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A kind of preparation method of microcapsule-embedded laver blood pressure lowering peptide

A blood pressure-lowering peptide and microcapsule technology are applied in the field of preparation of laver blood-pressure-lowering peptides, which can solve the problems of affecting vitality sustainability and the like, and achieve the effects of improving vitality sustainability, simple process and improving enzyme activity.

Inactive Publication Date: 2011-12-28
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After the laver hypotensive peptide is extracted, it is prone to oxidation and other conversion processes that affect biological activity when it contacts with air, seriously affecting the sustainability of vitality

Method used

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  • A kind of preparation method of microcapsule-embedded laver blood pressure lowering peptide
  • A kind of preparation method of microcapsule-embedded laver blood pressure lowering peptide
  • A kind of preparation method of microcapsule-embedded laver blood pressure lowering peptide

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preparation example Construction

[0035] Preparation of Yeast Display Polygalacturonase

[0036] Synthesize the polygalacturonase gene (Genbank No.: HQ446162.1) and the cell wall α-lectin gene of Pichia pastoris GS115 (Genbank No.: M28164) by artificial synthesis. At the same time, polygalacturonase gene C The connecting peptide sequence GSSGGSGGSGGSGGSGS(linker) is added to the end, and the nucleotide sequence PGA-linker-α-agglutinin is obtained after connection, and EcoR I and Not I enzyme cutting sites are added at both ends of the sequence, wherein PGA is polygalacturonic acid Enzyme gene, α-agglutinin is cell wall α lectin gene.

[0037] Using the above artificially synthesized sequence as a template, PCR amplification was performed using the following primer pair,

[0038] Upstream primer: 5'-ATGTTGTTATCGACACACAGTATTGTTCTG-3';

[0039] Downstream primer: 5'-TTTTCCTTTTGCGGCCGCTAATGAAACG-3'

[0040] The PCR reaction system is: 1 μl of template DNA, 0.5 μl of high-fidelity DNA polymerase, 0.4 μl of dNTP ...

Embodiment 1

[0053] Example 1 Preparation of Laver Hypotensive Peptide Embedded in Microcapsules

[0054] (1) Extraction of laver protein: get 20 grams of laver dry powder of particle diameter 60 purposes and add 1000 grams of water, mix well, be mixed with suspension; Stir and react at 35°C for 30 minutes, with a stirring rate of 100 rpm; perform ultrasonic crushing, with an ultrasonic power of 730W, an ultrasonic treatment time of 65 minutes, and an ultrasonic treatment temperature of 30°C; centrifuge at 5000g for 10 minutes, remove the precipitate, and obtain Contain the supernatant of laver content; Add ammonium sulfate in the supernatant, make the concentration of ammonium sulfate in the supernatant reach 20% (W / V), leave standstill 12 hours at 4 ℃; Through 5000g centrifugal 10 minutes, Take the precipitate to obtain laver protein.

[0055] (2) Preparation of laver hypotensive peptide: take the laver protein obtained in step (1) and add it to water, then add the yeast display-type pr...

Embodiment 2

[0059] Example 2 Preparation of Laver Hypotensive Peptide Embedded in Microcapsules

[0060] (1) Extraction of laver protein: get 20 grams of 100 order laver dry powders of particle diameter and add in 1000 grams of water, mix well, be mixed with suspension; Then add 1.6 grams of yeast display polygalacturonase prepared according to the above method, Stir and react at 35°C for 30 minutes, with a stirring rate of 200 rpm; carry out ultrasonic crushing, with ultrasonic power of 750W, ultrasonic treatment time of 65 minutes, and ultrasonic treatment temperature of 33°C; centrifuge at 5000g for 10 minutes, remove the precipitate, and obtain Contain the supernatant of laver content; Add ammonium sulfate in the supernatant, make the concentration of ammonium sulfate in the supernatant reach 20% (W / V), leave standstill 12 hours at 4 ℃; Through 5000g centrifugal 10 minutes, Take the precipitate to obtain laver protein.

[0061] (2) Preparation of laver hypotensive peptide: take the l...

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Abstract

The invention discloses a method for preparing micro capsulate embedded antihypertensive peptide derived from laver, which comprises: adding laver dry powder and yeast display type polygalacturonase into water, uniformly mixing, reacting at 35 to 37 DEG C for 30 to 45 minutes with stirring, crushing at 28 to 35 DEG C for 1 to 2 hour by ultrasonic waves with power of 730 to 750W, separating and purifying to obtain laver protein; adding laver protein and yeast display type polygalacturonase into water, uniformly mixing, reacting at 35 to 40 DEG C for 2 to 2.5 hours with stirring, separating andpurifying to obtain antihypertensive peptide derived from laver; and embedding the antihypertensive peptide derived from laver by using beta-cyclodextrin. In the invention, yeast display type polygalacturonase and ultrasonic waves are used in combination to make laver protein dissolve out, then the micro capsules of embedded antihypertensive peptide derived from laver are prepared by enzymolysis in presence of yeast display type protease and micro capsule embedding technology, and the obtained antihypertensive peptide derived from laver has high yield and high activity continuity.

Description

technical field [0001] The invention relates to the technical fields of bioengineering and microcapsules, in particular to a preparation method of microcapsule-embedded seaweed hypotensive peptide. Background technique [0002] Seaweed is rich in protein, which can be used to prepare active polypeptides with physiologically active effects such as blood pressure-lowering peptides. Porphyra cell wall is relatively hard, composed of polygalacturonic acid sulfate. In order to fully obtain laver protein for preparing laver hypotensive peptide, the cell wall structure of laver should be destroyed as much as possible to allow the cell content to flow out. The invention patent of CN100469894C discloses a method of preparing laver blood pressure-lowering peptide by using enzyme-membrane coupling technology and its application. Dried laver is crushed, degreased, dissolved, ultrasonically treated, proteolyzed, membrane separation coupling, and post-treated to obtain laver blood-loweri...

Claims

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Application Information

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IPC IPC(8): C12P21/06C07K17/10C07K17/04A61P9/12C12N15/81C12R1/84
Inventor 阮晖张延郭子堃徐娟周陈伟杜姗姗杨璐何国庆
Owner ZHEJIANG UNIV
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