Walnut meal anti-inflammatory peptide FPY and application thereof
A technology of anti-inflammatory peptides and anti-inflammatory drugs is applied in the field of anti-inflammatory peptides isolated from enzymatic hydrolysis products of walnut meal, which can solve the problems of lengthy, increased operating costs, environmental safety threats, etc.
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Embodiment 1
[0051] Example 1 Preparation, structure identification and molecular docking of walnut meal anti-inflammatory polypeptide
[0052] 1. Preparation of polypeptides with different molecular weights
[0053] Enzymatic hydrolysis of walnut dregs: Crushed walnut dregs, passed through a 200-mesh sieve, extracted with n-hexane at a ratio of 1:4 for 14 hours, then filtered, collected residues, and placed in a fume hood to volatilize the remaining organic solvents to obtain degreasing Walnut meal, stored at 4°C for later use.
[0054] Weigh a certain amount of defatted walnut meal, add 200mL distilled water with a solid-liquid ratio of 1:15 (w / v), adjust the pH value to 10.0 with 1mol / L NaOH solution, and perform ultrasonic treatment after magnetic stirring for 5min. The ultrasonic conditions are 50°C, 20min, after the end of ultrasonication, let stand in the room for 2h, centrifuge at a speed of 6000r / min for 15min, adjust the pH value of the supernatant to 4.5 with 1mol / L HCL, contin...
Embodiment 2
[0071] Example 2 Anti-inflammatory activity of walnut meal anti-inflammatory polypeptide
[0072] In order to further obtain anti-inflammatory active peptides, the top-ranked peptide FPY (Phe-Pro-Tyr, molecular weight 425.2Da) was artificially synthesized to verify its anti-inflammatory activity.
[0073] Polypeptide FPY was entrusted to Nanjing Yuanpeptide Biological Co., Ltd. to synthesize it through solid-phase synthesis, with a purity of >95%, which meets the requirements of related experiments for activity detection.
[0074] (1) Cytotoxicity
[0075] First, the safety of the polypeptide FPY was explored, such as Image 6 As shown, it was found that in the concentration range of 0-500 μM, the four polypeptides all had different degrees of promoting cell proliferation, that is, they had no toxicity ( Image 6 The first column is the blank group, the second column is the induction group, and there is no positive control group in the safety experiment).
[0076] Among the...
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