Gene expression and characteristics of a novel fibrinolytic enzyme (subtilisin DFE) in Escherichia coli.

Abstract:

AIMS:The objective of this study is to actively express a novel fibrinolytic enzyme, subtilisin DFE (douchi fibrinolytic enzyme), in Escherichia coli. METHODS AND RESULTS:The DNA fragments encoding pro-subtilisin DFE was amplified and cloned into the vector pET32a to obtain N-terminal Trx fusion expression plasmid. The recombinant subtilisin DFE was successfully expressed and processed in the soluble fraction of E. coli BL21(DE3) in a similar fashion as the endogenous one of Bacillus amyloliquefaciens DC-4, resulting in an active enzyme. Moreover, active enzyme can also be refolded from inclusion body. CONCLUSIONS:Active subtilisin DFE can be expressed and processed in E. coli. SIGNIFICANCE AND IMPACT OF THE STUDY:This study provides evidences that subtilisin DFE can be actively expressed in E. coli and the pro-peptide is essential for guiding the proper folding into the active conformation. As such, large quantities of recombinant subtilisin DFE can be produced for pharmacological and clinical research.

journal_name

Lett Appl Microbiol

authors

Zhang RH,Xiao L,Peng Y,Wang HY,Bai F,Zhang YZ

doi

10.1111/j.1472-765X.2005.01715.x

subject

Has Abstract

pub_date

2005-01-01 00:00:00

pages

190-5

issue

2

eissn

0266-8254

issn

1472-765X

pii

LAM1715

journal_volume

41

pub_type

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