[en] Different strains of Bacillus were screened for their ability to hydrolyse D-alanyl-p-nitroanilide. Activity was detected in Bacillus pumilus, Bacillus brevis, Bacillus licheniformis 749I and Bacillus subtilis 168. The last strain was the best producer and was selected for the production and purification of the enzyme. The determination of the N-terminal sequence identified the enzyme as the product of the dppA gene (previously named dciAA) belonging to the dipeptide ABC transport (dpp) operon expressed early during sporulation. Open reading frames (ORFs) encoding putative related proteins were found in the genomes of a variety of Archaea and both sporulating and non-sporulating bacteria. The enzyme behaves as a D-aminopeptidase and represents the prototype of a new peptidase family. Among the tested substrates, the highest activities were found with D-Ala-D-Ala and D-Ala-Gly-Gly. The active enzyme behaves as an octamer of identical 30 kDa subunits. It exhibits a broad pH optimum, extending between pH 9 and 11. It is reversibly inhibited in the presence of Zn2+ chelators, and the sequence comparisons highlight the conservation of potential Zn-binding residues. As it has been shown by others that null mutations in the dpp operon do not inhibit spore formation, the physiological role of DppA is probably an adaptation to nutrient deficiency.
Disciplines :
Microbiology
Author, co-author :
Cheggour, Abdelatif; Université de Liège - ULiège > Centre d'Ingénierie des Protéines
Fanuel, Laurence; Université de Liège - ULiège > CIP
Duez, Colette ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Joris, Bernard ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Bouillenne, Fabrice ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Devreese, Bart; RUG > Vakgroep Biochemie, Fysiologie en Microbiologie
Van Driessche, Gonzales; RUG > Vakgroep Biochemie, Fysiologie en Microbiologie
Van Beeumen, Josef; RUG > Vakgroep Biochemie, Fysiologie en Microbiologie
Frère, Jean-Marie ; Université de Liège - ULiège > Département des sciences de la vie > Département des sciences de la vie
Goffin, Colette ; Université de Liège - ULiège > Centre d'ingénierie des protéines
Language :
English
Title :
The Dppa Gene of Bacillus Subtilis Encodes a New D-Aminopeptidase
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