fibroin is used as a complete antigen to immunize animals, and after corresponding treatment, antibodies that can specifically bind to silk fibroin can be obtained, and these antibodies can be used to detect whether ancient textiles contain silk. However, the main comp...

Claims

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Application Information

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IPC IPC(8): C07K14/765C12P19/14C12P19/02
CPCC07K14/765C07K19/00C12P19/02C12P19/14C13K1/02
Inventor 王秉李津梁军龙陈茹茹何宇杰
Owner ZHEJIANG SCI-TECH UNIV
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