Recovery and separation of cell lysate proteins using hydrogels guided by aqueous two-phase extraction principles.
The addition of poly(ethylene glycol) and salts to clarified cell lysates of Thiosphaera pantotropha increases sorption of microbial proteins into dextran hydrogels, consistent with the thermodynamics of aqueous two-phase extraction. Addition of 12 wt% PEG-10,000 to the lysate increased total sorption of protein by the dextran gel from 5.2 mg/g dextran to 37 mg/g; addition of either 0.1 M potassium iodide or tetrabutylammonium fluoride along with PEG to the lysate increased protein sorption to more than 63 mg/g, a 12-fold increase. SDS-PAGE demonstrated that the type of salt added controls which proteins are absorbed by the gel. Previously demonstrated only with model solutions, these results suggest another approach to recovery and separation strategies for proteins produced by fermentation.
Putka
Christopher S
CS
University of Cincinnati, Department of Chemical Engineering, Cincinnati, OH 45221, USA. Chris_Putka@kinetico.com
Gehrke
Stevin H
SH
Willis
Mike
M
Stafford
David
D
Bryant
John
J
D000042:Absorption; D002458:Cell Fractionation; D002850:Chromatography, Gel / Q000379:methods*; D003911:Dextrans; D004798:Enzymes / Q000096:biosynthesis / Q000302:isolation & purification*; D020100:Hydrogels; D010230:Paracoccus / Q000201:enzymology; D011092:Polyethylene Glycols; D011506:Proteins / Q000302:isolation & purification*; D012996:Solutions
D003911:Dextrans; D004798:Enzymes; D020100:Hydrogels; D011506:Proteins; D012996:Solutions; D011092:Polyethylene Glycols