Preparation method of silk fibroin peptide with bacteriostatic activity

A technology of antibacterial activity and silk fibroin peptide, which is applied in the field of preparation of silk fibroin peptide with antibacterial activity, can solve the problems of destruction, tryptophan destruction, and reduction of nutritional value, so as to overcome low reaction efficiency, low cost, and environmental pollution. friendly effect

Inactive Publication Date: 2013-03-13
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Acid hydrolysis can fully hydrolyze silk fibroin, but tryptophan will be completely destroyed, and serine, tyrosine and threonine will also be partially destroyed; alkaline hydrolysis can also fully hydrolyze silk fibroin, but not enough The disadvantage is that it will cause racemization, thereby reducing its nutritional value; saline solution degradation can obtain silk fibroin with a molecular weight of tens of thousands, which is still a macromolecular protein and thus affects its absorption and nutritional value; The advantages of small damage to amino acids have become the most promising method for degrading silk fibroin
Therefore adopt enzymatic method to hydrolyze silk fibroin, can improve the output and quality of silk fibroin peptide greatly; There is no related report about the preparation of peptide with antibacterial activity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Add the cut cocoons to Na with a liquor ratio of 1:20 2 CO 3 (0.5%, g / mL) solution, degummed at 95 °C for 20 min, washed with distilled water and dried, added to CaCl 2 (1:10, g / mL) solution, dissolved in a temperature of 90 °C for 2 min, cooled to room temperature, and the obtained silk fibroin solution was adjusted to pH 8 with NaOH solution. %, the amount of enzyme added is 400 U / g, and neutral protease is used for enzymatic hydrolysis reaction, and the degree of hydrolysis (DH) = 4% can be controlled to obtain silk fibroin peptide products with antibacterial activity. The antibacterial activity of the product is characterized by its minimum inhibitory concentration (MIC) (the following other examples are also applicable): that is, the sample solution of a certain concentration is mixed with the medium and poured into the plate to inoculate Salmonella, Escherichia coli or Staphylococcus aureus. The beef extract peptone plate with the same solution was used as the ...

Embodiment 2

[0018] Add the cut cocoons to Na with a liquor ratio of 1:50 2 CO (0.5%, g / mL) 3 solution, degummed at a temperature of 100 °C for 50 min, washed with distilled water, filtered and dried, added to CaCl 2 (1:10, g / mL) solution, dissolved in the temperature of 100 ℃ for 30 min, cooled to room temperature, the obtained silk fibroin solution was adjusted to pH 8.5 with NaOH solution, and the temperature was 65 ℃, the substrate concentration was 10 %, the amount of enzyme added is 1200 U / g, and the enzymatic hydrolysis reaction is carried out with alkaline protease, and the antibacterial active silk fibroin peptide with the degree of hydrolysis (DH) = 12% can be obtained. The antibacterial activity of the product was 28.19%.

Embodiment 3

[0020] Add the cut cocoons to Na with a liquor ratio of 1:50 2 CO 3 (0.5%, g / mL) solution, degummed at 98 °C for 30 min, washed with distilled water, filtered and dried, added to CaCl 2 (1:10, g / mL) solution, dissolved at 98 °C for 25 min, cooled to room temperature, and adjusted the pH of the obtained silk fibroin solution to 8.5 with NaOH solution. %, the amount of enzyme added is 1000 U / g, and the enzymatic hydrolysis reaction is carried out with compound protease, and the silk fibroin peptide product with DH=14% has antibacterial activity. The antibacterial activity of the product was 42.11%.

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Abstract

The invention discloses a preparation method of silk fibroin peptide with bacteriostatic activity. The method comprises the following steps of: by using cut-off silkworm cocoons as a raw material, adding the raw material in a Na2CO3 solution, washing the raw material with distilled water after degumming and filtering the washed raw material to dry; adding the dried raw material in a CaCl2 solution, dissolving, and regulating the pH of the cooled silk fibroin with a NaOH solution; and carrying out enzyme hydrolysis reaction by protease, and controlling the degree of hydrolysis to obtain the silk fibroin peptide with the bacteriostatic activity. The preparation method overcomes the defect of low reaction efficiency in the prior art, and has the advantages of low cost, mild reaction conditions, simple and controllable reaction process and lower energy consumption and is environment-friendly, and industrial production is easy to realize.

Description

technical field [0001] The invention belongs to the technical field of preparation of silk peptides, and in particular relates to a method for preparing silk peptides with antibacterial activity. Background technique [0002] my country is a big country in silk production and processing, and has rich sources of silk fibroin. my country produces about 50,000 tons of silk every year, and it is roughly estimated that 56,000 tons of silk fibroin is produced in my country every year. In addition, due to the needs of seed production in silkworm breeding farms, a large number of cut cocoons are produced. Most of these cut cocoons cannot be reeled, and are usually used as raw materials for silk spinning or processed silk floss. A small part is used for cosmetics and pharmaceuticals, which is economical Low value. With the gradual deepening of people's understanding of the physical and chemical properties, structural characteristics and biological activities of silk fibroin, silk f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06
Inventor 李晓凤吴晖余以刚赖富饶
Owner SOUTH CHINA UNIV OF TECH
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