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Rapid protein release from Escherichia coli by chemical permeabilization under fermentation conditions
Author(s) -
Naglak Thomas J.,
Wang Henry Y.
Publication year - 1992
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.260390706
Subject(s) - guanidine , escherichia coli , lysis , fermentation , intracellular , cell growth , growth inhibition , mutant , chemistry , biochemistry , protein biosynthesis , cell , biology , gene
Overall protein release greater than 75% in less than 1 h can be attained by exposing exponentially growing Escherichia coli cells to 0.4 M guanidine plus 0.5% Triton X‐100 at 37°C in medium. Cell growth stops immediately upon addition of the chemicals, but the cells are not lysed. Guanidine concentrations lower than 0.2 M , in conjunction with 0.5% Triton X‐100, do not release significant intracellular protein, nor do they inhibit cell growth. Under these conditions, the cells undergo an adaptation that confers resistance to protein release by further treatment with guanidine and Triton X‐100. Cells treated with 0.2 M guanidine plus 0.5% Triton X‐100 display intermediate behavior. Protein release is approximately 35%, and growth is temporarily interrupted by an extended lag phase. Subsequent resumption of cell growth results in resistant cells and no additional protein release. This resistance is shown to be reversible and is most likely due to physiological adaptation rather than genetic mutation.