Servicio de Información Comunitario sobre Investigación y Desarrollo - CORDIS

A rapid screening method to determine endoxylanase substrate selectivity

The functionality of endoxylanases is affected by different parameters such as the endoxylanase substrate selectivity or the relative activity of endoxylanases towards water-unextractable (WU-AX) and water-extractable arabinoxylans (WE-AX). Previous research already showed the importance of substrate selectivity in biotechnological processes such as bread-making and gluten-starch separation.

A laborious and time-consuming method with native substrates was used to determine the substrate selectivity of endoxylanases. In this project, a rapid screening method using microtitre plates with chromogenic substrates was developed to determine this parameter.

The results were consistent with those obtained with the elaborate method. A high substrate selectivity factor indicates that the endoxylanase preferentially hydrolyses WU-AX over WE-AX, while endoxylanases with lower factors more readily hydrolyse WE-AX. The present rapid screening method is designed for use in routine applications by adaptation of the traditional enzyme assays in tubes to microtitre plate format, involving semi-automated multichannel pipettes and a microplate reader. The developed method is fast, accurate, reliable and reproducible and only small amounts of reagents and samples are needed.

The screening method can be used for both standard evaluations and scientific research. It can, for example, be used for establishment of the structure-function relationship of natural and engineered enzymes.

Reported by

Katholieke Universiteit Leuven, Laboratory of Food Chemistry
Kasteelpark Arenberg 20
3001 Heverlee
Belgium
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