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( Min Shen ),( Xin Ju ),( Xinqi Xu ),( Xuemei Yao ),( Liangzhi Li ),( Jiajia Chen ),( Cuiying Hu ),( Jiaolong Fu ),( Lishi Yan ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.7
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 (50oC, pH 8.0, and 1 mM Ca<sup>2+</sup>) were obtained to maximize the potential of OsRpiB in preparing L-rhamnulose. The catalytic properties of OsRpiB, including K<sub>m</sub>, K<sub>cat</sub>, and catalytic efficiency (K<sub>cat</sub>/K<sub>m</sub>), were determined as 43.47 mM, 129.4 sec<sup>-1</sup>, 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.