Method for purifying silk fibroin

A silk fibroin and purification method technology, applied in the field of biomaterials, can solve the problems of long time, weak degumming ability, high price, etc., and achieve the effect of improving performance and maintaining molecular weight

Active Publication Date: 2014-03-05
河南丝绸之宝卫生用品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the literature ([J] Journal of Liaocheng Normal University (Natural Science Edition), 1999, 01:43-47+62), it is introduced that boiling water degumming requires high temperature, long time, and degumming is not complete; soap degumming, soap consumption Large, accounting for about 20-30% of raw silk, and the requirements for soap are high, the effect of general soap is not ideal, the degumming ability is weak, the soap used in standard degumming is Marseille soap (a kind of soap prepared from olive oil) , the source is limited, needs to be imported, the price is expensive, and the sericin cannot be completely removed; the literature ([J] Japanese Sericulture Journal, 1955, 1 (6) 32~64) uses the organic acid tartaric acid as the degumming agent, and the degumming condition : The concentration of tartaric acid is 16%, the temperature is 93°C, and the degumming rate is 80% for 2 hours. The degumming rate becomes slower when the degumming time is extended, and tartaric acid is...

Method used

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  • Method for purifying silk fibroin
  • Method for purifying silk fibroin

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Take 40 grams of silkworm silk, soak it in 5% fatty alcohol polyoxyethylene ether at 40°C for 30 minutes, the bath ratio is 1:10, and then boil it with 0.01% sodium bicarbonate aqueous solution, the bath ratio is 1:50 , time 30min, repeat the above steps twice, after rinsing with deionized water, put it in a 60°C oven for drying, the dried silk is in phosphoric acid with a bath ratio of 1:10, a papain concentration of 3g / L, and a temperature of 65°C. Soak in disodium hydrogen-citric acid buffer solution (pH = 6) for 60 minutes to degumming, wash with deionized water, and dry in an oven at 60°C. The dried degummed silk is dissolved in a 9.3M lithium bromide solution, and dialyzed with deionized water for 3 to 4 days to obtain a pure silk fibroin solution.

Embodiment 2

[0019] Take 40 grams of silkworm cocoons, soak them in 2% fatty alcohol polyoxyethylene ether at 40°C for 30 minutes, with a bath ratio of 1:20, and then boil them with 0.5% aqueous sodium bicarbonate solution with a bath ratio of 1:50. After 30 minutes, rinse with deionized water and dry in a 60°C oven. The dried silk is buffered with disodium hydrogen phosphate-citric acid at a bath ratio of 1:10, a papain concentration of 5g / L, and a temperature of 60°C. Soak in solution (pH=6) for 60 minutes to degumming, wash with deionized water, and then dry in an oven at 60°C. The dried degummed silk is dissolved in a 9.3M lithium bromide solution, and dialyzed with deionized water for 3 to 4 days to obtain a pure silk fibroin solution.

Embodiment 3

[0021] Take 40 grams of silkworm cocoons, soak them in sodium salt of 5% glyceryl monostearate sulfate at 60°C for 30 minutes, with a bath ratio of 1:50, and then boil them with 0.01% aqueous sodium bicarbonate solution with a bath ratio of 1 :50, time 30min, repeat the above steps twice, rinse with deionized water, dry in a 60°C oven, dry silk at a bath ratio of 1:10, trypsin concentration 10g / L, temperature 37°C Soak in PBS buffer (pH = 8) for 60 min to degumming, wash with deionized water, and dry in an oven at 60°C. The dried degummed silk is dissolved in a 9.3M lithium bromide solution, and dialyzed with deionized water for 3 to 4 days to obtain a pure silk fibroin solution.

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Abstract

The invention discloses a method for purifying silk fibroin. According to the method, glycerin monostearate sulphate sodium salt, fatty alcohol-polyoxyethylene ether, and sodium diethylhexyl sulfosuccinate are taken as the penetrant. High molecular weight silk fibroin can be obtained through steps of steeping raw silks of silkworm or silkworm cocoon in the penetrant, degumming through weak base and enzyme so as to remove silk glue, then dissolving and dialyzing. The silk fibroin purified by the method does not contain silk glue, less molecular weight of silk fibroin is degraded, the chain H, chain L and P25 of the original silk fibroin can be reserved, the molecular weight of the original silk fibroin is reserved, and the materials such as silk fibroin membranes, fibroin gel, fibroin microsphere, and fibroin porous membranes prepared by the raw material silk fibroin are improved in properties.

Description

technical field [0001] The invention relates to a method for purifying protein, in particular to a method for purifying silk fibroin, and belongs to the technical field of biological materials. Background technique [0002] Silkworm silk is mainly composed of two proteins, silk fibroin and sericin, of which silk accounts for about 70% and sericin accounts for about 30%. Since pure silk fibroin has low antigenicity, while sericin has certain antigenicity, silk fibroin as a biomedical material needs to be purified to remove sericin. Sericin is a protein that constitutes the outer layer of silk fiber, and its protein chain has many amino acids with long side chains, such as arginine, lysine, glutamic acid, methionine, tryptophan, tyrosine etc., and many polar hydrophilic groups (such as hydroxyl, carboxyl, amino, imino, etc.). Sericin is water-soluble and easily soluble in hot water, acid, and alkali solutions, while silk fibroin is only water-swellable and insoluble in water...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/14
CPCC07K14/43586
Inventor 卢神州张小平邢铁玲
Owner 河南丝绸之宝卫生用品有限公司
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