Hydrolysis of wheat flour arabinoxylan, acid-debranched wheat flour arabinoxylan and arabino-xylo-oligosaccharides by β-xylanase, α-L-arabinofuranosidase and β-xylosidase.
A range of α-L-arabinofuranosyl-(1-4)-β-D-xylo-oligosaccharides (AXOS) were produced by hydrolysis of wheat flour arabinoxylan (WAX) and acid debranched arabinoxylan (ADWAX), in the presence and absence of an AXH-d3 α-L-arabinofuranosidase, by several GH10 and GH11 β-xylanases. The structures of the oligosaccharides were characterised by GC-MS and NMR and by hydrolysis by a range of α-L-arabinofuranosidases and β-xylosidase. The AXOS were purified and used to characterise the action patterns of the specific α-L-arabinofuranosidases. These enzymes, in combination with either Cellvibrio mixtus or Neocallimastix patriciarum β-xylanase, were used to produce elevated levels of specific AXOS on hydrolysis of WAX, such as 3(2)-α-L-Araf-(1-4)-β-D-xylobiose (A(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotriose (A(3)XX), 2(2)-α-L-Araf-(1-4)-β-D-xylotriose (XA(2)X), 3(2)-α-L-Araf (1-4)-β-D-xylotriose (XA(3)X), 2(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2)XX), 3(3)-α-L-Araf-(1-4)-β-D-xylotetraose (XA(3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotriose (A(2+3)XX), 2(3),3(3)-di-α-L-Araf-(1-4)-β-D-xylotetraose (XA(2+3)XX), 2(4),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(2+3)XXX) and 3(3),3(4)-di-α-L-Araf-(1-4)-β-D-xylopentaose (XA(3)A(3)XX), many of which have not previously been produced in sufficient quantities to allow their use as substrates in further enzymic studies. For A(2,3)XX, yields of approximately 16% of the starting material (wheat arabinoxylan) have been achieved. Mixtures of the α-L-arabinofuranosidases, with specific action on AXOS, have been combined with β-xylosidase and β-xylanase to obtain an optimal mixture for hydrolysis of arabinoxylan to L-arabinose and D-xylose.
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland. Electronic address: Barry@megazyme.com.
Barry V
BV
McCleary
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland.
Vincent A
VA
McKie
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland.
Anna
A
Draga
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland.
Edward
E
Rooney
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland.
David
D
Mangan
Megazyme International Ireland, Bray Business Park, Southern Cross Road, Bray, County Wicklow, Ireland.
Jennifer
J
Larkin
D002240:Carbohydrate Sequence; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism*; D006868:Hydrolysis; D009844:Oligosaccharides / Q000737:chemistry*; D013379:Substrate Specificity; D014908:Triticum / Q000737:chemistry*; D043325:Xylan Endo-1,3-beta-Xylosidase / Q000378:metabolism; D014990:Xylans / Q000737:chemistry*
D009844:Oligosaccharides; D014990:Xylans; C085118:arabinoxylan; D006026:Glycoside Hydrolases; D043325:Xylan Endo-1,3-beta-Xylosidase; C019713:alpha-N-arabinofuranosidase; D043364:Endo-1,4-beta Xylanases
Arabinoxylo-oligosaccharides; Enzymic hydrolysis; Wheat flour arabinoxylan; α-l-Arabinofuranosidase; β-Xylanase; β-Xylosidase