Method for crushing laver cells by using yeast display lipase cooperated with supersonic wave

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

Inactive Publication Date: 2012-01-18
ZHEJIANG UNIV
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, only by ultrasonic treatment, the porphyra cells are basically kept intact, and only a small amount of gaps are produced, which affects the breaking efficiency of porphyra cells and is not conducive to the extraction of porphyra polysaccharides and porphyrin proteins.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for crushing laver cells by using yeast display lipase cooperated with supersonic wave
  • Method for crushing laver cells by using yeast display lipase cooperated with supersonic wave
  • Method for crushing laver cells by using yeast display lipase cooperated with supersonic wave

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Preparation of yeast-displayed lipase and yeast-displayed protease

[0027] Synthesize lipase gene (Genbank No.: AF229435), protease gene (Genbank No.: NM_012708.2) and cell wall α-lectin gene of Pichia pastoris GS115 (Genbank No.: M28164) by artificial synthesis. (or protease gene) C-terminus plus linking peptide sequence GSSGGSGGSGGSGGSGS (linker), the nucleotide sequence LIP-linker-α-agglutinin (or PRO-linker-α-agglutinin) is obtained after connection, and EcoR I is added at both ends of the sequence and Not I restriction sites, wherein LIP is the lipase gene (PRO is the protease gene), and α-agglutinin is the cell wall α-lectin gene.

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

[0029] Primer pairs for lipase:

[0030] Upstream primer: 5'-AAGGAAAAAAAGAATTCGTTCCAGTTTCTGG~3';

[0031] Downstream primer: 5'-TTTTCCTTTTGCGGCCGCTAATGAAACG~3'

[0032] P...

Embodiment 2

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

example 1

[0040] Example 1: 1000 grams of water, 20 grams of dried seaweed powder, and 1 gram of yeast-displayed lipase were prepared into a mixed suspension. The stirring speed of the system was 100 rpm, the temperature was 35°C, and the treatment time was 30 minutes. 730W, ultrasonic treatment time of 65min, and treatment temperature of 30°C to obtain cell disruption liquid, centrifuge (4000g 30min) to remove the precipitate, and measure the yield of laver protein and laver polysaccharide in the supernatant.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a method for crushing laver cells by using yeast display lipase cooperated with supersonic wave, which comprises the following steps: adding laver dry powder, yeast display lipase into water and well mixing, performing an enzymatic hydrolysis reaction under the temperature of 35 DEG C-37 DEG C for 30-45 minutes, and then performing ultrasonic crushing by using the 730W-750W of power under the temperature of 30-32 DEG C for 65-70 minutes. The invention also discloses a method for extracting laver protein and laver amylase, which is characterized in that the cell crushed liquid of crushed laver cells is separated. The invention also discloses a method for preparing laver antihypertensive peptide, which is characterized in that the separated laver protein is performed the enzymatic hydrolysis reaction by using protease under the temperature of 36 DEG C-37 DEG C for 130-133 minutes. According to the invention, the yeast display lipase is used to degrade cell membranes of the laver cells and cooperated with the ultrasonic treatment, so that the method of the invention is capable of high efficiently crushing the laver cells, raising the laver cell crushing rate, increase the yields of laver amylase, laver protein and laver antihypertensive peptide.

Description

technical field [0001] The invention relates to the technical field of bioengineering, in particular to a method for disrupting laver cells with yeast display lipase 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 destroy...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & AuthorityApplications(China)
IPC IPC(8): C12N9/20C12N9/60C12N15/81C07K1/14C08B37/00C12P21/06C12R1/84
Inventor阮晖张延余璐徐娟周陈伟杜姗姗杨璐何国庆
OwnerZHEJIANG UNIV