Substrate selectivity and inhibitor sensitivity affect xylanase functionality in wheat flour gluten-starch separation

Addition of xylanases (EC 3.2.1.8) that varied in their substrate selectivities and/or wheat xylanase inhibitor sensitivities in dough batter gluten-starch separation of wheat flour showed the importance of these enzyme characteristics for their functionality in this process. A xylanase from Aspergillus aculeatus (XAA) with selectivity for hydrolysis of water extractable arabinoxylan (WE-AX;, which is not inhibited by wheat flour xylanase inhibitors decreased batter viscosity and improved gluten agglomeration behaviour. In contrast, a xylanase from Bacillus subtilis (XBSi) with selectivity for hydrolysis of water unextractable arabinoxylan (WU-AX), which is in vitro inhibited by wheat flour xylanase inhibitors had a negative effect on gluten agglomeration at low enzyme dosages. As expected, solubilisation of WU-AX increased batter viscosities. At higher dosages however, this enzyme also improved gluten agglomeration because of degradation of both WE-AX and enzymically solubilised AX. A mutated B. subtilis xylanase (XBSni) with selectivity for hydrolysis of WU-AX comparable to XBSi but which is not inhibited by wheat flour xylanase inhibitors, increased the level of large gluten aggregates as well as the total gluten protein recovery, even at lower dosages. Because of its inhibitor insensitivity, the solubilisation and degradation of AX proceeded further. An XBSni dosage approximately 4 times lower than XBSi performed as well as its inhibited counterpart. The degradation of both WE-AX and WU-AX by XBSni improved the gluten agglomeration behaviour to a larger extent than the XAA treatment which primarily resulted in hydrolysis of WE-AX. The results confirm the detrimental impact not only of WE-AX, but also of WU-AX, on gluten agglomeration in a dough batter gluten-starch separation process. At the same time, they provide firm evidence that xylanases are not only inhibited by xylanase inhibitors in vitro, but are also partly inhibited in the industrial process in which they are used. (C) 2004 Elsevier Ltd. All rights reserved.