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Biochim Biophys Acta. 2005 Mar 14;1747(2):179-87. Epub 2004 Dec 31.

Simultaneous refolding/purification of xylanase with a microwave treated smart polymer.

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  • 1Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.

Abstract

Affinity precipitation with a smart polymer, Eudragit S-100 (a methyl methacrylate polymer), was exploited for simultaneous refolding and purification of xylanase. Affinity precipitation consisted of this reversibly soluble-insoluble polymer-binding xylanase selectively. The complex was precipitated by lowering the pH and xylanase was eluted off the polymer using 1 M NaCl. For refolding experiments, the commercial preparation of Aspergillus niger xylanase was denatured with 8 M urea. Addition of microwave irradiated Eudragit S-100 and affinity precipitation led to recovery of 96% enzyme activity by refolding. Simultaneously, the enzyme was purified 45 times. Thermally inactivated preparation, when subjected to similar steps, led to 95% recovery of enzyme activity with 42-fold purification. The strategy has the potential for recovering pure proteins in active forms from overexpressed proteins, which generally form inclusion bodies in E. coli.

PMID:
15698952
[PubMed - indexed for MEDLINE]
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