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Sericin protein polypeptide and preparation and application thereof
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A technology of sericin and rubber powder, which is applied to peptide sources, animal/human peptides, cosmetic preparations, etc.
Inactive Publication Date: 2015-09-30
SUZHOU PULUODA BIOLOGICAL SCI & TECH
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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
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Embodiment 1
[0013] 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 pH5.0-6.0, stir and centrifuge at 10000rpm for 60 minutes to obtain the precipitate ; Redissolve the precipitate in pH 6.0-7.0 buffer, and use dialysis to separate the polypeptide with a molecular weight of 1600-3400Da. The sequence of the polypeptide was determined as SVISRAWDYVDDTDKSIAIL by a solid-phase automatic protein sequencer, and the purity was 85.3%.
Embodiment 2
[0015] 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 pH5.0-6.0, stir, and centrifuge at 15000rpm for 30 minutes to obtain the precipitate ; Redissolve the precipitate in pH 6.0-7.0 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 SVISRAWDYVDDTDKSIAIL, and the purity was 73.0%.
Embodiment 3
[0017] 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 solution increased to 34.89, 24.43, 32.65, 54.98% respectively.
[0018] 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 us...
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Abstract
The invention discloses a sericin protein polypeptide, and relates to the field of polypeptide preparation in biotechnology. The sequence of the sericin protein polypeptide is SVISRAWDYVDDTDKSIAIL. The sericin protein polypeptide is prepared according to the following steps: (1) taking 500 g of sericin powder to be added into deionized water to be dissolved according to the mass / volume ratio of the 1:(1-3); (2) quick-cooling a sericin solution to be 4-10 DEG C; (3) centrifuging at 4-10 DEG C and at 10000-15000 rpm for 30-60 min, and taking a supernatant; (4) adopting a rotary evaporimeter to concentrate the supernatant; (5) after the solution is cooled, adding hydrolase, and reacting at 20-40 DEG C for 30-60 min; (6) centrifuging at 2000-3000 rpm for 5-10 min and taking a supernatant; (7) adding the supernatant into a buffer solution with the pH of 5.0-6.0, stirring, and centrifuging at 10000-15000 rpm for 30-60 min to obtain sediment; (8) redissolving the sediment into a buffer solution with the pH of 6.0-7.0, and adopting a dialysis method to separate the polypeptide of which the molecular weight is 1600-3400 Da. The sericin protein polypeptide is applied to cosmetic preparation; the purity of the prepared sericin protein polypeptide is greater than 70%; experiments show that the polypeptide has the function of moisture preservation, and is equivalent to the original sericin protein in function.
Description
technical field [0001] The invention relates to the field of biotechnology polypeptide preparation, in particular to the field of sericin polypeptide preparation. [0002] technical background [0003] 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 mo...
Claims
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Application Information
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