Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography

John H.T. Luong; Keith B. Male; Alberto Mazza; Luke Masson; Roland Brousseau

doi:10.1002/elps.200406046

Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography

John H.T. Luong
, Keith B. Male
, Alberto Mazza
, Luke Masson
, Roland Brousseau

National Research Council of Canada

Research output: Contribution to journal › Article › peer-review

Abstract

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 mM borate, pH 11 separation buffer consisting of 25 mM sodium dodecyl sulfate (SDS) and 30 mM 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.

Original language	English
Pages (from-to)	3292-3299
Number of pages	8
Journal	Electrophoresis
Volume	25
Issue number	18-19
DOIs	https://doi.org/10.1002/elps.200406046
Publication status	Published - Oct 2004
Externally published	Yes

Keywords

Bacillus thuringiensis
Capillary electrophoresis
Phytic acid
Sodium dodecyl sulfate

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10.1002/elps.200406046

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title = "Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography",

abstract = "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 mM borate, pH 11 separation buffer consisting of 25 mM sodium dodecyl sulfate (SDS) and 30 mM 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.",

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doi = "10.1002/elps.200406046",

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TY - JOUR

T1 - Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography

AU - Luong, John H.T.

AU - Male, Keith B.

AU - Mazza, Alberto

AU - Masson, Luke

AU - Brousseau, Roland

PY - 2004/10

Y1 - 2004/10

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AB - 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 mM borate, pH 11 separation buffer consisting of 25 mM sodium dodecyl sulfate (SDS) and 30 mM 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.

KW - Bacillus thuringiensis

KW - Capillary electrophoresis

KW - Phytic acid

KW - Sodium dodecyl sulfate

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DO - 10.1002/elps.200406046

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SN - 0173-0835

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EP - 3299

JO - Electrophoresis

JF - Electrophoresis

IS - 18-19

ER -

Analysis of insecticidal proteins from Bacillus thuringiensis and recombinant Escherichia coli by capillary electrokinetic chromatography

Abstract

Keywords

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