A kind of walnut meal antioxidant peptide faw and its preparation method and application
An anti-oxidation peptide and anti-oxidation technology, applied in the field of food deep processing, can solve the problems of waste of walnut protein resources and hinder the development of walnut industry, etc., and achieve the effect of obvious anti-oxidation, low production cost and scavenging free radicals
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Embodiment 1
[0023] Embodiment 1: the extraction method of walnut meal polypeptide
[0024] (1) Degreasing of walnut meal:
[0025] The walnut meal was pulverized and passed through a 40-mesh sieve, added with petroleum ether at a ratio of 1:5 (w / v) to extract for 2 hours, filtered, the residue was collected, extracted three times continuously, and the residue was placed in a fume hood to evaporate the organic solvent. The walnut meal defatted powder was obtained, which was stored at 4°C for later use.
[0026] (2) Alkali-soluble acid precipitation to extract walnut protein
[0027] Accurately weigh 10g of walnut meal defatted powder into a 400mL beaker, add 200mL of distilled water, adjust the pH to 10.0, stir on a magnetic stirrer for 5min, then ultrasonicate for 20min, take it out immediately after the ultrasonication, let stand for 1h, 6000r / min Centrifuge for 20 min and take the supernatant for later use. Adjust pH to 4.5 with 1 mol / L HCl, stand for 1 h, centrifuge at 6000 r / min fo...
Embodiment 2
[0034] Example 2 ABTS free radical scavenging ability of different molecular weight walnut meal polypeptides after ultrafiltration
[0035] Weigh 5 mg of lyophilized powder and 1 mg of vitamin C of the walnut meal polypeptide solution with molecular weight >10KDa, 5-10KDa, 3-5KDa, and below 3KDa obtained after ultrafiltration in Example 1, respectively, and dissolve them in 5mL of water. , prepared into a polypeptide with a concentration of 1 mg / mL and a vitamin C solution with a concentration of 0.2 mg / mL.
[0036] Divided into 5 treatments:
[0037] Treatment 1: 0.1mL of polypeptide solution with molecular weight >10KDa;
[0038]Treatment 2: 0.1 mL of a polypeptide solution with a molecular weight of 5-10 KDa;
[0039] Treatment 3: 0.1 mL of a polypeptide solution with a molecular weight of 3-5KDa;
[0040] Treatment 4: 0.1 mL of a solution of a polypeptide whose molecular weight is below 3KDa;
[0041] Treatment 5: The positive control group was 0.1 mL of vitamin C solu...
Embodiment 3
[0048] Example 3 Sephadex chromatography on small molecule polypeptides of 0-3KDa
[0049] Sephadex G-25 was used to separate the ultrafiltration small molecule polypeptides of 0-3KDa. The separation conditions were as follows: the loading volume was 5 mg / mL, 5 mL, the size of the chromatographic column was 1.6 × 60 cm, the flow rate was 1 mL / min, distilled water was used as the eluent, and the UV detector was used for detection with a wavelength of 220 nm.
[0050] The ultra-filtered 0-3KDa small molecule polypeptide was subjected to Sephadex G-25, and three peaks of polypeptides were obtained: No. 1 peak F1, No. 2 peak F2 and No. 3 peak F3, the results are as follows figure 2 As shown, the ABTS free radical scavenging rate was measured, and the results were as follows image 3 shown. The ABTS free radical scavenging rate of component F2 was higher than that of component F1 and component F3 (P50 It can be seen that the activity of the purified fraction F2 is higher.
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