Separation and purification method of silkworm pupa protein polypeptide
A technology for the separation and purification of silkworm chrysalis protein polypeptides, which is applied in the field of polypeptide extraction, can solve the problems of poor separation and purification effect of silkworm chrysalis polypeptide 2, and achieve good separation effects, fast and efficient separation, and high resolution effects
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Embodiment 1
[0017] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:
[0018] 1) Preparation of silkworm chrysalis protein polypeptide crude solution: Dissolve the silkworm chrysalis protein enzymatic hydrolyzate in double distilled water according to the ratio of material to liquid 1:10 (g / mL), and then centrifuge in a centrifuge at a temperature of 5 °C and a speed of 8000 rpm for 10 min , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is ready for use. The components in the silkworm chrysalis protein crude polypeptide liquid are relatively complex, because their molecular weight and amino acid sequence are different, so they have different physical properties and biological activities. For more in-depth...
Embodiment 2
[0025] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:
[0026] 1) Preparation of silkworm chrysalis protein polypeptide crude liquid: according to the material-to-liquid ratio of 1:20 (g / mL), the enzymatic hydrolyzate of silkworm chrysalis protein was dissolved in double distilled water, and then centrifuged in a centrifuge at a temperature of 1°C and a speed of 10,000 rpm for 8 minutes , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is set aside;
[0027] 2) Ion-exchange resin chromatography: The silkworm chrysalis protein peptide liquid was prepared into a solution with a concentration of 55 mg / mL with double distilled water, and then centrifuged in a centrifuge at a temperature of 1°C a...
Embodiment 3
[0033] The method for separating and purifying silkworm chrysalis protein polypeptides comprises preparation of silkworm chrysalis protein polypeptide crude liquid, ion exchange resin chromatography, gel column chromatography, and high performance liquid chromatography purification, and its specific steps are:
[0034] 1) Preparation of silkworm chrysalis protein polypeptide crude solution: Dissolve silkworm chrysalis protein enzymatic hydrolyzate in double distilled water according to the ratio of material to liquid 1:15 (g / mL), and then centrifuge in a centrifuge at 4°C and 9000rpm for 9min , the supernatant is the crude silkworm chrysalis protein polypeptide liquid, which is set aside;
[0035] 2) Ion-exchange resin chromatography: the silkworm chrysalis protein peptide solution was prepared with double distilled water to a concentration of 50mg / mL, and then centrifuged in a centrifuge at a temperature of 4°C and a speed of 9000rpm for 9min to remove insoluble impurities, an...
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