The use of differently pre-treated lignocellulosic biomass (LCB), which contains varying levels of cellulose, hemicellulose and lignin content, is profitable during biofuel production but also require unique enzymatic preparations to convert these differently pre-treated LCB into their monomeric constituents. To address this question, this study employed simplex-lattice mixture design strategy to customize lignocellulolytic enzymatic cocktails for hydrolysis of different pre-treated substrates, obtained from PRAJ and IOCL Industries. In-house recombinant fungal auxiliary/accessory enzymes were evaluated to work in synergy with Cellic®CTec3. The mixtures of recombinant proteins derived from Scytalidium thermophilum CM-4T with Cellic®CTec3 were found to optimally hydrolyse diluted acid pre-treated rice straw slurries obtained from PRAJ and IOCL Industry using Simplex-lattice mixture designing (SLMD) and showed 70.39 % and 84.46 % saccharification efficiency, respectively.