PMID:25220241

Citation	Willias, SP, Chauhan, S and Motin, VL (2014) Functional characterization of Yersinia pestis aerobic glycerol metabolism. Microb. Pathog. 76:33-43
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.
Links	PubMed PMC4250381 Online version:10.1016/j.micpath.2014.08.010
Keywords	Animals; Aquaporins/genetics; Bacterial Proteins/genetics; Biofilms/growth & development; Disease Models, Animal; Fermentation; Fructose-Bisphosphatase/genetics; Glycerol/metabolism; Glycerolphosphate Dehydrogenase/genetics; Glycerophosphates/metabolism; Mice; Mutation; Plague/microbiology; Plague/pathology; Sequence Deletion; Temperature; Virulence; Yersinia pestis/enzymology; Yersinia pestis/genetics; Yersinia pestis/metabolism; Yersinia pestis/physiology
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
YERPE:Q93NB6		GO:0019662: non-glycolytic fermentation	ECO:0006049: genetic transformation evidence used in manual assertion		P		Table 2: Two different strains of Y. pestis, KIM6+, with a functional glpD gene, and CO92L, with a 93 bp inframe deletion of the glpD allele, were tested for glycerol fermentation under differing conditions. Glycerol fermentation was visualized by indicator plates. Unmodified strains were utilized as controls, in which KIM6+ was positive for glycerol fermentation and CO92L was negative for glycerol fermentation. Allelic exchange was used to exchange the functional glpD in KIM6+ with the nonfunctional glpD of CO92L. The nonfunctional glpD of CO92L was exchanged with the functional glpD of KIM6+. KIM6+ with nonfunctional glpD and CO92L with the functional glpD showed no indicators of glycerol fermentation. Glycerol fermentation was restored in the mutant KIM6+ with nonfunctional glpD when a plasmid vector of functional glpD derived from KIM6+ was added. This is evidence lack of glpD can disrupt glycerol fermentation. GO Term non-glycolytic fermentation was chosen due to the function of glpD. glpD in this experiment participates in aerobic glycerol fermentation to help form pyruvate from glycerol which does not involve the use of glycolysis as in glycolysis fermentation.	complete CACAO 13554

References

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