Role of the multiple efflux pump protein TolC on growth, morphology, and biofilm formation under nitric oxide stress in Cronobacter malonaticus

Nitric oxide (NO) is a biological signal molecule that can control and prevent the growth of most pathogens. Cronobacter species are a group of gram-negative foodborne pathogens that cause severe diseases, including neonatal meningitis, septicemia, and necrotizing enterocolitis, especially among newborns and infants consuming contaminated powdered infant formula. Cronobacter species might be tolerant to NO, resulting in severe infections. However, the specific mechanism of tolerance to NO in Cronobacter species is unclear. Here, we explore the effects of a key component, the protein TolC, of a multiple efflux pump on the growth, morphological changes, and biofilm formation of Cronobacter malonaticus under NO stress. We found that deletion of tolC resulted in a decreased growth rate under 100 mM sodium nitroprusside (NO donor) and led to more disruptive morphological injury to the bacterial cells. Furthermore, C. malonaticus lacking the TolC protein (ΔtolC mutant) showed weaker biofilm formation than the wild-type strain under normal or NO stress conditions. We have proved that TolC plays an important role in cell growth and biofilm formation of C. malonaticus. Therefore, our results may provide valuable theoretical basis for formulating clinical guidelines for treatment of disease caused by C. malonaticus and ensuring food safety.