Preparation method and application of malva seed active protein
A technology of active protein and mallow seed, applied in the field of preparation and application of plant protein, can solve the problems of no anti-tumor active protein of mallow seed, few and few reports of mallow seed, and achieve good anti-colon cancer cell activity Effect
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Embodiment 1
[0021] (1) Take 50g of traditional Chinese medicine mallow seeds, grind and pulverize them, add 250mL of pre-cooled protein extract (PBS buffer solution with pH 7.4, 1L of buffer solution contains 8g NaCI, 0.2g KCI, 3.63g NaCI 2 HPO4.12H 2 O, 0.24g KH 2 PO 4 ). Soak and stir in a refrigerator at 4°C for 12-16 hours, centrifuge at 11,000 rpm for 10 minutes at 4°C, and take the supernatant, which is the crude extract of mallow seed protein. Add 250mL protein extract to the filter residue after centrifugation again, operate as above, and combine the supernatant to be the crude protein extract of mallow seeds.
[0022] (2) Slowly add 500 mL of -20°C pre-cooled acetone to the crude mallow seed protein extract obtained in (1), stir while adding, and place it in a 4°C refrigerator for 1 hour to precipitate. Centrifuge at 11,000 rpm at 4°C for 10 minutes, discard the supernatant, collect the precipitate, and place the precipitate at -20°C for 30 minutes to completely evaporate the...
Embodiment 2
[0026] (1) Take 50g of traditional Chinese medicine mallow seeds, after grinding and pulverizing, add 200mL pre-cooled protein extract (PBS buffer solution with pH 7.4, 1L buffer solution contains 8g NaCI, 0.2g KCI, 3.63g NaCl 2 HPO4.12H 2 O, 0.24g KH 2 PO 4 ). Soak and stir in a refrigerator at 4°C for 12-16 hours, centrifuge at 11,000 rpm for 10 minutes at 4°C, and take the supernatant, which is the crude extract of mallow seed protein. Add 200mL of protein extract to the centrifuged filter residue again, operate as above, and combine the supernatants to obtain the crude protein extract of mallow seeds.
[0027] (2) Slowly add 400 mL of -20°C pre-cooled acetone to the crude mallow seed protein extract obtained in (1), stir while adding, and place it in a 4°C refrigerator for 1 hour to precipitate. Centrifuge at 11,000 rpm at 4°C for 10 minutes, discard the supernatant, collect the precipitate, and place the precipitate at -20°C for 30 minutes to completely evaporate the ...
Embodiment 3
[0031] (1) Take 50g of traditional Chinese medicine mallow seeds, grind and pulverize them, add 300mL of pre-cooled protein extract (PBS buffer solution with pH 7.4, 1L of buffer solution contains 8g NaCI, 0.2g KCI, 3.63g NaCl 2 HPO4.12H 2 O, 0.24g KH 2 PO 4 ). Soak and stir in a refrigerator at 4°C for 12-16 hours, centrifuge at 11,000 rpm for 10 minutes at 4°C, and take the supernatant, which is the crude extract of mallow seed protein. Add 300mL protein extract to the filter residue after centrifugation again, operate as above, and combine the supernatant to be the crude protein extract of mallow seeds.
[0032] (2) Slowly add 600 mL of -20°C pre-cooled acetone to the crude mallow seed protein extract obtained in (1), stir while adding, and place it in a 4°C refrigerator for 1 hour to precipitate. Centrifuge at 11,000 rpm at 4°C for 10 minutes, discard the supernatant, collect the precipitate, and place the precipitate at -20°C for 30 minutes to completely evaporate the...
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