Antitumor active peptide and its preparation method and application
A technology of anti-tumor activity and active peptides, applied in the field of anti-tumor active peptide substances and their preparation, can solve the problems of long incubation period, hidden disease, and difficulty in early diagnosis, and achieve the effects of low cost, wide application range, and simple process
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
preparation example Construction
[0022] The present invention also provides the preparation method of the anti-tumor active peptide, which is a method of enzymatically hydrolyzing pea protein, comprising the following steps: 1) thermal denaturation treatment of pea protein aqueous solution at 70°C-90°C for 10min-30min , to obtain a denatured pea protein aqueous solution; 2) enzymatically hydrolyze the denatured pea protein aqueous solution obtained in step 1) to obtain a pea protein hydrolyzate; 3) ultrafiltration fractionate the pea protein hydrolyzate obtained in step 2), and collect them separately Components with a molecular weight greater than or equal to 60KDa, [6-60) KDa, [1-6) KDa, and less than 1KDa; 4) Detect the anti-tumor activity of the above-mentioned collected components, and screen for components with anti-tumor activity; 5) The components with anti-tumor activity obtained in step 4) are subjected to molecular sieve chromatography, and each chromatographic peak component is collected according ...
Embodiment 1
[0040] Alkaline protease hydrolyzes pea protein to prepare anti-tumor active peptide, the steps are as follows:
[0041] (1) Preparation of proteolysis solution: Accurately weigh 6g of pea protein powder, add it into 100mL deionized water and mix well, heat treat pea protein at 70°C for 15min, add alkaline protease according to the enzyme-to-substrate ratio of 10000U / g, adjust The pH value was 8.5. After reacting in a water bath at 55°C for 5 hours, the enzyme was inactivated in a boiling water bath at 90°C for 10 minutes, and the degree of hydrolysis was determined to be 18.18%;
[0042] (2) Freeze-drying the pea protein hydrolyzate obtained after hydrolysis, freeze-drying conditions: temperature -50°C; time: 12h. The dry powder is dissolved in the cell culture medium DMEM, and the cell culture medium containing pea peptide is prepared to measure the inhibitory effect on the growth of cancer cells;
[0043] (3) The pea protein hydrolyzate obtained by enzymatic hydrolysis wit...
Embodiment 2
[0059] The MTT method is used to detect the inhibitory rate of anti-tumor active peptides on human cancer cells. The steps are as follows:
[0060] The anti-tumor active peptide is added to the cell culture medium DMEM at a certain concentration to detect the inhibitory effect of the anti-tumor active peptide on the growth of cancer cells in vitro. The operation steps are as follows:
[0061] a. Digest the cells in the logarithmic phase into a single cell suspension with trypsin. After counting, adjust the cell density to 5×10 4 cells / mL, inoculated into 96-well plate at 100 mL per well, and cultivated in 37°C, 5% CO2 incubator for 48 hours;
[0062] b. Add 200 μL of cell culture solution containing the corresponding concentration of anti-tumor active peptide to each well of the cells in the experimental group, add 200 μL of LDMEM culture solution to the blank control group, and use the culture solution containing the same concentration of pea protein powder as the negative co...
PUM
| Property | Measurement | Unit |
|---|---|---|
| degree of substitution | aaaaa | aaaaa |
Abstract
Description
Claims
Application Information
Login to View More