Please use this identifier to cite or link to this item: https://doi.org/10.1021/bp070304q
Title: Polyethyleneimine-mediated chemical extraction of cytoplasmic his-tagged inclusion body proteins from Escherichia coli
Authors: Nian, R.
Tan, L.
Choe, W.-S. 
Issue Date: Mar-2008
Source: Nian, R., Tan, L., Choe, W.-S. (2008-03). Polyethyleneimine-mediated chemical extraction of cytoplasmic his-tagged inclusion body proteins from Escherichia coli. Biotechnology Progress 24 (2) : 417-425. ScholarBank@NUS Repository. https://doi.org/10.1021/bp070304q
Abstract: The selectivity of polyethyleneimine (PEI) in DNA precipitation during chemical extraction was investigated. Chemical extraction was used to recover two His-tagged model proteins: gloshedobin, a thrombin-like enzyme from snake venom, and IbpA, a molecular chaperone, which were expressed mainly in the form of inclusion bodies. High DNA removal efficiency (more than 90%) was achieved at various cell densities (with OD600 ranging from 30 to 150) without affecting the solubility of host cell proteins. Compared to spermine-induced precipitation method reported elsewhere, PEI provided a higher DNA precipitation efficiency at a significantly lower cost. Moreover, PEI obviated the use of EDTA, which has been reported to be essential for the chemical extraction methods, hence exhibiting dual roles in replacing cost-prohibitive spermine and EDTA. The residual PEI in the post-extraction mixture was efficiently counteracted by addition of Mg2+, allowing the streamlined application of the extraction broth to immobilized metal affinity chromatography. Taken together, the PEI-mediated chemical extraction method provides a simpler and more economically viable processing route for the production of recombinant proteins whose expression is hampered by IB formation. © 2008 American Chemical Society and American Institute of Chemical Engineers.
Source Title: Biotechnology Progress
URI: http://scholarbank.nus.edu.sg/handle/10635/89839
ISSN: 87567938
DOI: 10.1021/bp070304q
Appears in Collections:Staff Publications

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

SCOPUSTM   
Citations

2
checked on Feb 21, 2018

WEB OF SCIENCETM
Citations

1
checked on Jan 15, 2018

Page view(s)

14
checked on Feb 18, 2018

Google ScholarTM

Check

Altmetric


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