AbstractXylan-degrading enzymes from Aspergillus niger and Hypocrea orientalis were characterized by enzyme activity assays and protein profiling with SDS-PAGE and LC-MS/MS. The hydrolysis of Miscanthus xylan by xylan-degrading enzymes from A. niger, H. orientalis, and Trichoderma reesei were comparatively studied by HPLC analysis. It was found that the glycoside hydrolase families 10 xylanase was the main xylanase secreted by H. orientalis and A. niger when using corn cob and wheat bran as inducers. Compared to the enzymes from T. reesei,the enzymes from A. niger showed better efficiency in the hydrolysis of Miscanthus xylan into monosaccharides. Nevertheless, the enzymes from H. orientalis were more preferable for the hydrolysis of Miscanthus xylan into xylo-oligosaccharides (XOS), especially xylobiose and xylotriose. Miscanthus xylan degradation was significantly influenced by the activities of β-xylosidase and α-L-arabinofuranosidase. Xylan-degrading enzymes with high ratios of β-xylosidase and α-L-arabinofuranosidase are necessary for the efficient conversion of Miscanthus xylan into monosaccharides. However, xylan-degrading enzymes with low β-xylosidase activity and high α-L-arabinofuranosidase activity were required for producing XOS.