期刊
FEBS LETTERS
卷 532, 期 3, 页码 432-436出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/S0014-5793(02)03725-0
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
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 mumol min(-1) mg(-1) at pH 7.5 and 55degreesC. The k(cat) value was 0.4 S-1 and the K-m value for HMBPP 0.42 mM. (C) 2002 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.
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