Detection and differentiation of pectic enzyme activity in vitro and in vivo by capillary electrophoresis of products from fluorescent-labeled substrate.

Published

Journal Article

A sensitive assay is described for the detection of pectate-depolymerizing enzymes using capillary electrophoresis of a fluorescent end-labeled pectate oligomer. The labeled oligomer is allowed to react with the enzyme either in vitro or in vivo, such as inside the intercellular spaces of a cotton cotyledon, and after an appropriate incubation time the products are analyzed by capillary electrophoresis. The site and mode of action of the pectate-depolymerizing activity can be inferred from the products. Both endo- and exopolygalacturonase activity, and lyase activity, were distinguished. Since only the fluorescent oligomer and products from its labeled reducing end are detected, there is no interference from other compounds; only pectic enzyme activity is detected. By this type of analysis we can show that there is considerable endo- and exo-polygalacturonase activity in the intercellular spaces of cotton cotyledons.

Full Text

Duke Authors

Cited Authors

  • Zhang, Z; Pierce, ML; Mort, AJ

Published Date

  • February 1996

Published In

Volume / Issue

  • 17 / 2

Start / End Page

  • 372 - 378

PubMed ID

  • 8900945

Pubmed Central ID

  • 8900945

International Standard Serial Number (ISSN)

  • 0173-0835

Digital Object Identifier (DOI)

  • 10.1002/elps.1150170214

Language

  • eng

Conference Location

  • Germany