title: Characterization of lactate utilization and its implication on the physiology of Haemophilus influenzae.
abstract: Haemophilus influenzae is a Gram-negative bacillus and a frequent commensal of the human nasopharynx. Earlier work demonstrated that in H. influenzae type b, l-lactate metabolism is associated with serum resistance and in vivo survival of the organism. To further gain insight into lactate utilization of the non-typeable (NTHi) isolate 2019 and laboratory prototype strain Rd KW20, deletion mutants of the l-lactate dehydrogenase (lctD) and permease (lctP) were generated and characterized. It is shown, that the apparent KM of l-lactate uptake is 20.1μM as determined for strain Rd KW20. Comparison of the COPD isolate NTHi 2019-R with the corresponding lctP knockout strain for survival in human serum revealed no lactate dependent serum resistance. In contrast, we observed a 4-fold attenuation of the mutant strain in a murine model of nasopharyngeal colonization. Characterization of lctP transcriptional control shows that the lactate utilization system in H. influenzae is not an inductor inducible system. Rather negative feedback regulation was observed in the presence of l-lactate and this is dependent on the ArcAB regulatory system. Additionally, for 2019 it was found that lactate may have signaling function leading to increased cell growth in late log phase under conditions where no l-lactate is metabolized. This effect seems to be ArcA independent and was not observed in strain Rd KW20. We conclude that l-lactate is an important carbon-source and may act as host specific signal substrate which fine tunes the globally acting ArcAB regulon and may additionally affect a yet unknown signaling system and thus may contribute to enhanced in vivo survival.
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Humboldtstr. 50, 8010 Graz, Austria.
authors.forename: Sabine
authors.initials: S
authors.lastname: Lichtenegger
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Humboldtstr. 50, 8010 Graz, Austria.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Bina
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Humboldtstr. 50, 8010 Graz, Austria.
authors.forename: Sandro
authors.initials: S
authors.lastname: Roier
authors.affiliation: Insitute of Hygiene and Microbiology, University of Wuerzburg, Josef-Schneider Str. 2 E1, Wuerzburg 97080, Germany.
authors.forename: Stilla
authors.initials: S
authors.lastname: Bauernfeind
authors.affiliation: Insitute of Hygiene and Microbiology, University of Wuerzburg, Josef-Schneider Str. 2 E1, Wuerzburg 97080, Germany.
authors.forename: Kristina
authors.initials: K
authors.lastname: Keidel
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Humboldtstr. 50, 8010 Graz, Austria.
authors.forename: Stefan
authors.initials: S
authors.lastname: Schild
authors.affiliation: Department of Paediatrics, Oxford University Hospitals, Headington, Oxford OX3 9DU, United Kingdom.
authors.forename: Mark
authors.initials: M
authors.lastname: Anthony
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Humboldtstr. 50, 8010 Graz, Austria. Electronic address: joachim.reidl@uni-graz.at.
authors.forename: Joachim
authors.initials: J
authors.lastname: Reidl
mesh_terms: D000818:Animals; D001770:Blood Bactericidal Activity; D004195:Disease Models, Animal; D004734:Energy Metabolism; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D006192:Haemophilus Infections / Q000382:microbiology; D006193:Haemophilus influenzae / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007770:L-Lactate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D008807:Mice, Inbred BALB C; D050296:Microbial Viability; D009305:Nasopharynx / Q000382:microbiology; D044967:Serum / Q000382:microbiology; D014774:Virulence
chemical_list: D026901:Membrane Transport Proteins; D019344:Lactic Acid; D007770:L-Lactate Dehydrogenase
keywords: Colonization; Haemophilus influenzae; l-Lactate utilization; lctP