title: Novel substrates for the automated and manual assay of endo-1,4-β-xylanase.
abstract: endo-1,4-β-Xylanase (EC 3.2.1.8) is employed across a broad range of industries including animal feed, brewing, baking, biofuels, detergents and pulp (paper). Despite its importance, a rapid, reliable, reproducible, automatable assay for this enzyme that is based on the use of a chemically defined substrate has not been described to date. Reported herein is a new enzyme coupled assay procedure, termed the XylX6 assay, that employs a novel substrate, namely 4,6-O-(3-ketobutylidene)-4-nitrophenyl-β-45-O-glucosyl-xylopentaoside. The development of the substrate and associated assay is discussed here and the relationship between the activity values obtained with the XylX6 assay versus traditional reducing sugar assays and its specificity and reproducibility were thoroughly investigated.
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland. Electronic address: david@megazyme.com.
authors.forename: David
authors.initials: D
authors.lastname: Mangan
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Claudio
authors.initials: C
authors.lastname: Cornaggia
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Agnija
authors.initials: A
authors.lastname: Liadova
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Niall
authors.initials: N
authors.lastname: McCormack
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Ruth
authors.initials: R
authors.lastname: Ivory
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Vincent A
authors.initials: VA
authors.lastname: McKie
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Aaron
authors.initials: A
authors.lastname: Ormerod
authors.affiliation: Megazyme, IDA Business Park, Southern Cross Road, Bray, Co. Wicklow, Ireland.
authors.forename: Barry V
authors.initials: BV
authors.lastname: McCleary
mesh_terms: D000818:Animals; D001331:Automation; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism*; D057075:Enzyme Assays / Q000379:methods* / Q000592:standards; D005947:Glucose / Q000737:chemistry / Q000378:metabolism; D012015:Reference Standards; D013379:Substrate Specificity
chemical_list: D043364:Endo-1,4-beta Xylanases; D005947:Glucose