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Endoxylanase-based affinity chromatography for efficient one-step purification of endoxylanase inhibitors

Initially, the isolation of barley endoxylanase inhibitors was based on an elaborate classical biochemical purification method. This, combined with the fact that the level of endoxylanase inhibitors in barley is approximately ten times lower than in wheat, resulted in difficulties when trying to obtain larger quantities of inhibitor for in-depth studies. Therefore, a new technique was developed for the purification of endoxylanase inhibitors based on affinity chromatography with immobilised endoxylanases from Bacillus subtilis and Aspergillus niger.

The main advantages of this procedure over the classical biochemical method are speed on the one hand and the possibility to isolate two types of barley endoxylanase inhibitors, i.e. Triticum aestivum endoxylanase inhibitor (TAXI)-type and endoxylanase inhibiting protein (XIP)-type inhibitors, on the other hand.

The choice of the endoxylanases that are immobilised on the affinity columns is such that TAXI-type inhibitors are retained at the first column (with B. subtilis endoxylanase) and XIP-type inhibitors are only retained on the second column (with A. niger endoxylanase).

Contacto

Jan DELCOUR, (Head of the Laboratory of Food Chemistry)
Tel.: +32-(0)16-321634
Fax: +32-(0)16-321997
Correo electrónico
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