Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/787
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Type: Journal article
Title: Chemical Treatment of Escherichia coli: 1. Extraction of Intracellular Protein from Uninduced Cells
Author: Falconer, R.
O'Neill, B.
Middelberg, A.
Citation: Biotechnology and Bioengineering, 1997; 53(5):453-458
Publisher: JOHN WILEY & SONS INC
Issue Date: 1997
ISSN: 0006-3592
1097-0290
Abstract: Extraction of intracellular protein from Escherichia coli is traditionally achieved by mechanical disruption. A chemical treatment that destroys the integrity of the bacterial cell wall and could provide an alternative technique is examined in this study. Treatment with a combination of the chelating agent ethylenediaminetet-raacetate (EDTA) (greater than 0.3 mM) and the chaotropic agent urea (6 M) is highly effective at releasing protein from uninduced E. coli. The 6 M urea in the presence of 3 mM EDTA can release cytoplasmic protein from both logarithmic-phase and stationary-phase E. coli cells at levels equivalent to mechanical disruption. The concentrations of the two chemical agents were the major variables affecting the maximum levels of protein release. Several minor variables and interactions were also identified. The kinetics of protein release is first order. For 2, 4, and 6 M urea with 3 mM EDTA, the time constant is approximately 2.5 min independent of urea concentration. Kinetics for 3 mM EDTA without urea is considerably slower, with a time constant of 12.3 min. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 453-458, 1997.
RMID: 0030003099
DOI: 10.1002/(SICI)1097-0290(19970305)53:5<453::AID-BIT2>3.0.CO;2-G
Appears in Collections:Chemical Engineering publications

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