Preparation method and application of Dendrobium officinale anti-tumor polypeptide
A Dendrobium officinale, anti-tumor technology, applied in the biological field
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Embodiment 1
[0031] The preparation method of Dendrobium officinale anti-tumor polypeptide, the steps are as follows:
[0032] (1) Weigh 100 g of Dendrobium officinale powder, grind it with liquid nitrogen, add 4 L of PBS and soak it for 20 h, then filter out the residue with gauze to obtain the PBS extract of Dendrobium officinale. Add 70% ammonium sulfate to the PBS extract, overnight, centrifuge (7000r / min) and remove the supernatant to obtain protein extract. Dissolve the protein extract with PBS, put it into a dialysis bag with a molecular weight cut-off range of 6000, dialyze for three days, and then freeze-dry to obtain protein powder;
[0033](2) Weigh 2g of protein powder and distilled water to make Dendrobium officinale protein liquid with a concentration of 3% by mass, and add Alcalase 2.4L (Novozymes), Alcalase 37017 (Novozymes) and trypsin (BioBasic Unit) respectively For hydrolysis, the hydrolysis conditions of alkaline protease Alcalase 2.4L were as follows: the enzyme-to-s...
Embodiment 2
[0037] The preparation method of Dendrobium officinale anti-tumor polypeptide, the steps are as follows:
[0038] (1) Weigh 120 g of Dendrobium officinale powder, grind it with liquid nitrogen, add 2 L of PBS and soak it for 30 h, and then filter out the residue with gauze to obtain the PBS extract of Dendrobium officinale. Add 60% ammonium sulfate to the PBS extract, overnight, centrifuge (9000r / min) and remove the supernatant to obtain protein extract. Dissolve the protein extract with PBS, put it into a dialysis bag with a molecular weight cut-off range of 5000, dialyze for three days, and then freeze-dry to obtain protein powder;
[0039] (2) Weigh 3g of protein powder and distilled water to make Dendrobium officinale protein solution with a concentration of 8% by mass, and add Alcalase 2.4L (Novozymes), alkaline protease 37017 (Novozymes) and trypsin (BioBasicUnit) for hydrolysis. The hydrolysis conditions of Alcalase 2.4L were as follows: the concentration ratio of enzy...
Embodiment 3
[0042] The preparation method of Dendrobium officinale anti-tumor polypeptide, the steps are as follows:
[0043] (1) Weigh 80 g of Dendrobium officinale powder, grind it with liquid nitrogen, add 1 L of PBS and soak it for 24 h, then filter out the residue with gauze to obtain the PBS extract of Dendrobium officinale. Add 70% ammonium sulfate to the PBS extract, overnight, centrifuge (7000 r / min) and remove the supernatant to obtain protein extract. Dissolve the protein extract with PBS, put it into a dialysis bag with a molecular weight cut-off range of 7000, dialyze for three days, and then freeze-dry to obtain protein powder;
[0044] (2) Weigh 4g of protein powder and distilled water to prepare Dendrobium officinale protein solution with a mass percentage concentration of 9%, and add Alcalase 2.4L (Novozymes), alkaline protease 37017 (Novozymes) and trypsin (BioBasicUnit) to hydrolyze. The enzymatic hydrolysis conditions of Alcalase 2.4L were as follows: the concentratio...
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