Precipitation of nucleic acids with poly(ethyleneimine).
Removal of nucleic acids from cell extracts is a common early step in downstream processing for protein recovery. We report on the precipitation of nucleic acids from a homogenate of Saccharomyces cerevisiae by addition of the cationic polyelectrolyte poly(ethyleneimine) (PEI), focusing on the effect of PEI dosage on particle size, protein loss, and extent of nucleic acid removal in both batch and continuous mode. Better than 95% removal of nucleic acids from yeast homogenates was achieved by means of precipitation with PEI with protein losses of approximately 15% with or without previous removal of cell debris. The coprecipitated protein is predominately large molecular weight material and exhibits both low and high isoelectric points. Such treatment does not aggregate the cell debris; size distribution of the precipitated particles from a continuous precipitator is very similar to that for protein precipitation.
Department of Chemical Engineering, Iowa State University, Ames 50011.
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Cordes
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Sims
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Glatz
D011232:Chemical Precipitation; D005656:Fungal Proteins / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D009696:Nucleic Acids / Q000737:chemistry; D011094:Polyethyleneimine; D012441:Saccharomyces cerevisiae / Q000032:analysis
D005656:Fungal Proteins; D009696:Nucleic Acids; D011094:Polyethyleneimine