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Expression, Purification, and Crystallization of D-Psicose 3-Epimerase from Agrobacterium tumefaciens  

Kim Kwang-Soo (School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Kim Hye-Jung (Department of Molecular Biotechnology, Konkuk University)
Oh Deok-Kun (Department of Molecular Biotechnology, Konkuk University)
Cheong Jong-Joo (School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Rhee Sang-Kee (School of Agricultural Biotechnology and Center for Agricultural Biomaterials, Seoul National University)
Publication Information
Journal of Microbiology and Biotechnology / v.16, no.4, 2006 , pp. 647-650 More about this Journal
Abstract
D-Psicose 3-epimerase (DPE) catalyzes the interconversion of D-fructose to D-psicose by epimerizing the carbon-3 position. The DPE from Agrobacterium tumefaciens was cloned and expressed in Escherichia coli. The expressed enzyme was purified by affinity chromatography on an IMAC, gel filtration chromatography on a Sephacryl S-300 HR, and anion-exchange chromatography on a RESOURCE Q. The molecular mass of the purified enzyme was estimated to be about 135 kDa by Superdex 200 gel filtration chromatography, corresponding to a homotetramer. The enzyme produced crystals suitable for X-ray diffraction to a $2.0{\AA}$ resolution at 100 K. The crystals were found to belong to the orthorhombic space group $P2_12_12_1$, with unit-cell parameters a=102.4, b=113.0, and $c=131.8{\AA}$. In addition, the calculated packing parameter $(V_m)$ was $2.79{\AA}^3/Da$, the solvent content was 55.92%, and an asymmetric unit consisted of four monomers.
Keywords
Agrobacterium tumefaciens; D-psicose 3-epimerase; crystallization; X-ray crystal structure;
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Times Cited By Web Of Science : 6  (Related Records In Web of Science)
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