TY - JOUR
T1 - Type 2 isopentenyl diphosphate isomerase from a thermoacidophilic archaeon Sulfolobus shibatae
AU - Yamashita, Satoshi
AU - Hemmi, Hisashi
AU - Ikeda, Yosuke
AU - Nakayama, Toru
AU - Nishino, Tokuzo
PY - 2004/3/1
Y1 - 2004/3/1
N2 - Although isopentenyl diphosphate-dimethylallyl diphosphate isomerase is thought to be essential for archaea because they use the mevalonate pathway, its corresponding activity has not been detected in any archaea. A novel type of the enzyme, which has no sequence similarity to the known, well-studied type of enzymes, was recently reported in some bacterial strains. In this study, we describe the cloning of a gene of a homologue of the novel bacterial isomerase from a thermoacidophilic archaeon Sulfolobus shibatae. The gene was heterologously expressed in Escherichia coli, and the recombinant enzyme was purified and characterized. The thermostable archaeal enzyme is tetrameric, and requires NAD(P)H and Mg2+ for activity, similar to its bacterial homologues. Using its apoenzyme, we were able to confirm that the archaeal enzyme is strictly dependent on FMN. Moreover, we provide evidence to show that the enzyme also has NADH dehydrogenase activity although it catalyzes the isomerase reaction without consuming any detectable amount of NADH.
AB - Although isopentenyl diphosphate-dimethylallyl diphosphate isomerase is thought to be essential for archaea because they use the mevalonate pathway, its corresponding activity has not been detected in any archaea. A novel type of the enzyme, which has no sequence similarity to the known, well-studied type of enzymes, was recently reported in some bacterial strains. In this study, we describe the cloning of a gene of a homologue of the novel bacterial isomerase from a thermoacidophilic archaeon Sulfolobus shibatae. The gene was heterologously expressed in Escherichia coli, and the recombinant enzyme was purified and characterized. The thermostable archaeal enzyme is tetrameric, and requires NAD(P)H and Mg2+ for activity, similar to its bacterial homologues. Using its apoenzyme, we were able to confirm that the archaeal enzyme is strictly dependent on FMN. Moreover, we provide evidence to show that the enzyme also has NADH dehydrogenase activity although it catalyzes the isomerase reaction without consuming any detectable amount of NADH.
KW - Archaea
KW - Flavoprotein
KW - Isopentenyl diphosphate-dimethylallyl diphosphate isomerase
KW - Isoprenoid
KW - NADH dehydrogenase
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U2 - 10.1111/j.1432-1033.2004.04010.x
DO - 10.1111/j.1432-1033.2004.04010.x
M3 - Article
C2 - 15009187
AN - SCOPUS:1642414342
VL - 271
SP - 1087
EP - 1093
JO - FEBS Journal
JF - FEBS Journal
SN - 1742-464X
IS - 6
ER -