Primary Structure, Partial Purification and Regulation of Key Enzymes of the Acetyl Cycle of Arginine Biosynthesis in Bacillus Stearothermophilus: Dual Function of Ornithine Acetyltransferase.
Sakanyan, Vehary and Charlier, Daniel and Legrain, Christianne and Kochikyan, Anahit and Mett, Igor and PiéRard, André and Glansdorff, Nicolas,, 139, 393-402 (1993),
doi = https://doi.org/10.1099/00221287-139-3-393,
publicationName = Microbiology Society,
issn = 1350-0872,
abstract= A 3·4 kb EcoRI fragment, cloned in E. coli, that carries part of a cluster of genes encoding arginine biosynthetic functions of the thermophilic bacterium Bacillus stearothermophilus, was sequenced on both strands. The sequence consists of a truncated argC gene, an argJ region encoding a polypeptide with both N-acetylglutamate synthase and ornithine acetyltransferase activities, the argB gene and the N-terminal part of argD. The argB gene encodes a 258-amino-acid polypeptide with a deduced M r of 26918. A very high and thermostable N-acetylglutamate 5-phosphotransferase activity was detected in extracts of E. coli argB mutants transformed with the 3·4 kb fragment on a plasmid. A polypeptide band of M r 27000 was detected by SDS-PAGE of heat-treated extract from such a strain. Both N-acetylglutamate synthase and ornithine acetyltransferase are encoded by the same 1290 bp open reading frame. The deduced sequence of 410 amino acids corresponds to a peptide of M r 43349. The subcloned B. stearothermophilus argJ can complement a double argA argE E. coli mutant to prototrophy. Gel-filtration of a heat-treated extract of the complemented double mutant E. coli host showed that N-acetylglutamate synthase and ornithine acetyltransferase activities co-elute in a single peak corresponding to M r 110000. Both activities were also heat-inactivated at the same temperature and strongly inhibited by ornithine. These results suggest that both activities can be ascribed to a single protein.,
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