Skip Navigation


PEDS Advance Access originally published online on August 2, 2009
Protein Engineering Design and Selection 2009 22(11):649-654; doi:10.1093/protein/gzp048
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
22/11/649    most recent
gzp048v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Casaite, V.
Right arrow Articles by Meskys, R.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Casaite, V.
Right arrow Articles by Meskys, R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© The Author 2009. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Expression and purification of active recombinant equine lysozyme in Escherichia coli

Vida Casaite1,4, Simona Bruzyte1, Virginijus Bukauskas2, Arunas Setkus2, Ludmilla A. Morozova-Roche3 and Rolandas Meskys1

1Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Mokslininku 12, Vilnius LT-08662, Lithuania 2 Semiconductor Physics Institute, A. Gostauto 11, Vilnius LT-01108, Lithuania 3Department of Medical Biochemistry and Biophysics, Umea University SE-90187, Umea, Sweden

4 To whom correspondence should be addressed. E-mail: vidac{at}bchi.lt

Equine lysozyme (EL) is a calcium (Ca)-binding lysozyme and is an intermediary link between non-Ca-binding C-type lysozyme and {alpha}-lactalbumin. The feature of lysozymes to assemble into the fibrils has recently gained considerable attention for the investigation of the functional properties of these proteins. To study the structural and functional properties of EL, a synthetic gene was cloned and EL was overexpressed in Escherichia coli as a fused protein. The His-tagged recombinant EL was accumulated as inclusion bodies. Up to 50 mg/l of the recombinant EL could be achieved after purification by Ni2+ affinity chromatography, refolding in the presence of arginine, CM-Sepharose column purification following TEV protease cleavage. The purified protein was functionally active, as determined by the lysozyme activity, proving the proper folding of protein. The purified lysozyme was used for the oligomerisation studies. The protein formed amyloid fibrils during incubation in acidic pH and elevated temperature. The recombinant EL forms two types of fibrils: ring shaped and linear, similar to the native EL.

Keywords: amyloid fibrils/equine lysozyme/heterologous expression/inclusion body

Received June 4, 2009; revised June 4, 2009; accepted July 8, 2009.


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.