title: Functional characterization of Yersinia pestis aerobic glycerol metabolism.
abstract: Yersinia pestis biovar Orientalis isolates have lost the capacity to ferment glycerol. Herein we provide experimental validation that a 93 bp in-frame deletion within the glpD gene encoding the glycerol-3-phosphate dehydrogenase present in all biovar Orientalis strains is sufficient to disrupt aerobic glycerol fermentation. Furthermore, the inability to ferment glycerol is often insured by a variety of additional mutations within the glpFKX operon which prevents glycerol internalization and conversion to glycerol-3-phosphate. The physiological impact of functional glpFKX in the presence of dysfunctional glpD was assessed. Results demonstrate no change in growth kinetics at 26 °C and 37 °C. Mutants deficient in glpD displayed decreased intracellular accumulation of glycerol-3-phosphate, a characterized inhibitor of cAMP receptor protein (CRP) activation. Since CRP is rigorously involved in global regulation Y. pestis virulence, we tested a possible influence of a single glpD mutation on virulence. Nonetheless, subcutaneous and intranasal murine challenge was not impacted by glycerol metabolism. As quantified by crystal violet assay, biofilm formation of the glpD-deficient KIM6+ mutant was mildly repressed; whereas, chromosomal restoration of glpD in CO92 resulted in a significant increase in biofilm formation.
authors.affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Stephan P
authors.initials: SP
authors.lastname: Willias
authors.affiliation: Department of Microbiology & Immunology, University of Texas Medical Branch, Galveston, TX, USA.
authors.forename: Sadhana
authors.initials: S
authors.lastname: Chauhan
authors.affiliation: Department of Pathology, University of Texas Medical Branch, Galveston, TX, USA; Department of Microbiology & Immunology, University of Texas Medical Branch, Galveston, TX, USA. Electronic address: vlmotin@utmb.edu.
authors.forename: Vladimir L
authors.initials: VL
authors.lastname: Motin
mesh_terms: D000818:Animals; D020346:Aquaporins / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D018441:Biofilms / Q000254:growth & development; D004195:Disease Models, Animal; D005285:Fermentation; D006597:Fructose-Bisphosphatase / Q000235:genetics; D005990:Glycerol / Q000378:metabolism*; D005993:Glycerolphosphate Dehydrogenase / Q000235:genetics*; D005994:Glycerophosphates / Q000378:metabolism; D051379:Mice; D009154:Mutation; D010930:Plague / Q000382:microbiology / Q000473:pathology; D017384:Sequence Deletion; D013696:Temperature; D014774:Virulence; D015010:Yersinia pestis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
chemical_list: D020346:Aquaporins; D001426:Bacterial Proteins; D005994:Glycerophosphates; C029620:alpha-glycerophosphoric acid; D005993:Glycerolphosphate Dehydrogenase; D006597:Fructose-Bisphosphatase; D005990:Glycerol
keywords: Biofilm; Glycerol; Yersinia pestis; glpD; glpFKX