Realization of the lowest possible cost of hydrolysis requires comprehensive management of the entire hydrolysis system to maximize enzyme cost performance. In this talk we will discuss how the interrelations and improvements of pretreatment, substrate conditioning, and enzymatic hydrolysis design parameters can be optimized to obtain the best tradeoff for a robust and low cost process. Furthermore, we will discuss how to leverage the latest advantages in enzyme development to minimize the cost of sugar production through hydrolysis process changes. Lab and pilot data from testing on various feedstocks will be evaluated through Novozymes’ advanced process cost model to demonstrate the Hydrolysis advances progress in reducing the Minimum Ethanol Selling Price.