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Preparation method for 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 problems such as loss of activity, and achieve the effects of reducing the amount of enzyme added, reducing costs, and simple operation

Active Publication Date: 2014-11-05
JIMEI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] In addition, the biologically active peptide prepared by the above invention is ingested by the human body

Method used

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  • Preparation method for phycoerythrin ACE inhibitory peptide
  • Preparation method for phycoerythrin ACE inhibitory peptide
  • Preparation method for phycoerythrin ACE inhibitory peptide

Examples

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Example Embodiment

[0028] Example 1:

[0029] 1) Phycoerythrin extraction: 200 g of red hair algae is used as raw material, washed and dried at 40 ℃, and crushed. Add 20 times the volume of distilled water to make a seaweed suspension, freeze and thaw it repeatedly at -20 ℃ and 4 ℃ 3 times to break the cells. After 20 minutes of homogenization, the tissues were sonicated for 10 minutes. After centrifugation, the supernatant was taken and subjected to 35-50% ammonium sulfate salting out. After salting out and dialysis, load the sample on DEAE-Sepharose anion exchange column, and use 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under low light conditions 2 PO 4 The solution (containing 200 mmol / L NaCl) is mixed for linear elution. Collect A565 / A280> The eluted fraction of 3.0 was freeze-dried to make phycoerythrin powder, and stored in the dark. Prepare phycoerythrin solutions of different concentrations to determine ACE inhibitory activity. Such as figure 1 As shown, ac...

Example Embodiment

[0033] Example 2:

[0034] 1) Phycoerythrin extraction: 200 g laver is used as raw material, washed and dried at 40 ℃ and crushed. Add 30 times the volume of distilled water to make a seaweed suspension, freeze and thaw it repeatedly at -20 ℃ and 4 ℃ 5 times to break the cells. After homogenizing the tissue for 20 minutes, it was sonicated for 20 minutes. After centrifugation, the supernatant was taken and subjected to 35-50% ammonium sulfate salting out. After salting out and dialysis, load the sample on DEAE-Sepharose anion exchange column, and use 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under low 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. Collect A565 / A280> The eluted fraction of 3.0 was freeze-dried to make phycoerythrin powder, and stored in the dark. Prepare phycoerythrin solutions of different concentrations to determine ACE inhibitory activity;

[0035] 2) Enzym...

Example Embodiment

[0037] Example 3:

[0038] 1) Phycoerythrin extraction: 1 kg of red hair algae is used as raw material, washed and dried at 40 ℃ and crushed. Add 20 times the volume of distilled water to make a seaweed suspension, and freeze and thaw it repeatedly at -20 ℃ and 4 ℃ for 5 times to break the cells. After 40 minutes of homogenization, the tissues were sonicated for 30 minutes. After centrifugation, the supernatant was taken and subjected to 35-50% ammonium sulfate salting out. After salting out and dialysis, load the sample on DEAE-Sepharose anion exchange column, and use 20 mmol / L PBS (pH 5.6, containing 50 mmol / L NaCl) and 20 mmol / L NaH under low 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. Collect A565 / A280> The eluted fraction of 3.0 was freeze-dried to make phycoerythrin powder, and stored in the dark. Prepare phycoerythrin solutions of different concentrations to determine ACE inhibitory activity...

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