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In this study, we found that biofilm formation is a critical factor affecting the activity of Aspergillus niger SJ1 xylanase. Xylanase activity increased 8.8% from 1046.88 to 1147.74 U/mL during A. niger SJ1 immobilized fermentation with biofilm formation. Therefore, we carried out the work of genomic analysis and biofilm-related time-series transcriptome analysis of A. niger SJ1 for better understanding of the ability of A. niger SJ to produce xylanase and biofilm formation. Genome annotation results revealed a complete biofilm polysaccharide component synthesis pathway in A. niger SJ1 and five proteins regarding xylanase synthesis. In addition, results of transcriptome analysis revealed that the genes involved in the synthesis of cell wall polysaccharides and amino acid anabolism were highly expressed in the biofilm. Furthermore, the expression levels of major genes in the gluconeogenesis pathway and mitogen-activated protein kinase pathway were examined.