Compatible Solute Synthesis and Import by the Moderate Halophile Spiribacter salinus: Physiology and Genomics

María J. León, Tamara Hoffmann, Cristina Sánchez-Porro, Johann Heider, Antonio Ventosa, Erhard Bremer

Members of the genusSpiribacterare found worldwide and are abundant in ecosystems possessing intermediate salinities between seawater and saturated salt concentrations.Spiribacter salinusM19-40 is the type species of this genus and its first cultivated representative. In the habitats ofS. salinusM19-40, high salinity is a key determinant for growth and we therefore focused on the cellular adjustment strategy to this persistent environmental challenge. We coupled these experimental studies to thein silicomining of the genome sequence of this moderate halophile with respect to systems allowing this bacterium to control its potassium and sodium pools, and its ability to import and synthesize compatible solutes.S. salinusM19-40 produces enhanced levels of the compatible solute ectoine, both under optimal and growth-challenging salt concentrations, but the genes encoding the corresponding biosynthetic enzymes are not organized in a canonicalectABCoperon. Instead, they are scrambled (ectAC;ectB) and are physically separated from each other on theS. salinusM19-40 genome. Genomes of many phylogenetically related bacteria also exhibit a non-canonical organization of theectgenes.S. salinusM19-40 also synthesizes trehalose, but this compatible solute seems to make only a minor contribution to the cytoplasmic solute pool under osmotic stress conditions. However, its cellular levels increase substantially in stationary phase cells grown under optimal salt concentrations.In silicogenome mining revealed thatS. salinusM19-40 possesses different types of uptake systems for compatible solutes. Among the set of compatible solutes tested in an osmostress protection growth assay, glycine betaine and arsenobetaine were the most effective. Transport studies with radiolabeled glycine betaine showed thatS. salinusM19-40 increases the pool size of this osmolyte in a fashion that is sensitively tied to the prevalent salinity of the growth medium. It was amassed in salt-stressed cells in unmodified form and suppressed the synthesis of ectoine. In conclusion, the data presented here allow us to derive a genome-scale picture of the cellular adjustment strategy of a species that represents an environmentally abundant group of ecophysiologically important halophilic microorganisms.