Two new aminopeptidases from Ochrobactrum anthropi active on D-alanyl-p-nitroanilide.
Two new enzymes which hydrolyse D-alanyl-p-nitroanilide have been detected in Ochrobactrum anthropi LMG7991 extracts. The first enzyme, DmpB, was purified to homogeneity and found to be homologous to the Dap protein produced by O. anthropi SCRC C1-38 (ATCC49237). The second enzyme, DmpA, exhibits a similar substrate profile when tested on p-nitroanilide derivatives of glycine and L/D-alanine, but the amounts produced by the Ochrobactrum strain were not sufficient to allow complete purification. Interestingly, the DmpA preparation also exhibited an L-aminopeptidase activity on the tripeptide L-Ala-Gly-Gly but it was not possible to be certain that the same protein was responsible for both p-nitroanilide and peptide hydrolysing activities. The gene encoding the DmpA protein was cloned and sequenced. The deduced protein sequence exhibits varying degrees of similarity with those corresponding to several open reading frames found in the genomes of other prokaryotic organisms, including Mycobacteria. None of these gene products has been isolated or characterised, but a tentative relationship can be proposed with the NylC amidase from Flavobacterium sp. K172.
Fanuel
L
L
Laboratoire d'Enzymologie et Centre d'Ingénierie des Protéines, Université de Liège, Sart Tilman, Belgium.
Thamm
I
I
Kostanjevecki
V
V
Samyn
B
B
Joris
B
B
Goffin
C
C
Brannigan
J
J
Van Beeumen
J
J
Frère
J M
JM
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*; D000814:Aniline Compounds; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D012230:Rhizobiaceae / Q000201:enzymology* / Q000235:genetics; D013379:Substrate Specificity
D000814:Aniline Compounds; D001426:Bacterial Proteins; D004269:DNA, Bacterial; C004813:alanine-4-nitroanilide; D000626:Aminopeptidases; C118321:DmpA protein, Ochrobactrum anthropi