Premium
An enhanced protocol for expression and purification of heat‐stable enterotoxin of enterotoxigenic Escherichia coli
Author(s) -
Aref NasrEldin M.,
Saeed A. Mahdi
Publication year - 2012
Publication title -
journal of biochemical and molecular toxicology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.526
H-Index - 58
eISSN - 1099-0461
pISSN - 1095-6670
DOI - 10.1002/jbt.21406
Subject(s) - enterotoxigenic escherichia coli , enterotoxin , chemistry , chromatography , escherichia coli , bioreactor , heat labile enterotoxin , microbiology and biotechnology , biochemistry , biology , organic chemistry , gene
We present an improved protocol for expression and purification of heat‐stable enterotoxin (STa) of enterotoxigenic Escherichia coli (ETEC). In this protocol, controlled growth conditions at different pHs (7.4, 8.0, and 8.6) were adopted using a bioreactor. In addition, specific adsorbent resins, methacrylate, were used for STa purification. The bioreactor provided optimal ETEC growth at pH 7.4 with high STa production. Furthermore, methacrylate bounded specifically to STa and dramatically enhanced the purification process of STa. The STa‐specific activity was high (8.9 × 10 6 units/mg protein), and the minimal effective dose of STa required for production of gut weight to remaining body weight ratio ≥ 0.083 was recorded as less than 0.2 ng in 2–3 days old suckling mice. The protocol presented, produces highly purified STa as documented by matrix‐assisted laser desorption ionization‐time of flight mass spectroscopy/. Also, as compared with the traditional methods, this procedure is trouble‐free and practical for scale‐up production and purification of STa peptides. © 2012 Wiley Periodicals, Inc. J Biochem Mol Toxicol 26:168–175, 2012; View this article online at wileyonlinelibrary.com . DOI 10.1002/jbt.21406