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
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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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