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Preparation method of silk fibroin oligopeptide for skin-care products

A technology of oligopeptides and skin care products, applied in the field of biopolymer materials, can solve the problems of silk fibroin such as large molecular weight, low controllability, and complicated process, and achieve high biological activity, easy absorption, and high output rate Effect

Active Publication Date: 2017-08-29
JIANGSU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The molecular weight of the obtained silk fibroin is above 100,000 Da, and the content of heavy metals is relatively high, so it can be directly used as raw materials for skin care products. Highly affect the safety of the product, such as "a preparation and identification method of silkworm silk fibroin solution" disclosed in Chinese patent CN201510071886.3, "a cationic silk fibroin and its preparation method" disclosed in Chinese patent CN201410352789.7 method, etc.
[0004] It has been reported to use protease to hydrolyze silk protein, mainly including alkaline protease, papain, pepsin, etc. (Food and Fermentation Industry, 2008, 6:28-29) reported that the optimum temperature for hydrolysis of silk fibroin by elastase is 45-55°C, the optimum pH is 9.0, and the relative molecular mass of the hydrolyzate is mainly above 1000D. Mainly macromolecular polypeptide substances, high technical requirements, complicated process, low degree of controllability

Method used

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  • Preparation method of silk fibroin oligopeptide for skin-care products

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Effect test

Embodiment 1

[0023] A method for preparing silk fibroin oligopeptides for skin care products, the preparation steps are as follows:

[0024] (1) Degumming: Cut silkworm cocoons into 1-2cm 2 , with 0.5-0.8% Na 2 CO 3 Degumming by soaking in the solution for 30 minutes, with a solid-to-liquid mass ratio of 1:200, then washing with water, repeating dipping and washing for 2-3 times, drying naturally or drying at 60°C to obtain degummed silk;

[0025] (2) Salt solution: put the degummed silk obtained in step (1) into 50-60% CaCl 2 The solution was heated to boiling and dissolved for 20 minutes, the solid-to-liquid mass ratio was 1:20, and after cooling, it was centrifuged at a speed of 3500 r / min for 20 minutes to obtain a salt solution;

[0026] (3) Dialysis: the salt solution obtained in step (2) was dialyzed for 2-3 days with a dialysis bag with a molecular weight cut-off of 15000-10000Da to obtain a low-molecular-weight silk fibroin solution;

[0027] (4) Double enzymolysis: mix the co...

Embodiment 2

[0032] A method for preparing silk fibroin oligopeptides for skin care products, the preparation steps are as follows:

[0033] (1) Degumming: Cut silkworm cocoons into 1-2cm 2 , with 0.5-0.8% Na 2 CO 3 Degumming by solution immersion for 35 minutes, with a solid-to-liquid mass ratio of 1:200, then washing with water, repeating the immersion and cleaning 2-3 times, and drying naturally or at 65°C to obtain degummed silk;

[0034] (2) Salt solution: put the degummed silk obtained in step (1) into 50-60% CaCl 2 The solution was heated to boiling and dissolved for 25 minutes, the solid-to-liquid mass ratio was 1:20, and after cooling, it was centrifuged at 3800 r / min for 25 minutes to obtain a salt solution;

[0035] (3) Dialysis: the salt solution obtained in step (2) was dialyzed for 2-3 days with a dialysis bag with a molecular weight cut-off of 15000-10000Da to obtain a low-molecular-weight silk fibroin solution;

[0036] (4) Double enzymolysis: mix the composite enzyme w...

Embodiment 3

[0041] A method for preparing silk fibroin oligopeptides for skin care products, the preparation steps are as follows:

[0042] (1) Degumming: cut the cocoon into 1-2cm 2 , with 0.5-0.8% Na 2 CO 3 Degumming by solution immersion for 40 minutes, with a solid-to-liquid mass ratio of 1:200, then washing with water, repeating the immersion and cleaning 2-3 times, and drying naturally or at 70°C to obtain degummed silk;

[0043] (2) Salt solution: put the degummed silk obtained in step (1) into 50-60% CaCl 2 The solution was heated to boiling and dissolved for 30 minutes, the solid-to-liquid mass ratio was 1:20, and after cooling, it was centrifuged at 4000 r / min for 30 minutes to obtain a salt solution;

[0044] (3) Dialysis: the salt solution obtained in step (2) was dialyzed for 2-3 days with a dialysis bag with a molecular weight cut-off of 15000-10000Da to obtain a low-molecular-weight silk fibroin solution;

[0045] (4) Double enzymolysis: mix the compound enzyme with the...

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Abstract

The invention discloses a preparation method of silk fibroin oligopeptide for skin-care products. The preparation method comprises the following steps: (1) degumming: steeping silkworm cocoons with 0.5-0.8% Na2CO3, rinsing and drying; (2) salt removing: dissolving in boiling water through 50-60% of CaCl2, cooling and then centrifuging, and dialyzing supernatant to obtain a small-molecular silk fibroin solution; (3) double-enzymolysis: performing double-enzymolysis with alkaline protease and actinozyme, centrifuging, and dialyzing supernatant to obtain low-molecular-weight silk fibroin solution; (4) membrane separation: separating with sephadex gel chromatographic column Sephadex G-15, and collecting components to obtain a silk fibroin oligopeptide solution; and (5) drying and storage: performing ultralow-temperature freeze drying on the silk fibroin oligopeptide solution to obtain silk fibroin oligopeptide powder. The preparation method of the silk fibroin oligopeptide is simple and convenient to operate, with extraction rate of more than 93%, and is applicable to addition of raw materials into skin-care products and cosmetics.

Description

technical field [0001] The invention relates to a method for preparing silk protein, in particular to a method for preparing a silk fibroin oligopeptide used in skin care products, and belongs to the technical field of biopolymer materials. Background technique [0002] Silk is a high-molecular animal protein (silk fibroin), rich in 18 kinds of amino acids, and has excellent properties such as moisturizing, breathable, antibacterial, anti-static, and anti-ultraviolet. As early as the Ming Dynasty, Li Shizhen recorded the beauty and skin care effects of silk in his medical masterpiece "Compendium of Materia Medica". With the advancement and development of science and technology, people's understanding of silk protein is getting deeper and deeper, and the research and development of new uses of silk have attracted people's attention. [0003] Natural silk fibroin protein has a relatively large molecular weight, about 350,000 Da (Dalton), exists in the form of fibers, and is n...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K1/34A61K8/64A61Q19/08A61Q17/00A61Q17/04A61Q19/00
CPCA61K8/64A61K2800/10A61Q17/005A61Q17/04A61Q19/00A61Q19/004A61Q19/08C07K1/34C12P21/06
Inventor 梁佳桂仲争翁渝洁赵山王佳贾俊强
Owner JIANGSU UNIV OF SCI & TECH
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