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Bamboo biomass is a potential source for the production of monomeric sugars containing high cellulose content with low amount of lignin. However, for efficient hydrolysis, the biomass treatment by effective pretreatment technique is required to minimize lignin content and other barrier components. During present study, the bamboo biomass was treated with different physical, chemical, biological and combined treatments to reduce the lignin content. Among all the pretreatments, the maximum lignin removal (14.5%) was obtained with the combined chemical and biological treatment under 2% NaOH+1% H2O2 +WDP2 fungal culture (5 plugs) conditions. In addition, lignolytic fungus and NaOH pretreatment was mainly effective in removing lignin, whereas the H2O2 pretreatment efficiently minimize cellulose crystallinity. To analyze structural changes of raw and treated biomass, we used scanning electron microscopy and fourier transform infrared spectroscopy. The structural analysis indicated that all treatments causes disruption in the biomass structure and loses the compactness of the biomass which facilitates the biomass conversion during hydrolysis process. The findings of the present study indicate effective pretreatment methods in breaching the recalcitrancy of the potential lignocellulosic biomass for maximum hydrolysis.