Unveiling the photoelectrocatalytic inactivation mechanism of Escherichia coli: Convincing evidence from responses of parent and anti-oxidation single gene knockout mutants

Abstract

This study investigated photoelectrocatalytic (PEC) inactivation mechanism of bacteria using parental Escherichia coli (E. coli) BW25113 and its isogenic mutants deficient in catalase HPI (katG−, JW3914-1) and Mn-SOD (sodA−, JW3879-1). BW25113 in the mid-log phase was less susceptible to PEC inactivation than those in early-log and stationary phases, consistent with the peak activities of catalase and superoxide dismutase (SOD) at mid-log phase (30.6 and 13.0 Unit/ml/OD600). For different strains all in mid-log phase, PEC inactivation efficiency followed the order katG− > sodA− > BW25113, with the duration of 60, 60 and 90 min for complete inactivation of ∼2 × 107 CFU mL−1 bacteria, respectively. Correspondingly, catalase and SOD levels of BW25113 were also higher than the mutants by 5.9 and 11.7 Unit/mL/OD600, respectively. Reactive oxygen species (ROSs) concentrations in PEC systems revealed that the inactivation performance coincided with H2O2 levels, rather than OH. Moreover, pre-incubation with H2O2 elevated catalase activities and PEC inactivation resistance of BW25113 were positively correlated. The above results indicated that H2O2 was the dominant PEC generated bactericide, and anti-oxidative enzymes especially catalase contributed greatly to the bacterial PEC resistance capacity. Further tests revealed that PEC treatment raised the intracellular ROSs concentration by more than 3 times, due to the permeated H2O2 and its intracellular derivative, OH. However, oxidative stress response of E. coli, such as increased catalase or SOD were not observed, perhaps because the ROSs overwhelmed the bacterial protective capacity. The accumulated ROSs subsequently caused oxidative damages to E. coli cells, including membrane damage, K+ leakage, and protein oxidation. Compared with BW25113, the mutants experienced damages earlier and at higher levels, confirming the essential roles of catalase and SOD in the bacterial PEC resistance.