A kind of crocodile antioxidant peptide complex and its preparation method and application

A technology of antioxidant peptides and complexes, applied in applications, cosmetic preparations, peptides, etc., can solve problems such as use restrictions, toxic and side effects, and achieve the effects of increasing added value, good thermal stability, and maintaining antioxidant activity

Active Publication Date: 2022-04-05
福建鼍龙实业有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, chemical antioxidants are the most produced, but their use is limited due to their obvious toxic and side effects

Method used

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  • A kind of crocodile antioxidant peptide complex and its preparation method and application
  • A kind of crocodile antioxidant peptide complex and its preparation method and application
  • A kind of crocodile antioxidant peptide complex and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] The preparation of crocodile antioxidant peptide complex, the steps are as follows:

[0110] After washing with 100g of ultrapure water and drying with filter paper, add 15-20 times the volume of ultrapure water (1500mL-2000mL), and cook at high temperature for 25min-35min. Using papain, under its optimal enzymolysis conditions (55°C, enzyme addition 3.00% (w / w), substrate concentration 6.5% (w / v)) enzymolysis 4h, boiling water bath inactivation 15min, and then Filter with 4 layers of gauze to obtain the enzymatic hydrolyzate of crocodile antioxidant peptide complex. The above enzymolysis solution was centrifuged at 12,000 rpm for 30 minutes at room temperature, and the supernatant was collected and freeze-dried to obtain crocodile antioxidant peptide complex powder.

Embodiment 2

[0112] Separation and purification of the crocodile antioxidant peptide complex obtained in Example 1

[0113] Ultrafiltration: Redissolve the crocodile antioxidant peptide complex powder in ultrapure water, and then pass through ultrafiltration membranes with molecular weight cut-offs of 100kDa, 5kDa, and 2kDa for classification, and determine the hydrolyzate of 100kDa hydrolyzate four components ABTS + · Free radical scavenging rate. The result is as Figure 5 As shown, it can be seen that the component crocodile antioxidant peptide-1 has the highest clearance rate, so the component is collected, freeze-dried, and subjected to the next step of anion exchange chromatography separation and purification.

[0114] Anion exchange chromatography: Capto TM Q-pillar (HiTrap TM , 5mL) fully equilibrated with 20mM Tris-HCl (pH9.0) buffer in advance; After mixing, use a 2mL syringe to load the sample, and the sample volume is 2.0mL; The buffer is eluted in steps. Elution profi...

Embodiment 3

[0122] Research on the antioxidant activity of the crocodile antioxidant peptide complex obtained in Example 1.

[0123] 3.1. The ability to scavenge DPPH free radicals

[0124] The antioxidant capacity in vitro was studied by measuring the free radical scavenging rate of 1,1-diphenyl-2-trinitrophenylhydrazyl (1-diphenyl-2-picrylhydrazyl, DPPH). Take 0.1ml of different concentrations of crocodile antioxidant peptide complex (0mg / mL, 0.5mg / mL, 1.0mg / mL, 1.5mg / mL, 2.0mg / mL, 2.5mg / mL, 3.0mg / mL), add 0.1 ml 0.2mM DPPH·solution, mix well, stand in the dark for 30min, then measure the absorbance at 517nm ultraviolet wavelength. The results are shown in Table 2 below:

[0125] DPPH free radical scavenging rate calculation formula:

[0126]

[0127] Among them, A c Absorbance value of 0.1mL ultrapure water + 0.1mL DPPH solution; A i It is the absorbance value of 0.1mL sample + 0.1mL DPPH solution; A j It is the absorbance value of 0.1mL ultrapure water + 0.1mL absolute ethano...

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Abstract

The present application discloses a crocodile antioxidant peptide complex, which includes: crocodile antioxidant peptide 1, crocodile antioxidant peptide 2, crocodile antioxidant peptide 3, crocodile antioxidant peptide 4 and crocodile antioxidant peptide 5; wherein, according to the mass fraction of the crocodile antioxidant peptide complex, the crocodile antioxidant peptide 1, crocodile antioxidant peptide 2, crocodile antioxidant peptide 3, crocodile antioxidant peptide 4 and crocodile antioxidant peptide 5 0.1%-2%, 0.5%-3%, 0.1%-2%, 0.1%-2% and 0.5%-5%, the molecular weight of the crocodile antioxidant peptide complex is in the range of 800Da-1500Da Inside. The crocodile antioxidant peptide complex can effectively scavenge DPPH free radicals, ABTS + · Free radicals and hydroxyl groups (OH ‑ ) free radicals; the crocodile antioxidant peptide complex has no toxic effect on Lo2 and HaCat cells, and has a repair function on HaCat cells induced by hydrogen peroxide.

Description

technical field [0001] The present application relates to the field of biological products, in particular to a crocodile antioxidant peptide complex and its preparation method and application. Background technique [0002] Crocodile (Crocodile) is a vertebrate amphibian reptile with a life history of more than 200 million years. It is one of the oldest and most primitive animals discovered so far, and is known as a "living fossil". Crocodiles are full of treasures, with high medicinal value, food value and economic value. For example, crocodile meat is rich in protein, and can reduce phlegm and relieve cough, nourish the heart and lungs; crocodile bone can be used to prevent and treat rheumatic bone pain, senile osteoporosis and children's rickets; crocodile gall has antipyretic, calming heat, and promotes metabolism It also has a significant effect on the treatment of fatty liver and cirrhosis; eating crocodile liver can detoxify eyesight, nourish blood and benefit the liv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K7/06
CPCC07K7/06C07K7/08A23L33/18A61K8/64A61Q19/08A23V2002/00A23V2200/318A23V2250/55
Inventor 张紫然叶沁陈清西李华亮熊佑熊
Owner 福建鼍龙实业有限责任公司
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