title: Direct detection of underivatized chitooligosaccharides produced through chitinase action using capillary zone electrophoresis.
abstract: Capillary electrophoresis with capacitively coupled contactless conductivity detection was successfully used to quantify N-acetylglucosamine and five N-acetyl-chitooligosaccharides (C2-C6) produced after reaction with a purified chitinase (TmChi) from Tenebrio molitor (Coleoptera). No derivatization process was necessary. The separation was developed using 10 mM NaOH with 10% (v/v) acetonitrile as background electrolyte and homemade equipment with a system that avoids the harmful effect of electrolysis. The limit of detection for all oligosaccharides was ca. 3microM, and the results indicated that the larger the oligosaccharide, the higher the sensitivity. Analysis of the chitooligosaccharides produced revealed that TmChi has an endolytic cleavage pattern with C5 as the best substrate (higher catalytic efficiency kcat/KM) releasing C2 and C3.
authors.affiliation: Departamento de Química, Instituto de Química, Universidade de São Paulo, C.P. 26077, 05513-970, São Paulo, Brazil.
authors.forename: Lucas
authors.initials: L
authors.lastname: Blanes
authors.forename: Renata M
authors.initials: RM
authors.lastname: Saito
authors.forename: Fernando A
authors.initials: FA
authors.lastname: Genta
authors.forename: Juliana
authors.initials: J
authors.lastname: Donegá
authors.forename: Walter R
authors.initials: WR
authors.lastname: Terra
authors.forename: Clélia
authors.initials: C
authors.lastname: Ferreira
authors.forename: Claudimir Lucio
authors.initials: CL
authors.lastname: do Lago
mesh_terms: D002384:Catalysis; D002688:Chitinases / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D009844:Oligosaccharides / Q000032:analysis*
chemical_list: D009844:Oligosaccharides; D002688:Chitinases