title: Capillary electrophoretic profiling of tryptic digests of water soluble proteins from Bacillus thuringiensis-transgenic and non-transgenic maize species.
abstract: Capillary zone electrophoresis (CZE) has been applied to separation and characterisation of enzymatic (tryptic) hydrolysates of water-soluble proteins from Bacillus thuringiensis (Bt)-transgenic (Aristis-Bt) and two native non-transgenic (Aristis and Coventry) maize varieties. Water-soluble proteins were extracted from the flour of these maize species and digested by bovine pancreatic trypsin immobilised on agarose gel in 100 mM ammonium hydrocarbonate buffer, pH 7.9. The yielded tryptic digests of proteins were analysed by CZE in four acidic background electrolytes (BGEs) (100 mM H₃PO₄, 50 mM Tris, pH 2.25; 500 mM acetic acid, pH 2.54; 200 mM formic acid, 200 mM acetic acid, pH 2.05; and 200 mM iminodiacetic acid, pH 2.26) using a lab-made CZE apparatus equipped with bare fused silica capillary and UV-absorption detector operating at 206 nm. Among the tested BGEs, the best resolution of the tryptic peptides of extracted proteins of the above three maize species was obtained in isoelectric BGE, 200 mM iminodiacetic acid, pH 2.26. Selected resolved tryptic peptides of proteins were characterised by effective electrophoretic mobilities and corrected (migration times normalised) peak areas. Some significant relative qualitative and quantitative differences in CZE-UV profiling of tryptic protein digests were found, which can be potentially used to differentiate transgenic Aristis Bt and non-transgenic Aristis varieties or two native non-transgenic varieties, Aristis and Coventry.
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovo n. 2, 166 10 Prague 6, Czech Republic.
authors.forename: Petra
authors.initials: P
authors.lastname: Sázelová
authors.affiliation: Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, v.v.i., Flemingovo n. 2, 166 10 Prague 6, Czech Republic.
authors.forename: Václav
authors.initials: V
authors.lastname: Kašička
authors.affiliation: Institute of Industrial Fermentations, CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Leon
authors.affiliation: Institute of Industrial Fermentations, CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Ibáñez
authors.affiliation: Institute of Industrial Fermentations, CSIC, Juan de la Cierva 3, 28006 Madrid, Spain.
authors.forename: Alejandro
authors.initials: A
authors.lastname: Cifuentes
mesh_terms: D000818:Animals; D001413:Bacillus thuringiensis / Q000737:chemistry*; D002417:Cattle; D019075:Electrophoresis, Capillary / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D003313:Zea mays / Q000737:chemistry*
chemical_list: D011506:Proteins