Method for separating and purifying sea-mussel mucin by using salting out and dialyzing

A mussel mucin, separation and purification technology is applied in the field of separation and purification of protein, which can solve the problems of high cost of mussel mucin equipment and materials, and achieve the effect of low cost

Active Publication Date: 2009-11-25
JIANGYIN USUN BIOCHEMICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The purpose of the present invention is to overcome the problem of high cost of equipment and materials for purifying mussel mucin by chromatographic methods, and provide a fast, low-cost, high-concentration mussel mucin separation and purification method by using salting out and dialysis

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1, isolating mussel mucin with perchloric acid extraction, salting out and dialysis

[0021] 1) Using 0.5% perchloric acid as the extraction solvent, add 300ml of extraction solvent to 100g of mussel foot silk, extract at 25°C for 10min, crush the frozen mussel foot silk at high speed with a stirrer to evenly suspend it in the extraction solvent;

[0022] 2) Centrifuge at 10000r / min for 30min to remove the residue and keep the supernatant;

[0023] 3) Use 2 times the volume of 6-12M sodium chloride to salt out at 15-25°C for 1-6h;

[0024] 4) Centrifuge at 10,000 rpm for 3 minutes at 4°C, collect the precipitate, and dissolve it with 2% acetic acid;

[0025] 5) Centrifuge at 10,000 rpm for 3 minutes at 4°C to remove insoluble components and dissolve them again with 2% acetic acid;

[0026] 6) Dialyze with 2% acetic acid at 4°C for 5 hours (molecular weight cut-off of the dialysis bag is 12000), and store at 4°C;

[0027] 7) Identification of mussel mucin f...

Embodiment 2

[0030] Embodiment 2, isolating mussel mucin with acetic acid extraction, salting out and dialysis

[0031] 1) Using 10% acetic acid as the extraction solvent, adding 1000 ml of extraction solvent to 100 g of mussel foot silk, leaching at 25 ° C for 20 minutes, crushing the frozen mussel foot silk at high speed with a stirrer to evenly suspend it in the extraction solvent;

[0032] 2) Remove the residue by filtering with a 45 μm filter membrane, and keep the supernatant;

[0033] 3) Use 3 times the volume of 10M sodium chloride to salt out at 25°C for 6 hours;

[0034] 4) Centrifuge at 15,000 rpm for 3 minutes at 4°C, collect the precipitate, and dissolve it with 3% acetic acid;

[0035] 5) Centrifuge at 15,000 rpm for 3 minutes at 4°C to remove insoluble components and dissolve them again with 3% acetic acid;

[0036] 6) Dialyze with 3% acetic acid at 4°C for 10 hours (molecular weight cut-off of the dialysis bag is 12000), and store at 4°C;

[0037] 7) Identification of mu...

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PUM

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Abstract

The invention relates to a method for separating and purifying the sea-mussel mucin by using salting out and dialyzing, the salting out and the dialyzing utilizes the proteinaceous physicochemical character to purify by a non- chromatograph method, and solves the problem of high apparatus and material cost in the chromatograph method for purifying the sea-mussel mucin, a quick, low-cost and high-concentration method for separating and purifying the sea-mussel mucin is provided. The strong acid is used for extracting, the salting out and the dialyzing are used for purifying, the sea-mussel mucin is determined by adopting the acetic acid-carbamide- polyacrylamide gel electrophoresis via the blue tetrazolium specificity colour development, and the purity is determined by the inversed phase chromatography quantitatively.

Description

technical field [0001] The invention relates to a method for separating and purifying protein, in particular to a method for separating and purifying mussel mucin by salting out and dialysis. Background technique [0002] Mussel adhesive protein (MAP), also called Mytilus edulisfoot protein (Mefp), comes from the marine shellfish Mytilus edulis, which has the ability to withstand the impact of waves in the offshore, it A protein glue is produced and stored in special glands, which is released by filaments onto solid surfaces such as rocks to form a water-resistant bond and fix itself. Studies on glass have shown that protein glue forms plaques, extending from them, with strong tensile strength (10 6 -10 7 newton meter -2 ), while the intraplaque material contains mussel mucin MAP. [0003] Mussel mucin contains L-3,4-dihydroxyphenylalanine (L-DOPA), which is formed by the action of tyrosinase on tyrosine residues. The L-DOPA residues are cross-linked to each other due t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C07K1/36C07K1/34C07K1/30
Inventor 孟桂凤邢思亮
Owner JIANGYIN USUN BIOCHEMICAL TECH CO LTD
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