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Recovery and separation of cell lysate proteins using hydrogels guided by aqueous two‐phase extraction principles
Author(s) -
Putka Christopher S.,
Gehrke Stevin H.,
Willis Mike,
Stafford David,
Bryant John
Publication year - 2002
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.10349
Subject(s) - lysis , chemistry , chromatography , dextran , sorption , extraction (chemistry) , self healing hydrogels , aqueous solution , ethylene glycol , cell disruption , aqueous two phase system , peg ratio , biochemistry , organic chemistry , adsorption , finance , economics
Abstract 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. © 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 80: 139–143, 2002.