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.

Active Publication Date: 2019-01-01
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The free radicals produced by oxidation in the human body are related to human aging and many diseases. Antioxidants BHA and BHT commonly used in the food industry have a good effect of scavenging free radicals, but they themselves have toxic and side effects

Method used

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  • Synthetic polypeptide with antioxidant effect and gene encoding same, and preparation method and application of synthetic polypeptide
  • Synthetic polypeptide with antioxidant effect and gene encoding same, and preparation method and application of synthetic polypeptide
  • Synthetic polypeptide with antioxidant effect and gene encoding same, and preparation method and application of synthetic polypeptide

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Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses a synthetic polypeptide with antioxidant effect and a gene encoding same, and a preparation method and application of the synthetic polypeptide. According to the invention, a Fmoc solid phase synthesis method is employed; solid-phase synthetic resin is used as a carrier; amino acids are coupled with the solid-phase synthetic resin one by one from the C terminal to the N terminal of the polypeptide according to the arrangement manner of an amino acid sequence under the action of a mixed coupling agent so as to synthesize resin containing a target polypeptide fragment; then outer-layer resin is pyrolyzed and a Fmoc protecting group at a side chain is removed under the action of a pyrolysis removing agent; and the target polypeptide is cut off from the solid-phase carrier and is subjected to separation and purification so as to obtain the polypeptide having antioxidant effect. The synthesis method of the invention is simple in operation, short in synthesis time, low in the amount of by-products and low in synthesis cost. The synthetic polypeptide of the invention has remarkable anti-oxidation activity in vitro and good effect, can be used for preparing health food and medicines related to oxidation resistance and aging alleviation, is high in safety and wide in an application scope, and has good market prospects.

Description

technical field [0001] The invention belongs to the technical field of polypeptides, and in particular relates to a synthetic polypeptide with antioxidant effect, a gene encoding the polypeptide, a preparation method and application thereof. Background technique [0002] Polypeptides are formed by connecting amino acids through amide bonds. They can be used as signaling factors such as neurotransmitters and hormones in organisms, and conduct information transmission and information exchange by interacting with receptors. Peptides have the characteristics of specific recognition of target molecules, and targeted drug delivery systems using peptides as molecular probes are widely used in the fields of medicine and materials. In vitro synthesis of peptides is an important method for in-depth study of their functions, biochemical properties and applications. Compared with the biological preparation method, the chemical synthesis method has a shorter preparation period and can r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C12N15/11C07K1/04C07K1/20A61K38/08A61P39/06A23L33/18A23L3/3526
CPCA23L3/3526A23V2002/00A61K38/00A23L33/18A61P39/06C07K7/06A23V2200/302A23V2250/55Y02P20/55
Inventor 任娇艳李宇娟
Owner SOUTH CHINA UNIV OF TECH
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