Penaeus japonicus anti-oxidative peptide and preparation method thereof
A technology of antioxidant peptide and bamboo shrimp, which is applied in the biological field to achieve strong antioxidant activity, increase yield, and increase hydrolysis rate
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Embodiment 1
[0039] Example 1: Preparation of bamboo shrimp antioxidant peptide
[0040] 1.1 Raw materials and reagents
[0041] Raw materials: bamboo shrimp head, purchased from Zhoushan Haisheng Biological Co., Ltd.
[0042] Reagents: trypsin, bromelain, pepsin, alkaline protease and neutral protease, all of which were purchased from Guangbo Nanning Pangbo Bioengineering Co., Ltd.; Gly, Gly-Gly-Gly, vitamin B12, aprotinin, cytochrome C, bovine serum albumin, acetonitrile (chromatographic grade), trifluoroacetic acid (chromatographic grade), formic acid (chromatographic grade), chromatographic column packing SephadexG-25 (U.S. Sigma company); absolute ethanol, 2,6-di-tert-butyl Base-4-methylphenol (BHT), hydrochloric acid, sodium hydroxide and other reagents were of analytical grade (Sinopharm Chemical Reagent Co., Ltd.).
[0043] 1.2 Instruments and equipment
[0044]WATERS SYNAPT ultra-high pressure liquid chromatography quadrupole tandem time-of-flight mass spectrometer (Waters, USA...
Embodiment 2
[0099] Example 2: Structural identification of antioxidant peptides
[0100] The anti-oxidant component SHP-D after separation and purification by reversed-phase high-performance liquid phase was identified by UPLC-TOF-MS / MS technology. Figure 17 shown. Depend on Figure 17 It can be seen that this component presents a single peak, so it can be considered that the peptide is relatively pure. The peak was analyzed by secondary mass spectrometry, and its structure was analyzed by peptidesequencing in Masslynx software. The results showed that the identification of the peptide from the SHP-D A polypeptide sequence with a reliability of more than 90% was obtained, and its structure was Gly-Asn-Gly-Leu-Pro (such as Figure 18 shown).
Embodiment 3
[0101] Example 3: Determination of biological stability of bamboo shrimp antioxidant peptide
[0102] Prepare artificial gastric juice and artificial intestinal juice for later use. The preparation process of artificial gastric juice is as follows: Measure 16.4mL of dilute hydrochloric acid and 800mL of distilled water respectively, mix them, then add 10g of pepsin, shake evenly, place in a volumetric flask, and constant volume to 1000mL. Preparation of artificial intestinal juice: Weigh 6.8g of potassium dihydrogen phosphate, dissolve it in 500mL of distilled water, adjust the pH value to 6.8 with 0.1mol / L NaOH solution, and take 10g of trypsin, dissolve it in a certain Amount of distilled water, shake evenly, and set the volume to 1000mL.
[0103] Prepare chicken ovalbumin solution and bamboo shrimp antioxidant peptide solution with the same concentration (10mg / ml), mix them in a ratio of (1:4, 1:5, 1:6), and then add the same amount of spare Artificial gastric juice or in...
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