I would like to purify proteins from a seaweed hydrolyzate. The problem is the large amount of polysaccharides and that proteins are linked in it. So, which method can be used to isolate these proteins without denaturating it?
IF the protein of your interest is a glycoproteins, then worth using the lectin binding matrices like Concovalin A matrix or other glycoprotein purification matrices.
if the hydrolysate is a mixture of protein and sugars than concentrate the protein and pass it through gel filtration columns of small size exclusions.
you can read the following article for polyssaharide extraction and purification https://www.researchgate.net/publication/260110789_Characterization_of_the_flocculating_agent_from_the_spontaneously_flocculating_microalga_Chlorella_vulgaris_JSC-7
Article Characterization of the flocculating agent from the spontane...
how you prepared the extract. Prepare separate extract. For polysaccharides prepare extract in 80% ethanol with pistal and mortar and for proteins prepare extract in buffer likely lysis buffer containing DTT. you may use following concentration for preparation of 100 ml buffer:10mM Tris HCl + 5mM EDTA + 1% Triton X 100 + 1mM PMSF + 2mM DTT. Adjust the pH to 8.
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