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A method for enzymatically preparing silk fibroin/elastin composite membrane material

An elastin and enzymatic preparation technology, which is applied in the field of textile biology, can solve the problems affecting the accessibility of enzymatic reactions and the grafting efficiency of silk fibroin enzymatic method needs to be improved, so as to achieve improved mechanical properties, good biocompatibility, The effect of broad application prospects

Active Publication Date: 2017-11-03
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the tyrosine residues in silk fibroin are distributed in the macromolecular chain composed of alanine and glycine with strong hydrophobicity, which affects the accessibility of enzymatic reactions, the efficiency of silk fibroin enzymatic grafting remains to be seen. promote

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] (1) Preparation of silk fibroin solution: add mulberry silk to lithium bromide-ethanol-water (mass ratio 45:45:10) solution, dissolve at 60°C, put the silk fibroin solution into a dialysis bag, and dialyze in deionized water for 24 hours. Change the water once an hour during this period, and filter to obtain 20g / L silk fibroin aqueous solution after dialysis;

[0022] (2) β-tyrosinase catalyzes amino acid conversion: in step (1) silk fibroin solution, add 20U / ml β-tyrosinase, 5g / L phenol, pyridoxal phosphate 2g / L and ammonium chloride 2g / L L, treated at 45°C and pH7.5 for 40 hours; after the treatment, the silk fibroin solution was dialyzed with deionized water for 24 hours;

[0023] (3) Tyrosinase catalyzes the cross-linking of silk fibroin and elastin: add 10U / ml tyrosinase and 10g / L elastin to the silk fibroin solution in step (2), at a temperature of 30°C and a pH of 7.0 12 hours of processing;

[0024] (4) Forming of silk fibroin / elastin composite film: take the ...

Embodiment 2

[0029] (1) Preparation of silk fibroin solution: add mulberry silk to calcium chloride-ethanol-water (molar ratio 1:1:7), dissolve at 70°C, put the silk fibroin solution into a dialysis bag, and dialyze in deionized water for 30 hours , during which the water was changed once an hour, and filtered to obtain a 15g / L silk fibroin aqueous solution after dialysis;

[0030] (2) β-tyrosinase catalyzes amino acid conversion: in step (1) silk fibroin solution, add 10U / ml β-tyrosinase, 4g / L phenol, pyridoxal phosphate 1g / L and ammonium chloride 1g / L L, treated at a temperature of 40°C and a pH of 8.0 for 30 hours; after the treatment, the silk fibroin solution was dialyzed with deionized water for 24 hours;

[0031] (3) Tyrosinase catalyzes the cross-linking of silk fibroin and elastin: in step (2) silk fibroin solution, add 8U / ml tyrosinase, 8g / L elastin, at a temperature of 30°C and a pH of 7.0 Under treatment for 20 hours;

[0032] (4) Formation of silk fibroin / elastin composite f...

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Abstract

The invention discloses a method for preparing a fibroin and elastin composite membrane material through an enzymic method. Beta-tyrosinase is applied to amino acid conversion in fibroin, the number of tyrosine residues in fibroin protein is increased, fibroin is catalytic oxidized to be crosslinked with elastin, and a fibroin and elastin composite membrane is prepared. The method comprises the steps that 1, a fibroin solution is prepared; 2, the beta-tyrosinase and phenol are added, and serine in the fibroin is catalyzed to be converted to tyrosine; 3, the tyrosinase is added in the fibroin solution, and the tyrosine residues on the fibroin are catalytic oxidized to be reacted with the elastin; 4, fibroin and elastin are dried through air to form the membrane or dried through refrigeration to form the membrane. Compared with a functional fibroin membrane prepared from traditional chemical cross-linking, according to the method for preparing the fibroin and elastin composite membrane material through the enzymic method, the enzyme catalysis efficiency is high, the enzyme treatment condition is mild, and the composite membrane performance can be obviously improved; the fibroin and elastin composite membrane which is built through the enzymic method is good in biocompatibility, and the membrane material mechanical property is improved.

Description

technical field [0001] The present invention relates to a method for enzymatic preparation of silk fibroin / elastin composite membrane material, in particular to a method for converting amino acids in silk fibroin by using β-tyrosinase, and catalyzing the exchange of silk fibroin and elastin by means of tyrosinase. The invention relates to a method for preparing a composite silk fibroin membrane material, which belongs to the field of textile biotechnology. Background technique [0002] Silk fibroin derived from mulberry silk has good biocompatibility, and the silk fibroin film processed from it is widely used in the construction of medical tissue engineering materials. Elastin is a highly cross-linked extracellular matrix protein that is extremely difficult to dissolve in water. It is mainly produced by smooth muscle cells, endothelial cells, fibroblasts and chondrocytes, and can be cross-linked with microfibrils to form elastic fibers in vivo , has the function of passive ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08J5/18C08J3/24C08L89/00C12P13/22
Inventor 王平余圆圆邓超王强范雪荣
Owner JIANGNAN UNIV
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