Proteomic changes in Escherichia coli TG1 after metabolic engineering for enhanced trichloroethene biodegradation.
Through metabolic engineering, new enzymatic pathways can be introduced into cells to enable or enhance production or biotransformation of chemicals. However, these changes have physiological consequences that can be important but are not well understood. Here we describe the use of two-dimensional gel electrophoresis (2-DE) to detect changes in the proteome of Escherichia coli cells that have been engineered to transform the pollutant trichloroethene (TCE) with the enzyme toluene o-monooxygenase (TOM). Comparison of 2-DE gels (isoelectric point range 4-7) for E. coli cells with and without the ability to synthesize TOM revealed 31 new proteins in TOM-containing cells as well as nine proteins not detected in those cells but present in the plasmid control strain. Exposure of TOM-containing cells to TCE led to the synthesis of four new proteins and the loss of only one protein. Thus, this example of metabolic engineering has a substantial and complex impact on the physiology of these cells that was clearly revealed using a proteomic approach.
Pferdeort
Valerie A
VA
Department of Chemical Engineering, Colorado State University, Fort Collins, CO 80523-1370, USA.
Wood
Thomas K
TK
Reardon
Kenneth F
KF
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism*; D005798:Genes, Bacterial; D007526:Isoelectric Point; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D015202:Protein Engineering; D020543:Proteome / Q000032:analysis*; D014241:Trichloroethylene / Q000378:metabolism*
D001426:Bacterial Proteins; D020543:Proteome; D014241:Trichloroethylene; D006899:Mixed Function Oxygenases; C093480:toluene ortho-monooxygenase