A kind of antioxidant peptide derived from casein and its application
A technology for antioxidant peptides and casein, applied in peptide/protein components, medical preparations containing active ingredients, peptides, etc., can solve problems affecting physiological activity and antioxidant peptides
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Embodiment 1
[0066] Casein antioxidant peptides were prepared by hydrolyzing casein with alkaline protease.
[0067] (1) Weigh 30 g of casein, immerse in 600 mL of deionized water with a pH of 8.0, preheat in a 60°C water bath for 10 min, and prepare a casein sample solution;
[0068] (2) Then add 0.6g of alkaline protease with an enzyme activity of 2.4U / g, and put it in a water bath at 60°C for 4 hours; h. At 4.0 h, the pH of the hydrolyzate was adjusted to 8.0 with 1M NaOH aqueous solution; a hydrolyzate rich in antioxidant peptides was obtained.
[0069] (3) After the hydrolysis ends, the hydrolyzed solution prepared in step (2) is heated in a boiling water bath at 100° C. for 15 minutes to passivate the enzyme, and after being cooled to room temperature, its pH is adjusted to 7.0 by using 1M hydrochloric acid solution; The final casein hydrolyzate was vacuum concentrated to a dry matter content of 40%.
[0070] (4) Dissolve the concentrated solution in step (3) in 1 mL of acetic acid...
Embodiment 2
[0072] Casein antioxidant peptides were prepared by hydrolyzing casein with papain.
[0073] (1) Weigh 30 g of casein, immerse in 450 mL of distilled water with a pH of 7.0, preheat in a water bath at 50°C for 10 min, and prepare a casein sample solution;
[0074] (2) Next, 0.6 g of papain was added, and put into a water bath at 50° C. for 3 hours for hydrolysis; a hydrolyzate rich in antioxidant peptides was obtained.
[0075] (3) After the hydrolysis ends, the hydrolyzed solution prepared in step (2) is heated in a boiling water bath at 100° C. for 15 minutes to passivate the enzyme, and after being cooled to room temperature, its pH is adjusted to 7.0 by using 1M hydrochloric acid solution; The final casein hydrolyzate was vacuum concentrated to a dry matter content of 30%.
[0076] (4) Dissolve the concentrated solution in step (3) in a Tris-HCl buffer solution with a pH of 8.3, filter through a 0.45 μm water filter, and separate on a DEAE-Sephadex A-25 ion exchange colum...
Embodiment 3
[0078] Casein antioxidant peptides were prepared by hydrolyzing casein with trypsin.
[0079] (1) Weigh 60 g of casein, immerse in 600 mL of pure water with a pH of 7.5, preheat in a 37°C water bath for 10 min, and prepare a casein sample solution;
[0080] (2) Next, 1.2 g of trypsin was added, and hydrolyzed in a water bath at 37°C for 3 hours; a hydrolyzate rich in antioxidant peptides was obtained.
[0081] (3) After the hydrolysis ends, the hydrolyzed solution prepared in step (2) is heated in a boiling water bath at 85°C for 25min, and after being cooled to the passivation room temperature of the enzyme, its pH is adjusted to 7.0 with 1M hydrochloric acid solution; The final casein hydrolyzate was vacuum concentrated to a dry matter content of 30%.
[0082] (4) Dissolve the concentrated solution in step (3) in 2 mL of acetic acid buffer solution with a concentration of 20 mM and a pH of 4.0. After fully dissolving, filter it with a 0.45 μm water filter membrane, and take...
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