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Method for immobilizing catalase with enzymatic silk fibroin film

A technology of catalase and silk fibroin, which is applied in the field of enzymatic silk fibroin film immobilization of catalase, can solve the problems of no reports, etc., and achieve the effect of promoting reaction, obvious performance and improved performance

Active Publication Date: 2014-03-26
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

With the help of tyrosinase catalyzed silk fibroin membrane grafted catalase for biosensor preparation method, there is no report at home and abroad at present

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] (1) Preparation of silk fibroin solution: Add the degummed silkworm silk into the lithium bromide-ethanol-water (mass ratio 45:45:10) solution, stir continuously at 65°C until dissolved (bath ratio 1:40), and obtain mixture. Put the silk fibroin solution into a dialysis bag, dialyze with deionized water for 30 hours, change the water every hour during this period, and filter to obtain silk fibroin aqueous solution after dialysis.

[0023] (2) Preparation of silk fibroin film: Pour a certain amount of silk fibroin solution into a PTFE circular vessel, spread it evenly by flowing, and dry at 85° C. for 3 hours to obtain a silk fibroin film.

[0024] (3) Enzymatic silk fibroin protein grafted catalase: soak the silk fibroin film in the solution of tyrosinase and catalase, wherein the dosage of tyrosinase is 50U / g silk fibroin, Catalase 100U / g silk fibroin, pH 6.5, temperature 40 ℃, treatment 6h.

[0025] (4) Post-washing treatment: after tyrosinase treatment, wash the si...

Embodiment 2

[0030] (1) Preparation of silk fibroin solution: add the degummed silkworm silk into the lithium bromide-ethanol-water (mass ratio 44:45:11) solution, stir continuously at 65°C until dissolved (bath ratio 1:50), and obtain mixture. Put the silk fibroin solution into a dialysis bag, dialyze with deionized water for 24 hours, change the water every hour during this period, and filter to obtain silk fibroin aqueous solution after dialysis.

[0031] (2) Preparation of silk fibroin film: Take a certain amount of silk fibroin solution and pour it into a PTFE circular vessel, spread it evenly by flowing, freeze at -4°C for 6 hours, and then vacuum at -50°C Dry to obtain silk fibroin film.

[0032] (3) Enzyme-promoting silk fibroin protein grafted catalase: soak the silk fibroin film in the solution of tyrosinase and catalase, wherein the dosage of tyrosinase is 100U / g silk fibroin, Catalase 200U / g silk fibroin, pH 6.5, temperature 30 ℃, treatment 8h.

[0033] (4) Post-washing trea...

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Abstract

The invention discloses a method for immobilizing catalase with an enzymatic silk fibroin film, belongs to the field of textile biotechnology, aims to provide an environmental-friendly method for immobilizing catalase with the silk fibroin film, and expands the application of an enzymatic silk fibroin film material serving as a biosensor. Due to the catalytic oxidation function of tyrosinase, the silk fibroin film is grafted with catalase, so that not only is the immobilization between enzyme molecules and silk fibroin realized, but also the application performance of the silk fibroin film serving as a catalase carrier is improved. The method particularly comprises the technological processes such as preparation of a silk fibroin solution and the silk fibroin film, grafting enzymatic silk fibroin with catalase, washing with water, and carrying out aftertreatment. The enzymatic silk fibroin film prepared by the invention is used with a dissolved oxygen meter in a combined manner, so as to not only evaluate catalase contents in different systems, but also ensure repeated circulatory use; the properties of strength, elasticity and the like of the silk fibroin film are improved.

Description

technical field [0001] The invention relates to an enzymatic method for immobilizing catalase on silk fibroin membrane, which is a method for immobilizing catalase on the surface of silk fibroin membrane by utilizing the catalytic oxidation of tyrosinase, and at the same time improving The invention relates to a method for the application performance of a membrane material, which belongs to the field of textile biotechnology. Background technique [0002] Silk fibroin has good biocompatibility and environmental friendliness. The membrane material processed with silk fibroin has the advantages of good biodegradability, non-toxic, non-irritating and good air permeability. It is a biological product with excellent performance and fixed transformation carrier. Using silk fibroin as a carrier can carry out the immobilization of various substances such as polysaccharide macromolecules, foreign proteins or biological enzymes, and the methods that can be used include blending embed...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N11/02C08J7/12
Inventor 王平丁雅静唐耿铁崔莉袁久刚王强范雪荣
Owner JIANGNAN UNIV
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