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Please use this identifier to cite or link to this item: http://hdl.handle.net/1942/4093

Title: Recombinant expression and characterization of XynD from Bacillus subtilis subsp subtilis ATCC 6051: a GH 43 arabinoxylan arabinofuranohydrolase
Authors: Bourgois, Tine M.
Van Craeyveld, Valerie
Van Campenhout, Steven
Courtin, Christophe M.
Delcour, Jan A.
Volckaert, Guido
Issue Date: 2007
Publisher: SPRINGER
Abstract: The complete genome sequence of Bacillus subtilis reveals that sequences encoding several hemicellulases are co-localised with a gene (xynD) encoding a putative family 43 glycoside hydrolase that has not yet been characterised. In this work, xynD has been isolated from genomic DNA of B. subtilis subsp. subtilis ATCC 6051 and cloned for cytoplasmatic expression in Escherichia coli. Recombinant XynD (rXynD) was purified using ion-exchange chromatography and gel permeation chromatography. The enzyme had a molecular mass of approximately 52 kDa, a pI above 9.0 and releases alpha-L-arabinose from arabinoxylo-oligosaccharides as well as arabinoxylan polymers with varying degree of substitution. Using para-nitrophenyl-alpha-L-arabinofuranoside as substrate, maximum activity was observed at pH 5.6 and 45 degrees C. The enzyme retained its activity over a large pH range, while activity was lost after pre-incubation above 50 degrees C. Gas-liquid chromatography and proton nuclear magnetic resonance spectrometry analysis indicated that rXynD specifically releases arabinofuranosyl groups from mono-substituted C-(O)-2 and C-(O)-3 xylopyranosyl residues on the xylan backbone. As rXynD did not display endoxylanase, xylosidase or arabinanase activity and was inactive on arabinan, we conclude that this enzyme is best described as an arabinoxylan arabinofuranohydrolase.
Notes: Katholieke Univ Leuven, Lab Gene Technol, B-3001 Louvain, Belgium. Katholieke Univ Leuven, Lab Food Chem & Biochem, B-3001 Louvain, Belgium. Univ Hasselt, Limburgs Univ Ctr, Biomed Res Inst, Sch Life Sci, B-3590 Diepenbeek, Belgium.Bourgois, TM, Katholieke Univ Leuven, Lab Gene Technol, Kasteelpk Arenberg 21, B-3001 Louvain, Belgium.tine.bourgois@biw.kuleuven.be
URI: http://hdl.handle.net/1942/4093
DOI: 10.1007/s00253-007-0956-2
ISI #: 000247463100011
ISSN: 0175-7598
Category: A1
Type: Journal Contribution
Validation: ecoom, 2008
Appears in Collections: Research publications

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