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One step purification and characterization of the pyrrolidone carboxyl peptidase of Streptococcus pyogenes over‐expressed in Escherichia coli
Author(s) -
Awadé A.,
Gonzalès Th.,
Cleuziat Ph.,
Robert-Baudouy J.
Publication year - 1992
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(92)81053-o
Subject(s) - escherichia coli , streptococcus pyogenes , enzyme , molecular mass , biochemistry , sodium dodecyl sulfate , chemistry , bacteria , chromatography , biology , gene , staphylococcus aureus , genetics
Pyrrolidone carboxyl peptidase (EC 3.4.11.8) (Pcp), an enzyme which selectively removes pyrrolidone carboxylic acid (PCA) from some PCA‐peptides and ‐proteins, was demonstrated in bacteria and in plant, animal and human tissues. In this paper we describe the purification to homogeneity of the enzyme of Streptococcus pyogenes , over‐expressed in Escherichia coli . This was achieved, for the first time in one step, by hydrophobic interaction chromatography. Analysis under non‐denaturing conditions revealed a molecular mass of 85 kDa and in the presence of sodium dodecyl sulfate gave a molecular mass of 23.5 kDa. Investigations on enzymatic properties showed that the Pcp over‐expressed in E. coli disclosed properties similar to those found for the enzyme extracted from S. pyogenes or for some other Pcps studied previously. Thus the over‐expressed enzyme should serve as a suitable source for N‐terminal unblocking prior to some PCA protein sequencing.