One step purification and characterization of the pyrrolidone carboxyl peptidase of Streptococcus pyogenes over‐expressed in Escherichia coli

FEBS Letters

Robert-Baudouy, J.¹  Awadé, A.¹  Cleuziat, Ph.¹  Gonzalès, Th.¹ 

[1] Laboratoire de Génétique Moléculaire des Microorganismes, Bâtiment 406, Institut National des Sciences Appliquées, 20 avenue Albert Einstein, 69621 Villeurbanne cedex, France

关键词: Peptidase;    Pcp protein;    Over-expression;    Purification;    Streptococcus pyogenes;    Pcp;    pyrrolidone carboxyl peptidase;    l-pyr-β-NA;    l-pyroglutamyl-β-naphthylamide;    l-pyr-p-NI;    l-pyroglutamyl-para-nitroanilide;    PCA;    pyrrolidone carboxylic acid;    OD;    optical density;   

DOI  :  10.1016/0014-5793(92)81053-O

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown