Enzymatic hydrolysis of hemicelluloses from Miscanthus to monosaccharides or xylo-oligosaccharides by recombinant hemicellulases

• Enzymatic hydrolysis of Miscanthus hemicelluloses by recombinant enzymes.
• AnXyn10C and HoXyn11A were able to directly act on holocellulose.
• Holocellulose was resistant to AnAxh62A and AnXln3D.
• The presence of a novel 2-O-β-d-Xylp-α-l-Araf side group in Miscanthus hemicelluloses.
• Hemicellulases show different synergistic effect on holocelluose and hemicelluloses.

Hemicelluloses isolated from holocellulose (MHOC) from Miscanthus were characterized by RP-HPLC-UV, FT-IR, and NMR. The hemicelluloses and recombinant hemicellulases, including endo-β-1,4-xylanases (HoXyn11A and AnXyn10C), β-xylosidases (AnXln3D), and α-l-arabinofuranosidases (AnAxh62A), as well as their interaction mechanisms were investigated by enzymatic hydrolysis. AnXyn10C released shorter end products than HoXyn11A from isolated hemicelluloses (IHEC). AnAxh62A was able to release all single-substituted α-l-arabinofuranosyl residues from IHEC. AnXyn10C and HoXyn11A were able to directly act on MHOC, whereas AnAxh62A and AnXln3D did not. The combination of HoXyn11A and AnAxh62A produced the highest xylo-oligosaccharides (XOS) yield from IHEC, whereas AnXyn10C alone produced the highest XOS yield from MHOC. The combination of HoXyn11A, AnAxh62A, and AnXln3D achieved the highest xylose yield from IHEC, whereas the combination of AnXyn10C, AnAxh62A, and AnXln3D achieved the highest xylose yield from MHOC. This study contributes to the development of efficient enzyme cocktails for the bioconversion of hemicelluloses from Miscanthus into monosaccharides and XOS.