Method for separating and purifying soybean agglutinin from soybean whey
A technology for soybean lectin and soybean whey, which is applied in the preparation methods of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of low purification efficiency and purity of guar gum, high cost, limited large-scale use, and cumbersome and complicated procedures. and other problems, to achieve the effect of low equipment requirements, high recovery rate and high purity
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Embodiment 1
[0020] Adjust the soybean whey to pH 4.5, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 8.0 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.5, measure 1L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add sulfuric acid to the supernatant Ammonium to 60% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 0.65g of the crude soybean lectin protein contained in it, dissolved it in 10mL deionized water, dialyzed and desalted it for 36 hours with a molecular weight cut-off of 3500, and vacuumed the retentate Freeze-dry to prepare a P-7S protein sample containing soybean lectin and β-amylase.
[0021] Accurately weigh 0.1g of the above primary isola...
Embodiment 2
[0025] Adjust the soybean whey to pH 4.7, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 8.5 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.5, measure 2L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add sulfuric acid to the supernatant Ammonium to 70% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 1.85g of crude soybean lectin protein, dissolved in 30mL of deionized water, dialyzed and desalted for 48 hours with a molecular weight cut-off of 3500, and vacuumed the retentate Freeze-dry to prepare a P-7S protein sample containing soybean lectin and β-amylase.
[0026] Accurately weigh 0.3g of the above-mentioned primary isolated P-7S prot...
Embodiment 3
[0030] Adjust the soybean whey to pH 4.8, centrifuge at 9500rpm for 30min to remove the precipitate; adjust to pH 9.0 again, centrifuge at 9500rpm for 30min, discard the precipitate, and collect the supernatant. Adjust the pH of the above supernatant to 4.7, measure 1.5L of the solution, add solid ammonium sulfate to 50% saturation according to the ammonium sulfate saturation table at 4-30°C, centrifuge at 9500rpm for 30min, collect the precipitate, and continue to add the supernatant Ammonium sulfate to 80% saturation, centrifuged again at 9500rpm for 30min, collected the precipitate, combined the two precipitates to obtain about 1.57g of crude soybean lectin protein, dissolved in 30mL of deionized water, dialyzed and desalted for 48 hours with a molecular weight cut-off of 3500, and vacuum freeze-dried , to prepare a P-7S protein sample containing soybean lectin and β-amylase.
[0031] Accurately weigh 0.4g of the above-mentioned primary isolated P-7S protein sample and 0.4g...
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