Reduced outer membrane permeability of Escherichia coli O157:H7: Suggested role of modified outer membrane porins and theoretical function in resistance to antimicrobial agents

M. B. Martinez, M. Flickinger, L. A. Higgins, T. Krick, G. L. Nelsestuen

Research output: Contribution to journalArticlepeer-review

27 Scopus citations

Abstract

Outer membrane permeability of Escherichia coli O157:H7 was determined by an in vivo kinetic model with the periplasmic enzyme alkaline phosphatase [Martinez et al. (1996) Biochemistry 35, 1179-1186]. p-Nitrophenyl phospate (PNPP) substrate, added to intact bacteria, must diffuse through the outer membrane to reach the enzyme. At low substrate concentration the bacterium was in the perfectly reactive state where all molecules that entered the periplasm were captured and converted to product. Transmembrane diffusion was rate limiting, and the permeability of the outer membrane was determined from kinetic properties. The O157:H7 strain grown at 30 °C showed one-sixth the permeability of wild-type E. coli grown at 30 °C. Wild-type bacteria grown at ≥37 °C show a physiological response with a shift in expression of outer membrane porins that lowered permeability to PNPP by approximately 70%. The O157:H7 strain did not display this temperature-sensitive shift in permeability even though a change in porin expression could be visualized by staining intensity of Omp F and Omp C on acrylamide gels. Altered behavior of the O157:H7 membrane was also indicated by a several thousand-fold lower response to transformation relative to wild-type E. coli. Matrix-assisted laser desorption ionization time of flight mass spectrometry and electrospray ionization mass spectrometry confirmed the expression of the Omp F and Omp C variants that are unique to E. coli O157:H7. This reduced outer membrane permeability can contribute to enhanced resistance of O157:H7 to antimicrobial agents.

Original languageEnglish (US)
Pages (from-to)11965-11974
Number of pages10
JournalBiochemistry
Volume40
Issue number40
DOIs
StatePublished - Oct 9 2001

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