Antioxidant peptide in perilla seeds and application thereof
An antioxidant peptide, perilla seed technology, applied in the fields of peptides, antitoxins, organic chemistry, etc., can solve the problems of less research on protein development and utilization, and no report on active peptide pharmacology research.
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Embodiment 1
[0026] The separation and purification of the antioxidant peptide of the present invention includes two steps of Sephadex G-25 gel filtration chromatography and reverse phase high performance liquid chromatography (RP-HPLC).
[0027] Preparation of perilla seed crude protein enzymatic hydrolysis product: first degrease perilla seed with petroleum ether, then use alkali extraction and acid precipitation method to obtain perilla seed crude protein, use alkaline protease to enzymatically hydrolyze perilla seed crude protein, use alkali Protease was hydrolyzed for 5.00h under the optimal hydrolysis conditions (pH 9.80, enzyme dosage 3000U / g, substrate concentration 3% (w / v)), inactivated in boiling water bath for 10min, centrifuged at 12000rpm for 10min, and taken The supernatant is freeze-dried, which is the enzymatic hydrolyzate of perilla seed protein;
[0028] Sephadex G-25 Gel Filtration Chromatography: Dissolve the freeze-dried powder of perilla seed enzymatic hydrolyzate in...
Embodiment 2
[0033] The activity of the natural antioxidant peptide obtained in Example 1 is studied:
[0034] ABTS free radical scavenging effect: prepare 7 mmol / L ABTS with distilled water + solution and 2.45 mmol / L potassium persulfate solution (must be placed at room temperature for 16 h 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 0.5 mL of samples with different concentrations and 0.5 mL of ABTS radical solution, mix and let stand for 10 min, then measure the absorbance at 734 nm. 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):
[0035] (1)
[0036] In the formula, A 0 : Absorbance value of blank control group; A s : Absorbance value of the s...
Embodiment 3
[0053] The natural antioxidant peptide obtained in embodiment 1 is carried out cytotoxicity and antioxidation research:
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