• Title/Summary/Keyword: Ochrobactrum sp. CSL1

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

  • Shen, Min;Ju, Xin;Xu, Xinqi;Yao, Xuemei;Li, Liangzhi;Chen, Jiajia;Hu, Cuiying;Fu, Jiaolong;Yan, Lishi
    • Journal of Microbiology and Biotechnology
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    • v.28 no.7
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    • pp.1122-1132
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    • 2018
  • 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 $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose, which was unconventional for RpiB. The optimal reaction conditions ($50^{\circ}C$, pH 8.0, and 1 mM $Ca^{2+}$) were obtained to maximize the potential of OsRpiB in preparing $\text\tiny{L}$-rhamnulose. The catalytic properties of OsRpiB, including $K_m$, $k_{cat}$, and catalytic efficiency ($k_{cat}/K_m$), were determined as 43.47 mM, $129.4sec^{-1}$, and 2.98 mM/sec. The highest conversion rate of $\text\tiny{L}$-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 $\text\tiny{L}$-rhamnose to $\text\tiny{L}$-rhamnulose by OsRpiB biocatalysis.