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Antioxidant peptide AOP-OM1 as well as preparation method and application thereof

An AOP-OM1, antioxidant peptide technology, applied in the field of medicine, can solve the problems of weak research on effector material basis and mechanism of action

Inactive Publication Date: 2015-04-22
YUNNAN MINZU UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the particularity of the source of medicinal materials, the research on the material basis and mechanism of its effect is still relatively weak, and there are almost no reports on its active ingredients.

Method used

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  • Antioxidant peptide AOP-OM1 as well as preparation method and application thereof
  • Antioxidant peptide AOP-OM1 as well as preparation method and application thereof
  • Antioxidant peptide AOP-OM1 as well as preparation method and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0022] The preparation method of the antioxidant polypeptide AOP-OM1 of the present invention includes the following steps:

[0023] A. Take 100mg of raw material big green smelly frog skin secretion, add 1ml of deionized water to dissolve it, load on Sephadex G-75 column, elution with Tris-HCl buffer at a flow rate of 1.5ml / min, collect once every 10min, respectively Measure the absorbance of the eluate at 280 nm, collect the eluate combined with active peak components, and concentrate and dry to obtain extract a;

[0024] B. Take 1 mg of extract a, add 1ml of deionized water to dissolve it, load it on a Hypersil ODS2 chromatographic column, elution with acetonitrile at 1ml / min, monitor the absorbance at 215nm, collect the eluent of combined active peak components, and concentrate After drying, the target is obtained.

[0025] The Sephadex G-75 column described in step A needs to be pre-equilibrated with 20 mmol / L, pH 7.8 Tris-HCl buffer for 24 hours.

[0026] The Hypersil ODS2 colu...

Embodiment 1

[0033] Example 1 Separation, purification and identification of the anti-oxidant peptide AOP-AM1 from the skin of a big green smelly frog

[0034] 1. Separation and purification

[0035] The big green smelly frog was collected from Zunyi, Guizhou, and was stimulated with 6V direct current for 6 seconds, then washed its skin with normal saline to obtain skin secretions, vacuum freeze-dried, and stored at -80° for later use.

[0036] The first step: Sephadex G-75 exclusion chromatography: Obtain the freeze-dried powder of skin secretions according to the above method, and dissolve it in deionized water. Take 1 ml (protein content of 100 mg) with 20 mM Tris-HCl buffer in advance (PH=7.8, containing 0.1 M NaCl) Equilibrate a Sephadex G-75 (GE Healthcare, ultrafine) column (length 30 cm, inner diameter width 1.5 cm) equilibrated for 24 hours, eluting with the same buffer at a flow rate of 1.5 ml / 10min, collect once every 10 min, measure its absorbance at 280 nm, the obtained separation ...

Embodiment 2

[0045] The antioxidant activity test of the anti-oxidant peptide AOP-AM1 from the skin of the big green smelly frog prepared in Example 1:

[0046] Methods: The free radical ABTS+ scavenging activity test was used to detect the antioxidant activity of the skin antioxidant peptide AOP-AM1 of the big green smelly frog. The results showed that the skin antioxidant peptide AOP-AM1 of the big green smelly frog had strong antioxidant activity.

[0047] Specific steps and results: Mix 7 mM ABTS in water and 2.8 mM potassium persulfate, and react for 6 hours in a dark environment at room temperature to make ABTS oxidized to ABTS + Free radicals, then diluted 50 times with deionized water, take 50 uL diluent and 50 uL sample mix, react for 30 minutes in the dark at room temperature, and then measure its absorbance at 415 nm wavelength. Free radical ABTS + The removal rate (%) is calculated with the following formula: (A blank -A sample ) ×100 / A blank . The antioxidant activity of the b...

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Abstract

The invention discloses an antioxidant peptide AOP-OM1 as well as a preparation method and an application thereof. The antioxidant peptide AOP-OM1 has amino acid sequences as shown in SEQ ID No. 1. The preparation method comprises the following steps of separating and purifying skin secretions of odorrana livida. A novel antioxidant is found in the skin of an amphibian odorrana livida, the chemical nature of the antioxidant is polypeptide, and the antioxidant, different from traditional two antioxidants, is encoded by a gene and but has very small molecular weight. The novel antioxidant peptide AOP-OM1, which is obtained from the skin of the odorrana livida growing in China, has very strong antioxidant activity, which is about 200 times of that of vitamin E.

Description

Technical field [0001] The invention belongs to the technical field of medicine, and specifically relates to an antioxidant polypeptide AOP-OM1 and a preparation method and application thereof. Background technique [0002] Antioxidants are substances that prevent the adverse effects of oxygen. They are a class of substances that can help capture and neutralize free radicals, thereby eliminating the damage of free radicals to the human body. A large number of antioxidants have been discovered. These antioxidants can be divided into the following two categories: 1) Non-gene-coded small molecules are helpless, such as vitamin E, vitamin C, glutathione, etc.; 2) genetically encoded Macromolecular substances, such as superoxide dismutase, catalase, thioredoxin, etc. These antioxidants have been widely used in various fields, including health, food, cosmetics, and various industries. For example, they can be used as stabilizers for fuels and lubricants to prevent oxidation, and they ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K7/06C07K1/16A61K38/08A61P39/06
Inventor 王滢梁醒财
Owner YUNNAN MINZU UNIV
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