title: A Mannose Family Phosphotransferase System Permease and Associated Enzymes Are Required for Utilization of Fructoselysine and Glucoselysine in Salmonella enterica Serovar Typhimurium.
abstract: Salmonella enteric serovar Typhimurium, a major cause of food-borne illness, is capable of using a variety of carbon and nitrogen sources. Fructoselysine and glucoselysine are Maillard reaction products formed by the reaction of glucose or fructose, respectively, with the ε-amine group of lysine. We report here that S. Typhimurium utilizes fructoselysine and glucoselysine as carbon and nitrogen sources via a mannose family phosphotransferase (PTS) encoded by gfrABCD (glucoselysine/fructoselysine PTS components EIIA, EIIB, EIIC, and EIID; locus numbers STM14_5449 to STM14_5454 in S. Typhimurium 14028s). Genes coding for two predicted deglycases within the gfr operon, gfrE and gfrF, were required for growth with glucoselysine and fructoselysine, respectively. GfrF demonstrated fructoselysine-6-phosphate deglycase activity in a coupled enzyme assay. The biochemical and genetic analyses were consistent with a pathway in which fructoselysine and glucoselysine are phosphorylated at the C-6 position of the sugar by the GfrABCD PTS as they are transported across the membrane. The resulting fructoselysine-6-phosphate and glucoselysine-6-phosphate subsequently are cleaved by GfrF and GfrE to form lysine and glucose-6-phosphate or fructose-6-phosphate. Interestingly, although S. Typhimurium can use lysine derived from fructoselysine or glucoselysine as a sole nitrogen source, it cannot use exogenous lysine as a nitrogen source to support growth. Expression of gfrABCDEF was dependent on the alternative sigma factor RpoN (σ(54)) and an RpoN-dependent LevR-like activator, which we designated GfrR.
  
  
  OBJECTIVE
  Salmonella physiology has been studied intensively, but there is much we do not know regarding the repertoire of nutrients these bacteria are able to use for growth. This study shows that a previously uncharacterized PTS and associated enzymes function together to transport and catabolize fructoselysine and glucoselysine. Knowledge of the range of nutrients that Salmonella utilizes is important, as it could lead to the development of new strategies for reducing the load of Salmonella in food animals, thereby mitigating its entry into the human food supply.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, Georgia, USA.
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Miller
authors.affiliation: Department of Chemistry, University of Georgia, Athens, Georgia, USA Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
authors.forename: Robert S
authors.initials: RS
authors.lastname: Phillips
authors.affiliation: Department of Microbiology, University of Georgia, Athens, Georgia, USA.
authors.forename: Paul B
authors.initials: PB
authors.lastname: Kilgore
authors.affiliation: Department of Microbiology, University of Georgia, Athens, Georgia, USA.
authors.forename: Grady L
authors.initials: GL
authors.lastname: Smith
authors.affiliation: Department of Microbiology, University of Georgia, Athens, Georgia, USA trhoover@uga.edu.
authors.forename: Timothy R
authors.initials: TR
authors.lastname: Hoover
mesh_terms: D000818:Animals; D002208:Caproates; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D005944:Glucosamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D015394:Molecular Structure; D010770:Phosphotransferases / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D051877:RNA Polymerase Sigma 54 / Q000235:genetics / Q000378:metabolism; D012486:Salmonella typhimurium / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: C518642:5-(acetylamino)-4-oxo-6-phenyl-2-hexenoic acid methyl ester; D002208:Caproates; D026901:Membrane Transport Proteins; C033186:fructosyl-lysine; D010770:Phosphotransferases; D051877:RNA Polymerase Sigma 54; D008239:Lysine; D005944:Glucosamine