2-deoxyribose-1-phosphate--(alpha-d-erythro)-isomer and mannose-1-phosphate

Four orthologous genes (TK1108, TK1404, TK1777, and TK2185) that can be annotated as phosphomannomutase (PMM) genes (COG1109) have been identified in the genome of the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. We previously found that TK1777 actually encodes a phosphopentomutase. In order to determine which of the remaining three orthologues encodes a phosphoglucomutase (PGM), we examined the PGM activity in T. kodakaraensis cells and identified the gene responsible for this activity. Heterologous gene expression and purification and characterization of the recombinant protein indicated that TK1108 encoded a protein with high levels of PGM activity (690 U mg(-1)), along with high levels of PMM activity (401 U mg(-1)). Similar analyses of the remaining two orthologues revealed that their protein products exhibited neither PGM nor PMM activity. PGM activity and transcription of TK1108 in T. kodakaraensis were found to be higher in cells grown on starch than in cells grown on pyruvate. Our results clearly indicate that, among the four PMM gene orthologues in T. kodakaraensis, only one gene, TK1108, actually encodes a protein with PGM and PMM activities.

Topics: Amino Acid Sequence; Archaeal Proteins; Cloning, Molecular; Coenzymes; DNA, Archaeal; Enzyme Stability; Genes, Archaeal; Glucosephosphates; Kinetics; Mannosephosphates; Molecular Sequence Data; Phosphoglucomutase; Phosphotransferases (Phosphomutases); Phylogeny; Pyruvic Acid; Recombinant Proteins; Ribosemonophosphates; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Starch; Substrate Specificity; Temperature; Thermococcus