Xaa-Pro aminopeptidases are peptidases responsible for the cleavage of any amino acid N-terminally adjacent to a proline residue. We identified a gene encoding a putative Xaa-Pro aminopeptidase in the genome of the filamentous fungus Aspergillus oryzae (genome database number: AO090701000720) and named this gene xpmA. We produced its enzyme in a C-terminally His(6)-tag-fused form in an Escherichia coli expression system and purified it. The purified re combinant XpmA (rXpmA) showed hydrolysis activity toward Xaa-Pro-oligopeptides, especially the two dipeptides Ala-Pro and Phe-Pro. The molecular weight of rXpmA was estimated to be 69 KDa by SDS-PAGE and 126 KDa by gel filtration, suggesting that it is a homodimer. The enzyme was activated by various divalent m etal ions such as Mn2+, Co2+, and Mg2+; in particular, the enzyme activity was increased 27.6-times relative to the no-addition control by 1 mM Mn2+. Additionally, 10 mM EDTA suppressed its activity to 0.26-times of the control level. Therefore, rXpmA was a metal-loprotease. Optimal hydrolytic activity of rXpmA was observed at 50 degrees C and pH 8.5-9.0. The enzyme was stable up to 50 degrees C and from pH 4.0 to 11.0. rXpmA showed substrate inhibition by Leu-Pro, Ser-Pro and Arg-Pro at concentrations over 4 mM, 10 mM, and 3 mM, respectively. NaCl increased the enzyme activity in the concentration range 0.5-3.0 M, suggesting that the enzyme is halophilic. (C) 2017, The Society for Biotechnology, Japan. All rights reserved.