The sustainable conversion of woody biomass (lignocellulose) to fermentable sugars currently constitutes one of the key challenges to the transition to an economy based on renewable resources. The aim of the enzyme hydrolysis cluster is to find biotechnological solutions, that are economically and environmentally sustainable, to the decomposition of underutilized agricultural substrates. Our approaches to discover, characterize, and engineer enzymes that efficiently hydrolyze untreated and chemically or biologically pretreated straws include: 1) detailed characterization of fungal pretreatment enzymes; 2) characterization of fungal mutants that are deficient in decomposition; 3) genomic analysis of fungal decomposers; 4) functional metagenomic and metatranscriptomic search for ruminant microbial enzymes; 5) transcriptomic and exo-proteomic analysis of lignocellulolytic fungi to facilitate the development of enzyme cocktails; 6) the use of directed evolution to improve targeted lignocellulolytic enzymes; and 7) the implementation and development of bioinformatic tools and infrastructure to support wet lab activities and analysis.  These research deliverables will be employed to support the establishment of a framework for a sustainable cellulosic-based Canadian biofuel industry.