Excessive damage to DNA and lipid membranes by reactive oxygen species reduces the viability of bacteria. In the present study, the proliferation of recA‑deficient Escherichia coli strains was revealed to be inhibited by 1% L‑histidine under aerobic conditions. This inhibition of proliferation was not observed under anaerobic conditions, indicating that L‑histidine enhances oxidative DNA damage to E. coli cells. Reverse transcription‑quantitative polymerase chain reaction analysis demonstrated that the expression of recA in E. coli MG1655 increased ~7‑fold following treatment with 10 mM hydrogen peroxide (H2O2) plus 1% L‑histidine, compared with that following exposure to H2O2 alone. L‑histidine increased the genomic fragmentation of E. coli MG1655 following exposure to H2O2. In addition, L‑histidine increased the generation of intracellular hydroxyl radicals in the presence of H2O2 in E. coli cells. Next, our group investigated the disinfection properties of the H2O2 and L‑histidine combination. The combination of 100 mM H2O2 and 1.0% L‑histidine significantly reduced the number of viable cells of extended‑spectrum‑β‑lactamase‑producing E. coli and multidrug‑resistant Pseudomonas aeruginosa, and this treatment was more effective than 100 mM H2O2 alone, but this effect was not observed in methicillin‑resistant Staphylococcus aureus or vancomycin‑resistant Enterococcus faecium. The combination of L‑histidine and H2O2 may be a useful strategy to selectively increase the microbicidal activity of oxidative agents against Gram‑negative bacteria.