Biotechnology and Bioengineering, Vol.90, No.4, 509-515, 2005
Electrochemical evaluation of cellular physiological status under stress in Escherichia coli with the rpoS-lacZ reporter gene
We developed an electrochemical detection method for evaluating cellular physiological status based on the stringent response as a means to monitor cell viability. A reporter plasmid was constructed by inserting the p-galactosidase gene (lacZ) under the control of the rpoS promoter, and then used to transform E. coli cells. Electro-chemical responses from the products catalyzed by β-galactosidase expressed by these E. cofi cells were detected using the chronoamperometric technique in a nondestructive manner. Comparisons of response currents between the relA-positive strain and relA-negative strain revealed that increases in these currents were caused by the stringent response due to the stressful alcoholic environment, and thus as a model of stressful cultivating conditions. The current was proportional to the β-galactosidase activity assayed by a conventional method that required the destruction of cells. The cellular physiological status, which depends on the stringent response as a viability marker, therefore, could then be evaluated online with a current using the rpoS-lacZ reporter gene in the relA-positive strain without pretreatment. © 2005 Wiley Periodicals, Inc.
Keywords:bioprocess monitoring;cellular physiological;status;electrochemical monitoring;rpoS promoter;stringent response