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Sericin polypeptide 2, and preparation and application thereof

A technology of sericin and rubber powder, which is applied in the fields of peptides, cosmetic preparations, dressing preparations, etc., and can solve the problems of small molecular weight and difficult coordination of sericin

Inactive Publication Date: 2015-11-04
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, how to make the decomposed sericin have a smaller molecular weight and have the effect of the original protein has always been a problem that is difficult to coordinate

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Take 500g of sericin powder and dissolve in 500ml of deionized water; quickly cool the sericin solution to 4°C; centrifuge at 4°C, 10,000rpm for 60 minutes, and take the supernatant; use a rotary evaporator to concentrate the supernatant solution; After cooling, add lysozyme and react at 37°C for 30 minutes; centrifuge at 3000rpm for 5 minutes and take the supernatant; add the supernatant to the buffer solution with pH5.7-6.3, stir and centrifuge at 10000rpm for 60 minutes to obtain the precipitate ; Redissolve the precipitate in pH 6.4-7.2 buffer, and use dialysis to separate the polypeptide with a molecular weight of 1600-3400Da. The sequence of the polypeptide determined by a solid-phase automatic protein sequencer was ACDSTTVNFIPSRATTEEQ, and the purity was 84.1%.

Embodiment 2

[0014] Take 500g of sericin powder and dissolve in 1000ml deionized water; quickly cool the sericin solution to 10°C; centrifuge at 10°C, 15000rpm, for 30 minutes, and take the supernatant; use a rotary evaporator to concentrate the supernatant solution; After cooling, add thrombin and react at 37°C for 30 minutes; centrifuge at 2000rpm for 10 minutes and take the supernatant; add the supernatant to the buffer solution with pH5.7-6.3, stir, and centrifuge at 15000rpm for 30 minutes to obtain a precipitate ; Redissolve the precipitate in pH 6.4-7.2 buffer, and use dialysis to separate the polypeptide with a molecular weight of 1600-3400Da. The sequence of the polypeptide determined by a solid-phase automatic protein sequencer was ACDSTTVNFIPSRATTEEQ, and the purity was 73.0%.

Embodiment 3

[0016] Add the polypeptide prepared in Example 1 into the cream base. Take different concentrations of sericin 2.0mg / ml and sericin polypeptide solution (0.5, 1.0, 2.0mg / ml) and apply it once for a short period of time at a coating density of 1.25μL / cm2, gently pat until absorbed, and measure 12 Moisture content of stratum corneum after hours. Data calculation: Change rate of skin hydration (%) = [skin hydration (test) - skin hydration (control)] / skin hydration (control) × 100%. The experiment was repeated three times, and the skin hydration rate of 2.0mg / ml sericin, 0.5, 1.0, 2.0, mg / ml sericin polypeptide solutions increased to 42.12, 29.61, 42.61, 53.02% respectively.

[0017] At the same time, take different concentrations of sericin 2.0mg / ml and sericin polypeptide solution (0.5, 1.0, 2.0mg / ml) and apply it once for a short period of time at a coating density of 1.25μL / cm2, and gently pat until absorbed. After 12 hours, the water loss of the epidermis before and after u...

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Abstract

The invention discloses a sericin polypeptide, relating to the field of polypeptide preparation in biotechnology. The sequence is ACDSTTVNFIPSRATTEEQ. The preparation method comprises the following steps: (1) taking 500g of sericin powder, and adding deionized water to dissolve the sericin powder according to the mass / volume ratio of 1:(1-3); (2) quickly cooling the sericin solution to 4-10 DEG C; (3) centrifuging at 4-10 DEG C at the rate of 10000-15000 rpm for 30-60 minutes, and taking the supernate; (4) concentrating the supernate solution by a rotary evaporator; (5) after the solution is cooled, adding hydrolase, and reacting at 20-40 DEG C for 30-60 minutes; (6) centrifuging at the rate of 2000-3000 rpm for 5-10 minutes, and taking the supernate; (7) adding the supernate into a buffer solution with the pH value of 5.7-6.3, stirring, and centrifuging at the rate of 10000-15000 rpm for 30-60 minutes to obtain a precipitate; and (8) redissolving the precipitate in a buffer solution with the pH value of 6.4-7.2, and separating out the polypeptide with the molecular weight of 1600-3400Da by dialysis, thereby obtaining the sericin polypeptide 2. The invention also discloses application of the sericin polypeptide in cosmetic preparation. The purity of the prepared sericin polypeptide is greater than 70%. The experiment indicates that the polypeptide can perform the moisturizing function and has equivalent actions to the original sericin.

Description

technical field [0001] The invention relates to the field of biotechnology polypeptide preparation, in particular to the field of sericin polypeptide preparation. technical background [0002] Sericin is a sericin-like protein, which is a globulin. Accounting for about 20% to 30% of silk, sericin is composed of Ⅰ, Ⅱ, Ⅲ, Ⅳ proteins from outside to inside. The overall molecular conformation is mainly random coil, and the molecular spatial structure is loose and disordered, including β structure, but no α helical structure. There are many amino acids with long side chains on the sericin chain, such as arginine, lysine, glutamic acid, methionine, tryptophan, tyrosine, etc., as well as many polar hydrophilic groups (such as -OH, -COOH, -NH2, > NH, etc.) are on the surface of the polypeptide chain, these hydrophilic groups can make the moisture in the body transfer to the stratum corneum of the skin and combine, so that the skin maintains a certain amount of moisture, so t...

Claims

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Application Information

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IPC IPC(8): C07K7/08C12P21/06A61K8/64A61Q19/00
Inventor 罗瑞雪
Owner SUZHOU PULUODA BIOLOGICAL SCI & TECH
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