Preparation method of active macromolecular keratin and application thereof as biological dressing

A technology of keratin and macromolecules, which is applied in the field of preparation of active macromolecular keratin, can solve the problems of difficult extraction and reuse of keratin, secondary environmental pollution, toxic reagent residues, etc., and achieve the extraction process of environmental protection, environmental protection, Effects that promote proliferation and migration

Active Publication Date: 2020-09-01
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003]Wool fiber has a complex three-dimensional structure that is highly intersected and connected by disulfide bonds, hydrogen bonds and van der Waals forces. It has strong resistance and is difficult to dissolve in water and weak acids Weak bases, organic solvents, etc. have brought great difficulties to the extraction and reuse of keratin
The traditional method of extracting active macromolecular keratin mainly relies on thiol-containing compounds, such as mercaptoethanol and thioglycolic acid; or sulfur-containing compounds, such as sulfite, EDTA, DTT and other toxic and harmful chemical reagents, which will cause secondary pollution to the environment and Toxic Reagent Residue
Keratinase is a recognized biological enzyme that can degrade keratin waste such as wool and feathers. However, the degradation products are currently mainly amino acids and low-molecular-weight small peptides, which are used in fertilizers and feeds with low added value.

Method used

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  • Preparation method of active macromolecular keratin and application thereof as biological dressing
  • Preparation method of active macromolecular keratin and application thereof as biological dressing
  • Preparation method of active macromolecular keratin and application thereof as biological dressing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] (1) Wash the wool fiber with deionized water, degrease it in a Soxhlet extractor with acetone 3 times the volume of wool for 24 hours, then wash off the residual acetone on the surface with 40°C deionized water, dry the obtained wool fiber shredded;

[0030] (2) Disperse 5 g of shredded degreased wool in 50 mL of water, adjust the pH value to 9 with 1 mol / L NaOH solution, seal it, raise the temperature to 65 ° C, and mechanically stir for 1 h;

[0031] (3) Lower the temperature to 50°C, add 100KU enzymatic keratinase solution for enzymolysis, and continue to degrade with mechanical stirring for 60 hours;

[0032] (4) centrifuging the mixed solution of wool and keratin obtained in step (3), and separating the supernatant keratin solution;

[0033] (5) Dialyzing the supernatant keratin solution obtained in step (4) for 72 hours;

[0034] (6) After the dialysis is completed, the dialysate was frozen at -80°C for 6 hours, and then dried on a freeze dryer for 72 hours to o...

Embodiment 2

[0036] (1) Wash the wool fiber with deionized water, degrease the wool fiber with 4 times the volume of wool in a Soxhlet extractor for 24 hours, then wash off the residual acetone on the surface with 50°C deionized water, dry the obtained wool fiber shredded;

[0037] (2) Disperse 5 g of shredded degreased wool in 100 mL of water, adjust the pH value to 10 with 1 mol / L NaOH solution, seal it, raise the temperature to 70 ° C, and stir it mechanically for 1 h;

[0038] (3) Lower the temperature to 50°C, add 150KU enzymatic keratinase solution for enzymolysis, and continue mechanical stirring for 54 hours to degrade;

[0039] (4) centrifuging the mixed solution of wool and keratin obtained in step (3), and separating the supernatant keratin solution;

[0040] (5) Dialyzing the supernatant keratin solution obtained in step (4) for 72 hours;

[0041] (6) After the dialysis is completed, the dialysate is frozen at -80°C for 8 hours, and then dried on a freeze dryer for 48 hours t...

Embodiment 3

[0043] (1) Wash the wool fiber with deionized water, degrease the wool fiber with 5 times the volume of wool in a Soxhlet extractor for 12 hours, then wash off the residual acetone on the surface with 50°C deionized water, dry the obtained wool fiber shredded;

[0044] (2) Disperse 5 g of shredded degreased wool in 150 mL of water, adjust the pH value to 11 with 1 mol / L NaOH solution, seal it, raise the temperature to 85 ° C, and mechanically stir for 1 h;

[0045] (3) Lower the temperature to 50°C, add 200KU enzymatic keratinase solution for enzymolysis, and continue mechanical stirring for 48 hours;

[0046] (4) centrifuging the mixed solution of wool and keratin obtained in step (3), and separating the supernatant keratin solution;

[0047] (5) Dialyzing the supernatant keratin solution obtained in step (4) for 72 hours;

[0048] (6) After the dialysis is completed, the dialysate is frozen at -80°C for 12 hours, and then dried on a freeze dryer for 48 hours to obtain soli...

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Abstract

The invention discloses a preparation method of active macromolecular keratin and application thereof as a biological dressing. According to the preparation method, macromolecular keratin with biological activity is extracted from waste wool by controlling the reaction conditions of keratinase, the molecular weight of the macromolecular keratin is mainly distributed between 45kDa and 28kDa, and the keratin can effectively promote proliferation and migration of fibroblasts and shows obvious oxidation resistance. The regenerated keratin powder can be self-assembled into hydrogel, can induce tissue regeneration and repair, effectively promotes wound healing, and is a good medical biological dressing. The method is simple in operation process, low in equipment cost and easy for industrial production; fibers, which cannot be used for textile processing, such as waste wool, cashmere, wool staple fibers, and the like, in the animal husbandry and the textile industry are used as raw materials,resources are saved, and the environment is protected. Keratinase is used for biological extraction, the extraction process is environmentally friendly, and pollution is avoided.

Description

technical field [0001] The invention relates to a preparation method of active macromolecular keratin and its application as a biological dressing, belonging to the field of biotechnology. Background technique [0002] Wool processing has already emerged in my country. After a long period of development, my country has become a big country for wool imports and wool products. However, domestic wool spinning enterprises produce hundreds of thousands of tons of wool waste every year. A large amount of waste wool is generated. If not utilized, these wastes will deepen environmental pollution and also be a huge waste. The protein content in wool fiber is as high as 99%, which is a high-quality protein resource. In recent years, with the in-depth study of keratin, researchers have found that keratin is a natural biomaterial with good biocompatibility and biodegradability. Therefore, keratin shows potential in tissue engineering and medical applications, such as promoting wound h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/47C12P21/06A61L26/00
CPCC07K14/4741C12P21/06A61L26/008A61L26/0047A61L26/0066A61L2300/252A61L2300/412C08L89/04
Inventor 史劲松龚劲松苏畅许正宏蒋敏叶金鹏
Owner JIANGNAN UNIV
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