Ionic liquid (IL) pretreatment represents an effective strategy for effective fractionation of lignocellulosic biomass (LB) to fermentable sugars in a biorefinery. Optimization of combinatorial pretreatment of sugarcane bagasse (SCB) with IL (1-butyl-3-methylimidazolium chloride [Bmim]Cl) and surfactant (PEG-8000) resulted in enhanced sugar yield (16.5%) upon enzymatic saccharification. The saccharification enzymes (cellulase and xylanase) used in the current study were in-house produced from a novel IL-tolerant fungal strain Aspergillus assiutensis VS34, isolated from chemically polluted soil, which produced adequately IL-stable enzymes. This is the first ever report of IL-stable cellulase/xylanase enzyme from Aspergillus assiutensis. To get the mechanistic insights of combinatorial pretreatment physicochemical analysis of variously pretreated biomass was executed using SEM, FT-IR, XRD, and ¹H NMR studies. The combined action of IL, surfactant and ultrasound had very severe and distinct effects on the ultrastructure of biomass that subsequently resulted in enhanced accessibility of saccharification enzymes to biomass, and increased sugar yield.