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dc.contributor.authorBiely, Peter
dc.contributor.authorHirsch, Jan
dc.contributor.authorLa Grange, Daniel C.
dc.contributor.authorVan Zyl, Willem H.
dc.contributor.authorPrior, Bernard A.
dc.date.accessioned2011-04-07T08:23:39Z
dc.date.available2011-04-07T08:23:39Z
dc.date.issued2000-08
dc.identifier.citationBiely, Peter, Hirsch, Jan, La Grange, Daniel C., Van Zyl, Willem H., Prior, Bernard A. 2000. A Chromogenic Substrate for a b-Xylosidase-Coupled Assay of a-Glucuronidase. Analytical Biochemistry 286:289-294.doi:10.1006/abio.2000.4810, http://www.sciencedirect.com.ez.sun.ac.za/science?_ob=MImg&_imagekey=B6W9V-45FK4RH-2H-3&_cdi=6692&_user=613892&_pii=S0003269700948103&_origin=search&_coverDate=11%2F15%2F2000&_sk=997139997&view=c&wchp=dGLbVtb-zSkzV&md5=97a7d694592b628ed93c14a774164170&ie=/sdarticle.pdfen_ZA
dc.identifier.issn0003-2697 (Print Version)
dc.identifier.issn1096-0309 (Online Version)
dc.identifier.otherdoi:10.1006/abio.2000.4810
dc.identifier.urihttp://hdl.handle.net/10019.1/8482
dc.descriptionThe original publication is available at www.elsevier.com.
dc.descriptionIncludes bibliography.
dc.description.abstract-Nitrophenyl 2-(4-O-methyl-α- -glucopyranuronosyl)-β- -xylopyranoside obtained on deesterification of 4-nitrophenyl 2-O-(methyl 4-O-methyl-α- -glucopyranosyluronate)-β- -xylopyranoside (Hirsch et al., Carbohydr. Res. 310, 145–149, 1998) was found to be an excellent substrate for the measurement of hemicellulolytic α-glucuronidase activity. A new precise α-glucuronidase assay was developed by coupling the α-glucuronidase-catalyzed formation of 4-nitrophenyl β- -xylopyranoside with its efficient hydrolysis by β-xylosidase. A recombinant strain of Saccharomyces cerevisiae, harboring and expressing the β-xylosidase gene xlnD of Aspergillus niger under control of the alcohol dehydrogenase II promoter on a multicopy plasmid, was used as a source of β-xylosidase. The activity values of β-xylosidase in the assay required to achieve a steady-state rate of 4-nitrophenol formation shortly after starting the α-glucuronidase reaction were obtained both experimentally and by calculation using the kinetics of coupled enzyme reactions.en_ZA
dc.format.extentp. 289-294 : ill.
dc.language.isoen_USen_ZA
dc.publisherElsevieren_ZA
dc.subjectα-Glucuronidaseen_ZA
dc.subjectXylosidaseen_ZA
dc.subjectEnzymesen_ZA
dc.subjectMicrobiological assayen_ZA
dc.subjectChromogenic substrateen_ZA
dc.subjectGlycosideen_ZA
dc.subjectAldobiuronic aciden_ZA
dc.subjectRecombinanten_ZA
dc.titleA Chromogenic substrate for a β-xylosidase-coupled assay of α-glucuronidaseen_ZA
dc.typeArticleen_ZA
dc.rights.holderElsevieren_ZA


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