Analysis of starch structure using fluorophore-assisted carbohydrate electrophoresis.
The analysis of the fine structure of starches is important to the investigation of linkages between starch structure and function and to the investigation of the properties and roles of starch biosynthetic, modifying and degradation enzymes. Fluorophore-assisted carbohydrate electrophoresis has recently been introduced as a method for the analysis of the oligosaccharide populations released by the enzymatic digestion of starches, which has advantages in resolution and sensitivity over previously used methods, and provides the capacity for the facile analysis of oligosaccharide populations on either a molar or mass basis. The use of fluorophore-assisted carbohydrate electrophoresis for the analysis of oligosaccharides is reviewed with particular reference to the choice of label, efficiency of labeling and separation techniques. Examples of separations using slab gel electrophoresis, DNA sequencer analysis and capillary electrophoresis are presented and we conclude that on the basis of resolution and reproducibility, capillary electrophoresis is the method of choice for the separation of oligosaccharides of degree of polymerization from 1 to 100. Examples of isoamylase-debranched starches and glycogens analyzed by capillary electrophoresis are presented. The capillary electrophoresis analysis of starch structure through the analysis of oligosaccharides released by the debranching of limit dextrins derived from starches and glycogens is introduced as a useful diagnostic of starch structure. The potential for future development of novel diagnostics for starch structure using fluorophore-assisted carbohydrate electrophoresis is discussed.
Morell
M K
MK
CSIRO Plant Industry, Camberra, ACT, Australia. morell@pi.csiro.au
Samuel
M S
MS
O'Shea
M G
MG
D000818:Animals; D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005456:Fluorescent Dyes; D006026:Glycoside Hydrolases / Q000378:metabolism; D007517:Isoamylase / Q000032:analysis; D009281:Naphthalenes; D009844:Oligosaccharides / Q000032:analysis; D011134:Polysaccharides / Q000032:analysis; D011721:Pyrenes; D017422:Sequence Analysis, DNA; D013213:Starch / Q000737:chemistry*; D000516:alpha-Amylases / Q000378:metabolism; D001614:beta-Amylase / Q000378:metabolism
C098210:1-aminopyrene-3,6,8-trisulfonic acid; D002241:Carbohydrates; D005456:Fluorescent Dyes; D009281:Naphthalenes; D009844:Oligosaccharides; D011134:Polysaccharides; D011721:Pyrenes; D013213:Starch; C046465:8-amino-1,3,6-naphthalenetrisulfonic acid; D006026:Glycoside Hydrolases; D000516:alpha-Amylases; D001614:beta-Amylase; C100181:pullulanase; D007517:Isoamylase