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Method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves

A technology of pectinase and ultrasound, applied in the field of bioengineering, can solve problems affecting the efficiency of laver cell crushing, unfavorable laver polysaccharide and laver protein extraction, etc.

Inactive Publication Date: 2013-07-24
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, with ultrasonic treatment alone, the polygalacturonic acid sulfate in the cell wall of laver remains basically intact, and only a small amount of voids are produced, which affects the breaking efficiency of laver cells and is not conducive to the extraction of laver polysaccharides and laver proteins.

Method used

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  • Method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves
  • Method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves
  • Method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 Preparation of Yeast Displayed Pectinase and Yeast Displayed Protease

[0027] By artificial synthesis, pectinase gene (Genbank number: GU277799.1), protease gene (Genbank number: NM_012708.2) and cell wall α-lectin gene of Pichia pastoris GS115 (Genbank number: M28164) were synthesized. The C-terminus of the pectinase gene (or protease gene) is added with the connecting peptide sequence GSSGGSGGSGGSGGSGS (linker), and the nucleotide sequence PEC-linker-α-agglutinin (or PRO-linker-α-agglutinin) is obtained after the connection, and the two Add EcoR I and Not I restriction sites at the end, where PEC is the pectinase gene (PRO is the protease gene), and α-agglutinin is the cell wall α-lectin gene.

[0028] Using the above artificially synthesized sequences as templates, PCR amplification was carried out using the following primer pairs respectively,

[0029] Primer pairs for pectinase:

[0030] Upstream primer: 5'-TGGCCCTGGACATGGAATCAGTGTTGGGAG-3';

[0031] ...

Embodiment 2

[0039] Example 2 Yeast Displayed Pectinase Collaborative Ultrasonic Disruption of Porphyra Cells

example 1

[0040] Example 1 Prepare a mixed suspension with 1000 grams of water, 20 grams of dried seaweed powder, and 1 gram of yeast-displayed pectinase. The stirring speed of the system is 100 rpm, the temperature is 35°C, and the treatment time is 30 minutes. The power was 730W, the ultrasonic treatment time was 65min, and the treatment temperature was 30°C to obtain a cell disruption solution, which was centrifuged (4000g for 30min) to remove the precipitate, and then the yield of laver protein and laver polysaccharide in the supernatant was measured.

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Abstract

The invention discloses a method for disrupting seaweed cells by using yeast display pectinase cooperated with ultrasonic waves, which comprises the following steps that: dry seaweed powder and yeast display pectinase are added into water, are uniformly mixed, are subjected to enzymatic reaction for 30 to 45min at the temperature of 35 to 37 DEG C, and are disrupted for 65 to 70min by utilizing ultrasonic waves with the power of 730 to 750W at the temperature of 30 to 32 DEG C. The invention also discloses a method for extracting seaweed proteins and porphyra polysaccharides, which is characterized in that: cell disruption liquid obtained after the seaweed cells are disrupted is separated. The invention also discloses a method for preparing seaweed antihypertensive peptides, which is characterized in that: the separated seaweed proteins are subjected to enzymatic reaction for 130 to 133min at the temperature of 36 to 37 DEG C by using protease. In the method, the seaweed cell walls are degraded by yeast display pectinase, and ultrasonic wave treatment is synergically carried out, so that the seaweed cells are efficiently disrupted, thereby the disruption rate of the seaweed cells is improved, and the yields of seaweed polysaccharides, seaweed proteins and seaweed antihypertensive peptides are increased.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for breaking laver cells with yeast display pectinase and ultrasonic waves. Background technique [0002] Laver is rich in polysaccharides and proteins, which can be used to prepare active polysaccharides and active polypeptides with physiologically active effects. Porphyra polysaccharide is polygalacturonic acid sulfate, which constitutes the main component of Porphyra cell wall. Porphyra cells are rich in protein, after destroying the porphyra cell wall and cell membrane, the contents of porphyra cells can flow out, and porphyra protein can be obtained. Therefore, to fully obtain Porphyra polysaccharides and Porphyra proteins, the Porphyra cell wall and cell membrane structure should be destroyed as much as possible. [0003] At present, laver cell disruption mainly adopts ultrasonic treatment. The cavitation effect produced by ultrasonic treatment can destro...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/26C12N9/60C12N15/81C07K1/14C08B37/00C12P21/06C12R1/84
Inventor 阮晖张延余璐徐娟周陈伟杜姗姗杨璐何国庆
Owner ZHEJIANG UNIV
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