Cardiff University | Prifysgol Caerdydd ORCA
Online Research @ Cardiff 
WelshClear Cookie - decide language by browser settings

Biochemical characterization of Bacillus subtilis type II isopentenyl diphosphate isomerase, and phylogenetic distribution of isoprenoid biosynthesis pathways

Laupitz, Ralf, Hecht, Stefan, Amslinger, Sabine, Zepeck, Ferdinand, Kaiser, Johannes, Richter, Gerald, Schramek, Nicholas, Steinbacher, Stefan, Huber, Robert, Arigoni, Duilio, Bacher, Adelbert, Eisenreich, Wolfgang and Rohdich, Felix 2004. Biochemical characterization of Bacillus subtilis type II isopentenyl diphosphate isomerase, and phylogenetic distribution of isoprenoid biosynthesis pathways. European Journal of Biochemistry 271 (13) , pp. 2658-2669. 10.1111/j.1432-1033.2004.04194.x

Full text not available from this repository.

Abstract

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 µmol·mg−1·min−1, respectively, as diagnosed by 1H 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)-α–hydroxyacid oxidizing enzymes including flavocytochrome b2 and l-lactate dehydrogenase and was shown to catalyse the formation of [2,3-13C2]lactate from [2,3-13C2]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.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Biosciences
Publisher: Wiley
ISSN: 0014-2956
Last Modified: 24 Jun 2017 11:00
URI: https://orca.cardiff.ac.uk/id/eprint/70160

Citation Data

Cited 59 times in Scopus. View in Scopus. Powered By Scopus® Data

Actions (repository staff only)

Edit Item Edit Item