Method for determining isolated soy protein digestibility in vitro
A technique of soybean protein isolate and in vitro digestibility, which is applied in biochemical equipment and methods, microbiological determination/inspection, biological testing, etc., and can solve the problem of large activity variation range, no comparison reference, and unstable in vivo digestibility measurement results and other problems, to achieve the effect of easy control and management, simple and convenient process operation
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Embodiment 1
[0081] Take the isolated soybean protein samples of brands A and B respectively, prepare a 0.3% soybean protein solution with double distilled water, and adjust the pH to 1.5. The sample was placed in a constant temperature shaking shaker at 37° C., and a solution of pepsin (Sigma, P6887) was added to make the amount reach 1%, and digestion was started. After 30 minutes, adjust the pH to 6.0 to stop the reaction. Then raise the temperature to 40°C and adjust the pH to 7.8. Add fresh trypsin (sigma company, P1625) solution, make its dosage reach 3%, after continuing to digest for 60min, add a certain amount of Na 2 CO 3 The solution was dissolved in the digest, and the digest was immediately cooled in an ice bath to terminate the reaction. In vitro digestibility determination of soybean protein digested samples by trichloroacetic acid-soluble nitrogen method. Each sample was repeated 6 times.
Embodiment 2
[0082] Embodiment 2 (comparative example)
[0083] Weigh 1.00g of soybean protein isolate samples of brand A and B respectively, add 10mL of 0.01g / mL (pH value 1.5) pepsin solution, seal with paragel, hydrolyze in a shaker at 35°C for 4h, and use Adjust to neutral with NaOH. When treating with trypsin, first tie up one end of the prepared dialysis tube with a nylon thread until there is no drip. Then transfer the entire amount of the neutralized sample to the dialysis tube (about 40 mL of phosphate buffer solution), add 10 mL of 0.025 g / mL (pH value 7.6) of trypsin solution, the other end is also tied tightly with a nylon thread until there is no drip, and placed in a 1000mL wide-mouth Erlenmeyer flask. Add 300mL of phosphate buffer solution of dialysate, and seal with paragel. Hydrolyze on a shaker for 24 hours. Then the dialysate and the residue in the tube are quantitatively analyzed. Each sample was repeated 6 times.
[0084] Digestibility (%) = (N d / N s )×100%
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