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Antioxidant peptide prepared from shark prolamin and preparation method and application thereof

A gliadin, antioxidant peptide technology, applied in the application, food preparation, peptides and other directions, can solve the problems of unreported shark gliadin peptide, application limitation, unsafe food safety of chemical antioxidant additives, etc. Monitoring, high activity, scientific and reasonable effect

Inactive Publication Date: 2014-01-22
ZHEJIANG OCEAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] In recent years, due to the unsafety of chemical antioxidant additives and food safety considerations, chemical antioxidant additives such as BHA (butyl hydroxyanisole), BHT (2,6-di-tert-hydroxy p-cresol) and TBHQ (tertiary butyl Hydroquinone) and other applications have been greatly restricted
However, there is no report on animal gliadin
[0005] The applicant found in experiments that scalloped hammerhead shark meat contains a large amount of gliadin, and the enzymatic hydrolyzate of gliadin has good antioxidant activity, but literature search found that shark meat was used as raw material to extract and prepare gliadin Protein, the preparation of prolamin antioxidant peptides by enzymatic hydrolysis and chromatographic techniques is in a blank stage, and the application of shark gliadin peptides in scavenging free radicals and inhibiting lipid peroxidation has not been reported.

Method used

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  • Antioxidant peptide prepared from shark prolamin and preparation method and application thereof
  • Antioxidant peptide prepared from shark prolamin and preparation method and application thereof
  • Antioxidant peptide prepared from shark prolamin and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] 1) Remove the skin and bones of the scalloped hammerhead shark, add 90% ethanol solution to the fish meat at a solid-to-liquid ratio of 1g:25mL, use a high-speed tissue masher to make a homogenate, and extract dynamically at 45°C for 90 minutes ;

[0043] 2) Adjust the pH value of the extract to 9.0 with 0.1mol / L NaOH, and adjust the ethanol concentration to 60% with distilled water. The diluted extract was placed at 25°C for 30min, centrifuged at 4500rpm for 20min, and the supernatant was collected;

[0044] 3) Add NaCl to the supernatant to a concentration of 5%, salt out for 12 hours, centrifuge at 4500 rpm for 15 minutes, and collect the precipitate;

[0045] 4) The precipitate was degreased with ether for 24 hours, and then centrifuged at 11,000 rpm for 12 minutes to remove the ether to obtain gliadin, which was freeze-dried to obtain a powder and stored at -20°C for later use.

[0046] 5) Using shark gliadin as a raw material, add phosphate buffer (pH5.8) accord...

Embodiment 2

[0057] 1) Remove the skin and bones of the scalloped hammerhead shark, add 85% ethanol solution to the fish meat at a solid-to-liquid ratio of 1g:20mL, use a high-speed tissue masher to make a homogenate, and extract dynamically at 30°C for 100min ;

[0058] 2) Adjust the pH value of the extract to 10.0 with 0.1mol / L NaOH, and adjust the ethanol concentration to 55% with distilled water. The diluted extract was placed at 25°C for 35min, centrifuged at 4000rpm for 30min, and the supernatant was collected;

[0059] 3) Add NaCl to the supernatant to a concentration of 7%, and after salting out for 10 hours, centrifuge at 4000 rpm for 20 minutes to collect the precipitate;

[0060] 4) Degrease the precipitate with ether for 20 hours, then centrifuge at 10,000 rpm for 15 minutes to remove the ether to obtain gliadin, freeze-dry to obtain powder, and store at -20°C for later use.

[0061] 5) Using shark gliadin as a raw material, add phosphate buffer (pH5.8) according to the solid...

Embodiment 3

[0065] 1) Remove the skin and bones of the scalloped hammerhead shark, add 95% ethanol solution to the fish meat at a solid-to-liquid ratio of 1g:30mL, use a high-speed tissue masher to make a homogenate, and extract dynamically at 50°C for 80min ;

[0066] 2) Adjust the pH value of the extract to 10.0 with 0.1mol / L NaOH, and adjust the ethanol concentration to 65% with distilled water. The diluted extract was placed at 25°C for 40 minutes, centrifuged at 5000 rpm for 20 minutes, and the supernatant was collected;

[0067] 3) Add NaCl to the supernatant to a concentration of 8%, and after salting out for 15 hours, centrifuge at 5000 rpm for 10 minutes to collect the precipitate;

[0068] 4) Degrease the precipitate with ether for 20 hours, and then centrifuge at 12,000 rpm for 10 minutes to remove the ether to obtain gliadin, freeze-dry to obtain powder, and store at -20°C for later use.

[0069] 5) Using shark gliadin as a raw material, add phosphate buffer (pH5.8) accordin...

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Abstract

The invention discloses an antioxidant peptide prepared from shark prolamin and a preparation method and application thereof. The amino acid sequence of the antioxidant peptide is Trp-Asp-Arg or / and Pro-Tyr-Phe-Asn-Lys; according to the ESI-MS detection, a molecular ion peak is m / z 476.40[M+H]<+> or / and m / z 668.60[M+H]<+>; when in preparation, the shark meat is used as a raw material, and the shark prolamin is extracted by use of ethanol; then, papain is used as an enzymolysis enzyme, and the antioxidant peptide is prepared by the enzymolysis technology in combination with macroporous resin desalination, ultrafiltration classification and chromatography refining. The preparation method disclosed by the invention is scientific and reasonable in process, the enzymolysis process is easy to monitor, and the prepared antioxidant peptide has relatively high activity; compared with a chemosynthetic antioxidant, the antioxidant peptide disclosed by the invention has the advantages of safety, no toxic or side effect, high antioxidant activity, easiness in digestion absorption and the like, and can be used as a medicine, a healthcare food, a food additive and the like.

Description

technical field [0001] The present invention relates to an antioxidant peptide prepared from shark gliadin and its preparation method and application. It is a method of extracting shark gliadin with ethanol and preparing alcohol with high antioxidant activity by using enzyme engineering technology and chromatographic technology. Methods of solubilizing peptides. Background technique [0002] In recent years, due to the unsafety of chemical antioxidant additives and food safety considerations, chemical antioxidant additives such as BHA (butyl hydroxyanisole), BHT (2,6-di-tert-hydroxy p-cresol) and TBHQ (tertiary butyl The application of base hydroquinone) has been greatly restricted. Therefore, the development of high-efficiency, low-toxic natural food antioxidants has become a research hotspot. [0003] At present, substances with remarkable antioxidant activity have been extracted from various plants and animal tissues, such as tea polyphenols in tea, isoflavones in soybe...

Claims

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Application Information

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IPC IPC(8): C07K5/097C07K7/06C07K1/20C07K1/18C07K1/16C12P21/06A61P39/06A23L1/29A23L1/305A23L33/18
Inventor 王斌罗红宇李忠瑞栗丽
Owner ZHEJIANG OCEAN UNIV
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