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Expression of a soluble and activatable form of bovine procarboxypeptidase A in Escherichia coli.

Abstract：

:Bovine pancreatic procarboxypeptidase A has been overexpressed in a soluble and activatable form in Escherichia coli. When the protein was expressed under the control of bacteriophage T7 promoter in E. coli ADA494 (a thioredoxin reductase deficient bacteria), a thioredoxin fusion protein was produced at relatively high level in the cytoplasm (4 mg/L culture medium). Although the recombinant protein essentially accumulated as inclusion bodies, as much as 30% of the fusion protein was recovered in a soluble form at low growth temperature and could therefore be purified to homogeneity in a single-step procedure by metal-affinity chromatography. The recombinant precursor form of bovine carboxypeptidase A was recognized by a monoclonal antibody directed against purified bovine pancreatic carboxypeptidase A. Moreover, upon tryptic activation it gave rise to an enzyme, the N-terminal sequence, molecular size,and specific activity of which were comparable to those of the enzyme derived from the native precursor purified from bovine pancreas.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Seddi R,Chaix JC,Puigserver A,Guo XJ

doi

10.1016/s1046-5928(02)00573-9

keywords:

subject

Has Abstract

pub_date

2003-02-01 00:00:00

pages

220-8

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046592802005739

journal_volume

27

pub_type

杂志文章

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