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Active polypeptide separated from anchovy

An active peptide and anchovy technology, applied in the field of peptide separation, can solve the problem of underutilization of resources and achieve the effects of increasing total antioxidant capacity, improving survival rate, and reducing nerve cell damage

Active Publication Date: 2014-07-23
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, as an important marine protein resource, anchovy and its processing by-products are very different from terrestrial biological protein resources in amino acid composition and sequence, and are far greater than terrestrial and freshwater protein resources in terms of variety and quantity. However, these resources are not fully utilized

Method used

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  • Active polypeptide separated from anchovy
  • Active polypeptide separated from anchovy
  • Active polypeptide separated from anchovy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] The preparation of embodiment 1 anchovy hair yeast liquid

[0035]Grind the by-products of anchovy with a mincer, perform coarse filtration to remove impurities, and then heat at 80°C for 2 hours to obtain a viscous semi-solid state to obtain an anchovy concentrate; the anchovy concentrate is prepared according to (1:3, w / w ) ratio into water, mixed evenly, added flavor protease with 3% weight of anchovy concentrate, and enzymolyzed at 60° C. for 5 hours to obtain anchovy enzymatic hydrolyzate. Add 5% bran by weight of the enzymatic hydrolyzate to the obtained anchovy hydrolyzate, sterilize at 121°C for 20 minutes, insert Aspergillus oryzae, ferment at 30°C, and the fermentation period is 3 days. After the fermentation is finished, the obtained fermented liquid is filtered with gauze, and sterilized at 100°C for 20 minutes to obtain the finished anchovy fermented yeast liquid.

[0036] The physical and chemical indicators of anchovy fermented yeast liquid and enzymatic...

Embodiment 2

[0041] Embodiment 2 active peptide G-15 gel chromatography

[0042] Freeze the finished anchovy yeast liquid into dry powder with a freeze dryer, dissolve it in ultrapure water to make a solution with a concentration of 0.1g / mL, pass through a 0.45μm inorganic water membrane, and perform G-15 gel chromatography The column was used for separation, the sample volume was 1 mL, distilled water was used as the eluent, the flow rate was 1 mL / min, and the detection wavelength was 220 nm to collect the components with the highest anti-nerve cell damage activity.

[0043] like figure 1 As shown, the sample is divided into three elution peaks after passing through the G-15 gel chromatography column, and the elution time is 16.5~18.5min respectively (P 1 peak), 27~30min(P 2 peak), 30~36min (P 3 Peak), the anti-neurocyte injury activity of the three peaks was determined, and it was found that P 2 The peak has the highest anti-nerve cell injury activity, and the collection of P 2 The ...

Embodiment 3

[0044] DEAE-Sepharose FF ion-exchange column chromatography of embodiment 3 active polypeptide

[0045] P 2 The peak was further separated and purified by DEAE-Sepharose FF ion-exchange column chromatography, and the DEAE-Sepharose FF ion-exchange column (18 × 80mm) equilibrated with 5mM Tris-HCl (pH8.5) buffer solution on the concentrated sample was used at a concentration of 0-2mol / L NaCl gradient elution, flow rate 1mL / min, detection wavelength 220nm, collect the components with the highest anti-nerve cell damage activity, dialyze with Tris-HCl solution, and freeze-dry. Active polypeptide DEAE-Sepharose FF ion exchange column chromatography results see figure 2 .

[0046] like figure 2 , P 2 After passing through DEAE-Sepharose FF, three elution peaks were formed, and the elution time was 3-6min (peak A), 13-20min (peak B), and 32-37min (peak C). The damage activity was measured, and it was found that peak A had the highest anti-nerve cell damage activity. A large am...

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Abstract

The invention aims to provide an active polypeptide separated from an anchovy fermentation liquid. The molecular weight of the nerve cell injury resistant polypeptide obtained by the invention is 2,181Da; and the polypeptide is composed of 11 amino acids such as aspartic acid, threonine, glutamic acid, glycine, alanine, isoleucine, tyrosine, phenylalanine, histidine, lysine, arginine and the like, and is a novel active polypeptide with activity. According to the polypeptide obtained by the invention, in a glutamine nerve cell injury model, the addition of the active polypeptide provided by the invention can obviously increase the survival rate of PC12 nerve cells; and when the final concentration is 50 mug / mL, the cell survival rate is increased by 26.4% in comparison with an injury group, thereby effectively relieving glutamine-induced nerve cell injury. Besides, the cell mechanism study on the nerve cell protective action of the polypeptide discovers that the addition of the active polypeptide obviously increases the intracellular total oxidation resistance and has a certain promoting action on increase of SOD (superoxide dismutase) activity.

Description

technical field [0001] The invention belongs to the technical field of polypeptide separation, in particular to an active polypeptide separated from anchovies, which has the function of resisting nerve cell damage. Background technique [0002] Anchovies are widely distributed in the temperate waters of the northern and southern hemispheres such as the Pacific Northwest, New Zealand, Australia, the Indian Ocean, the east coast of Africa, and the coast of South Africa. It is the fish species with the highest yield of a single fishery species in the world. Rich. Because anchovy is rich in fat, has high enzyme activity, and is easily oxidized and rotten, it cannot be stored for a long time. It is produced in the East my country Sea, Yellow Sea and Bohai Sea of ​​China, and is rich in resources. The annual catch can reach hundreds of thousands of tons. For a long time, due to insufficient understanding of its utilization value, it has not been developed and utilized on a large...

Claims

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Application Information

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IPC IPC(8): C12P21/06C12P21/02C07K14/46C07K1/18C07K1/16A61K38/17A61P25/00C12R1/69
Inventor 毛相朝于小航薛长湖
Owner OCEAN UNIV OF CHINA
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