Abstract
The gene coding for d-psicose 3-epimerase (DPEase) from Clostridium sp. BNL1100 was cloned and expressed in Escherichia coli. The recombinant enzyme was purified by Ni-affinity chromatography. It was a metal-dependent enzyme and required Co2+ as optimum cofactor. It displayed catalytic activity maximally at pH 8.0 and 65 °C (as measured over 5 min). The optimum substrate was d-psicose, and the K m, turnover number (k cat), and catalytic efficiency (k cat/K m) for d-psicose were 227 mM, 32,185 min−1, and 141 min−1 mM−1, respectively. At pH 8.0 and 55 °C, 120 g d-psicose l−1 was produced from 500 g d-fructose l−1 after 5 h.
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Acknowledgments
This work was supported by the NSFC Project (Nos. 31171705 and 21276001), the 973 Project (Nos. 2012CB720802), the 863 Project (Nos. 2011AA100904), the Fundamental Research Funds for the Central Universities (No. JUSRP51304A), and the Support Project of Jiangsu Province (Nos. BE2011622, BE2011766 and BE2010626).
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Wanmeng Mu and Wenli Zhang contributed equally to this work.
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Mu, W., Zhang, W., Fang, D. et al. Characterization of a d-psicose-producing enzyme, d-psicose 3-epimerase, from Clostridium sp.. Biotechnol Lett 35, 1481–1486 (2013). https://doi.org/10.1007/s10529-013-1230-6
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DOI: https://doi.org/10.1007/s10529-013-1230-6