Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Fibroin polypeptide and preparation and application thereof

A technology of silk protein and thistle protease, which is applied in the fields of peptides, cosmetic preparations, dressing preparations, etc., and can solve the problem of small molecular proteins losing their original protein functions

Inactive Publication Date: 2015-10-28
SUZHOU PULUODA BIOLOGICAL SCI & TECH
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, small proteins tend to lose the efficacy of the original protein
Therefore, how to make the decomposed silk protein small molecules have the effect of the original protein without causing allergies has always been a problem that is difficult to coordinate

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Fibroin polypeptide and preparation and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0013] Cut 500g of washed and dried silkworm cocoons into pieces, add 500-1000ml of deionized water; boil and ultrasonic for 1-2 hours while stirring slowly; after the solution is cooled, add bromelain and react at room temperature for 60 minutes; centrifuge at 2000rpm, 5-10 minutes, take the supernatant; add the supernatant to the acidic buffer solution of pH6.0-6.5, stir, centrifuge at 10000-15000rpm for 30-60 minutes to obtain the precipitate; redissolve the precipitate in pH5. 3-5.7 buffer solution, using Sephadex gel chromatography technique to separate the polypeptide with a molecular weight of 2-3KDa. The sequence of the polypeptide determined by a solid-phase protein automatic sequencer is DTNQTDKHGAILLLQEIV, and the purity is 80.2%.

Embodiment 2

[0015] Cut 500g of washed and dried silkworm cocoons into pieces, add 500-1000ml of deionized water; boil and sonicate for 1-2 hours while stirring slowly; after the solution is cooled, add ficin and react at 37°C for 30 minutes; centrifuge at 3000rpm , 5-10 minutes, take the supernatant; add the supernatant to the acidic buffer solution of pH 6.0-6.5, stir, and centrifuge at 10000-15000rpm for 30-60 minutes to obtain a precipitate; redissolve the precipitate in pH5 .3-5.7 buffer solution, using Sephadex gel chromatography technique to separate the polypeptide with a molecular weight of 2-3KDa. The sequence of the polypeptide determined by a solid-phase protein automatic sequencer is DTNQTDKHGAILLLQEIV, and the purity is 85.2%.

Embodiment 3

[0017] The peptide synthesized by Shanghai Jill was added to the cream base for immunogenicity testing. 6 BALB / c white mice, half male and half male, the back of the mice were depilated, and the skin was exposed. Take different concentrations of silk protein polypeptide solutions (0.5, 1.0, 2.0mg / ml) and apply them at a density of 1.25μL / cm2 for a short period of time, once a day for 8 consecutive weeks, and apply them on the orbital area of ​​1-12 weeks Blood was drawn from the venous plexus once a week, centrifuged at 12,000 rpm for 2 minutes, the plasma was separated to obtain the supernatant, and stored at -20°C for later use. After dissolving at room temperature, take 0.1ml of the supernatant to detect the antibody titer in the serum by indirect ELISA. The RA value is obtained by comparing the actual A value (OD450) of each sample with the A value of the negative control serum. RA ≥ 2.1 is positive. Blood bio-samples are tested after a series of dilutions. The highest di...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a fibroin polypeptide, and relates to the field of biotechnology polypeptide preparation. The sequence of the fibroin polypeptide is DTNQTDKHGAILLLQEIV. A preparation method of the fibroin polypeptide comprises the steps that 500 g of cleaned and dried silkworm cocoons are cut up and added into 500-1000 ml of deionized water; the water with the silkworm cocoons is boiled, ultrasound is conducted for 1-2 h, and moderate agitation is conducted simultaneously; after the solution is cooled, hydrolase is added into the solution, and reaction is conducted for 30-60 min at the temperature ranging from 20 DEG C to 40 DEG C; centrifugation is conducted for 5-10 min at the rotating speed ranging from 2000 rpm to 3000 rpm, and supernatant is obtained; the supernatant is added into a meta-acid buffer solution with pH ranging from 6.0 to 6.5, the agitation is conducted, and the centrifugation is conducted for 30-60 min at the rotating speed ranging from 10000 rpm to 15000 rpm to obtain sediments; the sediments are redissolved in a buffer solution with pH ranging from 5.3 to 5.7, the dextran gel chromatography technology is adopted to separate out polypeptide with the molecular weight ranging from 2 KDa to 3 KDa, and then the fibroin molecular weight is obtained. The fibroin polypeptide can be used as an additive to prepare cosmetics. An antigen experiment result shows that the fibroin polypeptide does not have antigenicity, and an experiment of in-vitro inhibitory of tyrosinase verifies that the fibroin polypeptide has the skin whitening function.

Description

technical field [0001] The invention relates to the field of biotechnology polypeptide preparation, in particular to the field of silk protein polypeptide preparation. technical background [0002] Silk protein (Fibroin) also known as: silk fibroin. Silk fibroin is a natural high-molecular fiber protein extracted from silk, and its content accounts for about 70% to 80% of silk. It contains 18 kinds of amino acids, of which glycine (gly), alanine (ala) and serine (ser) are about Accounting for more than 80% of the total composition. [0003] Silk protein is developed and utilized in the field of cosmetics. Because silk protein is very similar to human skin keratin protein, the two have excellent affinity; due to the special chemical structure of silk protein molecules, silk protein has good hygroscopic and hygroscopic properties, and can adapt to changes in the surrounding environment , play the role of natural humidity control, improve human skin nutrition, prevent skin w...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K7/08A61K8/64A61Q19/02A61Q19/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com