Synthetic polypeptide with antioxidant effect and gene encoding same, and preparation method and application of synthetic polypeptide
A technology for synthesizing peptides and anti-oxidation, applied in the field of peptides, can solve problems such as toxic and side effects, and achieve the effects of long shelf life, broad market development prospects and high safety.
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Embodiment 1
[0036] Embodiment 1, the Fmoc solid-phase synthesis of synthetic polypeptide:
[0037] In this test, the polypeptide was synthesized using a SYMPHONY 12-channel polypeptide synthesizer, using the Fmoc solid-phase synthesis strategy, using Fmco as the protecting group at the amino terminal of the amino acid, and using 2- CTC resin is used as a solid phase carrier resin, combined with 2-hydroxybenzotriazole, benzotriazole-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N-diisopropyl Base B's mixed coupling agent extends the polypeptide chain from the C-terminal to the N-terminal one by one. The specific operation scheme is as follows:
[0038] (1) Swelling of the resin: Add 1.5 g of 2-CTC resin to a clean peptide synthesis tube, stir with dichloromethane solution for 30 minutes to fully swell, and at the same time turn on nitrogen to protect it, and then use 10 mL N, N - Wash with dimethylformamide and drain.
[0039] (2) Amino acid connection: First, add 2mmol Fmoc-pro...
Embodiment 2
[0044] Embodiment 2, the structure and physicochemical property of synthetic polypeptide (PGACSN):
[0045] The molecular structural formula, molecular weight and LogP value of the synthetic polypeptide (PGACSN) are shown in Table 1. It can be seen from the table that the cysteine (Cys) of the synthetic polypeptide contains an -SH group, which can bind free radicals and remove them, thereby It has the effect of scavenging free radicals and anti-oxidation. The LogP value refers to the lipid-water partition coefficient of the compound. When LogP>0, the compound is a hydrophilic substance, and when LogP<0, the compound is a fat-soluble substance. Therefore, it can be seen from Table 1 that the synthetic polypeptide has good water solubility, which is conducive to the detection of its chemical properties, and is more convenient for the preparation of oral health-care functional foods and medicines.
[0046] Table 1 Molecular structural formula, molecular weight and LogP value o...
Embodiment 3
[0048] Embodiment 3, in vitro DPPH·scavenging activity assay of synthetic polypeptide:
[0049] DPPH·scavenging activity evaluation is a method for evaluating antioxidant capacity based on electron transfer mechanism. Use the 96 microplate reader detection plate to measure the change of the absorbance value of the system. Accurately absorb 100 μL of the samples to be tested at different concentrations into the well plate, and then add 100 μL DPPH solution to the well plate, and react in the dark for 30 minutes at room temperature. Determine the corresponding absorbance value, denoted as A Sample . Use 2mL distilled water and 2mL DPPH·solution as blank control, denoted as A control , 2mL distilled water and 95% ethanol mixed solution as a reference. Calculate DPPH·scavenging capacity according to the following formula:
[0050] DPPH·clearance capacity (%)=[(A control -A sample ) / A control ]×100%
[0051] like image 3 Shown is a graph of the in vitro DPPH·scavenging ac...
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