title: A chromogenic assay for limit dextrinase and pullulanase activity.
abstract: A new chromogenic substrate to assay the starch debranching enzymes limit dextrinase and pullulanase is described. The 2-chloro-4-nitrophenyl glycoside of a commercially available branched heptasaccharide (Glc-maltotriosyl-maltotriose) was found to be a suitable specific substrate for starch debranching enzymes and allows convenient assays of enzymatic activities in a format suited for high-throughput analysis. The kinetic parameters of these enzymes toward the synthesized substrate are determined, and the selectivity of the substrate in a complex cereal-based extract is established.
authors: Carlsberg Laboratory, Gamle Carlsberg vej 10, 1799 Copenhagen V, Denmark. Electronic address: marie.bojstrup@carlsberglab.dk.
authors: Marie
authors: M
authors: Bøjstrup
authors: Carlsberg Laboratory, Gamle Carlsberg vej 10, 1799 Copenhagen V, Denmark.
authors: Caspar Elo
authors: CE
authors: Christensen
authors: Carlsberg Laboratory, Gamle Carlsberg vej 10, 1799 Copenhagen V, Denmark.
authors: Michael Skovbo
authors: MS
authors: Windahl
authors: Carlsberg Laboratory, Gamle Carlsberg vej 10, 1799 Copenhagen V, Denmark.
authors: Anette
authors: A
authors: Henriksen
authors: Carlsberg Laboratory, Gamle Carlsberg vej 10, 1799 Copenhagen V, Denmark. Electronic address: ole.hindsgaul@carlsberglab.dk.
authors: Ole
authors: O
authors: Hindsgaul
mesh_terms: D057075:Enzyme Assays / Q000379:methods*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006027:Glycosides / Q000378:metabolism; D001467:Hordeum / Q000201:enzymology* / Q000378:metabolism; D007700:Kinetics; D009596:Nitrophenols / Q000378:metabolism; D013213:Starch / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D006027:Glycosides; D009596:Nitrophenols; C024719:4-nitrophenyl; D013213:Starch; D006026:Glycoside Hydrolases; C100181:pullulanase
keywords: Chromogenic substrate; Coupled assay; Limit dextrinase; Pullulanase