| Title : A chromogenic substrate for a beta-xylosidase-coupled assay of alpha-glucuronidase - Biely_2000_Anal.Biochem_286_289 |
| Author(s) : Biely P , Hirsch J , la Grange DC , van Zyl WH , Prior BA |
| Ref : Analytical Biochemistry , 286 :289 , 2000 |
| Abstract : 4-Nitrophenyl 2-(4-O-methyl-alpha-d-glucopyranuronosyl)-beta-d-xylopyranoside obtained on deesterification of 4-nitrophenyl 2-O-(methyl 4-O-methyl-alpha-d-glucopyranosyluronate)-beta-d-xylopyranoside (Hirsch et al., Carbohydr. Res. 310, 145-149, 1998) was found to be an excellent substrate for the measurement of hemicellulolytic alpha-glucuronidase activity. A new precise alpha-glucuronidase assay was developed by coupling the alpha-glucuronidase-catalyzed formation of 4-nitrophenyl beta-d-xylopyranoside with its efficient hydrolysis by beta-xylosidase. A recombinant strain of Saccharomyces cerevisiae, harboring and expressing the beta-xylosidase gene xlnD of Aspergillus niger under control of the alcohol dehydrogenase II promoter on a multicopy plasmid, was used as a source of beta-xylosidase. The activity values of beta-xylosidase in the assay required to achieve a steady-state rate of 4-nitrophenol formation shortly after starting the alpha-glucuronidase reaction were obtained both experimentally and by calculation using the kinetics of coupled enzyme reactions. |
| ESTHER : Biely_2000_Anal.Biochem_286_289 |
| PubMedSearch : Biely_2000_Anal.Biochem_286_289 |
| PubMedID: 11067752 |
Biely P, Hirsch J, la Grange DC, van Zyl WH, Prior BA (2000)
A chromogenic substrate for a beta-xylosidase-coupled assay of alpha-glucuronidase
Analytical Biochemistry
286 :289
Biely P, Hirsch J, la Grange DC, van Zyl WH, Prior BA (2000)
Analytical Biochemistry
286 :289