ABSTRACT Enterokinase (EK) is a heterodimeric serine protease which plays a key role in initiating the proteolytic digestion cascade in the mammalian duodenum. The enzyme acts by converting

trypsinogen to trypsin via a highly specific cleavage following the pentapeptide recognition sequence (Asp)4-Lys. This stringent site specificity gives EK great potential as a fusion protein

cleavage reagent. Recently, a cDNA encoding the catalytic (light) chain of bovine enterokinase (EKL) was identified, characterized, and transiently expressed in mammalian COS cells. We

report here the production of EKL in _Escherichia coli_ by a novel secretory expression system that utilizes _E. coli_ DsbA protein as an N-terminal fusion partner. The EKL cDNA was fused

the previously described COS-derived enzyme. Both forms of rEKL were capable of cleaving peptides, polypeptides and trypsinogen with the same specificity exhibited by the native

heterodimeric enzyme purified from bovine duodena. Interestingly, rEKL activated trypsinogen poorly relative to the native heterodimeric enzyme, but was superior in its ability to cleave

artificial fusion proteins containing the (Asp)4-Lys recognition sequence. Access through your institution Buy or subscribe This is a preview of subscription content, access via your

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OF BOVINE ENTEROKINASE IN _ESCHERICHIA COLI_ Article Open access 21 October 2022 RANDOM AND COMBINATORIAL MUTAGENESIS FOR IMPROVED TOTAL PRODUCTION OF SECRETORY TARGET PROTEIN IN

_ESCHERICHIA COLI_ Article Open access 05 March 2021 DEVELOPMENT OF HIGH-PERFORMANCE INDUCIBLE AND SECRETORY EXPRESSION VECTOR AND HOST SYSTEM FOR ENHANCED RECOMBINANT PROTEIN PRODUCTION

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AFFILIATIONS * Genetics Institute, Inc., 87 Cambridge Park Drive, Cambridge, MA, 02140 Lisa A. Collins-Racie, James M. McColgan, Kathleen L. Grant, Elizabeth A. DiBlasio-Smith, John M. McCoy

 & Edward R. LaVallie Authors * Lisa A. Collins-Racie View author publications You can also search for this author inPubMed Google Scholar * James M. McColgan View author publications

You can also search for this author inPubMed Google Scholar * Kathleen L. Grant View author publications You can also search for this author inPubMed Google Scholar * Elizabeth A.

DiBlasio-Smith View author publications You can also search for this author inPubMed Google Scholar * John M. McCoy View author publications You can also search for this author inPubMed 

Google Scholar * Edward R. LaVallie View author publications You can also search for this author inPubMed Google Scholar CORRESPONDING AUTHOR Correspondence to Edward R. LaVallie. RIGHTS AND

PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Collins-Racie, L., McColgan, J., Grant, K. _et al._ Production of Recombinant Bovine Enterokinase Catalytic Subunit

in _Escherichia coli_ Using the Novel Secretory Fusion Partner DsbA. _Nat Biotechnol_ 13, 982–987 (1995). https://doi.org/10.1038/nbt0995-982 Download citation * Received: 22 February 1995

* Accepted: 10 July 1995 * Issue Date: 01 September 1995 * DOI: https://doi.org/10.1038/nbt0995-982 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this

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