ABSTRACT
A newly isolated bacterium, Cellulomonas parahominis MB426, produced l-ribose isomerase (CeLRI) on a medium containing l-ribose as a sole carbon source. A 32 kDa protein isomerizing l-ribose to l-ribulose was purified to homogeneity from this bacterium. A set of degenerated primers were synthesized based on amino acid sequences of the purified CeLRI, and a 747 bp gene encoding CeLRI was cloned, sequenced and overexpressed in Escherichia coli. This gene encoded a 249 amino acid protein with a calculated molecular mass of 27,435. The deduced amino acid sequence of this gene showed the highest identity with l-ribose isomerase from Acinetobacter calcoaceticus DL-28 (71%). The recombinant l-ribose isomerase (rCeLRI) was optimally active at pH 9.0 and 40°C, and was stable up to 40°C for 1 h and not dependent for metallic ions for its activity. The rCeLRI showed widely substrate specificity for the rare sugar which involved l-erythro form such as l-ribose, d-lyxose, d-talose, d-mannose, l-gulose, and l-allose.
