Purification and characterization of dipeptidyl aminopeptidase from Aureobacterium sp. WO26.
We isolated a bacterial strain with an enzyme which releases dipeptide from Gly-Arg-p-nitroanilide. The bacterium was tentatively identified as Aureobacterium sp. The enzyme, named AuDAP, was purified and characterized. It was homogenous by SDS-PAGE and IEF, and had a molecular mass of 90,000 Da by SDS-PAGE and 88,000 Da by gel filtration, so it may be a monomer. The isoelectric point was 3.8 and the optimum pH was 10.0. The purified enzyme hydrolyzed Gly-Arg-pNA, a model substrate for DAP I, and Arg-Arg-MNA, a model substrate for DAP III. However, this enzyme did not hydrolyze Gly-Phe pNA, also a model substrate for DAP I. These results suggested that this enzyme did not fall under the classification of mammalian DAPs and was similar to DAP BI from Pseudomonas sp. WO24 and dDAP from Dictyostelium discoideum, although several differences were observed between them. The N-terminal amino acid sequence of this enzyme showed no significant homology to any enzyme and protein, except only for DAP BI.
Ogasawara
W
W
Department of Bioengineering, Nagaoka University of Technology, Japan.
Inanobe
N
N
Ochiai
K
K
Ando
K
K
Okada
H
H
Morikawa
Y
Y
D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D016987:Gram-Positive Rods / Q000737:chemistry / Q000201:enzymology*; D007526:Isoelectric Point; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity
D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases