Carrot seed antioxidant tripeptide as well as preparation method and application thereof
A carrot seed and oxidized tripeptide technology, applied in the field of health food, can solve the problems of carrot seed protein and active peptide exploration, and achieve the effects of high stability, strong acid resistance and strong alkali resistance
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Embodiment 1
[0020] The separation and purification of the antioxidant tripeptide in the present invention includes two steps of SephadexG-25 gel filtration chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC).
[0021] Preparation of enzymatic hydrolyzate of carrot seed protein: Carrot seed protein was obtained by extraction and acid precipitation method, and acidic protease was used to hydrolyze it under the optimal enzymatic hydrolysis conditions (enzyme addition 419.36U / g, substrate concentration 5.28% (w / v)) The enzymatic hydrolysis time is 3.50 hours, inactivated in a boiling water bath for 10 minutes, centrifuged at 12000 rpm for 20 minutes, and the supernatant is freeze-dried, which is the enzymatic hydrolyzate of leek seed protein;
[0022] SephadexG-25 gel filtration chromatography: redissolve the lyophilized powder of carrot seed protein hydrolyzate in deionized water, centrifuge at 12000rpm for 15min, and remove the supernatant. The SephadexG-25 ge...
Embodiment 2
[0027] The antioxidant tripeptide obtained in Example 1 was subjected to solid-phase synthesis, and the activity of the synthesized polypeptide was studied:
[0028] ABTS free radical scavenging effect: prepare 7mmol / L ABTS with distilled water + solution and 2.45mmol / L potassium persulfate solution (must be placed at room temperature for 16h before use), ABTS + Mix with potassium persulfate solution at a volume ratio of 1:1, and dilute the mixture to the absorbance value A with pH 7.4, 5 mmol / L phosphate buffer solution before use 734 is 0.70±0.02. Take 0.5mL of samples with different concentrations and 0.5mL of ABTS free radical solution, mix and let stand for 10min, then measure the absorbance value at 734nm. Deionized water and reduced glutathione were used instead of samples as blank control and positive control, respectively. ABTS free radical scavenging activity is calculated according to the following formula (1):
[0029]
[0030] In the formula, A 0 : Absorba...
Embodiment 3
[0033] Stability of the antioxidant tripeptide of the present invention:
[0034] Carrot seed antioxidant peptide YSL was heat-treated at 20°C-100°C for 1h-5h, and the ABTS free radical scavenging rate was measured every 1h. Such as Figure 5, it was determined that the free radical scavenging rate of YSL had no significant change after being treated at 20°C, 40°C and 60°C for 5 hours (p>0.05). The carrot seed antioxidant peptide YSL was treated with different pH deionized aqueous solutions (2.0, 4.0, 6.0, 8.0 and 10.0) for 1h~5h, and the ABTS free radical scavenging rate was measured every 1h. It has been determined that the tripeptide YSL has strong alkali resistance and strong acid resistance. Although the activity decreases under acidic conditions, its structure is not destroyed and can still maintain activity ( Image 6 ).
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