A Chromogenic substrate for a β-xylosidase-coupled assay of α-glucuronidase
| dc.contributor.author | Biely, Peter | |
| dc.contributor.author | Hirsch, Jan | |
| dc.contributor.author | La Grange, Daniel C. | |
| dc.contributor.author | Van Zyl, Willem H. | |
| dc.contributor.author | Prior, Bernard A. | |
| dc.date.accessioned | 2011-04-07T08:23:39Z | |
| dc.date.available | 2011-04-07T08:23:39Z | |
| dc.date.issued | 2000-08 | |
| dc.description | The original publication is available at www.elsevier.com. | |
| dc.description | Includes 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.extent | p. 289-294 : ill. | |
| dc.identifier.citation | Biely, 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.pdf | en_ZA |
| dc.identifier.issn | 0003-2697 (Print Version) | |
| dc.identifier.issn | 1096-0309 (Online Version) | |
| dc.identifier.other | doi:10.1006/abio.2000.4810 | |
| dc.identifier.uri | http://hdl.handle.net/10019.1/8482 | |
| dc.language.iso | en_US | en_ZA |
| dc.publisher | Elsevier | en_ZA |
| dc.rights.holder | Elsevier | en_ZA |
| dc.subject | α-Glucuronidase | en_ZA |
| dc.subject | Xylosidase | en_ZA |
| dc.subject | Enzymes | en_ZA |
| dc.subject | Microbiological assay | en_ZA |
| dc.subject | Chromogenic substrate | en_ZA |
| dc.subject | Glycoside | en_ZA |
| dc.subject | Aldobiuronic acid | en_ZA |
| dc.subject | Recombinant | en_ZA |
| dc.title | A Chromogenic substrate for a β-xylosidase-coupled assay of α-glucuronidase | en_ZA |
| dc.type | Article | en_ZA |