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Method for preparing jute cellulose enzymatic hydrolysis product conjugation proteins for detecting cultural relics

A technology of hemp cellulose and enzymatic hydrolyzate, which is applied in the field of preparation of hemp cellulose enzymatic hydrolyzate-coupled protein, can solve problems such as animal immunity, achieve the effects of improving purity, shortening the process flow, and saving production costs

Active Publication Date: 2017-11-21
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 hemp fiber is cellulose, which cannot be directly used as a complete antigen for animal immunization, so it is difficult to identify the hemp fabric in ancient textile cultural relics by using methods such as ELISA and Western Blotting

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] 1) Take 10 g of hemp 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 hemp fiber was washed 5 times with deionized water, and dried in an oven at 50 °C.

[0029] 2) Take 5 g of dried hemp fiber powder, add 100 mL of acetone and 50 mL of deionized water, stir and mix evenly, then add 10 mL of formic acid dropwise, soak for 2 h, and centrifuge the solution at 1000 r / min. Take the bottom precipitate and continue to treat it with the above solution and centrifuge, repeat the operation 4 times to obtain the final precipitate, wash it with deionized water 5 times, add 200 mL of 7% sodium hydroxide, heat it in a water bath at 75 °C for 12 h, and then filter it with suction , the precipitate was washed 5 times with deionized water.

[0030] 3.) Take the final precipitate o...

Embodiment 2

[0037] 1) Take 10 g of hemp fiber, grind it for 4 minutes, put it in a beaker, add 50 mL of benzene and 25 mL of ethanol, soak for 8 h to degrease the fiber, and evaporate the mixed solution under reduced pressure to recover benzene and Ethanol, the degreased hemp fiber was washed 5 times with deionized water, and dried in an oven at 50 °C.

[0038] 2) Take 5 g of dried hemp fiber powder, add 100 mL of acetone and 50 mL of deionized water, stir and mix evenly, then add 10 mL of formic acid dropwise, soak for 2 h, and centrifuge the solution at 1000 r / min. Take the bottom precipitate and continue to treat it with the above solution and centrifuge, repeat the operation 4 times to obtain the final precipitate, wash it with deionized water 5 times, add 200 mL of 8% sodium hydroxide, heat it in a water bath at 75 °C for 12 h, and then filter it with suction , the precipitate was washed 5 times with deionized water.

[0039] 3) Take the final precipitate obtained in step 2, add 10%...

Embodiment 3

[0046] 1) Take 10 g of hemp 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 hemp fiber was washed 5 times with deionized water, and dried in an oven at 50 °C.

[0047] 2) Take 5 g of dried hemp fiber powder, add 100 mL of acetone and 50 mL of deionized water, stir and mix evenly, then add 10 mL of formic acid dropwise, soak for 2 h, and centrifuge the solution at 1000 r / min. Take the bottom precipitate and continue to treat it with the above solution and centrifuge, repeat the operation 4 times to obtain the final precipitate, wash it with deionized water 5 times, add 200 mL of 9% sodium hydroxide, heat it in a water bath at 75 °C for 12 h, and then filter it with suction , the precipitate was washed 5 times with deionized water.

[0048] 3) Take the final precipitate o...

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Abstract

The invention relates to the field of cultural relic detection, and discloses a method for preparing jute cellulose enzymatic hydrolysis product conjugation proteins for detecting cultural relics. The method includes degreasing jute celluloses by the aid of benzene and ethyl alcohol; removing lignin by the aid of an acetone-formic acid-water mixed system; removing hemicelluloses by the aid of alkali liquor and then carrying out enzymatic hydrolysis on obtained celluloses by the aid of cellulase and beta-glucosidase; recycling enzymes by the aid of ultra-filtration technologies after enzymatic hydrolysis is carried out; carrying out freeze drying on filtrate to obtain glucose powder; dissolving the glucose powder and egg white proteins in deionized water and potassium bromide solution to obtain solution; placing the solution in a constant-temperature and constant-humidity tank; carrying out freeze drying to obtain glucose-egg white protein conjugates. The method has the advantages that conjugation macromolecules obtained by the aid of the method are stable and can be used as complete antigens, immune injection can be carried out on animals to obtain corresponding specific antibodies, and accordingly jute fabrics in the cultural relics which are ancient textiles can be feasibly authenticated.

Description

technical field [0001] The invention relates to the field of cultural relic detection, in particular to a method for preparing a hemp cellulose enzymatic hydrolyzate-coupled protein used for the detection of cultural relics. 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, th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/765C12P19/14C12P19/02
CPCC07K14/765C07K19/00C12P19/02C12P19/14C13K1/02
Inventor 王秉李津梁军龙陈茹茹陈博逸
Owner ZHEJIANG SCI-TECH UNIV
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