A kind of preparation method of phycoerythrin ACE inhibitory peptide

A technology of phycoerythrin and inhibitory peptides, which is applied in the field of preparation of phycoerythrin ACE inhibitory peptides, can solve the problems of loss of activity, complicated purification steps, unfavorable large-scale production, etc., and achieve reduced enzyme amount, low molecular weight, and technical parameters Effective and feasible effect

Active Publication Date: 2017-02-22
JIMEI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation process involves the combined use of multiple biotechnologies such as ultrafiltration, gel filtration, and ion exchange, and the purification steps are complicated, which is not conducive to large-scale production
[0007] In addition, the biologically active peptide prepared by the above invention is ingested by the human body, and after being digested by the gastrointestinal tract, it is likely to be degraded twice and lose its activity

Method used

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  • A kind of preparation method of phycoerythrin ACE inhibitory peptide
  • A kind of preparation method of phycoerythrin ACE inhibitory peptide
  • A kind of preparation method of phycoerythrin ACE inhibitory peptide

Examples

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Embodiment 1

[0029] 1) Extraction of phycoerythrin: take 200 g red algae as raw material, wash and dry at 40 ℃, and crush. Add 20 times the volume of distilled water to make a seaweed suspension, and freeze and thaw three times at -20 °C and 4 °C to break the cells. After 20 min of homogenization, the tissue was sonicated for 10 min. After centrifugation, take the supernatant and carry out 35~50% ammonium sulfate salting out. The salted-out precipitate was dialyzed and loaded onto a DEAE-Sepharose anion exchange column, and was washed with 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under weak light conditions 2 PO 4 Solution (containing 200 mmol / L NaCl) was mixed for linear elution. The eluted fractions with A565 / A280> 3.0 were collected, freeze-dried to make phycoerythrin powder, and stored away from light. Phycoerythrin solutions with different concentrations were prepared to measure the ACE inhibitory activity. like figure 1 As shown, the ACE inhibitory ac...

Embodiment 2

[0034] 1) Extraction of phycoerythrin: 200 g laver was used as raw material, washed and dried at 40 °C, and crushed. Add 30 times the volume of distilled water to make a seaweed suspension, and freeze and thaw five times at -20 °C and 4 °C to break the cells. After 20 min of homogenization, the tissue was sonicated for 20 min. After centrifugation, take the supernatant and carry out 35~50% ammonium sulfate salting out. The salted-out precipitate was dialyzed and loaded onto a DEAE-Sepharose anion exchange column, and was washed with 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under weak light conditions 2 PO 4 The solution (containing 200 mmol / L NaCl) was mixed for linear elution at a flow rate of 1 mL / min. The eluted fractions with A565 / A280> 3.0 were collected, freeze-dried to make phycoerythrin powder, and stored away from light. Prepare different concentrations of phycoerythrin solutions to measure ACE inhibitory activity;

[0035] 2) Enzymatic...

Embodiment 3

[0038] 1) Extraction of phycoerythrin: 1 kg of red hair algae was used as raw material, washed and dried at 40 ℃, and pulverized. Add 20 times the volume of distilled water to make a seaweed suspension, and freeze and thaw five times at -20 °C and 4 °C to break the cells. After 40 min of homogenization, the tissue was sonicated for 30 min. After centrifugation, take the supernatant and carry out 35~50% ammonium sulfate salting out. The salted-out precipitate was dialyzed and loaded onto a DEAE-Sepharose anion exchange column, and was washed with 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under weak light conditions 2 PO 4 The solution (containing 200 mmol / L NaCl) was mixed for linear elution at a flow rate of 1 mL / min. The eluted fractions with A565 / A280> 3.0 were collected, freeze-dried to make phycoerythrin powder, and stored away from light. Prepare different concentrations of phycoerythrin solutions to measure ACE inhibitory activity;

[0039]...

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Abstract

The invention discloses a preparation method for a phycoerythrin ACE inhibitory peptide. The method includes the steps of: extracting phycoerythrin; adding pepsin to conduct enzymolysis and then adding trypsin to perform enzymolysis; dissolving the freeze-dried powder subjected to enzymolysis in pure water, loading the obtained solution to a SephadexG-15 gel column, collecting the highest activity peak as a phycoerythrin ACE inhibitory peptide component, and further loading the obtained component to a high performance liquid chromatogram ZORBAX300SB-C18 to perform separation so as to detect phycoerythrin ACE inhibitory peptide fragments in the component and can realize preparation of high purity phycoerythrin ACE inhibitory peptide at the same time. The method provided by the invention employs pepsin and trypsin stepwise enzymolysis to prepare the ACE inhibitory peptide, can avoid inactivation of the active peptide due to degradation by gastrointestinal digestive fluid after intake by the human body, also reduces the cost, has a simple process, and can realize industrial production. The phycoerythrin ACE inhibitory peptide derives from natural vegetable protein, has a small molecular weight, is stable, safe, and easy to absorb by the human body.

Description

technical field [0001] The invention relates to a protein preparation method, in particular to a preparation method of phycoerythrin ACE inhibitory peptide. Background technique [0002] CN 200610097201 extracts oat protein from oats, hydrolyzes it with alkaline protease, and separates it through ion exchange chromatography, gel filtration chromatography and reversed-phase high performance liquid chromatography to prepare oat protein ACE inhibitory peptide. The patent application adopts ion exchange chromatography for separation, and the sample needs to be desalted before passing through the column, and a large amount of salt will be brought in during elution, which needs to be desalted again later, so the operation cost is high. [0003] CN 200310113446 prepares casein from fresh milk, and adds 3.5-6% protease to degrade the casein to obtain a milk-derived ACE inhibitory peptide. The preparation process requires a large amount of protease, which increases the production co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/36C07K14/405C12P21/06C07K1/30C07K1/18C07K1/16
Inventor 曹敏杰伍强蔡秋凤付晓苹翁凌刘光明
Owner JIMEI UNIV
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