Functional characterization of GcpE, an essential enzyme of the non‐mevalonate pathway of isoprenoid biosynthesis

FEBS Letters

Eberl, Matthias³  Kollas, Ann-Kristin³  Henne, Anke⁴  Steinbrecher, Irina²  Henschker, Dajana²  Altincicek, Boran³  Reichenberg, Armin³  Hintz, Martin³  Wiesner, Jochen³  Jomaa, Hassan³  Ostrovsky, Dmitry N⁵  Beck, Ewald³  Duin, Evert C¹  Hedderich, Reiner¹ 

[1] Max-Planck-Institut für terrestrische Mikrobiologie, Karl-von-Frisch-Strasse, 35043 Marburg, Germany;Jomaa Pharmaka GmbH, Frankfurter Strasse 50, 35392 Giessen, Germany;Biochemisches Institut, Justus-Liebig-Universität Giessen, Friedrichstrasse 24, 35392 Giessen, Germany;Institut für Mikrobiologie und Genetik, Georg-August-Universität Göttingen, Grisebachstrasse 8, 37077 Göttingen, Germany;Bakh Institute of Biochemistry, Leninsky prospect 33, 119071 Moscow, Russia

关键词: 2-C-Methyl-D-erythritol-2;    4-cyclodiphosphate;    GcpE;    (E)-4-Hydroxy-3-methyl-but-2-enyl diphosphate;    Isoprenoid biosynthesis;    LytB;    2-C-Methyl-D-erythritol-4-phosphate pathway;    DMAPP;    dimethylallyl diphosphate;    DOXP;    1-deoxy-D-xylulose-5-phosphate;    HMBPP;    (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate;    IPP;    isopentenyl diphosphate;    MEcPP;    2-C-methyl-D-erythritol-2;    4-cyclodiphosphate;    MEP;    2-C-methyl-D-erythritol-4-phosphate;    PBMC;    peripheral blood mononuclear cells;   

DOI  :  10.1016/S0014-5793(02)03725-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The gcpE gene product controls one of the terminal steps of isoprenoid biosynthesis via the mevalonate independent 2-C-methyl-D-erythritol-4-phosphate (MEP) pathway. This pathway is utilized by a variety of eubacteria, the plastids of algae and higher plants, and the plastid-like organelle of malaria parasites. Recombinant GcpE protein from the hyperthermophilic bacterium Thermus thermophilus was produced in Escherichia coli and purified under dioxygen-free conditions. The protein was enzymatically active in converting 2-C-methyl-D-erythritol-2,4-cyclodiphosphate (MEcPP) into (E)-4-hydroxy-3-methyl-but-2-enyl diphosphate (HMBPP) in the presence of dithionite as reductant. The maximal specific activity was 0.6 μmol min⁻¹ mg⁻¹ at pH 7.5 and 55°C. The k _cat value was 0.4 s⁻¹ and the K _m value for HMBPP 0.42 mM.

【 授权许可】

Unknown