Preparation and identification method of domestic silkworm silk fibroin solution

A technology of silk fibroin and protein solution, applied in the field of biomedicine, can solve the problems of weakening effect, unfavorable research and application of silk fibroin molecules, etc., and achieve the effects of reducing preparation cost, avoiding large-scale hydrolysis and clarifying appearance.

Active Publication Date: 2015-06-03
SUZHOU SIMEITE BIOTECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In Chinese patent CN103194068A, urea is added to destroy silk fibroin to stabilize the silk fibroin solution, which weakens the effect of hydrogen bonds in the silk fibroin molecule or between several silk fibroin molecules, and improves the stability of silk fibroin.

Method used

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  • Preparation and identification method of domestic silkworm silk fibroin solution
  • Preparation and identification method of domestic silkworm silk fibroin solution
  • Preparation and identification method of domestic silkworm silk fibroin solution

Examples

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Embodiment 1

[0038] The invention provides a method for preparing silkworm silk fibroin protein solution, which uses a ternary solution containing calcium chloride to dissolve the silk, comprising the following steps:

[0039] 1) Weigh 110 grams of anhydrous calcium chloride, dissolve it in 140 grams of pure water, and after it is fully dissolved and returned to room temperature, add 92 grams of absolute ethanol to prepare a calcium chloride-containing ternary solution as a solvent;

[0040] 2) Cut silkworm cocoons into pieces and weigh 30 grams, boil them in an aqueous solution of sodium carbonate with a molar concentration of 0.02 for 25-35 minutes to remove sericin, then wash them repeatedly with deionized water for 3 times, and put them in a fume hood after washing dry, ready to use;

[0041] 3) Dissolve the air-dried degummed silk in the ternary solution with a bath ratio of 1:25, and dissolve it at a constant temperature of 75-85°C for 0.5-2 hours, such as figure 1 As shown, it can ...

Embodiment 2

[0055] The present invention provides a method for preparing silkworm silk fibroin protein solution. The degumming, silk dissolving and renaturation treatment methods are basically the same as those described in Example 1, and will not be repeated here.

[0056] The difference is that in this example, the bath ratio is 1:10, the temperature is 60°C, and the time is 0.5 hours for silk-melting treatment, and then each step of dialysis is 1 hour.

[0057] The silk fibroin solution obtained through degumming, silk melting and renaturation treatment in this example: the silk melting time is shorter, the silk melting temperature is lower, and the molecular weight of silk fibroin is larger. In addition, due to the shorter dialysis time, the silk fibroin renaturation The effect is more general.

Embodiment 3

[0059] The present invention provides a method for preparing silkworm silk fibroin protein solution. The degumming, silk dissolving and renaturation treatment methods are basically the same as those described in Example 1, and will not be repeated here.

[0060] The difference is that in this embodiment, the bath ratio is 1:50, the temperature is 85° C., and the time is 24 hours for silk-melting treatment.

[0061] The silk fibroin solution obtained by degumming, silk melting and renaturation treatment in this example: the time of silk dissolution is longer, the temperature of silk dissolution is longer, and the molecular weight of silk fibroin is smaller. In addition, since the dialysis time exceeds 5 hours, the silk fibroin It is difficult to further improve the refolding effect.

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Abstract

The invention relates to the technical field of biomedicines, and in particular relates to a preparation and identification method of a domestic silkworm silk fibroin solution. The preparation method of the domestic silkworm silk fibroin solution sequentially comprises the following steps: (1) degumming, namely degumming domestic silkworm silks to remove silk sericin on the outer layer to obtain degummed silks; (2) dissolving the silks, namely dissolving the degummed silks by using a ternary solution containing calcium chloride to obtain a fibroin solution; (3) performing renaturation, namely sequentially performing dialysis treatment on the fibroin solution according to a concentration gradient from high to low by using a protein denaturant solution, and then performing water dialysis treatment to obtain a silk fibroin solution; and (4) storing, namely storing the silk fibroin solution. The invention also provides an identification method of the domestic silkworm silk fibroin solution. The method can be used for identifying the quality of the silk fibroin solution. The preparation method of the domestic silkworm silk fibroin solution disclosed by the invention is relatively high in economical performance, ensures that the molecular weight of the domestic silkworm silk fibroin solution is close to that of natural fibroin molecules, and is beneficial to the preparation of subsequent silk fibroin based biomaterials.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a method for preparing and identifying silkworm silk fibroin solution. Background technique [0002] Silk is a product of ancient Chinese civilization. It has been widely used as clothing material since ancient times due to its softness, good air permeability and soft texture. In recent decades, because silk protein is non-toxic, non-irritating, has excellent biocompatibility, can promote the growth of human cells, and has certain biodegradability, it has been widely used in biomedicine, environmental protection and daily use. Technological fields such as chemical engineering are also developing continuously. [0003] Silk is mainly composed of two proteins, sericin and silk fibroin. Mulberry silk core-shell structure, the outer layer is coated with sericin (Sericin), and the inner layer is silk fibroin (Silk Fibroin), in which the silk fibroin content is 70-80%. Silk fibro...

Claims

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Application Information

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IPC IPC(8): C07K14/435C07K1/34C07K1/113G01N33/68
CPCC07K14/43586G01N33/68
Inventor 王晓沁郑兆柱郭少哲
Owner SUZHOU SIMEITE BIOTECH CO LTD
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