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Purification and characterization of a post-proline dipeptidyl aminopeptidase from Streptococcus cremoris AM2

Published online by Cambridge University Press:  01 June 2009

Mary Booth ,
Ide Ni Fhaoláin ,
P. Vincent Jennings  and
Gerard O'Cuinn
Mary Booth
Affiliation:
Department of Biochemistry, University College, Galway and Department of Life Sciences, Regional Technical College, Galway, Ireland
Ide Ni Fhaoláin
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
P. Vincent Jennings
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
Gerard O'Cuinn
Affiliation:
Department of Life Sciences, Regional Technical College, Galway, Ireland
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Summary

The present study describes the purification of a post-proline dipeptidyl aminopeptidase from the cytoplasm of Streptococcus cremoris AM2. On the basis of its elution from a calibrated Sephadex G200 column, the enzyme had a molecular weight of 117000 and exhibited a broad pH optimum activity between 6·0 and 9·0. The activity was most comprehensively inhibited by phenylmethylsulphonylfluoride and more modestly inhibited by 1,10-phenanthroline and 8-hydroxyquinoline but not by EDTA. A range of peptides containing either proline or alanine as the penultimate amino acid residue could act as substrates. The presence of proline on the carboxy side of the scissile bond prevented hydrolysis. However the enzyme could release Pro-Pro from Pro-Pro-Gly-Phe-Ser-Pro. The significance of this substrate specificity is considered in the context of removal of either single proline residues or prolylproline sequences from oligopeptides during cheese ripening.

Type
Original Articles
Information
Journal of Dairy Research , Volume 57 , Issue 1 , February 1990 , pp. 89 - 99
Copyright
Copyright © Proprietors of Journal of Dairy Research 1990

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