Currently available enzyme technology is insufficient to economically degrade plant biomass, and presumably will remain so whilst fundamental questions are inadequately answered, the most evident being: “how do microbes and their enzymes interact with plant cell walls?” Compounding these difficulties is the “cultivability bottleneck”. The microbes that harbor the answers to these questions are largely irrecoverable in isolate form, which restricts access to their genetic and metabolic machinery.The present project will address these issues by applying a progressive interdisciplinary approach to study and compare natural and engineered digestive ecosystems that are linked together via overlapping phenotypic and functional traits (i.e. biomass degradation). The project aims to generate insight into diverse uncultured microbial lineages and uncover core enzyme systems for biomass degradation that are present in multiple environments. To achieve its objectives the project will employ a combination of predictive genome-reconstruction technologies, as well as metagenome-directed isolation strategies to target dominant and novel saccharolytic species. Furthermore we will develop and take advantage of advanced software for enzyme annotation and phylogenetic binning as it is being developed. Relevant genes identified from reconstructed genomes and/or transcriptome data for isolates will be cloned, over-expressed and their gene products tested using state-of-the-art carbohydrate microarray technologies, prior to being characterized in detail.The project will complement existing activities at the PI’s university on (1) polysaccharide converting enzymes in a biorefining context, (2) the impact of intestinal fiber deconstruction on satiety and (3) enhanced production of biogas. We expect to unravel novel aspects of the microbial ecology within these systems/processes. Furthermore, it is envisaged that novel isolates and enzymes will enter into live bioenergy projects.