Laupitz, R. and Hecht, S. and Amslinger, S. and Zepeck, F. and Kaiser, J. and Richter, G. and Schramek, N. and Steinbacher, S. and Huber, R. and Arigoni, D. and Bacher, A. and Eisenreich, W. and Rohdich, F. (2004) Biochemical characterization of Bacillus subtilis type II isopentenyl diphosphate isomerase, and phylogenetic distribution of isoprenoid biosynthesis pathways. Eur. J. Biochem. 271 (13), pp. 2658-2669.
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An open reading frame (Acc. no. P50740) on the Bacillus subtilis chromosome extending from bp 184 997-186 043 with similarity to the idi-2 gene of Streptomyces sp. CL190 specifying type II isopentenyl diphosphate isomerase was expressed in a recombinant Escherichia coli strain. The recombinant protein with a subunit mass of 39 kDa was purified to apparent homogeneity by column chromatography. The protein was shown to catalyse the conversion of dimethylallyl diphosphate into isopentenyl diphosphate and vice versa at rates of 0.23 and 0.63 mumol.mg(-1).min(-1), respectively, as diagnosed by H-1 spectroscopy. FMN and divalent cations are required for catalytic activity; the highest rates were found with Ca2+. NADPH is required under aerobic but not under anaerobic assay conditions. The enzyme is related to a widespread family of (S)-alpha-hydroxyacid oxidizing enzymes including flavocytochrome b(2) and L-lactate dehydrogenase and was shown to catalyse the formation of [2,3-C-13(2)]lactate from [2,3-C-13(2)]pyruvate, albeit at a low rate of 1 nmol.mg(-1).min(-1). Putative genes specifying type II isopentenyl diphosphate isomerases were found in the genomes of Archaea and of certain eubacteria but not in the genomes of fungi, animals and plants. The analysis of the occurrence of idi-1 and idi-2 genes in conjunction with the mevalonate and nonmevalonate pathway in 283 completed and unfinished prokaryotic genomes revealed 10 different classes. Type II isomerase is essential in some important human pathogens including Staphylococcus aureus and Enterococcus faecalis where it may represent a novel target for anti-infective therapy.
|Additional information (public):||Times Cited: 9|
|Institutions:||Chemistry and Pharmacy > Institut für Organische Chemie > Arbeitskreis Dr. Sabine Amslinger|
|Keywords:||isoprenoids mevalonate deoxyxylulose Idi-2 FMN Deoxyxylulose phosphate-pathway hmg-coa reductase escherichia-coli dimethylallyl diphosphate nonmevalonate pathway mevalonate pathway glycolate oxidase crystal-structure lytb protein active-site|
|Subjects:||500 Science > 540 Chemistry & allied sciences|
|Refereed:||Yes, this version has been refereed|
|Created at the University of Regensburg:||No|
|Owner:||Dr. Sabine Amslinger|
|Deposited On:||24 Nov 2009 14:26|
|Last Modified:||07 Nov 2012 11:09|