Please use this identifier to cite or link to this item:
Title: Effective reduction of truncated expression of gloshedobin in Escherichia coli using molecular chaperone ClpB
Authors: Nian, R.
Tan, L.
Choe, W.-S. 
Keywords: BL21(DE3)
Escherichia coli
Metal affinity chromatography
Thrombin-like enzyme
Truncated expression
Issue Date: Jun-2008
Source: Nian, R., Tan, L., Choe, W.-S. (2008-06). Effective reduction of truncated expression of gloshedobin in Escherichia coli using molecular chaperone ClpB. Chemical Engineering Science 63 (11) : 2875-2880. ScholarBank@NUS Repository.
Abstract: Snake venom thrombin-like enzymes (TLEs) have been widely studied for potential therapeutic applications as anti-coagulants in the treatment of blood clotting disorders. However, due to the cysteine-rich nature of these proteins, their expressions in Escherichia coli were often impeded by inclusion body (IB) formation. The formation of truncated expression products significantly complicated the production of gloshedobin, a recently isolated TLE from the snake venom of Gloydius shedaoensis [Yang, Q., Li, M., Xu, J.Q., Bao, Y.M., Lei, X.Y., An, L.J., 2003a. Expression of gloshedobin, a thrombin-like enzyme from the venom of Gloydius shedaoensis, in Escherichia coli. Biotechnology Letters 25, 101-104; Yang, Q., Xu, J.Q., Li, M., Lei, X.Y., An, L.J., 2003b. High-level expression of a soluble snake venom enzyme, gloshedobin, in E. coli in the presence of metal ions. Biotechnology Letters 25, 607-610]. In this work, it was found that the expression of gloshedobin was strongly dependent on the expression host. The truncated expression was reduced by 25% when the protein was expressed in E. coli BL21(DE3)pLysS instead of BL21(DE3). It was also demonstrated that co-expression of ClpB (a molecular chaperone) in BL21(DE3) enabled the expression of gloshedobin mostly in intact form without compromising expression level, while almost completely eliminating its truncation products. This suggests a new simple strategy to significantly improve the quality of protein expression in TLE production and may find its useful application for many other recombinant proteins whose expressions and/or purifications are hindered by the formation of truncation products. © 2008 Elsevier Ltd. All rights reserved.
Source Title: Chemical Engineering Science
ISSN: 00092509
DOI: 10.1016/j.ces.2008.03.003
Appears in Collections:Staff Publications

Show full item record
Files in This Item:
There are no files associated with this item.


checked on Mar 7, 2018


checked on Jan 31, 2018

Page view(s)

checked on Mar 11, 2018

Google ScholarTM



Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.