Prevention and induction of oxidative damage in E. coli cells by cationized proteins

Abstract

The successful prevention of oxidative damage in E. coli B cells by cationized catalase (cCAT), and the induction of oxidative stress by cationized glucose oxidase (cGO) and cationized superoxide dismutase (cSOD) is presented in this study. Exposure of E. coli cells to hydrogen peroxide and hydroxyl radical resulted in a rapid killing of the cells. Measurements of biochemical markers: cellular potassium levels and uptake and accumulation of leucin indicated membrane damage in some of the oxidants employed. Following incubation with native CAT or SOD, the cells were washed and exposed to oxidative stress. The results of this procedure did not protect the cells against the oxidative damage. In contrast, incubation of the cells with pretreated CAT with poly- l-histidine, followed by washing of the cells and the subsequent introduction of oxidative stress inducers, resulted in a pronounced protection of the cells against the oxidative stress. Employment of pretreated SOD, and exposure, after washing the cells, to oxidative stress, resulted in an enhancement of the oxidative damage in some cases. Exposure of the cells to cGO resulted in a marked killing of the cells as compared to the untreated enzyme. The use of E. coli cells as a model system for studying the effect of cationized enzymes on cell surfaces is discussed.