caa a22 4500 60550671X CHVBK 20210128100628.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6417-4 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6417-4 Essentiality of tetramer formation of Cellulomonas parahominis L-ribose isomerase involved in novel L-ribose metabolic pathway [Elektronische Daten] [Yuji Terami, Hiromi Yoshida, Keiko Uechi, Kenji Morimoto, Goro Takata, Shigehiro Kamitori] L-Ribose isomerase from Cellulomonas parahominis MB426 (CpL-RI) can catalyze the isomerization between L-ribose and L-ribulose, which are non-abundant in nature and called rare sugars. CpL-RI has a broad substrate specificity and can catalyze the isomerization between D-lyxose and D-xylulose, D-talose and D-tagatose, L-allose and L-psicose, L-gulose and L-sorbose, and D-mannose and D-fructose. To elucidate the molecular basis underlying the substrate recognition mechanism of CpL-RI, the crystal structures of CpL-RI alone and in complexes with L-ribose, L-allose, and L-psicose were determined. The structure of CpL-RI was very similar to that of L-ribose isomerase from Acinetobacter sp. strain DL-28, previously determined by us. CpL-RI had a cupin-type β-barrel structure, and the catalytic site was detected between two large β-sheets with a bound metal ion. The bound substrates coordinated to the metal ion, and Glu113 and Glu204 were shown to act as acid/base catalysts in the catalytic reaction via a cis-enediol intermediate. Glu211 and Arg243 were found to be responsible for the recognition of substrates with various configurations at 4- and 5-positions of sugar. CpL-RI formed a homo-tetramer in crystals, and the catalytic site independently consisted of residues within a subunit, suggesting that the catalytic site acted independently. Crystal structure and site-direct mutagenesis analyses showed that the tetramer structure is essential for the enzyme activity and that each subunit of CpL-RI could be structurally stabilized by intermolecular contacts with other subunits. The results of growth complementation assays suggest that CpL-RI is involved in a novel metabolic pathway using L-ribose as a carbon source. Springer-Verlag Berlin Heidelberg, 2015 Crystal structure nationallicence Cupin-type β-barrel nationallicence Rare sugar nationallicence L-Ribose isomerase nationallicence Terami Yuji Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut Yoshida Hiromi Life Science Research Center and Faculty of Medicine, Kagawa University, 1750-1, Ikenobe, Miki-cho, Kita-gun, 761-0793, Kagawa, Japan aut Uechi Keiko Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut Morimoto Kenji Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut Takata Goro Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut Kamitori Shigehiro Life Science Research Center and Faculty of Medicine, Kagawa University, 1750-1, Ikenobe, Miki-cho, Kita-gun, 761-0793, Kagawa, Japan aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6303-6313 0175-7598 99:15<6303 2015 99 253 https://doi.org/10.1007/s00253-015-6417-4 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6417-4 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Terami Yuji Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut NATIONALLICENCE 700 1- Yoshida Hiromi Life Science Research Center and Faculty of Medicine, Kagawa University, 1750-1, Ikenobe, Miki-cho, Kita-gun, 761-0793, Kagawa, Japan aut NATIONALLICENCE 700 1- Uechi Keiko Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut NATIONALLICENCE 700 1- Morimoto Kenji Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut NATIONALLICENCE 700 1- Takata Goro Rare Sugar Research Center and Faculty of Agriculture, Kagawa University, 2393, Ikenobe, Miki-cho, Kita-gun, 761-0795, Kagawa, Japan aut NATIONALLICENCE 700 1- Kamitori Shigehiro Life Science Research Center and Faculty of Medicine, Kagawa University, 1750-1, Ikenobe, Miki-cho, Kita-gun, 761-0793, Kagawa, Japan aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6303-6313 0175-7598 99:15<6303 2015 99 253