Characterization of Ribose-5-Phosphate Isomerase B from Newly Isolated Strain Ochrobactrum sp. CSL1 Producing...

In this study, we attempted to find new and efficient microbial enzymes for producing rare sugars. A ribose-5-phosphate isomerase B (OsRpiB) was cloned, overexpressed, and preliminarily purified successfully from a newly screened Ochrobactrum sp. CSL1, which could catalyze the isomerization reaction of rare sugars. A study of its substrate specificity showed that the cloned isomerase (OsRpiB) could effectively catalyze the conversion of _L-rhamnose to _L-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions (50°C, pH 8.0, and 1 mM Ca²⁺) were obtained to maximize the potential of OsRpiB in preparing _L-rhamnulose. The catalytic properties of OsRpiB, including K, k, and catalytic efficiency (k/K), were determined as 43.47 mM, 129.4 sec^-1, and 2.98 mM/sec. The highest conversion rate of _L-rhamnose under the optimized conditions by OsRpiB could reach 26% after 4.5 h. To the best of our knowledge, this is the first successful attempt of the novel biotransformation of _L-rhamnose to _L-rhamnulose by OsRpiB biocatalysis.