Determination of Cellular Carbohydrates in Peanut Fungal Pathogens and Baker's Yeast by Capillary Electrophoresis and Electrochromatography

Abstract

In this work, the quantitation of cellular carbohydrates, namely chitin and glucan, in peanut fungal pathogens and baker's yeast was carried out by capillary electrophoresis (CE) and capillary electrochromatography (CEC). The chitin and glucan of the fungi were hydrolyzed by the enzymes chitinase and glucanase, respectively, to their corresponding sugar monomers N-acetylglucosamine (GlcNAc) and glucose (Glc). These two monosaccharides were then tagged with 6-aminoquinoline (6-AQ) to allow their separation and detection in CE and CEC. The 6-AQ derivatives of GlcNAc and Glc formed the basis for the determination by CE and CEC of chitin and glucan in peanut fungi and baker's yeast. Several parameters affecting the separation of the 6-AQ derivatives of GlcNAc and Glc, including the separation voltage and the composition of the running electrolyte, were investigated. Under the optimized separation conditions, the contents of cellular carbohydrates including N-acetylglucosamine, chitin, glucose, and glucan in some fungi, such as Sclerotinia minor, Sclerotium rolfsii, and baker's yeast, were successfully determined. The method described here allowed the assessment of genetic differences in Sclerotium rolfsii isolates from various locations.

Sensitive determination of carbohydrates labelled with p‐nitroaniline by capillary electrophoresis with photometric detection using a 406 nm light‐emitting diode

“…Numerous reagents have been used in this fashion, such as 2-aminopyridine [24], aminonaphthalene sulphonic acid isomers [25,26], aminobenzene derivatives [27][28][29][30][31][32][33][34][35][36][37][38][39], aminopyrene trisulphonate derivatives [40], aminoacridone derivatives [41,42], 6-aminoquinoline [43][44][45], 7-amino-4-methylcoumarin [46,47], 4-amino-1,1-azobenzene-3,4-disulphonic acid [48] and 9-aminofluorene [49], as recently reviewed by Lamari et al [23]. Surprisingly, the use of a simple reagent like PNA for labelling of carbohydrates has so far been largely overlooked with only one report appearing in 1986 on labelling oligosaccharides [19].…”

Behavioural dynamics in the biological control of pests: role of silicon complexes

“…All the plants have been shown to produce chitinases and another compound, β-1,3-glucanase associated with the defence system of the plants against chitin-containing pathogens. [22,23] Therefore we expect at least the following regulatory processes to be operative:…”

The separation of biomolecules using capillary electrochromatography

“…PMP derivatives of aldopentose isomers were also excellently separated on a column prepared by in situ aminopropylation with 3-aminopropyltrimethoxsilane of a capillary packed with silica particles. Recently, quanti®cation of the cellular carbohydrates chitin and glucan in peanut fungal pathogens and baker's yeast was performed on CEC with ODS packing [35]. Separation of sucrose and saccharin on CEC was reported with highly sensitive detection using condensation nucleation light scattering detection [36].…”