Preparation method of regenerated-silk fibroin polypeptides
A protein polypeptide and regenerated silk technology, which is applied in the field of regenerated silk protein polypeptide preparation, can solve the problems of low strength, waste, and inability to weave silk, achieve stable structural properties, high raw material utilization, improve enzymatic hydrolysis efficiency and enzyme solution effect
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Embodiment 1
[0015] A method for preparing a regenerated silk protein polypeptide, the preparation method comprising:
[0016] 1) Pretreatment: Take 60 parts of regenerated silk and wash them, cut them into pieces with a shredder, and then add Na with a concentration of 4mol / L according to the mass ratio of 1:6 2 CO 3 Solution, mix well, stand still, get the pretreatment liquid. The preparation method has the advantages of simple operation, high reduction degree, high utilization rate of raw materials, low preparation cost, and the prepared protein polypeptide has excellent moisturizing and sun protection functions, low sensitization rate, and wide application range;
[0017] 2) Freezing and thawing: Take the pretreatment solution, freeze it at 73°C for 60 minutes, thaw it at 5°C, repeat the freezing and thawing process at least once, and then use a homogenizer to homogenize it for 10 minutes to obtain a homogenate solution. Repeated freezing and thawing can accelerate the cell rupture o...
Embodiment 2
[0025] A method for preparing regenerated silk protein polypeptides. The molecular weight of the prepared protein polypeptides is 2800 Da. The preparation method comprises: taking 55 parts of regenerated silk and washing them, cutting them into pieces with a shredder, and then adding them at a concentration of 3 mol according to the mass ratio of 1:3.7. / LNa 2 CO 3solution, mixed evenly, and stood still to obtain a pretreatment solution; take the pretreatment solution, freeze it at -68°C for 53 minutes, thaw it at 4°C, freeze and thaw it repeatedly for 3 minutes, and then use a homogenizer to homogenize it for 8 minutes to obtain a homogenized solution. Slurry; take the homogenate, add 2300U / g pepsin to hydrolyze for 20min, then add 2-hydroxyl-1,2,2-triphenylethyl acetate with a weight of 1.5‰ of pepsin, and continue to react for 14min. Frequency composite ultrasound and microwave radiation alternately assisted enzymolysis. After the enzymolysis was completed, the boiling wat...
Embodiment 3
[0028] A method for preparing a regenerated silk protein polypeptide, the preparation method comprising: taking 40 parts of regenerated silk and washing them, cutting them into pieces with a shredder, and then adding Na 2 CO 3 solution, mixed evenly, and stood still to obtain the pretreatment solution; take the pretreatment solution, freeze it at -55°C for 45 minutes, thaw it at 3°C, repeat the freeze-thaw at least once, and then use a homogenizer to homogenize it for 4 minutes to obtain Homogenate: Take the homogenate, add 2000U / g pepsin to hydrolyze it for 15min, then add 2-hydroxyl-1,2,2-triphenylethyl acetate with pepsin weight of 0.5‰, continue to react for 10min, during the enzymolysis process with Double-frequency composite ultrasound and microwave radiation alternately assisted enzymolysis. After enzymolysis was completed, boiling water bath inactivated for 10 minutes, and the supernatant was taken to obtain enzymolysis solution. The operation of dual-frequency composi...
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