Polyethyleneimine is an effective permeabilizer of Gram-negative bacteria

VTT Biotechnology and Food Research, PO Box 1501, FIN-02044 VTT, Espoo, Finland Department of Vaccine Development, KTL, Mannerheimintie 166, FIN-00300, Helsinki, Finland 1 Author for correspondence: IIkka M. Helander. Tel: +358 9 4564452. Fax: +3558 9 4552028. e-mail: Ilkka.Helander@vtt.fi ABSTRACT The effect of the polycation polyethyleneimine (PEI) on the permeability properties of the Gram-negative bacterial outer membrane was investigated using Escherichia coli, Pseudomonas aeruginosa and Salmonella typhimurium as target organisms. At concentrations of less than 20 µg ml -1 , PEI increased the bacterial uptake of 1- N -phenylnaphthylamine, which is a hydrophobic probe whose quantum yield is greatly increased in a lipid environment, indicating increased hydrophobic permeation of the outer membrane by PEI. The effect of PEI was comparable to that brought about by the well-known permeabilizer EDTA. Permeabilization by PEI was retarded but not completely inhibited by millimolar concentrations of MgCl 2 . PEI also increased the susceptibility of the test species to the hydrophobic antibiotics clindamycin, erythromycin, fucidin, novobiocin and rifampicin, without being directly bactericidal. PEI sensitized the bacteria to the lytic action of the detergent SDS in assays where the bacteria were pretreated with PEI. In assays where PEI and SDS were simultaneously present, no sensitization was observed, indicating that PEI and SDS were inactivating each other. In addition, a sensitizing effect to the nonionic detergent Triton X-100 was observed for P. aeruginosa. In conclusion, PEI was shown to be a potent permeabilizer of the outer membrane of Gram-negative bacteria. Keywords: Polyethyleneimine, Gram-negative bacteria, outer membrane, lipopolysaccharide