Chlorhexidine is a cationic bisbiguanide with broad antibacterial activity, and diverse activity encompassing gram-positive and gram-negative bacteria, yeasts, dermatophytes, and some lipophilic viruses. Chlorhexidine alters the permeability of the bacterial membrane. They also neutralize periodontal pathogens such as Streptococcus aureus, Porphyromonas gingivalis, and Prevotella intermedia. Porphyromonas gingivalis were cultured to isolate the Porphyromonas gingivalis outer membranes (OPG). Also, OPG were used to prepare large unilamellar liposomes with total lipids (OPGTL) extracted and prepared with the phospholipids mixture (PL). The effect of chlorhexidine digluconate on cell membrane kinetic changes was investigated using fluorescence polarization of n-(9-anthroyloxy) stearic acid with different rotational mobility depending on the probe substitution position (n) in the membrane phospholipid aliphatic chain. Chlorhexidine digluconate increased the hydrocarbon rotational mobility interior of the OPG, OPGTL and PL, but native and model membranes interfacial mobility were decreased. The sensitivity to increasing chlorhexidine digluconate effect on rotational mobility was proportional to the depth of probe position in descending order of 16-AP, 12-AS, 6-AS and 9-AS. Chlorhexidine digluconate disordering or ordering the effects on the membrane lipids may cause its bacteriostatic and bacteriocidal actions.