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Precipitation of nucleic acids with poly(ethyleneimine)
R M Cordes1, W B Sims, C E Glatz
1Department of Chemical Engineering, Iowa State University, Ames 50011.
Biotechnology Progress
|July 1, 1990
Summary
Poly(ethyleneimine) (PEI) effectively precipitates nucleic acids from yeast homogenates, achieving over 95% removal. This method results in minimal protein loss (around 15%) and does not aggregate cell debris.
Area of Science:
- Biotechnology
- Biochemistry
- Molecular Biology
Background:
- Nucleic acid removal is crucial for efficient protein recovery in downstream processing.
- Traditional methods can be inefficient or lead to significant protein loss.
Purpose of the Study:
- To evaluate the efficacy of poly(ethyleneimine) (PEI) for precipitating nucleic acids from Saccharomyces cerevisiae homogenates.
- To investigate the impact of PEI dosage on particle size, protein loss, and nucleic acid removal efficiency.
- To compare batch and continuous precipitation modes.
Main Methods:
- Precipitation of nucleic acids using varying dosages of PEI in yeast homogenates.
- Analysis of particle size distribution.
- Quantification of nucleic acid removal and protein loss.
- Comparison of batch and continuous precipitation processes.
Main Results:
- Achieved over 95% removal of nucleic acids from yeast homogenates using PEI precipitation.
- Reported approximately 15% protein loss, with minimal impact from prior cell debris removal.
- Coprecipitated protein was predominantly large molecular weight with diverse isoelectric points.
- PEI treatment did not aggregate cell debris, and particle size distribution was consistent across batch and continuous modes.
Conclusions:
- Poly(ethyleneimine) is an effective agent for nucleic acid removal in yeast homogenates.
- PEI precipitation offers a viable method for improving protein recovery efficiency.
- The process is robust, with consistent results in both batch and continuous operations.