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

Authors

Xing-Guang Hung, Department of Food Science, Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan
Ming-Yuan Yu, Department of Food Science, Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan
Yu-Chun Chen, Department of Food Science, Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, Taiwan
Tsuei-Yun Fang, Department of Food Science, Center of Excellence for the Oceans, National Taiwan Ocean University, Keelung, TaiwanFollow

Abstract

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°C 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°C for 2 h. Metal ions were not required for N-His-tagged L-RI activity to occur, but Hg2+ 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°C and 40°C 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.

Recommended Citation

Hung, Xing-Guang; Yu, Ming-Yuan; Chen, Yu-Chun; and Fang, Tsuei-Yun (2015) "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," Journal of Marine Science and Technology: Vol. 23: Iss. 4, Article 20.
DOI: 10.6119/JMST-014-0430-3
Available at: https://jmstt.ntou.edu.tw/journal/vol23/iss4/20