Water soluble pentosans (WSP) from doughs and breads made with different enzyme preparations are characterized according to extraction yield, sugar composition, xylose/arabinose ratio and molecular weight (MW) distribution. Extraction yield was greater for dough than for bread samples, ranging from 0.94 to 1.64%, but bread extracts had a higher purity. Percent of pentoses in purified WSP was greater in pentosanase supplemented samples (28-55%) than in control and amylase containing samples (23-32%). Major sugars were xylose and arabinose, but glucose and mannose also appeared in the extracts. The xylose/arabinose (Xyl/Ara) ratio was 1.3-1.6 and underwent small changes during processing. Enzyme addition caused an increase in Xyl/Ara ratio, attributable to a debranching of arabinoxylans (AX) with higher degree of Ara substitution by arabinofuranosidase. Addition of pentosanases had a significant effect in increasing WSP with MW over 39 000, whereas those of low MW changed only slightly. MW distribution depended on enzyme source, and whereas some enzymes showed activity during fermentation others increased their activity during baking. No synergistic effects were observed in studied variables due to the combination of amylases with pentosanases. Protein in WSP extracts eluted together with ferulic acid suggesting they were linked, but not associated with a determined carbohydrate fraction.

An extracellular alpha-glucuronidase was purified and characterized from a commercial Aspergillus preparation and from culture filtrate of Aspergillus tubingensis. The enzyme has a molecular mass of 107 KDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 112 kDa as determined by mass spectrometry, has a determined pI just below 5.2, and is stable at pH 6.0 for prolonged times. The pH optimum for the enzyme is between 4.5 and 6.0, and the temperature optimum is 70 degrees C. The alpha-glucuronidase is active mainly on small substituted xylo-oligomers but is also able to release a small amount of 4-O-methylglucuronic acid from birchwood xylan. The enzyme acts synergistically with endoxylanases and beta-xylosidase in the hydrolysis of xylan. The enzyme is N glycosylated and contains 14 putative N-glycosylation sites. The gene encoding this alpha-glucuronidase (aguA) was cloned from A. tubingensis. It consists of an open reading frame of 2,523 bp and contains no introns. The gene codes for a protein of 841 amino acids, containing a eukaryotic signal sequence of 20 amino acids. The mature protein has a predicted molecular mass of 91,790 Da and a calculated pI of 5.13. Multiple copies of the gene were introduced in A. tubingensis, and expression was studied in a highly overproducing transformant. The aguA gene was expressed on xylose, xylobiose, and xylan, similarly to genes encoding endoxylanases, suggesting a coordinate regulation of expression of xylanases and alpha-glucuronidase. Glucuronic acid did not induce the expression of aguA and also did not modulate the expression on xylose. Addition of glucose prevented expression of aguA on xylan but only reduced the expression on xylose.