The surface properties of bacteria play an essential role in their abilities to perform transmembrane communication, adherence, immobilization, flocculation, etc. However, the responsiveness of bacterial surfaces to elevated atmospheric CO 2 remains unknown. In this study, using the model bacteria, Paracoccus denitrificans , the effect of CO 2 on the primary bacterial surface properties, specifically hydrophobicity and surface charge, has been explored. We found that hydrophilicity and negative surface charge both rose in conjunction with increased atmospheric CO 2 concentrations. Studies of the potential mechanisms involved have illustrated that elevated CO 2 significantly increases the production of polysaccharides in extracellular polymeric substances (EPS). Various hydrophilic groups and negative charges in these polysaccharides prompt hydrophilicity and surface charge variations in bacteria. Further research has identified that elevations in CO 2 result in the accumulation of reactive species, specifically reactive nitrogen species (RNS). In this study, it was found that RNS damaged the permeability of bacterial membranes by inducing lipid peroxidation and then caused the leakage of intracellular substrate, which ultimately led to an increase in EPS polysaccharides. Our findings suggest that changes in bacterial surface properties due to atmospheric CO 2 elevation, as well as the reactions these trigger, merit widespread attention. [Display omitted] • Elevated CO 2 rose the hydrophilicity and negative surface charge of denitrifying bacteria. • CO 2 resulted in the accumulation of RNS during denitrification, inducing lipid peroxidation. • Lipid peroxidation increased the permeability of denitrifying bacteria. • Intracellular leakage raised due to the increase of bacterial membrane permeability. • The accumulation of intracellular leakage provided the polysaccharides in EPS. [ABSTRACT FROM AUTHOR]