Oligopeptidases involve in the latter process of protein hydrolysis and is practically valuable in the preparation of functional oligopeptides with different hydrolysis degrees. In this work, a new peptidase YpsA was cloned and identified from Aspergillus niger, which consisted of 492 amino acid residues, with a typical Sec signal peptide sequence of 18 amino acid residues in the N-terminal. Its primary structure was highly similar to that of aspartic peptidases; however, it shared only 33.05% the highest similarity with the functionally identified aspartic peptidases. By cloning and expressing, the recombinant YpsA was prepared and biochemically characterized. It had the highest activity at 40℃ and pH 7.0. Its activity was strongly inhibited by pepstatin, a specific inhibitor of aspartic peptidase, but was hardly inhibited by PMSF, E-64 and EDTA. YpsA preferred to hydrolyze five oligopeptide substrates tested and had very weak activities on whole protein and polypeptides. In conclusion, YpsA is a newly identified neutral aspartic oligopeptidase with hydrolysis preference for oligopeptide substrates. It has a specific potential of application in bioprocessing protein preparations with low molecular weight ranges.
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