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Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography
Author(s) -
Luong John H. T.,
Male Keith B.,
Mazza Alberto,
Masson Luke,
Brousseau Roland
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200406046
Subject(s) - micellar electrokinetic chromatography , chromatography , chemistry , electrokinetic phenomena , bacillus thuringiensis , sodium dodecyl sulfate , capillary electrophoresis , ion chromatography , high performance liquid chromatography , resolution (logic) , sodium , bacteria , biology , organic chemistry , artificial intelligence , computer science , genetics
Bacillus thuringiensis and recombinant Escherichia coli proteinaceous protoxins were subject to proteolysis and analyzed by capillary electrokinetic chromatography. Three resulting toxins (65 kDa) were baseline‐resolved within 22 min using a 10 m M borate, pH 11 separation buffer consisting of 25 m M sodium dodecyl sulfate (SDS) and 30 m M phytic acid. The toxins displayed differential interactions with the SDS and phytic acid phases to effect their separation. The ion‐pairing interaction between the analyte and phytic acid was also useful in preventing adsorption to the capillary walls and thus enhanced separation resolution and efficiency. The use of electrokinetic chromatography allows achievement of the separation in a significantly shorter time than conventional high‐performance liquid chromatography (HPLC) using a diethylaminoethyl (DEAE) weak‐anion exchanger.