- 期刊
Characterization of a Recombinant _L-RIBOSE Isomerase from ＂Geodermatophilus obscurus＂ DSM 43160 and Application of this Enzyme to the Production of L-Ribose From _L-ARABINOSE
並列摘要
_L-Ribose isomerase (_L-RI) catalyzes the aldose-ketose isomerization between _L-ribose and _L-ribulose. In this study, a putative L-RI gene of ＂Geodermatophilus obscurus＂ DSM 43160 was cloned by PCR into pET-15b and pET-21b, respectively. The cloned target gene was expressed in ＂Escherichia coli＂. The recombinant N-His-tagged and C-His-tagged proteins exhibited L-RI activity. Both N- and C-His-tagged L-RIs were purified from cell-free extracts by metal-affinity and ionexchange chromatography. The purified N-His-tagged L-RI demonstrated its optimal activity at 30-40℃ and pH of 9 (in glycine-NaOH buffer). The enzyme was stable at pH 7-9 and more than 90% activity was retained after incubation at 40℃ for 2 h. Metal ions were not required for N-His-tagged _L-RI activity to occur, but Hg^(2+) inhibited its activity completely. The conversion rates of _L-arabinose to _L-ribose by combining ＂Thermoanaerobacterium saccharolyticum＂ NTOU1 _L-arabinose isomerase and ＂G. obscurus＂ DSM43160 N-His-tagged _L-RI at 30℃ and 40℃ were 15.9% and 12.5% (mol mol^(-1)), respectively. Results obtained from this study suggest a potential application of this recombinant _L-RI for the _L-ribose production from _L-arabinose.
延伸閱讀
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