Antioxidant active peptide and preparation method thereof

An anti-oxidative active peptide and alkaline technology, applied in the field of anti-oxidative active peptide of marine low-value fish blue round trevally protein and its preparation, to achieve the effect of simple preparation method, cheap source and strong applicability

Active Publication Date: 2013-05-15
GUANGXI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These are only related to the enzymatic hydrolysis processing technology or the antioxidant activity and stability determination of the product, etc. The preparation method of separating and purifying a single antioxidant active peptide from the blue round trevally protein has not been reported yet.

Method used

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  • Antioxidant active peptide and preparation method thereof
  • Antioxidant active peptide and preparation method thereof
  • Antioxidant active peptide and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Mix the protein powder of blue trevally with water to make a reaction raw material solution with a protein content of 1wt%, then add 0.03wt% alkaline protease, hydrolyze at 50°C and pH 8.0 for 3 hours, and finally heat to 100°C to stop enzymatic reaction. Centrifuge the obtained mixture at 4°C and 8000r / min for 10-15min, take the supernatant of enzymatic hydrolysis and use an ultrafiltration membrane with a molecular weight of 5000Da to separate and collect the permeate, and use deionized water as the mobile phase to coagulate with Sephedex G-15. Gel chromatography separates the ultrafiltration permeate, and detects and measures the antioxidant activity of the eluent corresponding to each peak at 280nm with a protein and nucleic acid detector, and collects the No. 2 absorption peak with high activity (see figure 1 ) was further separated by reverse-phase HPLC, using an ODS column (Hedera ODS-C2, 10 × 250mm), adopting water and acetonitrile (5-60% acetonitrile in 55min) ...

Embodiment 2

[0028] Mix the protein powder of blue scorpion fish meat with water to make a reaction raw material solution with a protein content of 3wt%; then add 0.005wt% alkaline protease, hydrolyze at 55°C and pH 10 for 6 hours, and finally heat to 100°C to stop enzymatic reaction. Centrifuge the obtained mixture at 4°C and 8000r / min for 10-15min, take the supernatant of enzymatic hydrolysis and use an ultrafiltration membrane with a molecular weight of 5000Da to separate and collect the permeate, and use deionized water as the mobile phase to coagulate with Sephedex G-15. Gel chromatography separates the ultrafiltration permeate, and detects and measures the antioxidant activity of the eluent corresponding to each peak at 280nm with a protein and nucleic acid detector, and collects the No. 2 absorption peak with high activity (see figure 1 ) was further separated by reverse phase HPLC, using ODS column (Hedera ODS-C2, 10 × 250mm), adopting water and acetonitrile (0-60% acetonitrile wit...

Embodiment 3

[0030] Mix the protein powder of blue scorpion fish meat with water to make a reaction material solution with a protein content of 5wt%; then add 0.05wt% alkaline protease, hydrolyze at 45°C and pH 8.0 for 4.5 hours, and finally heat to 100°C to stop enzymatic reaction. Centrifuge the obtained mixture at 4°C and 8000r / min for 10-15min, take the supernatant of enzymatic hydrolysis and use an ultrafiltration membrane with a molecular weight of 5000Da to separate and collect the permeate, and use deionized water as the mobile phase to coagulate with Sephedex G-15. Gel chromatography separates the ultrafiltration permeate, and detects and measures the antioxidant activity of the eluent corresponding to each peak at 280nm with a protein and nucleic acid detector, and collects the No. 2 absorption peak with high activity (see figure 1 ) was further separated by reverse-phase HPLC, using an ODS column (Hedera ODS-C2, 10 × 250mm), adopting water and acetonitrile (5-55% acetonitrile wi...

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Abstract

The invention discloses an antioxidant active peptide and a preparation method thereof. The antioxidant active peptide is characterized by taking decapterus fish protein as the raw material, alkaline protease generated by bacillus subtilis 2709 is adopted for hydrolysis, after enzyme deactivation of the hydrolysate, the hydrolysate is separated through centrifugation, ultra-filtration, gel chromatography and reverse phase high performance liquid chromatography so as to determine the structures of two natural antioxidant peptide, the amino acid sequences are His-Asp-His-Pro-Val-Cys (histidine-aspartic acid-histidine-proline-valine-cysteine) and His-Glu-Lys-Val-Cys(histidine-glutamic acid-lysine-valine-cysteine), the concentration (EC50) for eliminating DPPH (1,1-Diphenyl-2-picrylhydrazyl) radical of 50% are 0.0310+ / -0.0011mu M and 0.0677+ / -0.0012mu M, the concentration for eliminating superoxide anion (O2-) radical of 50% are 0.3814+ / -0.0017mu M and 0.3744+ / -0.0021mu M, and the antioxidant active peptide has good antioxidant activity.

Description

technical field [0001] The invention belongs to the field of functional foods, and in particular relates to an antioxidative activity peptide of marine low-value fish, blue round trevally, meat protein and a preparation method thereof. Background technique [0002] Chemosynthetic antioxidants such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT), which are often used as food additives, are restricted in use due to their toxicity and side effects. Natural antioxidants can not only prevent the oxidation of the oil system in food, but also prevent the damage of free radicals in the human body, delay aging and various chronic degenerative diseases such as cancer, coronary heart disease, diabetes, etc. Peptides, which are between proteins and amino acids, have been widely studied due to their special structural characteristics, high safety and significant biological activity. In recent years, people have isolated a large number of natural antioxidant peptides...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/06C07K7/06C07K1/36C07K1/34C07K1/20C07K1/16
Inventor 廖丹葵蒋海萍孙建华韦藤幼童张法赵钟兴
Owner GUANGXI UNIV
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