An active polypeptide isolated from anchovy

An active polypeptide, anchovy technology, applied in the field of polypeptide separation, can solve problems such as underutilization of resources, and achieve the effects of increasing total antioxidant capacity, reducing nerve cell damage, and increasing SOD activity.

Active Publication Date: 2016-01-13
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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  • An active polypeptide isolated from anchovy
  • An active polypeptide isolated from anchovy
  • An active polypeptide isolated from anchovy

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Preparation of Anchovy Yeast Liquid

[0035] The by-products of the anchovy were minced with a mincer, and then coarsely filtered to remove impurities, and then heated at 80°C for 2 hours to a viscous semi-solid state to obtain an anchovy concentrate; the anchovy concentrate was subjected to (1:3, w / w) Add water in the proportion of ), mix well, add 3% flavor protease by weight of the anchovy concentrate, and hydrolyze at 60°C for 5 hours to obtain anchovy hydrolysate. The obtained anchovy enzymatic hydrolysate was added with 5% of the weight of the enzymatic hydrolysate of bran, sterilized at 121°C for 20 minutes, connected to Aspergillus oryzae, fermented at 30°C, and the fermentation period was 3 days. After the fermentation is finished, the obtained fermentation broth is filtered with gauze and sterilized at 100° C. for 20 min to obtain the finished product of anchovy yeast solution.

[0036] The physical and chemical indexes of the anchovy hair yeast liquid a...

Embodiment 2

[0041] Example 2 Active peptide G-15 gel chromatography

[0042] After the obtained anchovy yeast liquid is frozen into a dry powder with a freeze dryer, it is dissolved in ultrapure water to form a solution with a concentration of 0.1g / mL, passed through a 0.45μm inorganic water film, and applied to G-15 gel chromatography The column was separated, the loading volume was 1 mL, distilled water was used as the eluent, the flow rate was 1 mL / min, the detection wavelength was 220 nm, and the components with the highest anti-nerve cell damage activity were collected.

[0043] Such as 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 (P 1 Peak), 27~30min(P 2 Peak), 30~36min(P 3 Peak), the anti-neuronal damage activity of the three peaks was measured, and it was found that P 2 Peak has the highest activity against neuronal damage, collect P 2 Peaks, ready to be separated and puri...

Embodiment 3

[0044] Example 3 DEAE-SepharoseFF ion exchange column chromatography of active polypeptide

[0045] The P with the highest anti-nerve cell damage activity obtained in the previous step 2 The peak is further separated and purified by DEAE-SepharoseFF ion exchange column chromatography, and the concentrated sample is placed on a DEAE-SepharoseFF ion exchange column (18×80mm) equilibrated with 5mM Tris-HCl (pH8.5) buffer, with a concentration of 0~2mol / LNaCl gradient elution, the flow rate is 1mL / min, the detection wavelength is 220nm, the components with the highest anti-neuronal damage activity are collected, dialyzed with Tris-HCl solution, and freeze-dried. See the results of DEAE-SepharoseFF ion exchange column chromatography for active peptide figure 2 .

[0046] Such as figure 2 , P 2 After passing DEAE-SepharoseFF, three elution peaks are formed. The elution time is 3~6min (peak A), 13~20min (peak B), 32~37min (peak C). The three peaks are anti-neuronal cell damage The act...

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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, and specifically relates to an active polypeptide isolated from anchovy, which has the function of resisting nerve cell damage. Background technique [0002] Anchovy is 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 world’s largest fish species with the highest yield of a single fishery species. rich. Because anchovy is rich in fat, has high enzyme activity, and is easy to be oxidized and decayed, it cannot be stored for a long time. It is commonly known in my country as "Leshui Rotten". It is produced in the East my country Sea, the Yellow Sea and the Bohai Sea, with abundant resources, and the annual catch can reach several hundred thousand tons. For a long time, it has not been developed and utilized on a la...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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