FEBS Letters | |
One step purification and characterization of the pyrrolidone carboxyl peptidase of Streptococcus pyogenes over‐expressed in Escherichia coli | |
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
【 预 览 】
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