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Enzymatic hydrolysis of lignocellulosic biomass is limited by economic considerations arising from enzyme production costs and specific activities. The effect of six hemicellulases on raw sugarcane bagasse and two types of pretreated sugarcane bagasse was investigated using experimental designs. Our strategy was successful for developing more efficient and less expensive enzymatic mixture, and also revealed that hemicellulase mixtures with multiple activities could be less effective than expected. In this study, only two hemicelulases, the endo-1,4-xylanases (GH11) from Penicillium funiculosum (XynC11/CAC15487) and the feruloyl esterase (CE1) from Clostridium thermocellum (CtFAE/ATCC27405), effectively broke-down hemicellulose from pretreated sugarcane bagasse (up to 65%), along with the production of xylooligosaccharides (XOS). Our results also demonstrated that GH11 and CE1 can improve …